3. Slides: FluidFM nano-printing - FluidFM BOT Webinar

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9. SINGLE CELL EXPERIMENTS REINVENTED.

12. NANO - INJECTION .

3. FluidFM ® . WHAT IS IT?

19. NANO - EXTRACTION .

22. 31

23. ISOLATION OF ADHERENT CELLS .

26. 35

28. NANO - PRINTING .

5. FluidFM Chip Probe = Consumable Zoom in A CLOSE LOOK. F luid FM PROBES

13. A WORLD OF APPLICATIONS. 16 NANO - INJECTION INTO SINGLE CELLS

24. A WORLD OF APPLICATIONS. SINGLE CELL ISOLATION

2. 60 MINUTES OF FluidFM . What is FluidFM? FluidFM applications Live Demo 3 THE WEBINAR TODAY.

39. S WISS MADE TECHNOLOGY .

45.

1. FluidFM ® BOT WEBINAR. NANO - PRINTING. With live demo. June 2019. Dr. Paul Monnier, Dr. Pablo Dörig

14. 300 – 800 nm hole NANOSYRINGE NANO - INJECTION. F luid FM NANOSYRINGE

30. NANO - PRINTING. IN AIR OR LIQUID

42. UPCOMING. WEBINARS.

44. LIVE DEMO. NANO - PRINTING.

15. F luid FM ® BOT IN ACTION. INJECTING SINGLE CHO CELLS Link to Video: https://youtu.be/r2ZWAN9S8Ag

11. FULLY INTEGRATED SYSTEM SOLUTION. 14 F luid FM ® BOT

31. ANY INK. 51 FROM WATER TO HONEY Viscosity = 1 cP Viscosity = 10’000 cP Watery solutions Salty buffers Oils Acids Bases Gentle organic solvents and more

32. NANO - PRINTING . 54 SPOT ARRAYS With 300 nm FluidFM Nanopipette Glycerol in air. 100 x 100 um. Courtesy Nanosurf AG

4. 5 TECHNOLOGY • F luid FM : FLUIDIC FORCE MICROSCOPY • PATENTED KNOW - HOW

8. F luid FM. 10 MORE THAN JUST ONE TOOL MICROPIPETTE GRAB & MANIPULATE MICRO OBJECTS NANOSYRINGE INJECT AND EXTRACT

17. F luid FM ® NANO - INJECTION. 22 INJECTION INTO ADHERENT CELL LINES HepG2 Mouse primary hepatocytes HEK Pig neurons C2C12 MEFs hESCs mESCs

18. F luid FM ® NANO - INJECTION. 24 INJECTING SOLUBLE COMPOUNDS Lucifer Yellow UHRF1 proteins Antibodies CRISPR/Cas9 RNP complexes Plasmids Small RNAs

27. F luid FM ® CELL ISOLATION. 43 CELL VIABILITY UP TO 95% 12 CHO cells isolated in a 12 well - plate. 48 hours later.

29. F luid FM ® NANO - PRINTING. 49 DESIGN PRECISE PATTERNS 300 μ m 300 nm aperture FluidFM nanopipette DAPI on DNA coated glass 10 μ m

21. F luid FM ® B O T. 30 NANO - EXTRACTION. O. Guillaume - Gentil et al., Cell 2016, 166: 506 - 516 Anti - fouling coating Cell extract Oil Channel filled with oil

20. SINGLE CELL NANO - EXTRACTION. A fluorescent myoblast is contacted by FluidFM ® . Orane Guillaume - Gentil, Analytical Chemistry - 10.1021/acs.analchem.7b00367 A WORLD OF APPLICATIONS. NANO - EXTRACTION FROM SINGLE CELLS

10. F luid FM B O T. 12 SINGLE CELL EXPERIMENTS REINVENTED Adhesion of single cells Nano - extraction from single cells Nano - injection into single cells Spotting Nano - printing FluidFM micropipette 2, 4, 8 μm aperture FluidFM nanosyringe 300 - 800 nm aperture FluidFM nanopipette 300 nm aperture Cell isolation

34. NANO - PRINTING FOR NANOSCIENCE. 56 NANO - TO MICRO PATTERNS Ventrici et al. Physical Chemistry B 2017 • FluidFM Nanopipette • UV glue (10’000 cp) on glass • Nm to mm patterns

6. F luid FM ® – CUTTING EDGE & UNIQUE. 8 MICROCHANNELED PROBES 300 - 800 nm aperture FluidFM nanosyringe FluidFM nano - injection into single cells HIGH - QUALITY MICROFABRICATION PRECISE FEMTOLITER LIQUID VOLUMES DIVERSE APPLICATIONS

16. F luid FM ® NANO - INJECTION.  >300 cells injected per hour  10 minutes preparation time  >90% succesfull injection 21 NANO - INJECTION WITH THE FluidFM BOT

25. Viability > 95 % Same cantilever for trypsin release and cell displacement Serial isolation (>30 cells per probe) Guillaume - Gentil 2014 LabChip 14:402 20 μm LOCAL TRYPSINIZATION ISOLATION F luid FM ® B O T. 34 ISOLATION OF SINGLE ADHERENT CELLS

41. 63 QUALITY. CUSTOMER. WHERE WE FOCUS SUPPORT QC, QC, QC EVERY SINGLE PROBE CUSTOMER DRIVEN INNOVATION WORLDWIDE REMOTE SKYPE, TEAM VIEWER QUALITY PRODUCTION CUSTOMER FOCUS

35. NANO - PRINTING FOR NANOSCIENCE. 57 LAYER BY LAYER ON THE NANOSCALE Ventrici et al. Physical Chemistry B 2017 • FluidFM nanopipette • UV glue on glass • ~20 nm layers • 10 μ m pattern

36. NANOPRINTING AND BIOLOGY. 58 PATTERNING CELLS Saftics et al. Langmuir 2019 • FluidFM micropipette • CMD hydrogel background (repellent) • cRGDfK lines printed (attractive) • HeLa cells

37. NANO - PRINTING AND BIOLOGY. 59 NEURON PATTERNING + IN SITU GUIDANCE Dermutz et al. Langmuir 2015 • FluidFM Micropipette • PLL - g - PEG background (repellent) • PLL lines printed (attractive) • Primary neurons • PLL printed pathways guide neurons

33. VIRUS DEPOSITED CELL IS SICK 30 MIN LATER Stiefel et al. (2012) , Nano Lett 12 SINGLE PARTICLES. 55 PLACING A SINGLE VIRUS STUDY INFECTION STATISTICS • HeLa cells • GFP labeled Vaccinia Virus • Gentle contact • 5mbar – 1s

38. COMBINE IT. 60 HOW DO BIOCHEMICAL PATTERNS INFLUENCE CELLS? Distance Force Isolate & analyze • PCR • MS Design the substrate • Chemical patterns • Gradients • Structures Cultivate Measure biomechanics

7. FluidFM ® micropipette 0.3 – 4N/m, aperture sizes: 2, 4, 8 μm Isolation & adhesion of single cells , colloidal spectroscopy, local dispensing FluidFM ® nanopipette 0 .6 – 2N/m, aperture size: 300 nm N ano - printing, manipulation of sub μm particles, adhesion of single bacteria FluidFM ® nanosyringe 2 N/m, aperture size: 300 - 800 nm Nano - injection into & nano - extraction from single cells FluidFM ® prototyping probe A perture can be customized with Focused Ion Beam (FIB) A pplication depending on the customization CUSTOM SHAPES 300 nm to 8 μm STANDARD OPENING DIAMETERS THE HEART OF F luid FM. F luid FM ® PROBES PORTFOLIO PRECISE BATCH FABRICATION DOWN TO 30 nm POSSIBLE WITH FIB

40.  Founded 2009 in Zurich, Switzerland  Dr. Pascal Behr  Dr. Michael Gabi  ETH Zurich Spin - off  20+ employees in 2019  Prize winning company 62 THE COMPANY BEHIND F luid FM . CYTOSURGE AG 60+ PUBLICATIONS 2013 FIRST SALE 60+ SYSTEMS WORLDWIDE HIGH IMPACT JOURNALS Ø 8+ IMPACT FACTOR

43. FluidFM BOT WEBINAR:  September 2019  With live demo  What would you like to see? Tell us: [email protected]  June 26, 2019  Injection, extraction & deposition  O. Guillaume - Gentil will be available for questions  Author of pioneering FluidFM publications  Leading scientist developing FluidFM injection & extraction methods  Register here 71 STAY CURRENT. UPCOMING WEBINARS AFM Solutions Webinar: By Wiley &

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