November/December 2011 UK Issue 149

Plasma FIB Analysisof ThroughSilicon Vias p9

Secondary ElectronAtomic Imagingin S/TEM pS5

Variable PressureSEM ofPlant Stigmata p21

High ThroughputAutomatedS/TEM pS11

Ensure your slides are never again lost, damaged or forgotten.

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Keeping your slides, exactly as you know them.The Olympus VS120 virtual slide system. Digital slides at the highest resolution.

CIRCLE NO. 2 OR ONLINE: www.microscopy-analysis.com

REGULARS4 Dates for the Diary

23 People & Places

26 Literature Highlights

29 What’s New

41 Reader Enquiry form

MICROSCOPY AND ANALYSIS NOVEMBER 2011 3

CO N T E N T S

www.microscopy-analysis.com

See the Microscopy and Analysis online Diary for a full listing. Entries for the January issue are due 1 December. Email to: editor@microscopy-analysis.com

dates for the diaryDIARY

MICROSCOPY AND ANALYSIS NOVEMBER 20114

NOVEMBER 20118-11 Gatan EELS School, Austrian Centre for Electron Microscopy and

Nanoanalysis, TU Graz, Austriawww.felmi-zfe.tugraz.at www.gatan.com/company/news/news08291102.php

10 Electron Microscopy in Materials Science, Eindhoven University of Technology, The Netherlandshttp://nvvmmaterials2011.chem.tue.nl/

12-16 Neuroscience 2011: Washington DC, USAwww.sfn.org

14-18 NvVM European School on Nanobeams , Centre de Recherche Public - Gabriel Lippmann, Belvaux, Luxembourg www.nanobeams.org

16 Cryo Microscopy Group Annual Meeting, Boots Science Building, School of Pharmacy, University Park, Nottingham University, UKwww.cryomicroscopygroup.org.uk/CMG2011.html

22-23 Bruker Scanning Probe Microscopy Conference and Users’ Meeting, University of Manchester, UKwww.bruker-axs.com drew.murray@bruker-nano.com

28-2/12 2011 MRS Fall Meeting, Boston, Massachusetts, USAwww.mrs.org

DECEMBER 20113-4 Functional Optical Imaging, Univeristy of Nottingham Ningbo Campus,

Chinawww.nottingham.ac.uk/ibios/index.php?page=foi-11

3-7 American Society for Cell Biology 51st Annual Meeting, Denver, Colorado, USAwww.ascb.org

JANUARY 20124-7 Winter School on High-Resolution Electron Microscopy, Arizona State

University, Tempe, AZ, USAwintershcool@asu.edu http://le-csss.asu.edu/winterschool

FEBRUARY 20125-9 APMC 10: 10th Asia-Pacific Microscopy Conference;

ICONN 2012: International Conference on Nanoscience and Nanotechnology;

ACMM 22: Australian Conference on Microscopy and Microanalysis, Perth, Australiawww.apmc-10.org www.iconn-2012.org www.acmm-22.org

25-29 56th Biophysical Society Annual Meeting, San Diego, California, USAwww.biophysics.org/2012meeting/Main/tabid/2386/Default.aspx

MARCH 201211-16 Pittcon 2012, Orlando, FL, USA

www.pittcon.org18-23 Course in Cryotechniques for Electron Microscopy, Rothamsted

Research, Harpenden, UKwww.rms.org.uk/events/Forthcoming_Events/CoolRunnings

APRIL 20121-4 Focus on Microscopy 2012; 25th International Conference on 3D Image

Processing in Microscopy; 24th International Conference on Confocal Microscopy, Singaporewww.focusonmicroscopy.org

9-13 MRS 2012 Spring Meeting and Exhibition, San Francisco, CA, USAwww.mrs.org/spring2012

17-20 Analytica 2012: 23rd International Trade Fair for Laboratory Technology, Analysis and Biotechnology and analytica Conference, Munich Trade Fair Centre, Munich, Germanywww.analytica.de

MAY 20129-11 AMTC3: 3rd International Symposium on Advanced Microscopy and

Theoretical Calculations, Nagaragawa Convention Center, Japanwww.congre.co.jp/amtc3/

21-25 IFES2012: 53rd International Field Emission Symposium, University of Alabama, Tuscaloosa, AL, USAwww.ifes2012.au.edu

24-25 SCUR 2012: 39th Annual Meeting of Society for Cutaneous Ultrastructure Research, Lyon, Francehttp://orgs.dermis.net/content/e04scur/e03meetings/e770/e1074/index_ger.html

JUNE 20123-15 Microscopy School, Lehigh University, Bethlehem, PA, USA

Sharon Coe: sharon.coe@lehigh.edu www.lehigh.edu/microscopy

3 Introduction to SEM and EDS for the New Operator4-8 SEM and X-Ray Microanalysis11-14 Focused Ion Beam: Instrumentation and Applications11-15 Problem Solving with SEM, X-Ray Microanalysis and Electron Backscatter Patterns11-15 Quantitative X-Ray Microanalysis: Problem Solving with EDS and WDS Techniques11-15 Scanning Transmission Electron Microscopy: From Fundamentals to Advanced Applications

25-27 Abercrombie Cell Biology Symposium, Oxford, UKBSCB: www.bscb.org

27-30 CARS 2012: Computer Assisted Radiology and Surgery, 26th Intl Congress and Exhibition, Joint Congress of CAR / ISCAS / CAD / CMI / EuroPACS, Congress Palace, Pisa, Italywww.cars-int.org

JULY 20122-6 Electron Microscopy Summer School, University of Leeds, UK

RMS: www.rms.org/events 2-6 Light Microscopy Summer School, University of York, UK

RMS: www.rms.org/events 4-6 Optics Within Life Sciences (OWLS), Genoa, Italy

Organised by Italian Institute of Technology www.owls2012.org9-13 Inter/Micro: 63rd Annual Applied Microscopy Conference, Chicago, IL,

USAwww.mcri.org

11-13 CryoElectron Microscopy Short Course, University of Minnesota, MN, USANanostructural Materials and Processes Program of IPrime: mccormic@umn.edu www.iprime.umn.edu

29-2/8 Microscopy & Microanalysis 2012, Phoenix, Arizona, USAwww.microscopy.org

AUGUST 20126-10 ULTRAPATH XVI: Conference on Diagnostic Electron Microscopy, Basic

Research and Oncology, Regensburg, Germanywww.ultrapath.org

26-30 14th International Congress of Histochemistry and Cytochemistry, Kyoto, Japan www.acplan.jp/ichc2012/

SEPTEMBER 2012 16-21 European Microscopy Congress, Central Convention Complex,

Manchester, UKwww.rms.org.uk

OCTOBER 201213-17 Neuroscience 2012, New Orleans, Louisiana, USA

www.sfn.org

NOVEMBER 201226-30 2012 MRS Fall Meeting & Exhibition, Boston, Massachusetts, USA

www.mrs.org/fall2012

DECEMBER 201215-19 ASCB 52nd Annual Meeting, San Francisco, CA, USA

www.ascb.org

MARCH 201317-21 Pittcon 2013, Philadelphia, PA, USA

www.pittcon.org

APRIL 20131-5 2013 MRS Spring Meeting & Exhibition, San Francisco, CA, USA

www.mrs.org/spring2013

AUGUST 20134-8 Microscopy & Microanalysis 2013, Indianapolis, IN, USA

www.microscopy.org

NOVEMBER 20139-13 Neuroscience, San Diego, California, USA

www.sfn.org

LITERATURE HIGHL IGHTS

literature highlights�����������"�� �'!��� �������� ���"���

A large array of sub-10-nm sin-gle-grain gold nanodots for usein nanotechnology is describedby Nicolas Clément and col-leagues at the Institut d’Elec-tronique Microélectronique etNanotechnologie, CNRS, Univer-sity of Lille, France [Small7(18):2607-2613, 2011].A uniform array of single-

grain gold nanodots, as small as5-8 nm, was be formed on siliconusing e-beam lithography. Theas-fabricated nanodots wereamorphous, and thermal annealing converted themto pure Au single crystals covered with a thin SiO2layer. These findings were based on physical mea-surements by AFM, atomic-resolution STEM, andchemical techniques using energy dispersive X-rayspectroscopy. The authors demonstrated the forma-tion by e-beam lithography of sub-10-nm Au dotswith small dispersion and perfect alignment. Suchprecise formation of small dots enabled them to iden-

(a) STEM image showing the bulk silicon (Si), five annealed dots (Au), carbon layer (C), and platinum layers. (c) Coloured STEM image of a single annealed nanodot (260°C, 2 h). Reproduced with permission, Copyright ©2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

tify the critical size that determines whether a dot iscomposed of single or multiple crystal domains.Moreover, they showed that annealing at moderatetemperature can convert Au dots from amorphous tosingle-crystalline, and then they were covered with athin SiO2 layer. After easy removal of the SiO2 (diluteHF etching), these nanodots can be used as electrodesfor the characterization of organic self-assembledmonolayers (SAMs) with less than 200 molecules.

���"������ ���� ���&�����#� �!����������� ���'A miniature stage device to overcome resolutionanisotropy in fluorescence light microscopy isdescribed by Florian Staier and colleagues at theKirchhoff Institute for Physics, University of Heidel-berg, Germany [Rev. Sci. Instrum. 82:093701, 2011].To overcome the limitation of fluorescence micro-

scopes in anisotropic optical resolution or point local-ization precision micro-axial tomography was usedwhich allowed object tilting on the microscope stageand led to an improvement in localization precisionand spatial resolution. A glass fiber was placed in theobject space of the microscope lens and its rotationcontrolled by a miniaturized stepping motor. By Testparticles were fixed onto the glass fiber, opticallylocalized with high precision, and automaticallyrotated to obtain views from different perspectiveangles from which distances of corresponding pairs ofobjects were determined. From these angle depen-dent distance values, the real 3D distance was calcu-lated with a precision in the ten nanometer range(corresponding here to an optical resolution of 10-30nm) using standard microscopical equipment. As aproof of concept, the spindle apparatus of a maturemouse oocyte was imaged during metaphase II mei-otic arrest under different perspectives.

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A technique for the use of two fluorophores in stim-ulated emission depletion (STED) microscopy of livingcells is reported by Patrina Pellett and co-workers atthe Department of Cell Biology, Yale School of Medi-cine, CT [Biomedical Optics Express 2(8):2364-2371,2011]. Current applications of STED microscopy havebeen limited to single colour imaging of living cellsand multicolour imaging in fixed cells. However, tostudy active processes, such as protein interactions, atwo-colour STED imaging technique is needed in liv-ing cells. This was achieved for the first time by theauthors: the key to their success was in overcomingthe challenges in labeling target proteins in livingcells with dyes optimal for two-colour STEDmicroscopy. By incorporating fusion proteins, theresearchers were able to improve the targetingbetween the protein and the dye, effectively bridgingthe gap. This allowed the researchers to achieve reso-lutions of 78 nm and 82 nm for 22 sequential two-colour scans of epidermal growth factor and its recep-tor in living cells.

��� ���"��� �����#��"��������!��� ��"�������The three-dimensional point spread function of anaberration-corrected scanning transmission electronmicroscopy (STEM) has been simulated and experi-mentally tested by Andrew Lupini and Niels de Jongeat the Oak Ridge National Laboratory, TN [Microscopyand Microanalysis 17:817-826, 2011].Aberration correction reduces the depth of field in

STEM and thus allows three-dimensional imaging bydepth sectioning. This imaging mode offers thepotential for sub-Ångstrom lateral resolution andnanometer-scale depth sensitivity. For biological sam-ples, which may be many µm across and where highlateral resolution may not always be needed, opti-mizing the depth resolution even at the expense oflateral resolution may be desired, aiming to imagethrough thick specimens. Although there has beenextensive work examining and optimizing the probeformation in two dimensions, there is less knownabout the probe shape along the optical axis. The authors examined the probe shape in 3D in an

attempt to better understand the depth resolution in

���������'!�!����������"�������"����������"!A fractal dimension analysis and mathematical mor-phology of structural changes in actin filamentsimaged by electron microscopy is reported by Yoshi-taka Kimori et al at the National Institutes of NaturalSciences, in Tokyo [Journal of Structural Biology176(1):1-8, Oct 2011].The authors examined structural changes of actin

filaments interacting with myosin visualized by quickfreeze deep-etch replica EM by using a new methodof image processing and analysis based on mathe-matical morphology. To quantify the degree of struc-tural changes, two characteristic patterns wereextracted from the EM images: the winding patternof the filament shape (WP) reflecting flexibility of the

�"��� ����������������!����#��"#������!�A. Beltrán and colleagues at the Department of Mate-rials Sciences, University of Cadiz, Spain, report thatthree dimensional atom tomography resolves thequantum ring morphology of self-assembled GaSbburied nanostructures [Ultramicroscopy 111(8):1073-1076, July 2011].Unambiguous evidence of ring-shaped self-assem-

bled GaSb nanostructures grown by molecular beamepitaxy is presented on the basis of atom-probetomography reconstructions and darkfield transmis-sion electron microscopy imaging. From atom-probetomography compositional distribution has beenobtained. The GaAs capping process causes a strongsegregation of Sb out of the center of GaSb quantumdots, leading to the self-assembled GaAsxSb1-x quan-tum rings of 20-30 nm in diameter with x~0.33.

filament, and the surface pattern of the filament (SP)reflecting intramolecular domain mobility of the actinmonomers in the filament. EM images were processedby morphological filtering followed by box-countingto calculate the fractal dimensions for WP (DWP) andSP (DSP). The result indicated that DWP was largerthan DSP irrespective of the state of the filament(myosin-free or bound) and that both parameters formyosin-bound filaments were significantly largerthan those for myosin-free filaments. This work is thefirst quantitative insight into how conformational dis-order of actin monomers is correlated with themyosin-induced increase in flexibility of actin fila-ments along their length.

this mode. They present examples of how aberrationschange the probe shape in three dimensions, and itwas found that off-axial aberrations may need to beconsidered for focal series of large areas. It was shownthat oversized or annular apertures theoreticallyimprove the vertical resolution for three-dimensionalimaging of nanoparticles. When imaging nanoparti-cles of several nanometers in size, regular scanningtransmission electron microscopy can thereby be opti-mized such that the vertical full-width at half-maxi-mum approaches that of the aberration-correctedSTEM with a standard aperture.

MICROSCOPY AND ANALYSIS NOVEMBER 201126

CIRC

LENO. 1 O

RONLINEwww.microscop

y-analysis.com

The cover shows a confocal image (top) of aDrosophila embryo at stage 11, expressing the trachealmarker trh-LacZ (Cy3, red) and the cell membranemarker Dlg (Alexa 488, green). The enlarged view(below) shows invaginating tracheal placodes in X-Y(left) and Y-Z (right) projections. (Courtesy of DrTakefumi Kondo and Dr Shigeo Hayashi at theLaboratory for Morphogenetic Signalling, RIKEN Centrefor Developmental Biology, Japan.)

The new UPLSAPO30xS and UPLSAPO60xS siliconoil-immersion objectives from Olympus are ideal for livecell experiments investigating thick samples or requiringlong-term imaging. Users can generate bright images athigher resolutions, since silicon oil significantly improvesoptical performance compared to water-based methodsand offers larger working distances than oil immersionobjectives. This maximises the effectiveness ofbrightfield, differential interference contrast,fluorescence, confocal laser scanning and multiphotonstudies. Silicon oil allows users to capture bright, highresolution images of living samples, even deep into cellsand tissues. This can be achieved because the refractiveindex of silicon oil (n = 1.40) is almost identical to thatof living biological samples (n =1.38 on average) sosilicon oil-immersion objectives minimise refractive indexmismatch and spherical aberration. This is a key factorfor improving the focal spot, which can be even furtheroptimised to correct for temperature changes byadjusting the available correction collar. The propertiesof silicon oil also make it ideal for long-term, time-lapsestudies since it is stable, viscous and of low volatility; itwill not dry out or absorb moisture over time.

With its 30x magnification and high NA of 1.05, theUPLSAPO30xS delivers highly resolved images of anextensive sample area. The Olympus UPSLAPO60xSprovides increased magnification (60x) and NA (1.30),allowing researchers to produce highly detailed livesample images using fluorescence, confocal laserscanning and multiphoton excitation techniques. Thisobjective is also ideal for high resolution 3D imagingapplications by offering improved spherical aberrationcorrection and a shorter working distance of 0.3 mm.

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15 Fracture Surface, Impact Energy and Hardness of Ni-FreeHigh-Mn Steels W Sha, H Haji Talib, E Wilson, R Rajendran,S Malinov, H Charlesworth, L Ibbitson

21 Variable Pressure Scanning Electron Microscopy of Viciafaba Stigmatic Papillae W Chen, F Stoddard and T Baldwin

S5 Atomic Resolution Secondary Electron Imaging in AberrationCorrected STEM H Inada, M Konno, K Tamura,Y Suzuki, K Nakamura, Y Zhu

S11 Development of a High Throughput Electron Microscope forNanoscale Analysis M Matsushita, S Kawai, T Iwama, K Tanaka,T Kuba, N Endo, T Isabell

S19 Low Beam-Energy Energy-Dispersive X-Ray Spectroscopy forNanotechnology P Camus

S23 Product Focus - Nanotechnology and Electron Microscopy

C O V E R S T O R Y

MICROSCOPY & ANALYSISISSN 0958-1952 - UKISSN 2043-0655 - Europe© 2011 John Wiley & Sons, LtdIssued in: January, March, May, July, September, NovemberPublished by: John Wiley & Sons, Ltd, The Atrium, Southern Gate, Chichester, West Sussex PO19 8SQ, UK Tel: +44 (0)1243 770257 Fax: +44 (0)1243 770432Email: editor@microscopy-analysis.com Website: www.microscopy-analysis.com

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EDITORIAL BOARDJim Bentley, Oak Ridge National Lab, TN, USAEd Boyes, University of York, UKRay Carpenter, Arizona State University, AZ, USAChristian Colliex, CNRS Lab.de Physique des Solides, FranceAlby Diaspro, University of Genoa, ItalyPeter Hawkes, CNRS, Toulouse, FranceColin Humphreys, University of Cambridge, UKCornelis van Noorden, University of Amsterdam, The NetherlandsJim Pawley, University of Wisconsin, WI, USAJohn Spence, Arizona State University, AZ, USANestor Zaluzec, Argonne National Lab, IL, USA

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See the Microscopy and Analysis online Diary for a full listing. Entries for the January issue are due 1 December. Email to: editor@microscopy-analysis.com

dates for the diaryDIARY

MICROSCOPY AND ANALYSIS NOVEMBER 20114

NOVEMBER 20118-11 Gatan EELS School, Austrian Centre for Electron Microscopy and

Nanoanalysis, TU Graz, Austriawww.felmi-zfe.tugraz.at www.gatan.com/company/news/news08291102.php

10 Electron Microscopy in Materials Science, Eindhoven University of Technology, The Netherlandshttp://nvvmmaterials2011.chem.tue.nl/

12-16 Neuroscience 2011: Washington DC, USAwww.sfn.org

14-18 NvVM European School on Nanobeams , Centre de Recherche Public - Gabriel Lippmann, Belvaux, Luxembourg www.nanobeams.org

16 Cryo Microscopy Group Annual Meeting, Boots Science Building, School of Pharmacy, University Park, Nottingham University, UKwww.cryomicroscopygroup.org.uk/CMG2011.html

22-23 Bruker Scanning Probe Microscopy Conference and Users’ Meeting, University of Manchester, UKwww.bruker-axs.com drew.murray@bruker-nano.com

28-2/12 2011 MRS Fall Meeting, Boston, Massachusetts, USAwww.mrs.org

DECEMBER 20113-4 Functional Optical Imaging, Univeristy of Nottingham Ningbo Campus,

Chinawww.nottingham.ac.uk/ibios/index.php?page=foi-11

3-7 American Society for Cell Biology 51st Annual Meeting, Denver, Colorado, USAwww.ascb.org

JANUARY 20124-7 Winter School on High-Resolution Electron Microscopy, Arizona State

University, Tempe, AZ, USAwintershcool@asu.edu http://le-csss.asu.edu/winterschool

FEBRUARY 20125-9 APMC 10: 10th Asia-Pacific Microscopy Conference;

ICONN 2012: International Conference on Nanoscience and Nanotechnology;

ACMM 22: Australian Conference on Microscopy and Microanalysis, Perth, Australiawww.apmc-10.org www.iconn-2012.org www.acmm-22.org

25-29 56th Biophysical Society Annual Meeting, San Diego, California, USAwww.biophysics.org/2012meeting/Main/tabid/2386/Default.aspx

MARCH 201211-16 Pittcon 2012, Orlando, FL, USA

www.pittcon.org18-23 Course in Cryotechniques for Electron Microscopy, Rothamsted

Research, Harpenden, UKwww.rms.org.uk/events/Forthcoming_Events/CoolRunnings

APRIL 20121-4 Focus on Microscopy 2012; 25th International Conference on 3D Image

Processing in Microscopy; 24th International Conference on Confocal Microscopy, Singaporewww.focusonmicroscopy.org

9-13 MRS 2012 Spring Meeting and Exhibition, San Francisco, CA, USAwww.mrs.org/spring2012

17-20 Analytica 2012: 23rd International Trade Fair for Laboratory Technology, Analysis and Biotechnology and analytica Conference, Munich Trade Fair Centre, Munich, Germanywww.analytica.de

MAY 20129-11 AMTC3: 3rd International Symposium on Advanced Microscopy and

Theoretical Calculations, Nagaragawa Convention Center, Japanwww.congre.co.jp/amtc3/

21-25 IFES2012: 53rd International Field Emission Symposium, University of Alabama, Tuscaloosa, AL, USAwww.ifes2012.au.edu

24-25 SCUR 2012: 39th Annual Meeting of Society for Cutaneous Ultrastructure Research, Lyon, Francehttp://orgs.dermis.net/content/e04scur/e03meetings/e770/e1074/index_ger.html

JUNE 20123-15 Microscopy School, Lehigh University, Bethlehem, PA, USA

Sharon Coe: sharon.coe@lehigh.edu www.lehigh.edu/microscopy

3 Introduction to SEM and EDS for the New Operator4-8 SEM and X-Ray Microanalysis11-14 Focused Ion Beam: Instrumentation and Applications11-15 Problem Solving with SEM, X-Ray Microanalysis and Electron Backscatter Patterns11-15 Quantitative X-Ray Microanalysis: Problem Solving with EDS and WDS Techniques11-15 Scanning Transmission Electron Microscopy: From Fundamentals to Advanced Applications

25-27 Abercrombie Cell Biology Symposium, Oxford, UKBSCB: www.bscb.org

27-30 CARS 2012: Computer Assisted Radiology and Surgery, 26th Intl Congress and Exhibition, Joint Congress of CAR / ISCAS / CAD / CMI / EuroPACS, Congress Palace, Pisa, Italywww.cars-int.org

JULY 20122-6 Electron Microscopy Summer School, University of Leeds, UK

RMS: www.rms.org/events 2-6 Light Microscopy Summer School, University of York, UK

RMS: www.rms.org/events 4-6 Optics Within Life Sciences (OWLS), Genoa, Italy

Organised by Italian Institute of Technology www.owls2012.org9-13 Inter/Micro: 63rd Annual Applied Microscopy Conference, Chicago, IL,

USAwww.mcri.org

11-13 CryoElectron Microscopy Short Course, University of Minnesota, MN, USANanostructural Materials and Processes Program of IPrime: mccormic@umn.edu www.iprime.umn.edu

29-2/8 Microscopy & Microanalysis 2012, Phoenix, Arizona, USAwww.microscopy.org

AUGUST 20126-10 ULTRAPATH XVI: Conference on Diagnostic Electron Microscopy, Basic

Research and Oncology, Regensburg, Germanywww.ultrapath.org

26-30 14th International Congress of Histochemistry and Cytochemistry, Kyoto, Japan www.acplan.jp/ichc2012/

SEPTEMBER 2012 16-21 European Microscopy Congress, Central Convention Complex,

Manchester, UKwww.rms.org.uk

OCTOBER 201213-17 Neuroscience 2012, New Orleans, Louisiana, USA

www.sfn.org

NOVEMBER 201226-30 2012 MRS Fall Meeting & Exhibition, Boston, Massachusetts, USA

www.mrs.org/fall2012

DECEMBER 201215-19 ASCB 52nd Annual Meeting, San Francisco, CA, USA

www.ascb.org

MARCH 201317-21 Pittcon 2013, Philadelphia, PA, USA

www.pittcon.org

APRIL 20131-5 2013 MRS Spring Meeting & Exhibition, San Francisco, CA, USA

www.mrs.org/spring2013

AUGUST 20134-8 Microscopy & Microanalysis 2013, Indianapolis, IN, USA

www.microscopy.org

NOVEMBER 20139-13 Neuroscience, San Diego, California, USA

www.sfn.org

CIRCLE NO. 3 OR ONLINE: www.microscopy-analysis.com

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Top left: Dislocations and growth zonation revealed in free standing GaN wafer by panchromatic imaging.Top right: Surface plasmon resonance modes along vertices and at apex of pyramid structures revealed using monochromatic imaging. Pyramids formed by 200nm Au film deposited on patterned substrate. Overlay of secondary electron image (grey) and, 650nm (green) and 750nm (red) monochromatic images. Bottom left: Irregularity in emission of GaAs nanowires.  In-lens secondary electron image overlaid with panchromatic CL image (green); images acquired simultaneously.Bottom right: Four AlGaN concentrations revealed in graded AlxGa1-xN on GaN film using spectrum-linescan of sample cross section. Blue shift in spectrum from 365 to 320nm associated with change in Al fraction. Spectra acquired at rate of 20 spectra/s.

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Top: A 3D reconstruction of a dendrite from a 15,625 μm³ (25 x 25 x 25 μm) volumetric data set containing 500 serial images of mouse cerebellum generated by Gatan 3View®. Dendrite structure (green), buttons (yellow), and vesicles (red). Bottom Left: Confocal image of a dendrite. Middle left: 3View® image showing wire frame traces. Middle right: Wire frame traces rendered into a volumetric model. Bottom right: Ultra resolution dendritic spine model with synapses. Sample courtesy of Tom Deerinck and Dr. Mark Ellisman, National Center for Microscopy and Imaging Research, University of California, San Diego. Serial images were segmented using Imaris to create a 3D model of a neuron of interest.

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ANALYTICAL TEMDIGITAL IMAGINGSPECIMEN PREPARATIONTEM SPECIMEN HOLDERSSEM PRODUCTSSOFTWAREX-RAY MICROSCOPY

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Top left: Dislocations and growth zonation revealed in free standing GaN wafer by panchromatic imaging.Top right: Surface plasmon resonance modes along vertices and at apex of pyramid structures revealed using monochromatic imaging. Pyramids formed by 200nm Au film deposited on patterned substrate. Overlay of secondary electron image (grey) and, 650nm (green) and 750nm (red) monochromatic images. Bottom left: Irregularity in emission of GaAs nanowires.  In-lens secondary electron image overlaid with panchromatic CL image (green); images acquired simultaneously.Bottom right: Four AlGaN concentrations revealed in graded AlxGa1-xN on GaN film using spectrum-linescan of sample cross section. Blue shift in spectrum from 365 to 320nm associated with change in Al fraction. Spectra acquired at rate of 20 spectra/s.

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solutions for the fabrication of high value-added heterogeneous components and sys-tems including memories, logic, sensors, actu-ators and wireless communications. Partici-pants in the program include materialproviders, laboratories, research centers andmanufacturers of equipment, componentsand systems.The project is structured around five themes:· Methodology and evaluation tools to inte-

grate elementary components in 3D systems.· 3D technologies and integration processesonto materials and non-silicon substrates.· 3D technologies and integration processesonto silicon, using processes closely derivingfrom microelectronics.· Reliability methodologies and analysis forintegrated 3D systems.· Performance evaluation and equipment val-idation for volume production equipment andgeneric manufacturing.SINTEF, The Fraunhofer Institute and FEI are

participants in JEMSiP_3D, and the work pre-sented here was funded in part by the project.

3 D I N T E G R AT I O NWhile the performance and productivity ofmicroelectronics have increased continuouslyover more than four decades due to the enor-mous advances in lithography and device tech-nology, it has now become questionable ifadvances in these areas alone will be able to

B I O G R A P H YMaaike M. V. Takloreceived her PhD in phys-ical electronics from theUniversity of Oslo in 2002for her thesis entitled‘Wafer bonding forMEMS’. From 1998 until2010 she was employed at the Departmentof Microsystems and Nanotechnologywithin SINTEF ICT in Norway where sheworked on MEMS processing and wasresponsible for their wafer level bondingactivities. Maaike is now a senior scientist atSINTEF ICT at the Department of Instrumen-tation and is the group leader for ‘AdvancedPackaging and Interconnects’.

A B S T R A C T3D integration schemes connect stackedintegrated circuits using through silicon vias(TSV) and special bonding techniques. Phys-ical characterization of these TSVs andbonds is essential, but their relatively largesize (tens or hundreds of micrometers)requires prohibitively long milling times inthe conventional focused ion beam (FIB) sys-tems typically used for this work. A newplasma-based FIB system can remove mater-ial more than 20 times faster, providing thespeed and precision required to ensurerobust processes and reliable products.

K E Y W O R D Sfocused ion beam, scanning electronmicroscopy, plasma ion source, ion beammilling, 3D integration, through silicon vias

A C K N O W L E D G E M E N T SA part of this work has been performed inthe project JEMSiP_3D, which is funded bythe public authorities in France, Germany,Hungary, The Netherlands, Norway andSweden, as well as by the ENIAC JointUndertaking.

A U T H O R D E TA I L SDr Maaike M. Visser Taklo, SINTEF ICT, Department of Instrumentation,PO Box 124 Blindern, N-0314 Oslo, NorwayTel: +47 2206 7300Email: MaaikeMargrete.VisserTaklo@sintef.nowww.sintef.no

Microscopy and Analysis 25(7):9-12 (EU), 2011

PFIB IN MICROELECTRONICS

I N T R O D U C T I O N3D integration schemes, which stack inte-grated circuits and other microelectronic orMEMS devices and interconnect them usingthrough silicon vias (TSV), are likely to be thenext revolution in electronic fabrication. Theycan be used to continue the increases in speedand density of microelectronic systemsdescribed by Moore’s Law (More Moore), butthey may offer even greater benefits whenused to connect devices of different technolo-gies (More than Moore), packing more perfor-mance and functionality into smaller volumes. In either case, the ability to physically char-

acterize the TSVs and mechanical bonds usedin 3D integration is essential for developingrobust manufacturing processes and fabricat-ing reliable products. Focused ion beam (FIB)systems have long provided physical analysisin the manufacture of integrated circuits, butconventional FIB cannot remove material fastenough to analyze these relatively large struc-tures used in 3D integration. The launch of anew plasma-based FIB system now providesthe speed and precision needed to developand deploy these exciting new technologies.

J E M S i P _ 3 DThe Joint Equipment and Materials for System-in-Package and 3D Integration (JEMSiP_ 3D) isa project undertaken by a consortium of Euro-pean manufacturers to validate technological

Bonding and TSV in 3D IC Integration:Physical Analysis with a Plasma FIBMaaike M. V. Taklo,1 Armin Klumpp,2 Peter Ramm,2 Laurens Kwakman3 and German Franz 3

1. SINTEF, Oslo, Norway. 2. Fraunhofer EMFT, Munich, Germany. 3. FEI Company, Eindhoven, The Netherlands

Figure 1a: Schematic of a xenon plasma focused ion beam (PFIB) system. A PFIB uses an inductively coupled plasma to deliver high beam current. The source islarger than a liquid metal ion source (LIMS), but delivers a more collimated beam, enabling better beam spot performance at high beam currents.

MICROSCOPY AND ANALYSIS NOVEMBER 2011 9

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overcome the predicted performance and costproblems of future IC fabrication. The ITRSroadmap predicts 3D integration as a key tech-nology to overcome this so-called ‘wiring crisis’and the solution will most likely be based onTSV technology.

The most promising 3D integration schemescurrently under consideration involve the ver-tical stacking of integrated circuits and otherdevices. These schemes vary in their details butall must solve two central problems: how tobond the integrated layers together and howto create electrical connections among them.Bonding and TSV technologies each have theirown unique set of considerations which oftencenter around how the structure will hold upduring subsequent processing, such as theaddition of another layer:·Will the stresses induced by additional ther-mal processing cause debonding or shifting ofthe existing bonds? · Will the stress and strain cause cracks ordelamination in the TSVs? · What are the best materials and processesto use to minimize these negative effects?

P LASMA FOCUSED ION BEAMFIB systems, which use an ion beam to cut andimage cross sections through subsurface struc-tures with nanoscale precision and imagingresolution, have long been a mainstay of phys-ical analysis for integrated circuits. Althoughthe structures used in 3D integration can beexpected to decrease in size as the technolo-gies evolve, they are much larger than thedimensions of the transistors and intercon-nects used in current integrated circuits, andthe cutting speed of FIBs designed for ICs isgenerally inadequate for TSVs and bondingstructures. A typical 10 µm � 10 µm IC cross-section requires the removal of 1000 µm3 ofmaterial and takes a few minutes. A 100 µm �100 µm TSV cross-section requires the removalof 1,000,000 µm3 of material and would takemost of a day with conventional FIB.

The Vion PFIB system (FEI Company, Hills-boro, Oregon, USA) uses an inductively cou-pled plasma source [1-3] (Figure 1) to providematerial removal rates 20� faster than con-ventional FIBs that use liquid metal ion sources

(LMIS). A LMIS is essentially a point source 50nm in diameter with a low angular intensity.The Vion system’s plasma source is larger, 15µm, but has a much higher angular intensity.Because of its small virtual size, the LMIS is easyto focus into a small spot at low beam currents,but at beam currents above 10 nA sphericalaberration effects severely degrade perfor-mance. The plasma source can deliver currentsin excess of a µA (>20� greater than a typicalLMIS based system) while still maintaining awell focused beam. Since material removalrates are primarily a function of beam current,the PFIB has an advantage of 20� or moreover conventional FIB at high currents, whilestill preserving excellent milling precision andimaging resolution at low beam currents.

The xenon ion beam emitted by the plasmasource has high sputtering yield, high bright-ness and low energy spread. In addition, byintroducing various gases, the PFIB can selec-tively etch specific materials or deposit pat-terned conductors and insulators (similar toconventional FIB systems). The plasma sourcealso offers the potential to use different ionspecies to enhance performance in specificapplications.

CURTA IN INGThe difference in FIB milling rates of the vari-ous materials present in a device (Cu, Si, Sn,dielectrics, polyimides and mold compounds)can cause ‘curtaining’ when milling cross-sec-tions. This milling artifact can make detailed

Figure 2: Curtaining artifacts (upper left), caused by variations in milling rate for different materials, can be effectively suppressed (right) by rocking the sampleto mill in a sequence of alternating angles (lower left).

MICROSCOPY AND ANALYSIS NOVEMBER 201110

Figure 1b: The PFIB maintains excellent spot size performance over a broad range of beam currents.

Figure 1c: At high beam currents the PFIB can remove material twenty times faster than a liquid metal ion source.

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PFIB I N MI C R O E L E C T R O N I C S

analysis of the structures difficult or evenimpossible.

Figure 2 shows typical curtaining effects onthe silicon substrate as well as the TSV itself,caused by milling through the overlying roughpoly crystalline metal film. These curtainingeffects can be effectively suppressed by rock-ing the sample during the FIB milling process.Milling in a sequence of alternating incidenceangles creates a clean cross section free of cur-taining artifacts without the need for time-consuming low current polish steps.

EXAMPLES OF APPL ICATIONS OFPLASMA FOCUSED ION BEAM

Through Silicon ViasTSVs are themselves subject to a number ofeffects that can result in defects and failures.For example, the large differences in thermalexpansion between copper via fill and the sur-rounding silicon substrate can cause crackingwithin the copper and delamination from thevia sidewall during thermal processing. ‘Key-holing’ results from incomplete filling of vias(Figure 3).

Solid Liquid Interdiffusion BondingOne of the most difficult issues to address isthe behavior of bonds between chips duringsubsequent processes (Figure 4). For example,it is critical that a bond between the first chipsin the stack not be disturbed by the subse-quent bonding of an additional chip. Solid-liquid-interdiffusion (SLID) [4] is a uniquedirect metal bonding technology that avoidsremelting of existing bonds during the forma-tion of new bonds by using high melting inter-metallic phases. During bond formation, solidmetal diffuses into the liquid phase of a lowermelting metal resulting in high melting pointfinal phase that remains solid during subse-quent bond forming processes.

Anisotropic Conductive AdhesivesAnisotropic conductive adhesives (ACA) can beused [5] to bond wafers together physicallyand electrically using an organic bonding com-pound (benzocyclobutene, BCB) filled with 4-µm sized metal covered polymer spheres(MPS). The BCB assures mechanical strengthwhereas the MPS provide the required electri-cal conductivity at interconnection points. Theconcentration of MPS must be high enough toensure good electrical contact betweenopposed pads and at the same time lowenough to guarantee electrical insulationwhere pads are not present.

To study the bonding in detail, samples werecleaved, then milled with the plasma-FIB toreveal the bonding region and finallyinspected with plasma-FIB imaging. Theplasma-FIB milling speed makes it possible toprepare the sample (~200 � 50 � 600 µm3

material removed) within 30 minutes. Themetal layer covering the polymer spherescould be observed at the bonding interfacewith sufficient resolution to estimate both thelocal MPS density and their compression statebetween the bond pad metal layers. In Figure5 the bonding process is illustrated in the top

Figure 4: The void between these pads is the result of an incomplete SLID bonding process. The various intermetallic phases are clearly visible above, below andto the right of the void.

MICROSCOPY AND ANALYSIS NOVEMBER 2011 11

Figure 3: (a, b) Differing thermal expansionbetween copper via fill and silicon substrate caused delamination shownin this via before (a) and after (b)annealing. (c) Keyholing occurred when this viawas not filled completely with tung-sten.

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MICROSCOPY AND ANALYSIS NOVEMBER 201112

images and the bottom plasma-FIB imagesshow details of the bonding interface andcompressed spheres.

3D Test ChipThe 3D integrated reliability test chip shown inFigure 6 is a 3-level-stack with a modular lay-out designed to permit evaluation of assemblyprocesses between two initial layers and, sub-sequently, the effects of adding a third layer[6]. The PFIB can mill a cross section throughthe entire three layer stack showing criticaldetails of both upper and lower bondingregions and the complete TSV through themiddle layer.

CONCLUS IONSBy combining high-speed milling and deposi-tion with precise control and high qualityimaging, the plasma focused ion beam pro-vides critically needed physical analysis for TSVand bonding processes that are essential tocurrent 3D integration schemes. At high beamcurrents, cross-sections with dimensions ofhundreds of micrometers can be completed inless than an hour, fast enough to provideeffective feedback on process performance. Atlow beam currents, the same system delivershigh resolution imaging for accurate structuralanalysis.

The PFIB provides an effective, practical toolfor a variety of 3D integration applications,including failure analysis of bumps, wirebonds, TSVs, and stacked die; site specificremoval of package and other materials toenable failure analysis and fault isolation onburied die; circuit and package modificationsto test design changes without repeating thefabrication process or creating new masks;process monitoring and development at thepackage level; and defect analysis of packagedparts and MEMS devices.

REFERENCES1. Smith, N. S., Skoczylas, W. P., Kellogg, S.M., Kinion, D.E.,

Tesch, P.P., Sutherland, O., Aanesland, A., Boswell, R.W. HighBrightness Inductively Coupled Plasma Source for HighCurrent Focused Ion Beam Applications. J. Vac. Sci. Technol.B24(6):2902-2906, 2006.

2. Kellogg, M., Schampers, R., Zhang, S.Y., Graupera, A.A.,Miller, T., Laur, W.D., Dirriwachter, A.B. High ThroughputSample Preparation and Analysis using an InductivelyCoupled Plasma (ICP) Focused Ion Beam Source. Microsc.Microanal. 16(Suppl 2):222-223, 2010.

3. Kwakman, L., Franz, G., Taklo, M. M. V., Klumpp, A., Ramm,P. Characterization and Failure Analysis of 3D IntegratedSystems using a novel plasma-FIB system. Proc. InternationalConference on Frontiers of Characterization and Metrologyfor Nanoelectronics, Grenoble, France, 2011.

4. Ramm, P. Method of making a three-dimensional integratedcircuit. US Patent 5,563,084; P. Ramm, A. Klumpp. Methodof vertically integrating microelectronic components. USPatent 6,548,391.

5. Taklo, M. et al. Anisotropic Conductive Adhesive for Wafer-to-Wafer Bonding, Proceedings of 7th Intl Conference andExhibition on Device Packaging, March 2011.

6. Ramm, P., Klumpp, A., Franz, G., Kwakman, L. FailureAnalysis and Reliability of 3D Integrated Systems. Proc.IMAPS Device Packaging Conf., Scottsdale, Arizona, 2011.

©2011 John Wiley & Sons, Ltd

Figure 6: The high milling speed of PFIB permits cross-sections through the full three layer stack of the test chip, revealing both upper and lower bonding regionsand the entire TSV.

Figure 5: Anisotropic conductive adhesives provide mechanical bonding and electrical conductivity. (a,b) Images show metal coated spheres before mixing withBCB (a), and a schematic of how TSVs can be electrically connected to pads on another wafer using BCB filled with such spheres (b). (c-f) The lower four images are a clockwise sequence of increasing magnification with the compressed metal coated spheres clearly visible in the twobottom images.

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site and austenite, as measured by opticalmicroscopy. Nikbakht et al. [7] showed that astabilized alloy 193 (Fe-8%Mn with 0.17%Ti,0.18%Al and 0.018%C) exhibited brittle cleav-age on ice brine quenching from 900°C, basedon clear fracture surface examination. This is inagreement with a much earlier but convincingfracture profile analysis [8] and thermody-namic calculations on alloy 193 showed thatthere is less than 0.03 ppm N in solid solution.On tempering alloy 193 for 6 min at 450°C, theDBTT rose from 27 to 125°C, giving intergran-ular failure. This indicated that embrittlementis due to segregation of Mn per se to prioraustenite grain boundaries.In the last five years, there have been many

creditable studies in related subjects. Forexample, Vaynman et al. [9] have found thatthe increase in strength in a low-carbon, Fe-Cu–based steel was derived from a large num-ber density of copper-iron-nickel-aluminum-manganese precipitates, characterized bystate-of-the-art atom-probe tomography.Sathiya et al. [10] found using cross-sectionimaging that the shape of the fusion zone,generally characterized by a few geometricalfeatures, namely bead width, bead height anddepth of penetration, depended upon a num-ber of parameters such as gas flow rate, volt-age, travel speed and wire feed rate. Recentresearch in ferrous materials in general hasbeen well reviewed in a recent book by Bernsand Theisen [11]. However, there has been lit-tle research reported on the steels studiedhere.

MATER IALS AND METHODS

SteelsIn the present research, two cryogenic steelswith around 20% manganese were investi-gated: Fe-19.7%Mn (VM339A) and Fe-19.7%Mn stabilized with 0.056%C, 0.19%Ti

B IOGRAPHYWei Sha obtained a BEngat Tsinghua University in1986. He was awarded in1992 a PhD by OxfordUniversity and in 2009 aDSc by Queen’s Univer-sity Belfast. He previouslyworked at Imperial College and CambridgeUniversity. He is presently Professor of Mate-rials Science, with research interests in phasetransitions and SEM.

ABSTRACTTwo manganese steels were investigated:Fe-19.7%Mn (VM339A) and Fe-19.7%Mnstabilized with 0.056%C, 0.19%Ti and0.083%Al (VM339B). The toughness ofVM339A was higher than VM339B, butVM339B had higher hardness. Temperingdoes not affect the toughness of the alloys.SEM images of the fracture surface for boththe alloys revealed ductile fractures. A fur-ther alloy with a lower manganese content,Fe-8.46%Mn-0.24%Nb-0.038%C, and thuseven lower cost than the conventional 3.5Nicryogenic steel, was tested for its impacttoughness after heat treatment at 600°C,giving promising results.

KEYWORDSscanning electron microscopy, hardnessmeasurement, mechanical characterization,steel, fracture

AUTHOR DETA I L SProfessor Wei Sha, School of Planning, Architecture and CivilEngineering, Queen’s University Belfast, Belfast BT7 1NN, UKTel: +44 28 90974017Email: w.sha@qub.ac.uk

Microscopy and Analysis 25(7):15-19 (EU), 2011

ANALYS I S OF STEELS

I N TRODUCT IONNickel raises the yield strength of iron, asfound many decades ago by Roberts andOwen [1] by measuring the strength of manyFe-Ni alloys. Nickel also lowers the ductile brit-tle transition temperature (DBTT, cleavage) ofiron, as proved by Floreen et al. [2] by measur-ing the impact properties of Fe-Ni alloys withdifferent Ni contents. Both results have beenverified in the extensive steel research that fol-lowed, so nickel is an established alloy methodof raising strength and increasing toughness. As shown by Avery and Parsons [3] with

some real examples, nickel has been widelyused in cryogenic steels, but the high cost ofnickel now demands a second thought on theactual amounts of this element required inthese steels. Oshima et al. [4] have vividly illus-trated the sharp rise of nickel price on the Lon-don Metal Exchange. Oshima et al. [4] alsoexplored the ways of nickel saving, by adetailed property comparison between lownickel stainless steels and conventional stain-less steels. Based on their findings, nickel-freehigh-manganese steels could have a combina-tion of good tensile strength and ductility andso could provide a great potential in applica-tions for structural components in industry. In recent years, numerous attempts have

been made to improve the performance ofmanganese steels by modifying their composi-tion and by applying heat or thermomechani-cal treatments. For example, by examining themicrostructure and the performance [5] it wasconcluded that the capability of work harden-ing and impact abrasion resistance wereenhanced greatly after rolling. The cold asyn-chronous rolling technique taken was only oneway of achieving potential property improve-ment.In their classical work, Holden et al. [6]

reported that 15-20%Mn steels, with around0.02% carbon, contain mostly epsilon marten-

Fracture Surface, Impact Energy andHardness of Ni-Free High-Mn SteelsWei Sha,1 H. H. Haji Talib,1 Eric A. Wilson,2 Raj Rajendran,3 Savko Malinov,4 Harvey R. Charlesworth,5 Lee Ibbitson6

1. School of Planning, Architecture and Civil Engineering, Queen’s University Belfast, UK. 2. Faculty of Arts, Computing, Engi-neering and Sciences, Sheffield Hallam University, UK. 3. School of Mechanical and Building Sciences, B. S. Abdur RahmanUniversity, India. 4. School of Mechanical and Aerospace Engineering, Queen’s University Belfast, UK. 5. London & Scandi-navian Metallurgical Co. Ltd., UK. 6. Tata Steel Speciality, UK

Table 1: Hardness (HV1) of theexperimental alloysunder various heattreatment conditions.

MICROSCOPY AND ANALYSIS NOVEMBER 2011 15

Heat Treatment VM339A VM339B

850oC 1h, air cooled 222±2 281±12

850oC 1h, water quenched (WQ) 226±2 288±2

WQ + 450oC 0.1 h 230±2 299±11

WQ + 450oC 1 h 263±17 –

WQ + 450oC 10 h 270±7 294±9

WQ + 450oC 100 h 274±9 –

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The traditional scientific programs of each Conference will be run in parallel, with some joint sessions.Delegates may attend any they choose through the single registration process. The Exhibition and social events will be fully shared, to enable our communities to network extensively.

The Event is being conducted under the auspices of the Council of Asia-Pacific Societies for Microscopy (CAPSM), The Australian Nanotechnology Network (ANN), the Australian Microscopy and Microanalysis Society Inc (AMMS) and the International Federation of Societies for Microscopy (IFSM). ANN /ICONN are funding registrations for eligible ANN students and ECR; AMMS and IFSM have travel bursaries for microscopy students and ECR.Confirmed plenary speakers include Professor Knut Urban, Juelich, Germany, joint winner of the 2011 Wolf Prize in Physics. Professor Urban will give the inaugural “Cockayne Lecture”, Professor Frank Caruso, Melbourne, Aus-tralia , a 2010 Top 100 most-cited researcher in nanotechnology, and Zhores Alferov , Ioffe Institute, Russia, the winner of the Nobel Prize in Physics, 2000.

The Conference is dedicated to the memory of Professor David Cockayne FRS.

Brendan Griffin & Lorenzo FaraoneCo-Chairs

For more information see: www.apmc-10.org or www.iconn-2012.org

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2012

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On behalf of the Organising Committee it is our pleasure to extend an invitation to the 10th Asia-Pacific Microscopy Conference (APMC-10), the International Conference on Na-noscience and Nanotechnology (ICONN 2012) and the 22nd Australian Conference on Microscopy and Microanalysis (ACMM-22) to be held in Perth, Western Australia, 5th – 9th February 2012. The Event will include Short Courses, Workshops and a con-current major Equipment Exhibition. The combined event will be the largest microscopy and nanotechnology-related event in Australiaʼs history. Over 2,000 delegates, from more than 30 nations, are expected to provide a unique science and technology forum.

CRITICAL DATES

APMC AD.indd 1 25/08/2011 10:44

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and 0.083%Al (VM339B). The Mn content wasselected based around commercial highermanganese content steels, summarised fromindustrial data by Subramanyam et al. [12], butits particular value was not thought to be crit-ical, if it was, say, 1% higher or lower. Carbonwas kept low to improve toughness. Ti and Alwere added to tie up residual C, N and O.These alloys cost about half as much as a3.5%Ni steel. The alloys were austenitised at 850oC, water

quenched or air cooled and also tempered for0.1, 1, 10 or 100 h at 450oC. Tempering in therange of 300-500oC increases the ductile-brit-tle transition temperature (DBTT), as found byBolton et al. [13] when measuring this prop-erty in a series of Fe-Mn alloys, albeit withoutdetailed microstructural characterisation workback then. Nasim et al. [14] noted a rapid grainboundary segregation of Mn, P, and N, on age-ing, based on Auger electron spectroscopymeasurements. The control group was in awater-quenched condition. Embrittlementwas the most severe at 450oC.Further, as a continuation of a previous

work, where Nikbakht et al. [7] identified theembrittlement mechanism in Fe-8%Mn as wellas Fe-8%Mn stabilised with Ti and Al, a thirdalloy was studied, to investigate the effect ofNb. A Fe-8.46%Mn-0.24%Nb-0.038%C alloywas vacuum-melted as a 20 kg cast and upsetforged at 1200°C into a 32�76 mm plate. Thisalloy costs about one third of a 3.5%Ni steel.The heat treatment at 600°C on thehomogenised alloy was carried out to intro-duce reverted austenite to improve impacttoughness. Excess C over stoichiometry wasabout 70 ppm, which lowers the DBTT (see Fig-ure 9 of ref. [15]). The alloy was homogenisedas two blocks 32�60�76 mm for 50 h at1100°C, air cooled, and followed by two dif-ferent heat treatment routes for each block.The first consisted of 1.5 h at 850°C, air-cool-ing, and then 4 h at 600°C, air-cooling. The sec-ond consisted of 1.5 h at 850°C, water quench-ing, and then 4 h at 600°C, water quenching.Longitudinal V notch Charpy specimens5�10�55 mm were machined from the heat-treated blocks.

Vickers Hardness and Impact TestingVickers hardness was measured using a load of1 kg, at room temperature in the unstrainedpart of the impact-tested specimens. Impacttests were conducted on a Charpy machinewith maximum impact energy of 300 J. Sam-ples of size 5�10�55 mm with a V-notch wereused. The temperatures of impact tests were20°C, -20°C, -100°C, and -196°C. Testing tem-peratures below the room temperature wereobtained by using a chamber cooled by flow ofliquid nitrogen.

Scanning Electron MicroscopyThe steel specimens were polished using a 0.05µm colloidal suspension of silica after mechan-ical polishing down to 1 µm. The microstructure of the steels was exam-

ined in a Jeol 6500 FEG scanning electronmicroscope. The SEM was operated at 5-20 kV.The fracture surfaces of the samples were

examined under the SEM to determinewhether the sample surface had brittle cleav-age type or ductile type failure.

RESULTS AND D I SCUSS IONTable 1 shows the hardness values of theexperimental alloys VM339A and VM339Bunder various heat treatment conditions. AlloyVM339B is harder than VM339A because itcontains 0.056%C, 0.19%Ti and 0.083%Al. Itwas found that deep quenching the alloysdown to -20, -100 and -196°C does not changethe hardness after the specimens werereturned to room temperature before thehardness measurements were taken.Table 1 includes the hardness values for both

alloys tempered at 450°C with increasing timeand then impact tested. In the case ofVM339A, the hardness increased significantlyafter tempering for 1-100 hours, compared towithout tempering or tempering for 0.1 h. Onthe other hand, little change of hardness hasbeen found when the tempering time variesfrom 1 to 100 h. However, there exists an obvi-ous difference between tempering for 0.1 hand 1 h. There could be two reasons con-tributing to this effect:1. The effect of tempering reaches a satura-

tion point after around one hour or some timebefore one hour. Here, a saturation pointmeans no further increase in hardness withincreasing tempering time.2. Experimental limitations meant that heat-

ing for 6 minutes did not lead to effective tem-pering, because the specimens were sealed insilica tubes in vacuum, inserted into the pre-heated furnace, and kept for the specifiedlength of time. There might be a heating-upperiod that could take minutes.Impact testing was carried out on both alloysto estimate the ductile-brittle transition tem-perature, which gives the change in behaviourfrom ductile at high temperature to brittle atlower temperature (Table 2). As a generalstatement, ductile fracture initiates at a par-ticular toughness valueMicrographs of fracture surface of alloy

VM339A and VM339B (1 h at 850°C, WQ)impact tested at 20°C are shown in Figures 1and 2, respectively. The uneven or rough sur-face of the fracture can be seen at low magni-fication (Figures 1a and 2a). High magnifica-tion shows a ductile type failure for both alloys(Figures 1b and 2b). Tearing and cone shapeddimples were observed in alloy VM339A (Fig-ure 1), which had the higher impact toughnessindicating it is more ductile than alloyVM339B. Micrographs of fracture surfaces forboth alloys at different conditions and tem-pering time were examined which showedductile type fracture with no obvious differ-ence.Micrographs of the surface structure for

both alloys at different conditions and tem-pering time were examined but there were novisible differences in the structures or grain-size.It is possible in Fe-20%Mn to get peculiar

stress-strain curves at room temperature, dueto gamma and epsilon phases, like in the ther-mally cycled Fe-8%Mn alloy [16]. Nikbakht etal. [7] summarised the formation of thesephases from relevant literature.The Charpy impact test results of the heat

treated Fe-8.46%Mn-0.24%Nb-0.038%C alloyare shown in Figure 3. It should be noted thatthe linear regression lines are used to onlyillustrate the difference between specimensafter the two cooling treatments. The linearregression may or may not represent the realvariation of the impact energy with testingtemperature.These impact test results show that the

water quenched alloy has higher impactenergy. The reason may be that air coolingresulted in the formation of precipitates dur-ing the slower cooling process, which increasesstrength but decreases toughness.In the quenched condition, this third alloy

had a higher impact energy than bothVM339A and VM339B, showing the beneficialeffect of Nb stabilisation. In the air cooled con-dition, however, the Nb-containing alloy hadcomparable impact energy with VM339A and

Table 2: Impact energy (in joules J) absorbed by the experimental alloys under various heat treatment and impact test temperatures.

Alloys Heat Treatment 20oC -20oC -100oC -196oC

VM339A 850oC 1h, air cooled 68 58 8 –

850oC 1h, water quenched (WQ) 71 61 23 8

WQ + 450oC 0.1 h 68 60 11 –

WQ + 450oC 1 h 62 61 12 5

WQ + 450oC 10 h 75 38 9 –

WQ + 450oC 100 h 66 45 7 –

VM339B 850oC 1h, air cooled 43 56 19 8

850oC 1h, WQ 54 50 20 8

WQ + 450oC 0.1 h 54 49 15 –

WQ + 450oC 1 h 52 42 12 5

ANALYS I S OF STEELS

MICROSCOPY AND ANALYSIS NOVEMBER 2011 17

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Applications, Labeling Strategies and Fluorophores for Super-Resolution Great insights can be obtained from conventional fluorescence microscopy, but studying the architecture and protein dynamics of sub-cellular compartments can be challenging, since a major portion of information concerning the structural organization is lost due to the light diffraction limit. Several approaches to overcome this limitation have been developed and super-resolution has proven an extremely valuable tool.

Speakers:Dr. Marko Lampe, Leica Microsystems CMS GmbH, Wetzlar, GermanyDr. Wernher Fouquet, Leica Microsystems CMS GmbH, Mannheim, Germany

Online Seminar/Webinar on Applications for Super-Resolution Register at http://www.microscopy-analysis.com/leicawebinars and join on Tuesday 15th November 2011 at 17:00h (CET), 16:00h (UK)

Recently, super-resolution microscopy has made its way into research labs and is addressing more and more scientific questions. Today, we will highlight STED and GSDIM and show how these methods have been optimized in the tried and proven commercially available solutions from Leica Microsystems.

In addition to a review of the principles of method and system architecture, we will expand on specific guidelines for sample prepa-ration, suitable fluorophores and labeling strategies. We will also highlight selected application examples, including super-resolution imaging of intracellular substructures such as cytoskeleton elements and associated proteins, nuclear pore complexes and cellular compartments. In particular for STED imaging, we will consider applications requiring multi-color and live imaging.

Register at http://www.microscopy-analysis.com/leicawebinars

This webinar will be available on demand under the registration link above.

Image taken with a Leica TCS STED CW microscope of wildtype HeLa cells stained against Histone H3/Chromeo505 and Tubulin/V500. Courtesy: Samples were kindly provided by Active Motif Chromeon

2 color STED GSD application image

Ptk2-cells. NPC-staining: anti-NUP153/Alexa Fluor® 532 | Microtubule-staining: anti-ß-tubulin/Alexa Fluor® 647.Courtesy: Wernher Fouquet, Leica Microsystems in collaboration with Anna Szymborska and Jan Ellenberg, EMBL, Heidelberg, Germany

Applications, Labeling Strategies and Fluorophores for Super-Resolution Great insights can be obtained from conventional fluorescence microscopy, but studying the architecture and protein dynamics of sub-cellular compartments can be challenging, since a major portion of information concerning the structural organization is lost due to the light diffraction limit. Several approaches to overcome this limitation have been developed and super-resolution has proven an extremely valuable tool.

Speakers:Dr. Marko Lampe, Leica Microsystems CMS GmbH, Wetzlar, GermanyDr. Wernher Fouquet, Leica Microsystems CMS GmbH, Mannheim, Germany

Online Seminar/Webinar on Applications for Super-Resolution Register at http://www.microscopy-analysis.com/leicawebinars and join on Tuesday 15th November 2011 at 17:00h (CET), 16:00h (UK)

Recently, super-resolution microscopy has made its way into research labs and is addressing more and more scientific questions. Today, we will highlight STED and GSDIM and show how these methods have been optimized in the tried and proven commercially available solutions from Leica Microsystems.

In addition to a review of the principles of method and system architecture, we will expand on specific guidelines for sample prepa-ration, suitable fluorophores and labeling strategies. We will also highlight selected application examples, including super-resolution imaging of intracellular substructures such as cytoskeleton elements and associated proteins, nuclear pore complexes and cellular compartments. In particular for STED imaging, we will consider applications requiring multi-color and live imaging.

Register at http://www.microscopy-analysis.com/leicawebinars

This webinar will be available on demand under the registration link above.

Image taken with a Leica TCS STED CW microscope of wildtype HeLa cells stained against Histone H3/Chromeo505 and Tubulin/V500. Courtesy: Samples were kindly provided by Active Motif Chromeon

2 color STED GSD application image

Ptk2-cells. NPC-staining: anti-NUP153/Alexa Fluor® 532 | Microtubule-staining: anti-ß-tubulin/Alexa Fluor® 647.Courtesy: Wernher Fouquet, Leica Microsystems in collaboration with Anna Szymborska and Jan Ellenberg, EMBL, Heidelberg, Germany

CIRCLE NO. 9 OR ONLINE: www.microscopy-analysis.com

ANALYS I S OF STEELS

VM339B, depending on the testing tempera-ture. The reason is perhaps, as explained ear-lier, air cooling has more detrimental effect ontoughness, most likely related to Nb precipita-tion.

CONCLUS IONSIn conclusion, the mechanical properties andmicrostructure were investigated to find outthe suitability of the nickel-free manganesesteels to be used at low temperatures. Theexperimental results are summarised below.1. It is clear that VM339A is tougher than

VM339B. It has higher impact resistance, forexample 61 J at -20°C in the as quenched con-dition. The alloying elements such as carbon,titanium and aluminium in alloy VM339Bmake the alloy more brittle. These elementshave significant effect on the hardness of thealloy.2. The tempering at 450oC up to 100 hours

does not affect the toughness of the alloys sig-nificantly. This is an added advantage whencompared with other cryogenic alloys thatneed a precise heat treatment and tempering.A further study is required to substantiate thatthe nickel free steels form a potential candi-date for cryogenic applications. These alloysare cheaper than the present 9% nickel alloys.It is found from this research that temperinghas no effect on the alloys, which may be use-ful for welding works. Therefore, weldingeffects can be investigated under hot and coldworking condition to study the suitability forthe applications.

REFERENCES1. Roberts, M. J., Owen, W. S. The strength of martensitic iron-

nickel alloys. ASM Trans. Quart. 60:687-692, 1967.2. Floreen, S., Haynes, H. W., Devine, T. M. Cleavage initiation

in Fe-Ni alloys. Metall. Trans. 2:1403-1406, 1971.3. Avery, R. E., Parsons, D. Welding stainless and 9% nickel

steel cryogenic vessels. Welding Journal 74(11):45-50, 1995.4. Oshima, T., Habara, Y., Kuroda, K. Efforts to save nickel in

austenitic stainless steels. ISIJ Int. 47, 359-364, 2007.5. Qiu, C. M., Wang, Y. F., Yu, J. Effect of asynchronous rolling

on wear-resisting performance of high manganese steel.Adv. Mater. Res. 146-147:340-344, 2011.

6. Holden, A., Bolton, J. D., Petty, E. R. Structure and propertiesof iron-manganese alloys. J. Iron Steel Inst. 209, 721-728,1971.

7. Nikbakht, F., Nasim, M., Davies, C., Wilson, E. A., Adrian, H.Isothermal embrittlement of Fe–8Mn alloys at 450°C. Mater.Sci. Technol. 26:552-558, 2010.

8. Roberts, M. J. Effect of transformation substructure on thestrength and toughness of Fe-Mn alloys. Metall. Trans.1:3287–3294, 1970.

9. Vaynman, S., Isheim, D., Kolli, R.P., Bhat, S.P., Seidman, D.N.,Fine, M.E., 2008. High-strength low-carbon ferritic steelcontaining Cu-Fe-Ni-Al-Mn precipitates. Metall. Mater.Trans. A 39:363-373, 2008.

10. Sathiya, P., Aravindan, S., Ajith, P. M., Arivazhagan, B., Haq,A. N. Microstructural characteristics on bead on platewelding of AISI 904 L super austenitic stainless steel usinggas metal arc welding process. Int. J. Eng. Sci. Technol.2(6):189-199, 2010.

11. Berns, H., Theisen, W. Ferrous Materials: Steel and Cast Iron,Springer, Berlin, pp. 1-418, 2008.

12. Subramanyam, D. K., Swansiger, A. E., Avery, H. S. Austeniticmanganese steels. In Properties and Selection: Irons, Steels,and High-Performance Alloys, vol. 1, ASM Handbook. ASM

Figure 1: Scanning electron microscope images of the fracture surface of alloy VM339A, 850oC for1 h, water quenched, and then impact tested at 20oC: mainlyductile failure, impact energy 71 joules. (a) Low magnification. (b) High magnification.

MICROSCOPY AND ANALYSIS NOVEMBER 2011 19

Figure 2: Scanning electron microscope images of the fracture surface of alloy VM339B, 850oC for1 h, water quenched, and then impact tested at 20oC: mainlyductile failure, impact energy 54 joules. (a) Low magnification. (b) High magnification.

Figure 3: Charpy impact energy as a function of specimen temperature at testing for the Fe-8.46%Mn-0.24%Nb-0.038%C alloy.

International, Materials Park, OH, pp. 822-840, 1990.13. Bolton, J. D., Petty, E. R., Allen, G. B. The mechanical

properties of ��-phase low-carbon Fe-Mn alloys. Metall.Trans. 2:2915-2923, 1971.

14. Nasim, M., Edwards, B. C., Wilson E. A. A study of grainboundary embrittlement in an Fe–8%Mn alloy. Mater. Sci.Eng. A 281:56-67, 2000.

15. Wilson, E. A., Ghosh, S. K., Scott, P. G., Hazeldine, T. A.,

Mistry, D. C., Chong, S. H. Low cost grain refined steels asalternative to conventional maraging grades. Mater.Technol. 23:1-8, 2008.

16. Nasim, M., Wilson E. A. The effect of thermal cycling on theimpact toughness of an Fe-8Mn alloy. In PhaseTransformations, Vol. 1, The Institution of Metallurgists,York, pp. v21-v25, 1979.

©2011 John Wiley & Sons, Ltd

a b

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cone-shaped dimple

tearing

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Aligned with your application on any TEM TEM camera systems for Life and Materials Sciences

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MATER IALS AND METHODSPreparation of Plant TissuesStigmas were harvested from both autosterile(D07) and autofertile (K25) lines of Vicia fabain the days leading to anthesis. Samples wererapidly frozen in liquid nitrogen and attachedto the cold stage (kept at -20oC) with carbonsticky pads.

Scanning Electron MicroscopyPlant specimens were examined in a Carl ZeissEVO LS 15 SEM with variable pressure capabil-ity at 20 kV and imaged using a variable pres-sure secondary electron (VPSE) detector.In addition, the SEM was equipped with a

Coolstage from Deben UK Ltd, Suffolk, and theimages presented in this note were obtainedwith the stigmas cooled to approximately -20°C. In order to reduce charging artifacts, alow pressure of about 30 Pa of air was used.This very low pressure is sufficient to compen-sate for specimen charging and to provide agas phase scintillation signal for the VPSEdetector.The Coolstage, shown in Figure 3, is a Peltier

device that is able to reduce the temperatureof the specimen. Extended periods of SEMimaging are then possible as hydration of thespecimen is maintained. This approach offersan alternative to the full environments SEM ifdynamic processes involving water are notrequired.

B IOGRAPHYTimothy Baldwin has aPhD in botany from theUniversity of Reading.After a post-doctoralposition at the JohnInnes Centre, Norwich,he became a lecturer inthe Botany Department at Universiti SainsMalaysia. In 1999 he returned to the UK andwas appointed a post-doctoral researchassociate in the Department of Biochemistryat the University of Cambridge. Since 2001,Tim has been senior lecturer in plant sci-ences at the University of Wolverhampton.His research interests include the role of theplant cell wall in plant growth and develop-ment, pollination biology, orchid conserva-tion and plants used in traditional Chinesemedicine.

ABSTRACTThe cell walls of the stigma and style areimportant zones for cell-cell recognition,nutrition, guidance and protection of thepollen tube along the transmitting tract.The SEM data presented here was a compo-nent of a larger study the main objective ofwhich was to investigate pistil developmentand pollination in the crop species Vicia fabaL. (the faba bean). The data demonstratethat there is a developmentally regulateddifference in the structural integrity of thestigmatic surface in autofertile (K25) andautosterile (D07) lines of the faba bean. Inthe autofertile lines the stigmatic surfaceruptures two days prior to flower opening(anthesis), whereas in the autosterile plantsthe stigma remains intact, until anthesis.The VPSEM technique used in the currentstudy shows this difference to great effectand as such is an invaluable tool for use inplant cell biology.

KEYWORDSscanning electron microscopy, variable pres-sure, cryo electron microscopy, plant sci-ences, pollination

AUTHOR DETA I L SDr Timothy C. Baldwin, Senior Lecturer –Plant Sciences, School of Applied Sciences, University of Wolverhampton, Wolverhampton WV1 1SB, UKTel: +44 (0) 1902 322142Email: t.baldwin@wlv.ac.uk

Microscopy and Analysis 25(7):21-22 (EU), 2011

VPSEM IN PLANT SC IENCES

I N TRODUCT IONGrain legumes are extremely important inworld agriculture yet information on the struc-ture, composition, and functioning of theirsolid stigma and open style is limited. Globalfood production is underpinned by plantbreeding programmes centred on importantcrops including the faba bean (Vicia faba L.)(Figure 1), soybean, pea, and the commonbean. Understanding the mechanism of polli-nation in such species enhances the effective-ness of these programmes.The faba bean is particularly suitable for

study because it produces numerous largeflowers with straight styles that are easy to dis-sect and is partially dependent upon bees forpollination.For the faba bean there are both autofertile

and autosterile lines. Autofertile lines do notrequire a pollinating insect but do not havevery large yields. Autosterile lines do need apollinating insect and have relatively high cropyields.The aim for plant scientists is to compare

autosterile and autofertile lines so that crossesbetween the two lines can be developed. Theoverall aim is increased food production.In this article the development of the faba

bean stigmatic papillae was observed in thedays leading up to anthesis (flower opening)using low-temperature variable pressure scan-ning electron microscopy (VPSEM).

Variable Pressure Scanning Electron Micro-scopy of Vicia faba Stigmatic PapillaeWen Chen, Fred Stoddard and Timothy C. BaldwinSchool of Applied Sciences, University of Wolverhampton, UK

Figure 1: Vicia faba. From Prof. Dr Otto Wilhelm Thomé Flora von Deutschland,Österreich und der Schweiz 1885, Gera, Germany. www.biolib.de

MICROSCOPY AND ANALYSIS NOVEMBER 2011 21

Figure 2: Photomicrograph showing the general anatomy of the faba beanpistil. Key: (b) – stigma, (c) – apical region, (d) – middle region, (e) –basal region, ov – ovary, sh – stylar hairs, stg – stigma, sty – style.Scale bar = 500 µm.

Baldwin_Layout_Final_Euro_CMYK_ArticleTemplate_Jan2008.qxd 10/25/2011 9:59 AM Page 21

RESULTS AND D I SCUSS IONFigure 2 shows the general anatomy of thefaba bean pistil and the positions of thestigma, style, stylar hairs and ovary.Figure 4 shows the structure of faba beanstigmas from autofertile K25 (a-e) andautosterile D07 (f-j) lines at d-4 (a, f), d-3 (b, g),d-2 (c, h), d-1 (d, i) and anthesis (e, j). The stig-matic cuticle started to become ruptured(arrowed) at 2 days pre-anthesis in K25 whilstremaining intact until anthesis in D07.Figure 5 shows close ups of faba bean stig-matic papillae.These data show the differences in stigmaticmorphology in autofertile and autosterilelines of faba bean in the days prior to floweropening and pollination which clearly indicatewhy each line is autofertile or autosterilerespectively. They also demonstrate the utilityof VPSEM for similar studies in other plantsspecies where changes in surface morphologycorrelate with biological function.

REFERENCES1. Chen, W., Stoddard, F. L. and Baldwin, T. C. Int. J. Plant Sci.

167(5): 919-932, 2006.2. Chen W. and Baldwin T. C. An improved method for the

fixation, embedding and immunofluorescence labelling ofresin-embedded plant tissue. Plant Molecular BiologyReporter, 25:25-37, 2007.

ACKNOWLEDGEMENTSWe are very grateful to Professor W. Link, Uni-versity of Göttingen, for supplying the seeds ofthe inbred lines; to Mr Robert Hooton, Univer-sity of Wolverhampton, who assisted with thecultivation of the faba beans and to Ms Bar-bara Hodson for help and advice with the SEM.W. C. was financially supported by a Universityof Wolverhampton PhD studentship, grantnumber RS 328, ‘A molecular and structuralinvestigation of autofertility and autosterilityin Vicia faba (faba bean)’. The project wasassociated with European Commission grantQLK5-CT-2002-02307, ‘Faba bean breeding forsustainable agriculture in Europe’ (acronymEU-Faba).

©2011 John Wiley & Sons, Ltd

Figure 3 (above): The cold stage used on the VP SEM. Courtesy of Carl Zeiss NTS.

MICROSCOPY AND ANALYSIS NOVEMBER 201122

Figure 4 (right):Structure of stigma from K25 (a-e) and D07 (f-j) at d-4 (a, f), d-3 (b, g),d-2 (c, h), d-1 (d, i) and anthesis (e, j). The stigmatic cuticle started tobecome ruptured (arrowed) at 2 days pre-anthesis in K25 whilst remain-ing intact until anthesis in D07. Scale bars = 40 µm.

Figure 5:Close up of faba bean stigmatic papillae (a) and (b).Scale bars = 20 µm.

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people and placesPEOPLE AND PLACES

MICROSCOPY AND ANALYSIS NOVEMBER 2011 23

The Royal Swedish Acad-emy of Sciences hasawarded the Nobel Prize inChemistry for 2011 toDaniel Shechtman Technion– Israel Institute of Technol-ogy, Haifa, Israel “for thediscovery of quasicrystals”.On 8 April 1982, an

image counter to the lawsof nature appeared inDaniel Shechtman’s elec-tron microscope. In all solid matter, atoms were believed to be packed inside crys-tals in symmetrical patterns that were repeated periodically over and over again.For scientists, this repetition was required in order to obtain a crystal. Shechtman’simage, however, showed that the atoms in his crystal were packed in a pattern thatcould not be repeated. Such a pattern was considered just as impossible as creat-ing a football using only six-cornered polygons, when a sphere needs both five- andsix-cornered polygons. Aperiodic mosaics, such as those found in medieval Islamicmosaics have helped scientists understand what quasicrystals look like at the atomiclevel. In those mosaics, as in quasicrystals, the patterns are regular – they followmathematical rules – but they never repeat themselves. Following Shechtman’s dis-covery, scientists have produced other kinds of quasicrystals in the lab and discov-ered naturally occurring quasicrystals in mineral samples from a Russian river. ASwedish company has also found quasicrystals in a certain form of steel, where thecrystals reinforce the material like armor. Scientists are currently experimentingwith using quasicrystals in different products such as frying pans and diesel engines.Daniel Shechtman was born 1941 in Tel Aviv, Israel. He has a PhD 1972 from the

Technion - Israel Institute of Technology in Haifa, Israel, and is now a distinguishedprofessor holding the Philip Tobias Chair, at that university.

Nobel Prize for Shechtman

The National Institute of Materials Physics (NIMP) in Magurele, near Bucharest,Romania, has opened new labs equipped with state-of-the-art microscopes for thecomplex examination and characterization of the microstructure of materials. Thesuppliers of these systems were JEOL (Europe), Shimadzu Handelsgesellschaft andTESCAN. Together they offered their best instruments to the lab: a TESCAN LYRA3field-emission scanning electron microscope equipped with a focused ion beamand a JEOL JEM-ARM 200F Cs-corrected field emission atomic resolution analyticaltransmission electron microscope.The inauguration programme included a workshop with presentations by rep-

resentatives of several important European scientific and academic institutionssuch as the University of Antwerp, Belgium, the University of Caen, France, ClaudeBernard University in Lyon, France, and the Institute of Physics and Chemistry ofMaterials in Strasbourg, France. TESCAN and JEOL representatives then followedwith presentations of their most advanced equipment including the two instru-ments delivered to NIMP in Magurele.

Romanian National Institute

Left: Dr Zadrazil of Tescan. Right: TESCAN Lyra 3 at NIMP.

Following the successful completion of a two-year evaluation phase, the Universityof Ulm, the Heidelberg-based company CEOS GmbH and Carl Zeiss Nano Technol-ogy Systems have signed an agreement to embark on the next phase of the SALVE(sub-angstrom low-voltage electron) microscopy project. SALVE is one of the most ambitious research projects in the field of electron

microscopy to be undertaken in Germany in recent years. The objective of the pro-ject is to develop and build a transmission electron microscope capable of imaging

samples with atomic resolution at very lowacceleration voltages. The advantages offeredby this approach are clear: Unlike the currentgeneration of TEMs with accelerating voltagesof between 200 and 300 kV, which destroy radi-ation-sensitive samples before researchers canrecord usable images or perform material analy-sis, the SALVE project will keep specimens stablelong enough to perform experimental work.The the first phase of the co-operation project

conducted between 2009 and 2011 – in whichresearchers analyzed the feasibility of the keyprinciples involved – has produced some spec-tacular results, with the scientists successfullygenerating atomic-resolution images at acceler-ating voltages well below 80 kV. During celebrations to mark the start of the

project’s second phase, Project Manager Professor Ute Kaiser from the Universityof Ulm and a number of guest speakers, including Nobel Prize winner Klaus vonKlitzing, presented some of the fascinating ways in which the SALVE system couldpotentially be used. Ranging from studies of superconductors and semiconductorsto research into lithium-ion batteries, plastics and biological materials, some of theexamples they highlighted have already yielded preliminary results. While Carl Zeiss presses ahead with development of the system itself, the Uni-

versity of Ulm will be working on application development and conductingresearch into sample preparation methods. Meanwhile, the third project partner,CEOS, having a lot of expertise in the development of advanced electron opticalsystems, is focusing its efforts on a new optimized corrector to compensate thechromatic and the spherical aberration for low voltages.

SALVE Project enters Phase II

Synoptics has set up the Advanced Technology Group, a new division which willwork with life science companies and academic clients to deliver bespoke imagingequipment to improve bench-based research, quality control or clinical develop-ment processes.Synoptics, comprising the Syngene, Synbiosis and Syncroscopy divisions, provides

innovative products that life scientists need to enhance the quality and speed oftheir research. Syngene produces the G:BOX image analyser to analyse gels andblots in the fields of genomics and proteomics. Synbiosis provides the ProtoCOL 2,a colony counting and zone sizing system used in many major pharmaceutical com-panies, and Syncroscopy offers Auto-Montage software for producing focusedimages of 3D samples, which is marketed by major microscope manufacturer, Leica.Richard Maskell, Synoptics’ new head of the Advanced Technology Group

explained: “We are increasingly being approached to develop novel systems andsoftware to bring about a step change in biological quality control and researchprocesses. We have experience in solving challenges associated with imaging awide variety of materials, as well as taking those solutions to market and aredelighted to offer access to this expertise via our new Advanced TechnologyGroup.” Paul Ellwood, Managing Director of Synoptics added: “We encourage academic

researchers experiencing issues with their biological imaging processes, or tech-nology providers that are looking to design an imaging system to bring to market,to contact us today to discuss how working with our new Advanced TechnologyGroup could help them make a strategic impact on their research or quality con-trol objectives.”

Synoptics Technology Group

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CIRCLE NO. 40 OR ONLINE: www.microscopy-analysis.comMICROSCOPY AND ANALYSIS NOVEMBER 2011 25

PEOPLE AND PLACES

Andor Technology, a world leader in scientific imag-ing and spectroscopy solutions, has announced thattwo visually stunning and scientifically captivatingentries have won the Andor Insight Awards ScientificImaging Competition. The winning entry in the Phys-ical Sciences category was submitted by Dr RobertMarshall of Boston University and the winning entryin the Life Sciences category by Dr Satoshi Nishimuraof the University of Tokyo.Dr Satoshi Nishimura of the Department of Cardio-

vascular Medicine at the University of Tokyo entereda series of confocal captures entitled ‘Inflammatorycellular dynamics in obese adipose tissue revealed byin vivo imaging technique’. This imaging techniqueallows scientists to work at a cellular level and willprovide the basis for future clinical usage of in-vivoimaging for humans. It was captured using an AndoriXon3 EMCCD camera and a Confocal Spinning DiskUnit.

Andor Insight Imaging Awards

According to a recently published report fromResearch and Markets Ltd in Dublin, the world micro-scopes market, encompassing light, confocal, electronand scanning probe microscopes, generated an esti-mated $5.6 billion in revenues in 2010. The market isexpected to grow to over $9 billion by 2017. Increased government and corporate funding in

life sciences, materials research and nanotechnologyis leading to sustained growth in microscopy. Lightmicroscopes currently account for the majority of the

World Microscopy Market Growsmicroscopy market but will lose market share to elec-tron and scanning probe microscopes in the comingyears. The largest end-user market for microscopy issemiconductors, followed by life sciences, and nano-technology and nanomaterials research. The industrywas adversely affected the global recession overall,especially in the semiconductors end-user sector butwitnessed growth in the life sciences and healthcare.Electron and scanning probe are the fastest growingmicroscopy markets.

At the Institute of Photonic Technology (IPHT) in Jena,Germany, light is the central focus of research anddevelopment. The new IPHT sees photonics as themost important key technology of the 21st century. Itis guaranteed to play a leading role in the fields ofinformation technology and communications, secu-rity, material science, life science and health.Dr Volker Deckert is the head of the Nanoscopy

department which utilizes instrumental methods inthe development of molecular spectroscopic meth-ods with the highest spatial resolution. Central to thisprogram has been the use of tip-enhanced Ramanscattering, TERS, where the NanoWizard systems andTip-Assisted Optics module from JPK Instrumentshave provided a platform in the development ofthese experimental methods. In many cases, the struc-tural sizes of components are below the capabilitiesof normal optical microscopic or spectroscopic tech-niques. Optical near-field microscopy in combinationwith Raman spectroscopy pushes the achievable res-olution significantly below the diffraction limit ofstandard instruments. The goal of their work is the

Institute of Photonic Technology in Jena

advancement of TERS to become an accessible andsensitive tool for the analysis of surfaces and bound-aries under ambient conditions. Applications are care-fully selected. For example, heterogeneous catalyticreactions are studied because such application-ori-ented experiments may be used to verify and improvethe functionality of the instrument and to demon-strate its practicality for ‘real’ problems.

LITERATURE HIGHL IGHTS

literature highlightsGold Nanodot Arrays for Nanofabrication

A large array of sub-10-nm sin-gle-grain gold nanodots for usein nanotechnology is describedby Nicolas Clément and col-leagues at the Institut d’Elec-tronique Microélectronique etNanotechnologie, CNRS, Univer-sity of Lille, France [Small7(18):2607-2613, 2011].A uniform array of single-

grain gold nanodots, as small as5-8 nm, was be formed on siliconusing e-beam lithography. Theas-fabricated nanodots wereamorphous, and thermal annealing converted themto pure Au single crystals covered with a thin SiO2layer. These findings were based on physical mea-surements by AFM, atomic-resolution STEM, andchemical techniques using energy dispersive X-rayspectroscopy. The authors demonstrated the forma-tion by e-beam lithography of sub-10-nm Au dotswith small dispersion and perfect alignment. Suchprecise formation of small dots enabled them to iden-

(a) STEM image showing the bulk silicon (Si), five annealed dots (Au), carbon layer (C), and platinum layers. (c) Coloured STEM image of a single annealed nanodot (260°C, 2 h). Reproduced with permission, Copyright ©2011 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

tify the critical size that determines whether a dot iscomposed of single or multiple crystal domains.Moreover, they showed that annealing at moderatetemperature can convert Au dots from amorphous tosingle-crystalline, and then they were covered with athin SiO2 layer. After easy removal of the SiO2 (diluteHF etching), these nanodots can be used as electrodesfor the characterization of organic self-assembledmonolayers (SAMs) with less than 200 molecules.

A Stage for Micro-AxialFluorescence TomographyA miniature stage device to overcome resolutionanisotropy in fluorescence light microscopy isdescribed by Florian Staier and colleagues at theKirchhoff Institute for Physics, University of Heidel-berg, Germany [Rev. Sci. Instrum. 82:093701, 2011].To overcome the limitation of fluorescence micro-

scopes in anisotropic optical resolution or point local-ization precision micro-axial tomography was usedwhich allowed object tilting on the microscope stageand led to an improvement in localization precisionand spatial resolution. A glass fiber was placed in theobject space of the microscope lens and its rotationcontrolled by a miniaturized stepping motor. By Testparticles were fixed onto the glass fiber, opticallylocalized with high precision, and automaticallyrotated to obtain views from different perspectiveangles from which distances of corresponding pairs ofobjects were determined. From these angle depen-dent distance values, the real 3D distance was calcu-lated with a precision in the ten nanometer range(corresponding here to an optical resolution of 10-30nm) using standard microscopical equipment. As aproof of concept, the spindle apparatus of a maturemouse oocyte was imaged during metaphase II mei-otic arrest under different perspectives.

Enabling Two-ColourSTED of Live Cells

A technique for the use of two fluorophores in stim-ulated emission depletion (STED) microscopy of livingcells is reported by Patrina Pellett and co-workers atthe Department of Cell Biology, Yale School of Medi-cine, CT [Biomedical Optics Express 2(8):2364-2371,2011]. Current applications of STED microscopy havebeen limited to single colour imaging of living cellsand multicolour imaging in fixed cells. However, tostudy active processes, such as protein interactions, atwo-colour STED imaging technique is needed in liv-ing cells. This was achieved for the first time by theauthors: the key to their success was in overcomingthe challenges in labeling target proteins in livingcells with dyes optimal for two-colour STEDmicroscopy. By incorporating fusion proteins, theresearchers were able to improve the targetingbetween the protein and the dye, effectively bridgingthe gap. This allowed the researchers to achieve reso-lutions of 78 nm and 82 nm for 22 sequential two-colour scans of epidermal growth factor and its recep-tor in living cells.

3D Point Spread Function in Cs-Corrected STEMThe three-dimensional point spread function of anaberration-corrected scanning transmission electronmicroscopy (STEM) has been simulated and experi-mentally tested by Andrew Lupini and Niels de Jongeat the Oak Ridge National Laboratory, TN [Microscopyand Microanalysis 17:817-826, 2011].Aberration correction reduces the depth of field in

STEM and thus allows three-dimensional imaging bydepth sectioning. This imaging mode offers thepotential for sub-Ångstrom lateral resolution andnanometer-scale depth sensitivity. For biological sam-ples, which may be many µm across and where highlateral resolution may not always be needed, opti-mizing the depth resolution even at the expense oflateral resolution may be desired, aiming to imagethrough thick specimens. Although there has beenextensive work examining and optimizing the probeformation in two dimensions, there is less knownabout the probe shape along the optical axis. The authors examined the probe shape in 3D in an

attempt to better understand the depth resolution in

Image Analysis of Deep Etched Actin FilamentsA fractal dimension analysis and mathematical mor-phology of structural changes in actin filamentsimaged by electron microscopy is reported by Yoshi-taka Kimori et al at the National Institutes of NaturalSciences, in Tokyo [Journal of Structural Biology176(1):1-8, Oct 2011].The authors examined structural changes of actin

filaments interacting with myosin visualized by quickfreeze deep-etch replica EM by using a new methodof image processing and analysis based on mathe-matical morphology. To quantify the degree of struc-tural changes, two characteristic patterns wereextracted from the EM images: the winding patternof the filament shape (WP) reflecting flexibility of the

Atom Probe Imaging ofGaAsSb Quantum Rings A. Beltrán and colleagues at the Department of Mate-rials Sciences, University of Cadiz, Spain, report thatthree dimensional atom tomography resolves thequantum ring morphology of self-assembled GaSbburied nanostructures [Ultramicroscopy 111(8):1073-1076, July 2011].Unambiguous evidence of ring-shaped self-assem-

bled GaSb nanostructures grown by molecular beamepitaxy is presented on the basis of atom-probetomography reconstructions and darkfield transmis-sion electron microscopy imaging. From atom-probetomography compositional distribution has beenobtained. The GaAs capping process causes a strongsegregation of Sb out of the center of GaSb quantumdots, leading to the self-assembled GaAsxSb1-x quan-tum rings of 20-30 nm in diameter with x~0.33.

filament, and the surface pattern of the filament (SP)reflecting intramolecular domain mobility of the actinmonomers in the filament. EM images were processedby morphological filtering followed by box-countingto calculate the fractal dimensions for WP (DWP) andSP (DSP). The result indicated that DWP was largerthan DSP irrespective of the state of the filament(myosin-free or bound) and that both parameters formyosin-bound filaments were significantly largerthan those for myosin-free filaments. This work is thefirst quantitative insight into how conformational dis-order of actin monomers is correlated with themyosin-induced increase in flexibility of actin fila-ments along their length.

this mode. They present examples of how aberrationschange the probe shape in three dimensions, and itwas found that off-axial aberrations may need to beconsidered for focal series of large areas. It was shownthat oversized or annular apertures theoreticallyimprove the vertical resolution for three-dimensionalimaging of nanoparticles. When imaging nanoparti-cles of several nanometers in size, regular scanningtransmission electron microscopy can thereby be opti-mized such that the vertical full-width at half-maxi-mum approaches that of the aberration-correctedSTEM with a standard aperture.

MICROSCOPY AND ANALYSIS NOVEMBER 201126

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Free Educational Webinar: Sample Preparation for Scanning Electron Microscopy (SEM) State of the art Critical Point Drying with the New fully automated Leica EM CPD300Speaker: Dr. Ruwin Pandithage, Leica Mikrosysteme GmbH, Vienna, Austria

Online Seminar/Webinar on Sample Preparation for SEM Register at http://www.microscopy-analysis.com/leicawebinars and join on Tuesday 25th October 2011 at 17:00 h (CET), 16:00 h (UK)

Watch this webinar if you are responsible for

• EM sample preparation of delicate biological specimens such as pollen, tissue, plants or insects• EM sample preparation of industrial samples for MEMS (Micro Electro Mechanical Systems) applications

Specimens that can be damaged due to surface tension when changing from the liquid to gaseous state, need special treatment during sample preparation. Critical point drying is an efficient method for drying such delicate samples for SEM applications because it preserves the surface structure. Before drying, many biological samples are additionally prepared through fixation and dehydration and then coated after drying with a metal such as gold, platinum or palladium to make their surfaces electrically conductive for the SEM analysis. If the surface structure is altered during the drying process the results of the following SEM application will provide incorrect results.

In the past, critical point drying was a time consuming process with low sample reproducibility due to manual operation.

This webinar will show you how the New Leica EM CPD300 dries delicate biological or industrial samples in a fully automated and controlled process to preserve your samples for subsequent treatment and analysis.

Register at http://www.microscopy-analysis.com/leicawebinars

This webinar is available on demand under the above registration link.

Critical Point Drying with Leica EM CPD300

Free Educational Webinar: Sample Preparation for Scanning Electron Microscopy (SEM) State of the art Critical Point Drying with the New fully automated Leica EM CPD300Speaker: Dr. Ruwin Pandithage, Leica Mikrosysteme GmbH, Vienna, Austria

Online Seminar/Webinar on Sample Preparation for SEM Register at http://www.microscopy-analysis.com/leicawebinars and join on Tuesday 25th October 2011 at 17:00 h (CET), 16:00 h (UK)

Watch this webinar if you are responsible for

• EM sample preparation of delicate biological specimens such as pollen, tissue, plants or insects• EM sample preparation of industrial samples for MEMS (Micro Electro Mechanical Systems) applications

Specimens that can be damaged due to surface tension when changing from the liquid to gaseous state, need special treatment during sample preparation. Critical point drying is an efficient method for drying such delicate samples for SEM applications because it preserves the surface structure. Before drying, many biological samples are additionally prepared through fixation and dehydration and then coated after drying with a metal such as gold, platinum or palladium to make their surfaces electrically conductive for the SEM analysis. If the surface structure is altered during the drying process the results of the following SEM application will provide incorrect results.

In the past, critical point drying was a time consuming process with low sample reproducibility due to manual operation.

This webinar will show you how the New Leica EM CPD300 dries delicate biological or industrial samples in a fully automated and controlled process to preserve your samples for subsequent treatment and analysis.

Register at http://www.microscopy-analysis.com/leicawebinars

This webinar is available on demand under the above registration link.

Critical Point Drying with Leica EM CPD300

Online Seminar/Webinar on Sample Preparation for SEMRegister at http://www.microscopy-analysis.com/leicawebinars

Free Educational Webinar: Sample Preparation for Scanning Electron Microscopy (SEM) State of the art Critical Point Drying with the New fully automated Leica EM CPD300Speaker: Dr. Ruwin Pandithage, Leica Mikrosysteme GmbH, Vienna, Austria

Online Seminar/Webinar on Sample Preparation for SEM Register at http://www.microscopy-analysis.com/leicawebinars and join on Tuesday 25th October 2011 at 17:00 h (CET), 16:00 h (UK)

Watch this webinar if you are responsible for

• EM sample preparation of delicate biological specimens such as pollen, tissue, plants or insects• EM sample preparation of industrial samples for MEMS (Micro Electro Mechanical Systems) applications

Specimens that can be damaged due to surface tension when changing from the liquid to gaseous state, need special treatment during sample preparation. Critical point drying is an efficient method for drying such delicate samples for SEM applications because it preserves the surface structure. Before drying, many biological samples are additionally prepared through fixation and dehydration and then coated after drying with a metal such as gold, platinum or palladium to make their surfaces electrically conductive for the SEM analysis. If the surface structure is altered during the drying process the results of the following SEM application will provide incorrect results.

In the past, critical point drying was a time consuming process with low sample reproducibility due to manual operation.

This webinar will show you how the New Leica EM CPD300 dries delicate biological or industrial samples in a fully automated and controlled process to preserve your samples for subsequent treatment and analysis.

Register at http://www.microscopy-analysis.com/leicawebinars

This webinar is available on demand under the above registration link.

Critical Point Drying with Leica EM CPD300

LEICA WEBINAR.indd 1 24/10/2011 15:40

CIRCLE NO. 25 OR ONLINE: www.microscopy-analysis.com

Discover how to make every electron count at fei.com/chemistem.

Introducing the newest member of the Titan family: Titan™ G2 80-200 with ChemiSTEM™ Technology. Combining FEI’s groundbreaking ChemiSTEM Technology with Titan’s market-leading sub-atomic resolution

imaging, Titan G2 80-200 offers highly sensitive, fast elemental mapping, superior ultra-low concentration

detection and the highest analytical probe current available. Even EDX tomography is easily achieved with

stunning results.

Make Every Electron Count ChemiSTEMTM Technology

GaAs Atomic EDX showing 1.4 Angstroms dumbbells with Titan G2 80-200 with ChemiSTEM Technology

Sample courtesy of Ionela Vrejoiu and Eckhard Pippel, Max Planck Institute of Microstructure Physics, Halle/Saale, Germany.

La1-xSrxMnO3/SrRuO3 multilayer/quantum well systems in [100] projection

CIRCLE NO. 26 OR ONLINE: www.microscopy-analysis.com

FOR FREE PRODUCT INFORMATION VISIT OUR WEBSITE: www.microscopy-analysis.com

what’s newWHAT’S NEW

MICROSCOPY AND ANALYSIS NOVEMBER 2011 29

3D Imaging Dual BeamFEI, a leading instrumenta-tion company providingelectron microscope systemsfor applications in researchand industry, has released itsnew Versa 3D DualBeam sys-tem, which provides high-resolution, three-dimen-sional (3D) imaging andanalysis on a wide range ofsample types. The Versa 3D’shighly configurable plat-form allows customers toadapt the system’s capabili-ties to their specific require-ments. “The flexible configura-

tion of the Versa 3D meetsthe demands of today’sresearchers who study awide variety of materials,”said Trisha Rice, vice presi-dent and general managerof FEI’s Research BusinessUnit. “FEI’s pioneering leadership in ion beam and electron beam techniques andmethodologies are well matched to give researchers information from even themost challenging samples. Last year, FEI introduced the latest generation HeliosNanoLab, the highest resolution DualBeam in the world that incorporates indus-try-leading electron and ion beam technologies, and today we are unveiling themost flexible DualBeam, the Versa 3D.”Versa 3D is available with either high vacuum-only or high and low vacuum

electron imaging hardware. Low vacuum electron imaging capabilities allows thesystem to accommodate contaminating or outgassing samples that are incom-patible with high vacuum operation. Low vacuum also provides the ability tocompensate for charge build up in non conductive samples even at the high cur-rents required for analysis techniques, such as energy dispersive (X-ray) spec-troscopy (EDS) and electron backscatter diffraction (EBSD).The Versa 3D combines FEI’s leadership in Schottky field emission electron

beam and high throughput ion beam technologies into a configurable Dual-Beam system, setting a new standard for 3D characterization and analysis, site-specific sample modification and advanced sample preparation for transmissionelectron microscopes (TEMs) and atom probes. The high-performance platformcan also be configured with FEI’s impressive low vacuum capabilities and evenenvironmental scanning electron microscopy (ESEM) for in situ analysis. Advanced SEM scanning and FIB patterning yield powerful imaging and

milling performance. New features, such as FEI’s SmartSCAN and Drift CorrectedFrame Integration (DCFI), facilitate electron beam imaging of sample types witha range of different properties. Advanced backscattered electron, as well as sec-ondary electron and ion detectors, collect a wide variety of topographic, ele-mental and compositional information ‘from every angle’. The combination of the latest AutoSlice & View G3 software option, the ver-

satile electron imaging hardware and high throughput ion column enablesresearchers to capitalize on the charge balancing capabilities of ions and elec-trons. Milling (with positive ions) and imaging or drift suppression (with elec-trons) provides a unique synergy for automation of 3D serial slicing, imaging andanalysis of both electrically conductive and non-conductive samples. When com-bined with EDS or EBSD, FEI’s EDS3 and EBS3 software options can also be usedto reconstruct elemental maps or crystallographic orientation data in 3D. The Versa 3D addresses the diverse needs in materials research, life sciences,

electronics and geosciences. It is available for ordering immediately. Contact: FEI Company www.fei.com

MagneticFieldCancellation

Technical Manufacturing Corporation(TMC) has improved upon the originaldesign of its Mag-NetX magnetic fieldcancellation system to allow for easierinstallation and service and to makeMag-NetX more adaptable to cus-tomers’ changing needs. The stainlesssteel struts are now modular and inter-changeable and available in a varietyof lengths to accommodate virtuallyevery commercial SEM. In addition, anew modular corner piece is designedwith integrated electronics for easyinstallation and service. TMC developed Mag-NetX to

actively compensate for magnetic fieldfluctuations caused by nearby machin-ery, elevators, power lines, and otherexternal sources. Such sources candiminish the performance of scanningand transmission electron microscopes,electron beam lithography systems,focused ion beam instruments, andother tools that incorporate a chargedbeam.The open cube-shaped Mag-NetX

detects magnetic fields and sends outan equal and opposite field to cancelthe interference. The system consists ofa dedicated controller with automatedcalibration and self-test features, ACand DC magnetic sensor, andHelmholtz coils in a structural casing. Itcan be floor-,wall-, or tool-mountedand sized to user-specific require-ments. The system is dynamic, continu-ously monitoring and achieves 35-40dB of attenuation. Contact: Technical ManufacturingCorporation www.techmfg.com

Sample Vice

The Micro Vice sample holder from STJapan is a simple yet versatile tool toadjust and hold samples accurately foranalysis with light microscopes, stereo-microscopes and FTIR- and Ramanmicroscopes. It efficiently supportsmicroscope analysis of difficult shapedsamples, fibers and films and all kind ofsamples up to a diameter of 40 mm.The Micro Vice: holds round and

unevenly shaped samples such astablets, minerals, etc. in the desiredposition; stretches accurately and holdspolymer films, fibers, hair, etc.; and tiltssamples for correcting oblique sampleorientation.In microscopical analysis it is often

challenging to securely place your sam-ples, such as tablets, gem stones, filmsor fibers in the exactly desired position.With the MicroVice it is easy to holdround samples such as tablets undercompression in the desired position,hold films and other flexible materialsunder tension and even stretch them.The MicroVice sample holder isdesigned to be easily attached to thestages of light microscopes, stereo-scopes and also on infrared and Ramanmicroscopes.You can place your sample in the

exactly desired position and tightenthe two screw handles. The sample willbe hold securely under compression.Even round samples can be easily heldand won’t slip. The clamps are cush-ioned and protect your sample fromdamage. The sample holder is supplied with

two small plates that are used forclamping taut and holding films, fibers,yarns, etc. By using the two screw han-dles you can stretch the samples usingtension by user defined increments.Due to the design of the MicroVice,exact parallel stretching of the samplesis ensured.Additionally, the fixed sample can

also be tilted up to a certain angle forcorrecting oblique sample orientation. Contact: S.T. Japan-Europe GmbHwww.stjapan.de

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MICROSCOPY AND ANALYSIS NOVEMBER 201130

Quantitative CathodoluminescenceAttolight AG debuted the first-ever quantitativecathodoluminescence microscope featuringnanoscale resolution and picosecond timing in aneasy-to-use platform at the recent M&M meeting inNashville, TN.“This is really the beginning of a whole new revo-

lution in cathodoluminescence,” said Attolight CEO,Dr Samuel Sonderegger. “There is no need to com-promise with this system: we run the full spectrumfrom UV to IR while maintaining 10 nm spatial reso-lution, and the full temperature range from 15K to300K, all with 100� larger field of view and up to100� more collection efficiency than any prior CLtechnology. Scientists can now have nanoscale imag-ing and pico-second time resolved spectroscopy, all inone instrument, with no compromises.” At the heart of the Attolight CL line is a newly

designed scanning electron microscope (SEM) con-taining an embedded optical microscope, a 9-axis cryonano-stage, and a fully integrated (not merely inter-faced) cathodoluminescence system. Available in two versions, the continuous wave CL

10-Infinity can be field-upgraded to the picosecond,time-resolved CL10-10.

The specifications are: spatial resolution: 10 nmacross the full spectrum; spectral range: UV to IR; fieldof view: 300 µm; optical NA of 0.78 with 100� the col-lection efficiency; positioning: 9-axis cryo stage withpivot point lock; pulsed operation: picosecond speedwith no electron dispersion; drift and vibration: min-imized; beam blanking: proprietary laser driven, psphotoelectron gun; no compromise in spatial (10 nm)or temporal (ps) resolution across the spectrum fromUV to IR; thermal range: 20K to 300K.Contact: Attolight AG www.attolight.com

Plasma FIB-FESEM WorkstationTESCAN, a world leading manufacturer of scanningelectron microscopes and focused ion beam worksta-tions has introduced the FERA3 XMH – a high-resolu-tion Schottky field-emission scanning electron micro-scope with a fully entegrated plasma source focusedion beam. The system has been developed in co-oper-ation with the French company Orsay Physics. In addition to electron and ion columns, the FERA3

XMH Plasma FIB-FESEM can be configured with gasinjection systems, nanomanipulators, and a wide vari-ety of detectors including SE detector, BSE detector, SI(secondary ion) detector, CL (cathodoluminescence)detector, EDX and EBSD microanalyzers, etc. The use of a xenon plasma source for the focused

ion beam allows the FERA3 to satisfy high-resolutionFIB requirements (imaging, fine milling/polishing), aswell as achieving high ion currents needed for ultra-fast material removal rates. The resolution of theplasma ion beam is <100 nm and the maximum Xe ioncurrent is >1 µA. Compared to existing FIB technolo-gies with gallium sources, the material removal rateachievable for silicon with the PFIB (plasma FIB) is over30x faster. For this reason the FERA 3 XMH is wellsuited for applications requiring the removal of largevolumes of material, particularly in the semiconduc-tor packaging corridor where TSV technology is beingutilized.

TESCAN will deliver the first system to the MiQroInnovation Collaborative Centre (C2MI) in Canada thisyear. The system will be used for the inspection ofintegrated circuit (IC) packaging.The FERA 3 FIB-SEM workstation’s integration of

both an electron and a focused ion beam places thistool in a class all its own, affording the end user thebenefits of electron beam analysis and characteriza-tion. Generally, systems of this kind will be used forcircuit editing, 3D metrology, defect analysis and fail-ure analysis.Contact: TESCAN www.tescan.com

Thermo Fisher Scientific has announced that its newThermo Scientific STP 420ES Tissue Processor for high-throughput tissue processing has been independentlyvalidated for xylene-free protocols. Scientists at the UK’sNewcastle upon Tyne Hospitals NHS Foundation Trustconfirmed the successful application of a standardxylene-free protocol on the STP 420ES. The tissue proces-sor delivered quality results using existing protocols, withthe exception of the requirement for a lower tempera-ture of 65°C, not 85°C, for the initial wax step. Elimination of xylene from tissue processing can cut costs, save time and improve the laboratory environment.

Using isopropyl alcohol (IPA) as an alternative wax miscible dehydrant removes the risks of cumulative exposureand the high disposal costs associated with xylene, which is a hazardous chemical. Using IPA also shortens cycletimes and enables leaner workflows so laboratories can deliver patient results faster. Contact: Thermo Fisher Scientific www.thermoscientific.com/pathology

Xylene-Free Tissue Processor

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EBSD/EDS

FREEWEBINARREGISTER

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Advanced Phase ID Using Combined EBSD & EDS on SEM Date: 11 January 2012, 16.00 GMT, 17.00 CEST, 11.00 EDT

Duration: 1 hour

Presenters: Dr. Daniel Goran, EBSD Application Scientist Dr. Laurie Palasse, EBSD Application Scientist

Electron backscatter diffraction (EBSD) and energy dispersive X-ray spectrometry (EDS) are common analytical methods used on the scanning electron microscope (SEM). They are complementary techniques and provide structural and compositional information respectively. Building on its recent developments in integrating EBSD and EDS, Bruker is now releasing an advanced phase identification feature. This new method significantly increases efficiency when dealing with multiphase materials and allows experts as well as less experienced users to acquire the best quality results.

The webinar will focus on describing the new phase identifi cation procedure and its advantages compared to the common phase identifi cation method.

Our experts will present numerous materials and earth science application examples.

Who should attend? Researchers working in electron microscopy labs studying crystalline

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Innovation with Integrity

Volcanic rock - phase map

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CIRCLE NO. 28 OR ONLINE: www.microscopy-analysis.com

The Dimension FastScan AFM: SEM like user experience, True 3D nanometrology, and quantitative material property mapping in ambient, fl uid, and controlled environments

Innovation with Integrity Atomic Force Microscopy

In many applications Atomic Force Microscopy (AFM) can provide unique and preferred sample information, however its slow speed and high complexity have often offset these benefi ts in favor of Electron Microscopy.

The latest generation of Bruker’s AFM, the Dimension FastScan™, enables nanometer resolution imaging, in a fraction of a minute, on a large-sample, fully automatable stage. The included ScanAsyst™ algorithm provides robust, intuitive, self-optimizing work-fl ow based operation. Combined these two capabilities create a highly productive nano-imaging solution akin to current Scanning Electron Microscopes (SEM).

While both techniques provide surface imaging on the nano scale, the insights gained from each technique are also complimentary:

• AFM can be performed, at nm-resolution, in ambient and fl uid environments, and typically requires no alterations of the sample surface chemistry prior to imaging. This enables non-destructive sample prep, convenient (multi-) sample loading, easy sample access, and imaging of dynamic sample changes over time.

• AFM provides true nano-metrological information in all three sample dimensions

• While SEM techniques can provide contrast based on elemental analysis, the latest-generation AFM mode, Bruker’s proprietary PeakForce-QNM, provides quantitative nanoscale mapping of surface mechanical properties, such as modulus, adhesion, or dissipation, in addition to the standard topographical information

Please join us for this focused review of the recent advances of the “other” nanoscale surface imaging technology.

DATE: December 7th 4 pm BST, 5 pm CET, 11:00 am EDT

Presenting: Dr. Johannes Kindt

The Dimension FastScan AFM: SEM like user experience, True 3D nanometrology, and quantitative material property mapping in ambient, fl uid, and controlled environments

DATE: December 7th

FREEWEBINARREGISTERTODAY!

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BRUKER NANO WEBINAR REV.indd 1 24/10/2011 10:36

CIRCLE NO. 29 OR ONLINE: www.microscopy-analysis.com

MICROSCOPY AND ANALYSIS NOVEMBER 2011 33

WHAT’S NEW

CIRCLE NO. 30 ORONLINE: www.microscopy-analysis.com

Hybrid Light and Electron MicroscopeTopcon Positioning Systems (TPS), a globalleader in precision optical, GPS, and automa-tion systems for geodetic, engineering and con-struction applications, has introduced theAquila hybrid microscope to the North Ameri-can market.The microscope magnifies from 30� in opti-

cal mode, to 50,000� as an SEM. This uniquehybrid design provides key advantages thatshould expand the use of SEM technology.Ray O’Connor, TPS president and CEO, said,

“The Aquila hybrid scope puts the versatility ofan optical microscope and the power of anscanning electron microscope (SEM) into a sim-ple, portable, and affordable package.” Thesimplicity of the Aquila, both in design andfunctionality “creates an instrument with virtu-ally no learning curve,” he said. “With an array

Mini-SEMHolders

In close co-operation with its preferreddevelopment partner Deben, Phenom-World has developed a Temperature Con-trolled Sample Holder to study vacuum-sensitive and vulnerable samples on itsPhenom G2 desktop SEMs. This active sample holder is designed to

control the temperature of the samplebetween -25°C and +50°C. With the use ofthe temperature controlled holder, thetemperature of the sample is manipu-lated, therefore the humidity around itcan be controlled. This enables imaging ofmoistures and water containing samplesas well as reducing the effect the electronbeam has on beam sensitive samples. Thisresults in an extended viewing time, with-out noticeable vacuum artifacts. Theholder can be retrofitted to all versions ofthe Phenom G2 system.The improved Fibermetric application

allows measurements and analysis oncomplicated fiber structures, ranging fromspunbond and electrospun fibers to themelt blown type of fibers. Fibermetric pro-vides accurate size information from microand nano fiber samples. Through furtherautomization of several important fea-tures, the Fibermetric has become evenmore user-friendly and guarantees a fastreturn on investment. The automated fea-tures that have the most effect on this arethe high number of measurements (max.1000 per image), the automated featureand fiber size detection, and the analysisof the data points.Contact: Phenom-World BV www.phenom-world.com

of features that make it attractive to both theexperienced and new SEM user, it is the mostversatile and easy-to-use microscope on themarket.”“Topcon has a long and respected history in

the optical market,” O’Connor said. “Eventhough Aquila is not our typical positioninginstrument, it has incredible application oppor-tunities in several markets TPS already serves –mining, forensics, education, and water man-agement in particular.” In addition to the areaspointed out by O’Connor, he noted the instru-ment has practical usage in air and water mon-itoring, metallurgy and metal analysis, materi-als analysis, food industry, quality control andmicro-mechanical systems. Contact: Topcon Positioning Systems, Inc. www.topconsem.com

MICROSCOPY AND ANALYSIS NOVEMBER 201134

Hitachi High-Technologies has launched the SU8000 family of ultrahigh-resolutionfield-emission scanning electron microscopes (FESEMs) for investigating the finesurface structure of materials in a wide range of nanotechnology fields. The newSU8000 series features a common, high performance electron optical platform toprovide excellent imaging performance, and offers a variety of stages, chambersand signal detection systems. The new SU8010, SU8020 and SU8030 join the existing SU8040 to form this com-

prehensive family of ultrahigh-resolution microscopes. A high-brightness cold cath-ode field-emission source is used in combination with the latest generation ofHitachi’s patented super ExB in-lens detection systems for energy filtering, chargesuppression, and contrast control. All microscopes offer excellent imaging perfor-mance at low accelerating voltage to minimize sample damage, and enhancedelectron deceleration technology has improved resolution at ultralow landing volt-ages to just 1.3 nm at 1.0 kV. The SU8010 is the entry level model with dual (upper and lower) secondary elec-

tron detectors with secondary and backscattered electron signal mixing capabili-ties for versatile imaging. A three-axis motorized stage is provided as standard,capable of accommodating samples up to 100 mm in diameter.The SU8020 offers the same sample handling capabilities using a 5-axis motor-

ized stage as standard but benefits from Hitachi’s unique triple detector system toextend the capability to collect secondary electrons and low energy backscatteredelectrons. The SU8030 features a large chamber with large specimen stage for sam-ples up to 150 mm in diameter. Contact: Hitachi High-Technologies Corporation www.hht-eu.com

High Resolution FE-SEMs

PicoQuant GmbH hasannounced the successfulcombination of Pico-Quant’s time-resolved con-focal fluorescence micro-scope MicroTime 200 withBruker’s BioScope Catalystatomic force microscope.The combination of thesetwo systems enables simul-taneous recordings of AFMand optical images of thesame sample region and makes new investigation schemes in the field of live-cellimaging feasible.The combined setup of the MicroTime 200 and the Bioscope Catalyst is straight-

forward without the need of larger modifications of the two systems. The syn-chronized data acquisition enables scientists to analyze, e.g., the impact of proteinchanges on cell shape and structure. It also allows high-resolution imaging bymerging of sub-nm topography with optically encoded functionality as well asinvestigations of inter- and intramolecular distances using force spectroscopy.The Bioscope Catalyst AFM including its sample stage is mounted onto the

inverted microscope body of the MicroTime 200, which is configured for objectivescanning. In this way, precise overlay of the confocal volume and the AFM tip canbe realized. Electronic communication between the sample-scanner of the AFMand the data acquisition electronics of the MicroTime 200 enables simultaneousrecordings with the two instruments. Contact: PicoQuant GmbH www.picoquant.com

Combined Confocal and AFM

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Lift-Out Shuttle for TEMJPK Instruments hasexpanded its family ofhigh performanceresearch AFM systemswith the announcementof the availability of theNanoWizard 3 Nano-Optics AFM system.Over the past decade,

optical phenomena onthe nanoscale havedeveloped into an excit-ing area of research. Tostudy light on the nanoscale and especially its interaction with matter, researcherslook for methods with nanometer spatial resolution. The combination of lightmicroscopy-derived techniques and scanning probe microscopy is a powerful solu-tion. This so-called near-field optical microscopy delivers optical information fromsample surfaces with sub-wavelength resolution.JPK strongly believes in combining techniques, in particular AFM, with optics.

This has opened up a field of new applications including TERS/SERS, tip-enhancedfluorescence, nanomanipulation with light, chemical surface analysis and com-pound detection, metamaterials, developments of optically active componentssuch as dyes, markers, light sources and switches. A large number of user publica-tions underscore the success of this technology approach. Now, JPK introducestheir latest platform for AFM and optics - the NanoWizard3 NanoOptics system.The NanoWizard NanoOptics head comes with excellent physical and optical

access to the sample from top and bottom as well as from front and side, evenwhen the head and condenser are in place. Additionally, it has an integrated portfor fiber SNOM applications.The new system is ready for a broad range of applications from nanoscale opti-

cal imaging by aperture and scattering-type SNOM to experiments involving inter-actions of light with the sample such as absorption, excitation, nonlinear effectsand quenching; these include aperture fiber SNOM experiments using an inte-grated fiber SNOM port in the NanoOptics head and the tuning fork module. Contact: JPK Instruments AG www.jpk.com

Nano Optics on AFMWorking with leadingGerman manufacturerof precision manipula-tion products, KleindiekNanotechnik, Agar Sci-entific are pleased tooffer the Lift-Out Shut-tle for the UK and Irishmarkets.In-situ lift-out tech-

niques have becomemore reliable methodsfor preparation of samples requiring TEM and atom probe inspection. However,despite their new-found popularity, they remain considerably more expensive thanex-situ lift-out techniques and require lots of valuable time in the focused ionbeam (FIB). Time and cost factors call for a faster, simpler procedure while furtherimproving the reliability of the technique. By combining a precision sub-stage with a microgripper system and their novel

SemGlu, Kleindiek have developed a new, fast, easy and efficient tool: The Lift-OutShuttle. This device is small enough to fit through the load lock of most commonelectron microscopes. Mounting the sample and the TEM-grid that has beentreated with a small amount of SemGlu to a single microscope stub puts all thepieces necessary for lift-out in place. The stub is then attached to the four-axis(XYZR) sub-stage. Additionally, the Microgripper is fixed to same mounting plateon which the sub-stage is positioned. In this configuration, the gripper remains sta-tionary above the sub-stage and therefore is always centred in the field-of-view ofthe microscope. The stub containing the sample and grid is positioned under thegripper using the sub-stage.With a minimal amount of practice, a sample can be fixed to a grid in five min-

utes or less by gripping the previously cut sample, lifting out of the substrate, mov-ing the stage to the TEM-grid (which has been pre-treated with SEMGLU by simplyapplying some glue to the grid using a needle), and mounting the sample to thegrid by making contact between the two and focussing the e-beam on the contactpoint.Contact: Agar Scientific Limited www.agarscientific.com

CIRCLENO. 33 ORONLINE: www.microscopy-analysis.com

SURFACECOMPOSITION

ANALYSISFOR ION MICROSCOPYHiden’s EQS and MAXIM SIMS analysers provide:

� chemical surface compositionanalysis for ion probemicroscopy

� depth profiling andsurface imaging atthe nano scale

� interface toexisting systems

for further details of Hiden Analytical products contact:

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MICROSCOPY AND ANALYSIS NOVEMBER 201136

Boeckeler RMC Productsrecently celebrated thesuccessful launch of itsnew environmentalchamber for ultramicro-tomy during a trainingdemonstration at theSociété Française Mu(SFmu) 2011 conferenceheld in Strasbourg,France last June. Accord-ing to Boeckeler President Pat Brey, the demonstration included an impressivearray of sectioning methods including sectioning for low voltage TEM (transmissionelectron microscopy), Tokuyasu immunolabeling and CEMOVIS (cryo-electronmicroscopy of vitreous sections) sectioning.

The environmental chamber was critical to creating the parameters necessary forproducing a continuous ribbon of 25 sections of a frozen hydrated sample 50 nmthick that had been flash frozen under high pressure. Such high-pressure freezingof biological specimens retains as much cell structure as possible for viewing underan electron microscope. The CEMOVIS method of sample preparation is experi-encing resurgence in popularity among cell biologists since it was introduced in the1980s, due in part to advances in overcoming some of the difficulties first encoun-tered in the method.

“These tiny ribbons are so thin, they could be free-floating,” Mr Brey says. “So,that means that the environment around the RMC ultramicrotome had to be espe-cially free of air movement and static charges, which the chamber and ionizer suc-cessfully provided.”

The work was performed on samples frozen in the lab of Daniele Spehner fromthe Institute of Genetics and Molecular and Cellular Biology. Colleague CarolineKizilyaprak from Bruno Humbel’s lab at the University of Lausanne, and GregBecker of RMC performed the sample trimming and sectioning, using a Diatome35-degree diamond knife. Contact: Boeckeler Instruments Inc. www.rmcproducts.com

Environmental Cryo Chamber

CEMOVIS 50 nm serial sections of yeast cut on an RMC PT-PC system

The new phoenix x|aminer from GE’s Inspection Technologies business is a 5-axis,microfocus X-ray inspection system, which has been developed for quality controlapplications in the manufacture of electronics sub-assemblies. It is particularly suit-able for the reliable and accurate inspection of soldered joints. The phoenixx|aminer features two megapixel high resolution and high magnification. In addi-tion, its powerful imaging software permits intuitive programming and compo-nent manipulation is precise and easy using a computer mouse or joystick.

An important design feature of the new system is its OVHM (oblique view highmagnification) module, which allows an oblique angle view of up to 70° as well asa total magnification of up to >23,000�. This ensures best possible quality of defectinformation in the vertical direction. Ease of maintenance is also permitted with thesystem’s open, 160 kV microfocus tube design, whose easy cathode replacementensures unlimited operating life, while its 20 W of tube power at target can pene-trate even the most radiation-absorbing components. The new x|aminer is suppliedwith the recently launched phoenix x|act software, which is designed specially forelectronics inspection. Contact: GE Energy Services www.ge-mcs.com

X-Ray Inspection Solution

Backscatter Diffraction

Oxford Instruments NanoAnalysis, a world leader in microanalysis systems,launched a new EBSD detector at M&M 2011 in Nashville. The new NordlysNanoaddresses the growing requirements of nanoscale applications: it is 60% more sen-sitive than the previous generation of EBSD detector, the NordlysS.

Increased sensitivity offers a number of benefits. Firstly, accurate EBSD data canbe collected at lower beam energy, including low beam currents (<0.1 nA) and lowkV. Using lower energy beams is essential for applications where spatial resolutionis required, for example nanoscale applications. It is also important when lookingat beam sensitive samples, or non-conducting samples. In addition, increased sen-sitivity enables faster data acquisition under comparable beam conditions.

According to EBSD Business Manager, Dr Jenny Goulden, “As a business, OxfordInstruments aims to use innovation to turn smart science into world class productsthat delight our customers, and the NordlysNano is a classic example of achievingjust that!” The new NordlysNano EBSD detector is 60% more sensitive than its pre-decessor, the NordlysS. It enables faster and higher resolution analysis of a crystal structure.Contact: Oxford Instruments www.oxford-instruments.com

Hitachi High-Technologies has launched the IM4000 Ion Milling system. Used toprepare specimens for scanning electron microscope (SEM) imaging and analyticalstudies such as EDX and EBSP, the versatile IM4000 is capable of both pin-pointcross-section and flat surface ion milling.

Cross-section milling provides smooth cross-section specimens for high resolutionimaging of subsurface structures, with the cross-section position accurately con-trollable by fine positioning of a beam-shielding mask edge. Flat milling providesuniform polishing of surfaces of 5 mm in diameter or more with variable anglemilling, to either flatten surfaces or to selectively enhance specimen surface fea-tures (relief milling). The two IM4000 applications – cross-section and flat surfacemilling – are realised via two different removable sample stage units, allowing forconvenient specimen setting and cutting edge definition using an external opticalmicroscope.

The IM4000 ion milling system features a new high-current argon ion gun thatdelivers cross-sectional milling rates of 300 um/hr in silicon for dramatically reducedcross-sectioning times. The wide Argon ion beam can define sharp cross sectionseven on samples of dissimilar materials with different hardness that can not be cutor broken without causing material deformations or dislocations. Sensitive mate-rials like polymers or papers can be processed by freely selecting proper lower ionbeam energies between 0 and 6 kV, without need for special sample cooling.

Wide and smooth flat milled surfaces of approximately 5mm in diameter ormore can be achieved within minutes by shifting the centre of the defocused ionbeam from the sample rotation or swinging centre. The beam irradiation angle tothe specimen surface is selectable from 0 to 90 degrees. Contact: Hitachi High-Technologies Corporation www.hht-eu.com

Argon Ion Milling System

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Web Microscopy 2.0We live in the midst of Web 2.0 and Cloud computingrevolution. Today you can store documents, imagesand music on the Cloud and access them from anyweb browser; you can share data instantly and col-laborate in real time. If we can do so many thingsonline, why not image analysis? With proliferation ofhigh-speed internet and digital imaging, may be it istime to bring web 2.0 technology to microscopy?

One aspiring technology startup thinks so: SmartImaging Technologies, a VC-backed technology ven-ture from Texas, pledges to bring Web 2.0 technologyto microscopy with Simagis Live cloud software. “Wewant to bring power and convenience of web appli-cations to microscopy and microanalysis, and makeweb technology available to millions of microscopeusers around the globe. If you have digital cameraand Internet- you are ready for Web Microscopy,”says company founder Vitali Khvatkov.

With Web Microscopy, a small software utility onyour desktop will automatically upload your digitalimages to a cloud server so you can work with themonline via a web interface. If you have automatedmicroscope stage, you can stream individual image

200 kV Transmission Electron MicroscopeA new 200 kV transmission electron microscope fromJEOL delivers high throughput nano-analysis forprocess and quality control of mass produced semi-conductor and materials samples. The multi-functionJEOL JEM-2800 features high resolution imaging inTEM, STEM, and SE modes; ultrasensitive elementalmapping with a large angle Energy Dispersive Spec-trometer (EDS); Electron Energy Loss Spectroscopy(EELS) for chemical analysis; critical dimension analy-sis; tomography; and in situ observation of samples.The all-new TEM functions without use of the tradi-tional fluorescent screen on the electron column.

The JEM-2800 speeds specimen observationthrough fully automatic functions including adjust-ment of focus, astigmatism, contrast, brightness, crys-tal zone axis alignment, and height. Switchingbetween analysis modes is seamless, and quick datacollection shortens turnaround time between sam-ples. An operator navigation system and on-screenoperating guide make the JEM-2800 a high through-put, user-friendly TEM for any skill level.

Additional features and key specifications of theJEM-2800 include a Schottky field-emission electrongun, highly stable eucentric side-entry goniometerstage, a magnification range of 100� to

Widefield Superresolution MicroscopeWith the new Leica SR GSD from LeicaMicrosystems, scientists can now achieve res-olutions far below the limit of diffraction thathave never been attained before in widefieldfluorescence microscopy. The system is capa-ble of resolving details as small as 20 nm.

The SR GSD is based on GSDIM (groundstate depletion followed by individual mole-cule return) technology exclusively licensedfrom MPI Goettingen. It has already delivered amaz-ing results in scientific experiments during its testphase. One of the key advantages of the GSDIMmethod is that it can be used with conventional fluo-rescence labels routinely applied in fluorescenceimaging applications. GSDIM provides the highest res-olution possible with a light microscope today, almostequalling that of an electron microscope.

The Leica SR GSD is based on a fully automated TIRF

150,000,000�using STEM, 0.1 nm TEM resolution and0.20 nm BF/DF STEM resolution.

The JEM-2800 is JEOL’s latest addition to its com-prehensive lineup of 100-300kV TEMs and mostrecently the ARM200F atomic resolution TEM. Thefirst US customer, a global semiconductor manufac-turer, will take delivery of the new JEM-2800 withlarge solid angle EDS this summer. Contact: JEOL USA, Inc. www.jeolusa.com

tiles to the server for stitching and creating mosaicimages or virtual slides, to view your entire specimenat full resolution online. Once your digital images orvirtual slides are uploaded, they can be viewed, anno-tated and measured by any number of people at thesame time from any computer or web tablet. You canupload composite images with several fluorescencechannels and view them online with overlays.

Web Viewers utilize zoom technology similar toGoogle Maps and allow panning and zooming on vir-tual slides of unlimited size. Contact: Smart Imaging Technologies Companywww.live.simagis.com

system. Combining the benefits of super-resolutionwith TIRF microscopy is one of many options. The sys-tem can also be used for a wide range of applicationsin all areas of live cell microscopy and high-end fluo-rescence microscopy. As a flexible, multifunctional sys-tem, the Leica SR GSD gives researchers the freedomto tailor the system exactly to their needs.Contact: Leica Microsystems www.leica-microsystems.com

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Bruker has introduced the XFlash 5060 T, thelatest addition to the XFlash family of detec-tors for X-ray micro- and nanoanalysis (EDS) inelectron microscopy. The XFlash 5060 T is oneof two detectors available for use on (scan-ning) transmission electron microscopes. Pro-viding 60 mm² active area, it guarantees opti-mum solid angle for the analysis at low beamcurrents and of samples with low X-ray yield.The slender detector end cap and microscope-specific collimator design allow shortestdetector-sample distances and provide a hightake-off angle without requiring sample tilt. Compared to Si(Li) detectors, still commonly used

for EDS on TEM, the XFlash 5060 T exhibits superiorspeed and drastically lower dead times, providing sig-nificant advantages in collection efficiency, even inlow count rate situations. Additionally, the XFlash5060 T can operate with good energy resolution atcount rates far beyond what any Si(Li) or even com-peting SDD can handle on TEM. The detector is fullyoperational at input count rates of up to 750,000 cps,a big advantage for low-mag high count rate STEMmapping. Also, there is no danger of ‘locking up’ thespectrometer for minutes when hitting a supportgrid.The superb sensitivity of the XFlash 5060 T allows

detection of high energy radiation. In combinationwith high-end signal processing electronics andsophisticated software, reliable quantitative analysisof element peaks at 40 kV and above is possible.

Large Area EDS Detector for S/TEM

Bruker‘s many years of experience in SDD and signalprocessing electronics design, have led to the out-standing energy resolution of the XFlash 5060 T atonly moderate cooling temperatures, enabling reli-able and efficient light element analysis of elementsdown to boron.The XFlash 5060 T, including electronics, is designed

to cause minimum interference with all compatibletransmission electron microscopes, conventional oraberration-corrected. Light-weight and with passivecooling, this detector causes minimal strain on the col-umn and introduces no vibrations. The low tempera-ture gradient provides stable measurement condi-tions and the completely non-magnetic detectorhead minimizes beam shift, when moving the detec-tor in or out during TEM operation.Contact: Bruker Nano GmbH www.bruker-nano.com

Chemical Analysis in the S/TEM

FEI, a leading instrumentation company providingelectron microscope systems for applications inresearch and industry, has announced the release ofits Titan G2 80-200 with ChemiSTEM technology, anew member of the Titan G2 series of S/TEM (scan-ning / transmission electron microscopes). “By combining the Titan platform’s latest genera-

tion of electron optics with the revolutionary analyt-ical sensitivity of ChemiSTEM Technology, we havecreated a microscope which can deliver atomic reso-lution elemental maps in minutes and adds new capa-bilities in addressing our customer’s applications inmaterials science, chemistry and nanotechnology,”said Trisha Rice, vice president and general managerof FEI’s Research Business Unit.Dr Paul Kotula of Sandia National Laboratories,

said: “Our institute chose the FEI Titan G2 80-200 dueto its innovative combination of the latest in probe-correction technology and large solid-angle, win-dowless silicon-drift X-ray detectors (SDDs). We esti-

GaAs Atomic EDX in the [110] crystal projection showing 1.4 Angstroms Ga-As dumbbells obtained with Titan G2 80-200 with ChemiSTEM technology. TheGaAs [110] dumbbell splitting of 0.14 nm is clearly resolved by chemical mapping using energy dispersive X-ray analysis with a Titan G2 80-200 with ChemiS-TEM technology and a probe Cs-corrector at 200 kV acceleration voltage, using a 200 pA probe current. On the right hand side the atomic structure of GaAsthe [110] projection is shown along with the grayscale HAADF-STEM image. This represents the highest resolution ever obtained in atomic elemental mappingby any technique using an S/TEM.

mate that we willgain a factor of 50 to100 in terms of ana-lytical sensitivity,speed, and spatialresolution com-bined, over our exist-ing FEG analyticalelectron microscope.It is already clear thatatomic resolution X-ray microanalysis is not only possible but practicalwith this new microscope. Once the domain only ofelectron energy loss spectrometry, atomic resolutionmicroanalysis with x-rays gives us access to more ofthe periodic table and the possibility to use existingquantification methods to routinely analyze manymaterials at the highest resolution and sensitivityneeded.” Contact: FEI Company www.fei.com/chemistem

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MICROSCOPY AND ANALYSIS NOVEMBER 201140

For LM, SPM, EM, Bold entries are the standard types of instruments; other entries arethe modes in which they are used. Use this also for web page cross referencing.

A N D A N A L Y S I S

C00 COMPOSITIONAL ANALYSIS­C02 Calibration devices and standardsC03 CathodoluminescenceC04 Electron backscatter diffraction

(EBSD)C05 Electron beam induced current

(EBIC)C06 Electron diffraction, convergent

beam electron diffraction (ED,CBED)C07 Electron energy loss spectroscopy,

electron spectroscopic imaging(EELS, ESI)

C08 Energy and wavelengthdispersive X-ray spectroscopy(EDX, WDX)

C09 Mass spectrometry (MS, SIMS)C10 X-ray diffraction, X-ray fluorescence,

X-ray photoelectron spectroscopy(XRD, XRF, XPS)

D00 DIGITAL IMAGING ANDANALYSIS

D01 CCD camerasD02 CMOS camerasD04 Frame grabbersD05 Image analysis hardwareD06 Image analysis softwareD07 Image archiving and reportingD09 Image intensifiersD10 Machine vision systemsD11 MetrologyD12 Particle countingD13 Stereoscopic displayD14 Video camerasD15 Video processors

P00 PHOTOGRAPHIC EQUIPMENTAND SUPPLIES

P01 Chemicals, film and paper

F00 SPECIMEN PREPARATIONEQUIPMENT AND SUPPLIES

F01 Coating unitsF02 Critical point dryersF03 Cryofixation and cryosubstitution

devicesF05 CryostatsF07 Cutting, grinding, polishing and

thinning devicesF08 Cytochemical, immunochemical

and in-situ probesF09 Electrolytic thinningF11 Fixatives, stains and chemicalsF12 Freeze drying equipmentF13 Freeze etch and fracture unitsF14 Glass/steel/diamond knivesF15 Gold probesF16 Grids F17 Histology equipment and suppliesF18 Ion beam etching and thinningF19 Ion beam millingF20 Ion beam sputter coatingF21 KnifemakersF22 MicrotomesF23 Microwave processingF24 Plasma cleaningF25 Plasma etchingF26 Reactive ion beam etchingF27 Section stainersF28 Tissue processorsF29 UltramicrotomesF30 Vibratomes

G00 GENERAL MICROSCOPY EQUIPMENT,SUPPLIES AND SERVICES

G01 Anti-vibration systemsG04 Image analysis serviceG05 Maintenance contracts and servicingG06 Microhardness and failure testingG07 Thermal analysisG08 TrainingG09 Used equipmentG10 Workshops for microscopists

H00 SOCIETIESH01 CONFERENCES, COURSES, AND

EXHIBITIONSH03 RECRUITMENT SERVICES

BUYER’S GUIDE

www. microscopy-analysis.com

L00 LIGHT MICROSCOPYL01 Transmitted light microscopesL02 Reflected light microscopesL03 Stereomicroscopes L04 AcousticL05 Confocal L06 Differential interference contrastL08 Field and mobile microscopesL10 Fluorescence lifetimeL11 Fluorescence resonance

energy transferL12 Fluorescence spectroscopyL13 High resolution imaging (4Pi, STED)L14 InfraredL15 InterferenceL16 Modulation contrastL17 MultiphotonL18 Multispectral imaging L19 Phase contrastL20 PolarizationL21 Profilometry and metrologyL22 RamanL23 Total internal reflection fluorescenceL24 UltravioletL30 LM ACCESSORIES AND SUPPLIESL31 Autofocusing devicesL32 Calibration devices and standardsL33 CondensersL35 Environmental chambersL36 EyepiecesL37 Filters, beamsplitters, polarizersL38 Lamps, IlluminatorsL39 Lasers, LEDsL40 Micromanipulators and

microinjectorsL41 ObjectivesL42 OpticsL43 Scanning headsL44 StagesL45 Stands

E00 ELECTRON, ION AND X-RAYMICROSCOPES

E01 Transmission electron microscopesE02 Scanning electron microscopesE03 Ion beam microscopesE05 X-ray microscopesE07 Analytical TEME08 Atom probesE09 Auger microscopesE10 Cryoelectron microscopesE11 Dual beam microscopesE12 Energy filtering TEME13 Environmental and variable

pressure SEME14 Helium ion microscopesE15 High and intermediate voltage

electron microscopesE17 Low energy electron microscopesE18 MicroprobesE19 Nanofabrication,nanolithographyE21 Secondary ion microscopesE22 TomographyE23 X-ray microtomographyE30 EM ACCESSORIES AND SUPPLIESE32 Anti-contamination systemsE33 Anti-vibration systemsE34 Apertures and filamentsE35 Calibration standardsE36 Cryotransfer systemsE37 Energy filtersE38 Magnetic field cancellationE39 Specimen holders for TEME40 Spectrometers (EDX, WDX, EELS)E41 Stages for SEME42 Vacuum equipmentE44 Plasma cleanersE45 Anti-contaminators

M00 SCANNING PROBE MICROSCOPESM01 Scanning tunnelling microscopes M02 Atomic force microscopes M03 Nearfield scanning optical

microscopesM04 Specialized SPMsM05 Dual mode (LM/SEM/TEM) SPMsM06 SPM NanolithographyM10 SPM ACCESSORIES AND SUPPLIESM11 Calibration devices and standardsM12 Cantilevers M13 ScannersM14 StagesM15 Tips

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S2 Agilent 128 Andor 516 APMC 2012 835 Applied Scientific Instrumentation 325 Asylum Research 3S15 Bruker GmbH 1731 Bruker GmbH 2832 Bruker Nano 2930 Cargille 27S9 E2V 14S1 FEI Company 1128 FEI Company 2643 FEI Company 386 Gatan 425 Herzan 4035 Hiden 3314 Hitachi Europe GmbH 7S4 Hitachi Europe GmbH 613 Jeol UK 16S10 Jeol UK 16S9 John Wiley & Sons Ltd. 1524 John Wiley & Sons Ltd. 2440 John Wiley & Sons Ltd. 3738 JPK Instruments 3518 Leica 927 Leica 25S22 Märzhäuser Wetzlar GmbH 20S31 NT-MDT 221 Olympus Europa Holding GmbH 12 Olympus Europa Holding GmbH 220 Olympus Soft Imaging Solutions GmbH 10S16 Oxford Instruments 1834 Physik Instrumente 3133 Picoquant 3037 Prior Scientific 3437 Quorum Technologies 4039 Schaefer-Tec 3644 Skyscan 39S18 Thermo Scientific 19S22 XEI Scientific, Inc. 21S32 Carl Zeiss 23

RESPONSIBILITYApprove equipment Specify equipmentPurchase equipment None of the above

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Austenic-ferritic duplex steel, 16 x 12 x 18 μm3

volume acquired with the AutoSlice and View™ application. A series of top-down high energy, high angle SEM-BSE images were collected automatically. The distance between each slice is 30 nm. Courtesy of FEI NanoPort.

Surface of uncoated pollen, imaged using SEM at very low kV (50 V).

The horizontal field width is 51 μm. Courtesy of

FEI NanoPort.

• Ultra-stable, contamination and damage free imaging of uncoated charging or beam-sensitive samples

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Platinum nanowire deposited and milled to about 50 nm diameter for use as a gas sensorCourtesy of Peter Heard,

Bristol University,

United Kingdom.

Helios NanoLab™ 50 Series

Learn more at www.fei.com/research

View, Analyze, & Create in 3Dwith the most powerful FIB and SEM

Austenic-ferritic duplex steel, 16 x 12 x 18 μm3

volume acquired with the AutoSlice and View™ application. A series of top-down high energy, high angle SEM-BSE images were collected automatically. The distance between each slice is 30 nm. Courtesy of FEI NanoPort.

Surface of uncoated pollen, imaged using SEM at very low kV (50 V).

The horizontal field width is 51 μm. Courtesy of

FEI NanoPort.

• Ultra-stable, contamination and damage free imaging of uncoated charging or beam-sensitive samples

• Best in class sample preparation: precise milling of large volume, very low kV polishing, process monitoring

• Most complete and integrated suite of prototyping capabilities with SEM, FIB and beam chemistries

• Robust, reproducible and versatile multi-signal 3D Slice and View automation

• Accurate and flexible sample positioning and handling

• Outstanding application and service support

Platinum nanowire deposited and milled to about 50 nm diameter for use as a gas sensorCourtesy of Peter Heard,

Bristol University,

United Kingdom.

Helios NanoLab™ 50 Series

NANO AD CORRECT SIZE.indd 1 24/08/2011 15:21FEI View Analize.indd 1 25/08/2011 13:12

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