Nanofair Gesim
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www.gesim.de µ-Contact Printing Method to create µm and nm Patterns on Cell Based Bio-Chips Steffen Howitz, Frank Baudisch, Felix Franz, GeSiM mbH Großerkmannsdorf Michael Gepp and Heiko Zimmermann, Fraunhofer IBMT St.Ingbert Stefan Fiedler and Michael Zwanzig, Fraunhofer IZM-Berlin
description
Nanotech technology capabilities
Transcript of Nanofair Gesim
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µ-Contact Printing
Method to create µm and nm Patterns on Cell Based Bio-Chips
Steffen Howitz, Frank Baudisch, Felix Franz, GeSiM mbH Großerkmannsdorf
Michael Gepp and Heiko Zimmermann, Fraunhofer IBMT St.Ingbert
Stefan Fiedler and Michael Zwanzig, Fraunhofer IZM-Berlin
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• motivation
• comparison of print technologies
• new PDMS based stamp concept
• µCP2.1 for printing and imprinting
• first results
• conclusion
Content
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1. motivation
• rough overview of surface functionalization methods
• research platform µCP2.1 for nano surface
programming and imprinting
• explain new PDMS-stamp concept
• first results
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soluble
+
immobilised
differentiation using + signalfactors
2. print technologies for surface functionalizationhow we can bring surface relevant factors to biochips?
Cell Programming byNanoscaled Devices
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2. comparison of print technologies
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Stamp correlatedPin-Tool headcorrelated
programmable
Array Desing
one sampleper Ink Step
Well-Plate correlatedprogrammableSample Access
parallelparallelserialPrint Mode
contactcontactnon contactSurface Contact
~ 5000 x 5000 spots~ 5x5 spots~ 5x5 spotsArray Densityper mm²
< 50nm50…120µm60...150µmFoot Print of asingle spot
µ-Contact-PrintingµCP2.1
Pin-Tool PrinterNP2.1
Piezo-PrintingNP2.1
Feature
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2. piezo-non-contact-printing at NP2.1
Ref.: IBMT- R. Strelow
1. Piezo-Pipette non-contact printing of DNA and Proteins2. Integrated Imaging System3. Windows Software NPC164. Customized Work-Plate5. Coolable Plate-Holder and Slidetray6. Humidifyer , 40....80% rel. Humidity
System Features
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2. pin-tool-contact-printing at NP2.1
System Features• Silicon-Pin Tools from Parallel Synthesis Technologies• Integrated Imaging System• Windows Software NPC16• Customized Work-Plate• Coolable Plate-Holder and Slidetray• Humidifyer , 40....80% rel. Humidity
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2. µ-contact-printing at µCP2.1
• (a) inking station with 4 ink pads
• (b) drying nozzles, two per stamp
• (c) stamping unit
a
b
c
µCP- print process
• inking for 30…60sec
• drying at 1bar/N2 and 60sec
• stamping at 0,2bar and 60sec
1 print cycle 3 minutes
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3. new stamp design for system µCP2.1
compressed air
a) Basic Mode: PDMS-membran planar b) Print-Mode: PDMS-membran deflected
Stamp Chamber
PDMS-Membran
Stampframe
Stampholder Stamp Chamber
nm/µm Patterns
SEM picture Si-Master Master in Casting Staion Casting of PDMS-Stamp PDMS-Stamp µCP-Stamping Unit
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3. different stamps made in PDMS
25µm40µm
50µmbevel position
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3. stamp gets in contact to a glass slide
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Stamping Unit
Slide Tray
Microscope
4. µ-contact-printing at µCP2.1
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4. 3d-imprinting at µCP2.1
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µCP2.1 with UV-light source
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soluble
+
immobilised
differentiation using + signalfactors
5. first results of surface functionalization
Cell Programming byNanoscaled Devices
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5. µ-contact-printing with µm-scaled stamps
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25x25µm²Quadrate
2µm Stege
Fig.1: first µ-Contact-Print of CY3-labeled fibronectin on hydrophilic glass
slide, stamp area 1x1cm²
Fig.2: second print without new inking Fig.3: third print without new inking
method:a) inking of stamp 1min,b) drying of stamps withcompressed air 2 bar/30sec,c) contact print applying 0,25barstamp pressure for 60sec.
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5. µ-contact-printing with a defined parallel shift
AØ
A2
xA
AØ
A
2x
A
Pattern: A=100
Fig.1: µ-contact-print of CY3-labeled fibronectinon glass slide, stamp area 1x1cm²
Method:
• first print after inking the stamp
• second print realised with a parallel shift and without new inking
Fig.2: µ-contact-print like figure 1with larger patterns, stamp area 1x1cm²
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5. Cell Adhesion Assay with L929 Mouse FibroblastsECM Stamping on hydrophilic Glass Slides
AØ
A2
xA
AØ
A
2x
A
Pattern:A=100
1) 1..5 h µCP-pattern related cell adhesion visible,
2) 5…20 h a general cell adhesion occurs, patterns are then invisble
Next Experiments: ECM µ-ContactPrints on cell repulsive surfaces like Teflon!
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L929 fibroblast on silicon substrate50 – 100 nm wide fibronectin lines were printed
5. Cell surface interaction with nano-structured surfaces
Images courtesy AMO and FhG-IBMT
Cell Programming byNanoscaled Devices
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5. µ-contact-printing to transfer nano-particles
3µm 8µm
µ-contact-printing of spherical gold nano-particles (Ø=37nm, diluted in water) ona hydrophilic glass slide
25x25µm²Quadrat e
2µm Stege
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5. imprinting of spin-coated photo restist films
Fig.1: Imprinted photo resist on glassslides, thickness 8µm
50µm 50µm
IMPRINTING – A fast way to fabricate simple fluidics on glass slides!
Fig.2: Imprinted photo resist on glass slides,thickness 500nm
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5. MagnaLab – Non-Contact and magnetic handling of glassbased cell-carriers in micro fluidic channels
Cell Programming byNanoscaled Devices
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5. Demands for Cell-Carrier,functional layers
Cell Programming byNanoscaled Devices
sliding
shielding
magnetic
glass basis
NanoScapeTMcell repulsive
tech
nolo
gybi
olog
y
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5. cell carrier with NanoScapes
Cell-carrier chip processedcompletely
Cell Programming byNanoscaled Devices
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6. conclusion
• µCP technology close the gap from µm to nmfor surface programming for printing and imprinting
• status quo of µCP2.1 - semi automatically research tool• future oportunities enlarge print area and increase automation• µCP reasonable tool for 3D-surface patterning
• potental applicationbasic material researchbio-sensor technologycell-bacteria-virus researchsubstrates for cell storing e.g. in cryo-biotechnologydisposable micro-fluidic systemscombination with MEA-technolgy e.g. for neuronal networks combinedwith fluidics …..
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Thank you for your attention!
