1. Making Coverslips by Steve Smith, Cheng-Zhong Zhang and Amy Xu

2. Inspect coverslip for flatness Interference fringes most visible in fluorescent light through a magnifying glass.

3. Interference fringes Each fringe reflects a ½ λ change in thickness. Few stripes are best- this coverslip is OK. Beware of coverslips with so many fringes they appear invisible.

4. Place coverslips on jig

5. Place jig in laser engraver Index position to upper left corner

6. Focus Laser Beam Position laser lens proper distance from coverslips by raising and lowering the table.

7. Close lid and engrave! Set printer for 5 copies for deeper engraving

8. Poke out holes Use sharp probe and metal backup plate

9. Making a Gasket by Steve Smith, Cheng-Zhong Zhang and Amy Xu

10. Nescofilm with empty roller

11. Thread Nescofilm through rollers Suspend Nescofilm flat and taut, far away from any surface (including paper backing)!

12. Place roller in Laser Engraver

13. Focus Laser Beam

14. Close lid and engrave! Smoke is sucked up the vent if your fan is working properly

15. Remove stray Nescofilm

16. Advance Nescofilm roller

17. Pulling a Pipette by Steve Smith, Cheng-Zhong Zhang and Amy Xu

18. Clamp Glass Tubing Carefully clamp pipette at top and bottom, threading through the filament. Don’t touch glass tubing in the center. Fingerprints = bad.

19. Centering the tube

20. Shield from air currents and PULL! Cover pipette puller with a plastic beaker and turn on power supply to heat the filament. Weight should drop in ~10 seconds. Faster heating = smaller tips.

21. Remove pipette

22. Inspect pipette tip

23. Pipette tip

24. Assembling a Microchamber by Steve Smith, Cheng-Zhong Zhang and Amy Xu

25. Trim the gaskets

26. Lay gasket on coverslip with no holes

27. Lay gasket on coverslip with 6 holes Align holes

28. Flatten gaskets

29. Place pipette in middle

30. Pipette tip placement

31. Cut dispenser tubes Cut ~2.5 mm pieces from long tube. Secure on double sided tape.

32. Place dispenser tubes

33. Dispenser tube placement

34. Make a sandwich

35. Press ends together Gently

36. Place glass slide on top of sandwich

37. Press on glass slide to flatten sandwich With considerable force

38. Pick up sandwich and slide together

39. Place on heated plate

40. Place heated weight on glass slide

41. Place microchamber on frame

42. Tighten microchamber on frame Press down in the middle of the clamp while gently tightening the screws.

43. Check alignment of chamber and frame holes Adjust clamps as necessary

44. Thread PE10 tubing onto pipette Tighten bottom clamp with pipette slot facing down.

45. Dispense small amount of epoxy Norland UV cure optical adhesive

46. Touch epoxy to end of tubing

47. Epoxy enters tube by capillary action Be careful epoxy does not run too far and clog back end of pipette!

48. UV crosslink epoxy 2-5 minutes in crosslinking chamber oven

49. Screw in nylon tube fittings Rinse out fittings first

50. Completed microchamber Congratulations!