µ-Contact Printing

Method to create µm and nm Patterns on Cell Based Bio-Chips


           Steffen Howitz, Frank Baudisc...
Content

•   motivation
•   comparison of print technologies
•   new PDMS based stamp concept
•   µCP2.1 for printing and ...
1. motivation



• rough overview of surface functionalization methods


• research platform µCP2.1 for nano surface
 prog...
2. print technologies for surface functionalization
                        how we can bring surface relevant factors to b...
2. comparison of print technologies


Feature           Piezo-Printing    Pin-Tool Printer        µ-Contact-Printing
     ...
2.   piezo-non-contact-printing at NP2.1




                                                 Ref.: IBMT- R. Strelow




S...
2.     pin-tool-contact-printing at NP2.1




System Features
•   Silicon-Pin Tools from Parallel Synthesis Technologies
•...
2. µ-contact-printing at µCP2.1

                                      c

                    b




                     a...
3. new stamp design for system µCP2.1


                                                                                  ...
3. different stamps made in PDMS




                                      40µm
 25µm




                                ...
3.   stamp gets in contact to a glass slide




                                              www.gesim.de
4.   µ-contact-printing at µCP2.1




                                        Stamping Unit




                          ...
4.   3d-imprinting at µCP2.1




                    µCP2.1 with UV-light source



                                      ...
5. first results of surface functionalization




                              soluble

differentiation using   ++ signal...
5.        µ-contact-printing with µm-scaled stamps




Fig.1: first µ-Contact-Print of CY3-       Fig.2: second print with...
5. µ-contact-printing with a defined parallel shift

                                           Pattern: A=100




       ...
5. Cell Adhesion Assay with L929 Mouse Fibroblasts
      ECM Stamping on hydrophilic Glass Slides
                        ...
5. Cell surface interaction with nano-structured surfaces


    L929 fibroblast on silicon substrate
    50 – 100 nm wide ...
5. µ-contact-printing to transfer nano-particles



                                                                      ...
5. imprinting of spin-coated photo restist films




  50µm                                       50µm



Fig.1: Imprinted...
5.               MagnaLab – Non-Contact and magnetic handling of glass
                      based cell-carriers in micro ...
5. Demands for Cell-Carrier,functional layers
biology
                                cell repulsive
                     ...
5. cell carrier with NanoScapes




                                            Cell-carrier chip processed
              ...
6. conclusion

•   µCP technology close the gap from µm to nm
    for surface programming for printing and imprinting
•   ...
Thank you for your attention!




                                www.gesim.de
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  1. 1. 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 www.gesim.de
  2. 2. Content • motivation • comparison of print technologies • new PDMS based stamp concept • µCP2.1 for printing and imprinting • first results • conclusion www.gesim.de
  3. 3. 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 www.gesim.de
  4. 4. 2. print technologies for surface functionalization how we can bring surface relevant factors to biochips? soluble differentiation using ++ signal factors immobilised Cell Programming by Nanoscaled Devices www.gesim.de
  5. 5. 2. comparison of print technologies Feature Piezo-Printing Pin-Tool Printer µ-Contact-Printing NP2.1 NP2.1 µCP2.1 Foot Print of a 60...150µm 50…120µm < 50nm single spot Array Density ~ 5x5 spots ~ 5x5 spots ~ 5000 x 5000 spots per mm² Surface Contact non contact contact contact Print Mode serial parallel parallel Sample Access programmable Well-Plate correlated one sample per Ink Step programmable Pin-Tool head Stamp correlated correlated Array Desing www.gesim.de
  6. 6. 2. piezo-non-contact-printing at NP2.1 Ref.: IBMT- R. Strelow System Features 1. Piezo-Pipette non-contact printing of DNA and Proteins 2. Integrated Imaging System 3. Windows Software NPC16 4. Customized Work-Plate 5. Coolable Plate-Holder and Slidetray 6. Humidifyer , 40....80% rel. Humidity www.gesim.de
  7. 7. 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 www.gesim.de
  8. 8. 2. µ-contact-printing at µCP2.1 c b a • (a) inking station with 4 ink pads • (b) drying nozzles, two per stamp • (c) stamping unit µCP- print process • inking for 30…60sec • drying at 1bar/N2 and 60sec • stamping at 0,2bar and 60sec 1 print cycle 3 minutes www.gesim.de
  9. 9. 3. new stamp design for system µCP2.1 compressed air Stamp Chamber Stampholder Stamp Chamber Stampframe PDMS-Membran nm/µm Patterns a) Basic Mode: PDMS-membran planar b) Print-Mode: PDMS-membran deflected SEM picture Si-Master Master in Casting Staion Casting of PDMS-Stamp PDMS-Stamp µCP-Stamping Unit www.gesim.de
  10. 10. 3. different stamps made in PDMS 40µm 25µm 50µm bevel position www.gesim.de
  11. 11. 3. stamp gets in contact to a glass slide www.gesim.de
  12. 12. 4. µ-contact-printing at µCP2.1 Stamping Unit Slide Tray Microscope www.gesim.de
  13. 13. 4. 3d-imprinting at µCP2.1 µCP2.1 with UV-light source www.gesim.de
  14. 14. 5. first results of surface functionalization soluble differentiation using ++ signal factors immobilised Cell Programming by Nanoscaled Devices www.gesim.de
  15. 15. 5. µ-contact-printing with µm-scaled stamps Fig.1: first µ-Contact-Print of CY3- Fig.2: second print without new inking Fig.3: third print without new inking labeled fibronectin on hydrophilic glass slide, stamp area 1x1cm² method: a) inking of stamp 1min, b) drying of stamps with 25x25µm² compressed air 2 bar/30sec, Quadrate c) contact print applying 0,25bar stamp pressure for 60sec. 2µm Stege www.gesim.de
  16. 16. 5. µ-contact-printing with a defined parallel shift Pattern: A=100 Ø A A Ø 2xA A A 2xA Fig.1: µ-contact-print of CY3-labeled fibronectin Fig.2: µ-contact-print like figure 1 on glass slide, stamp area 1x1cm² with larger patterns, stamp area 1x1cm² Method: • first print after inking the stamp • second print realised with a parallel shift and without new inking www.gesim.de
  17. 17. 5. Cell Adhesion Assay with L929 Mouse Fibroblasts ECM Stamping on hydrophilic Glass Slides Patter : A=100 n ØA ØA 2x A A A 2xA 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! www.gesim.de
  18. 18. 5. Cell surface interaction with nano-structured surfaces L929 fibroblast on silicon substrate 50 – 100 nm wide fibronectin lines were printed Images courtesy AMO and FhG-IBMT Cell Programming by Nanoscaled Devices www.gesim.de
  19. 19. 5. µ-contact-printing to transfer nano-particles 25x25µm² Quadrat e 2µm Stege 3µm 8µm µ-contact-printing of spherical gold nano-particles (Ø=37nm, diluted in water) on a hydrophilic glass slide www.gesim.de
  20. 20. 5. imprinting of spin-coated photo restist films 50µm 50µm Fig.1: Imprinted photo resist on glass Fig.2: Imprinted photo resist on glass slides, slides, thickness 8µm thickness 500nm IMPRINTING – A fast way to fabricate simple fluidics on glass slides! www.gesim.de
  21. 21. 5. MagnaLab – Non-Contact and magnetic handling of glass based cell-carriers in micro fluidic channels Cell Programming by Nanoscaled Devices www.gesim.de
  22. 22. 5. Demands for Cell-Carrier,functional layers biology cell repulsive NanoScapeTM glass basis technology magnetic shielding sliding Cell Programming by Nanoscaled Devices www.gesim.de
  23. 23. 5. cell carrier with NanoScapes Cell-carrier chip processed completely Cell Programming by Nanoscaled Devices www.gesim.de
  24. 24. 6. conclusion • µCP technology close the gap from µm to nm for 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 application basic material research bio-sensor technology cell-bacteria-virus research substrates for cell storing e.g. in cryo-biotechnology disposable micro-fluidic systems combination with MEA-technolgy e.g. for neuronal networks combined with fluidics ….. www.gesim.de
  25. 25. Thank you for your attention! www.gesim.de

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