Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob...
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18-12-2017 Ben N. G. Giepmans Dept. Cell Biology, UMCG Microscopy & Imaging Center (UMIC) UMC Groningen, The Netherlands [email protected] www.cellbiology.nl Nanotomy & High-Resolution Color-EM
Transcript of Nanotomy & High-Resolution Color-EM · PDF fileNanotomy & High-Resolution Color-EM Jacob...
18-12-2017
Ben N. G. Giepmans Dept. Cell Biology, UMCG Microscopy & Imaging Center (UMIC) UMC Groningen, The Netherlands [email protected] www.cellbiology.nl
Nanotomy &
High-Resolution Color-EM
Nanotomy &
High-Resolution Color-EM
Jacob Hoogenboom
Marijke Scotuzzi
Kees Hagen
Dasha Wensveen
Ben Giepmans
Jeroen Kuipers
Pascal de Boer
Probes & techniques for microscopy
- Identify and quantify -
How do molecules, organelles & cells regulate (patho)physiology Need probes: Resolution, dynamics, identification, quantification Large scale EM / nano-anatomy (“nanotomy”): More identification/quantification at high resolution Big data sharing New probes & techniques allow nm-scale “color-EM”: Proof-of-concept and limitations
Large scale EM (Nanotomy)
5 rbv3.com/blog/wp-content/uploads/2014/03/Google.jpg nanotomy.org
Nano-anatomy: anatomy of tissues, cells, organelles at the macromolecular level, “Google-tissue” Major benefits of Nanotomy: - Overview and high resolution - Rare events may be imaged - Unbiased examination of tissue, cells, organelles and molecules - Open access raw data sharing - Routine technique - Grey scale images www.nanotomy.org
175 µm 64 µm 16 µm 4 µm 1 µm
From Islet to Insulin granule
7 https://pdb101.rcsb.org/motm/14
Insulin
8 http://chemistry.tutorvista.com/biochemistry/cysteine.html
Cysteine
9 Shutterstock
Sulphur
Energy dispersive X-rays (EDX)
10 Thermo Scientific
Incident electrons
Image by: Thermo Scientific
11 Bushberg et al. (2011)
Energy dispersive X-rays (EDX)
Energy dispersive X-rays (EDX)
12 pinterest.com
X-ray detection
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X-ray detector
Hoogenboom et al., TU-Delft
15 www.bpc.edu
Periodic table of elements
X-ray spectrum of gold versus cadmium
16 www.xrfresearch.com
Detect Au in immunogold and Cd in QDs?
Scotuzzi et al. Scientific Reports 2017
EDX reveals Cd in QDs labeling insulin
18 Scotuzzi et al. Scientific Reports 2017
Au /G4 at P-rich heterochromatin
19 Scotuzzi et al. Scientific Reports 2017
Multi-colour nm-scale EM (G4 and insulin)
Scotuzzi et al. Scientific Reports 2017
Multi-colour nm-scale EM (G4 and insulin)
Scotuzzi et al. Scientific Reports 2017
Energy-Dispersive X-ray Spectra (EDX / EDS)
22 Scotuzzi et al. Scientific Reports 2017
Elemental maps
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Au Ca Cd Cl Fe
K Mg N Na Os
P S Se U Zn
Scotuzzi et al. Scientific Reports 2017
Contrast /morphology: Distinct granules?
24 Scotuzzi et al. Scientific Reports 2017
Distinct granules based on elements!
25 Scotuzzi et al. Scientific Reports 2017
Distinct granules based on elements!
26 Scotuzzi et al. Scientific Reports 2017
18-12-2017
Scotuzzi et al. Scientific Reports 2017
Nanotomy, EDX & ColorEM
“Superdepuper” resolution colorEM with EDX
Endogenous elements (S; P; distinct vesicles)
Paint organelles (Os, W)
Identify labels (Cd, Au)
Revolution in computer sciences allow data
acquisition and data sharing
Large-scale EM
www.nanotomy.org
Scie
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Acknowledgements
University Medical Center Groningen
Jeroen Kuipers
Pascal de Boer
Anouk Wolters
Marit de Beer
Nicole Pirozzi
Klaas Sjollema
Delft University of Technology
Marijke Scotuzzi
Dasha Wensveen
Lennard Voortman
Kees Hagen
Jacob Hoogenboom
