1. Design of an integrated liquid flow cell for correlative microscopy
Diederik Morsink First Msc. presentation

2. Outline Crash course in Life science imaging Introducing Delmic
Previous research
My Project
Progress
To do
Aimed results
Planning

3. Crash course in Life science imaging
Highest goal in life science imaging:
High resolution imaging of living cells in their natural environment
~1675
2013

4. Crash course in Life science imaging
Fluorescence Microscopy
Scanning Electron Microscopy
Correlative Microscopy

5. Crash course in Life science imaging
Fluorescence microscopy
Advantages:
Color contrast
No damage to sample
Functional information
Disadvantage:
Diffraction limited

6. Crash course in Life science imaging
Fluorescence microscopy
Advantages:
Color contrast
No damage to sample
Functional information
Disadvantage:
Diffraction limited
Image courtesy: Nalan Liv

7. Crash course in Life science imaging
Scanning electron microscopy
(SEM)
Advantage:
High resolution
Structural information
Disadvantage:
Needs vacuum environment
Can damage the sample

8. Crash course in Life science imaging
Scanning electron microscopy
(SEM)
Advantage:
High resolution
Structural information
Disadvantage:
Needs vacuum environment
Can damage the sample
Image courtesy: Nalan Liv

9. Crash course in Life science imaging
Correlative Light and Electron Microscopy
(CLEM)
Combining the information
of from SEM and Fluorescence imaging
Structural and functional information

10. Crash course in Life science imaging
Correlative Light and Electron Microscopy
(CLEM)
Combining the information
of from SEM and Fluorescence imaging
Structural and functional information
3μm
Image courtesy: Nalan Liv

11. Delmic Spinoff TNW SECOM platform Correlative microscopy
Image courtesy: Delmic and Ruud van Tol

12. Previous research
Liquid cell
Liquid flow cell

13. Previous research Liquid cell Liquid Flow cell
Image courtesy: Daan van Oosten Slingeland

14. Previous research Liquid cell Liquid Flow cell
Image courtesy: Daan van Oosten Slingeland

15. My project Liquid cell Liquid Flow cell ....
Image courtesy: Daan van Oosten Slingeland

16. My project
Integrating pump, imaging area and reservoirs in a single MEMS device.
Simultaneous imaging and manipulation
Living cells
Working prototype

17. Progress So Far: Literature study on Life science imaging techniques
Literature study on Micropumps
Requirements
Selecting micropump designs
Conceptual design of how to integrate pump, imaging area and reservoirs in a single device

18. Integrated Liquid Flow cell
Requirements
Integrated Liquid Flow cell
General
Vacuum compatible
Biocompatible
Control interface
Fit SECOM platform
Imaging area
Optical window: 170 µm thick
Electron beam window: 50 nm thick
Gap between windows: <50µm
Flow velocity 0-1mm/s
Cells must be trapped at membrane
Microfluidic pump
Self priming
Reservoirs: 5-10µl

19. Conceptual design:

20. Conceptual design: Microfluidic pump
Based on designs by Linnemann (1998) and Kang (2008)
Stack of 3 Si wafers
pumpchamber
Pump diaphragm
Piezo element
Inlet
Outlet

21. Conceptual design: Imaging area
Separate chip with silicon nitride membrane (orange)
Silicon nitride membrane
Imaging area
Silicon spacer
Microchannel
Glass coverslip

22. Conceptual design: Integration in a single device

23. Conceptual design: Integration in a single device

24. Conceptual design: Integration in a single device

25. Conceptual design: Integration in a single device
Outlet
Inlet

26. Conceptual design: Integration in a single device
Al. bottom
Si3N4 Chip
Si wafer 1
Sii wafer 2
Si wafer 3
O-ring
Al. top
Coverslip

27. Conceptual design: Integration in a single device
Al. bottom
Si3N4 Chip
Si wafer 1
Sii wafer 2
Si wafer 3
O-ring
Al. top
Coverslip

28. Design overview Two microfluidic pumps and reservoirs
Imaging area with separate silicon nitride chip
Stack of 3 silicon wafers on glass substrate

29. Fabrication

30. Fabrication
Manufacturing complete device not feasible (time restriction)

31. Fabrication
Manufacturing complete device not feasible (time restriction)
Two critical steps:
1. Bonding coverslip-silicon
2. Clamping chip with silicon nitride membrane F 1. 2.

32. To do
Gain experience with bonding coverslip-silicon techniques for this purpose
Build a simple microfluidic device that demonstrates that the chosen bonding technique is appropriate

33. To do
Gain experience with bonding coverslip-silicon techniques for this purpose
Build a simple microfluidic device that demonstrates that the chosen bonding technique is appropriate
If time permits:
Develop a suitable way for clamping silicon nitride chip to silicon spacer
Build a simple microfluidic device that demonstrates successful clamping of silicon nitride chip

34. Final results aimed for
Conceptual design for an integrated
liquid flow cell for correlative microscopy
Simple microfluidic device demonstrating
successful manufacturing of a critical step
in the process

35. Planning

36. Design of an integrated liquid flow cell for correlative microscopy
Diederik Morsink First Msc. presentation