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

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    20-Dec-2015
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Transcript of Making Coverslips by Steve Smith, Cheng-Zhong Zhang and Amy Xu

  • Slide 1
  • Making Coverslips by Steve Smith, Cheng-Zhong Zhang and Amy Xu
  • Slide 2
  • Inspect coverslip for flatness Interference fringes most visible in fluorescent light through a magnifying glass.
  • Slide 3
  • Interference fringes Each fringe reflects a change in thickness. Few stripes are best- this coverslip is OK. Beware of coverslips with so many fringes they appear invisible.
  • Slide 4
  • Place coverslips on jig
  • Slide 5
  • Place jig in laser engraver Index position to upper left corner
  • Slide 6
  • Focus Laser Beam Position laser lens proper distance from coverslips by raising and lowering the table.
  • Slide 7
  • Close lid and engrave! Set printer for 5 copies for deeper engraving
  • Slide 8
  • Poke out holes Use sharp probe and metal backup plate
  • Slide 9
  • Making a Gasket by Steve Smith, Cheng-Zhong Zhang and Amy Xu
  • Slide 10
  • Nescofilm with empty roller
  • Slide 11
  • Thread Nescofilm through rollers Suspend Nescofilm flat and taut, far away from any surface (including paper backing)!
  • Slide 12
  • Place roller in Laser Engraver
  • Slide 13
  • Focus Laser Beam
  • Slide 14
  • Close lid and engrave! Smoke is sucked up the vent if your fan is working properly
  • Slide 15
  • Remove stray Nescofilm
  • Slide 16
  • Advance Nescofilm roller
  • Slide 17
  • Pulling a Pipette by Steve Smith, Cheng-Zhong Zhang and Amy Xu
  • Slide 18
  • Clamp Glass Tubing Carefully clamp pipette at top and bottom, threading through the filament. Dont touch glass tubing in the center. Fingerprints = bad.
  • Slide 19
  • Centering the tube
  • Slide 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.
  • Slide 21
  • Remove pipette
  • Slide 22
  • Inspect pipette tip
  • Slide 23
  • Pipette tip
  • Slide 24
  • Assembling a Microchamber by Steve Smith, Cheng-Zhong Zhang and Amy Xu
  • Slide 25
  • Trim the gaskets
  • Slide 26
  • Lay gasket on coverslip with no holes
  • Slide 27
  • Lay gasket on coverslip with 6 holes Align holes
  • Slide 28
  • Flatten gaskets
  • Slide 29
  • Place pipette in middle
  • Slide 30
  • Pipette tip placement
  • Slide 31
  • Cut dispenser tubes Cut ~2.5 mm pieces from long tube. Secure on double sided tape.
  • Slide 32
  • Place dispenser tubes
  • Slide 33
  • Dispenser tube placement
  • Slide 34
  • Make a sandwich
  • Slide 35
  • Press ends together Gently
  • Slide 36
  • Place glass slide on top of sandwich
  • Slide 37
  • Press on glass slide to flatten sandwich With considerable force
  • Slide 38
  • Pick up sandwich and slide together
  • Slide 39
  • Place on heated plate
  • Slide 40
  • Place heated weight on glass slide
  • Slide 41
  • Place microchamber on frame
  • Slide 42
  • Tighten microchamber on frame Press down in the middle of the clamp while gently tightening the screws.
  • Slide 43
  • Check alignment of chamber and frame holes Adjust clamps as necessary
  • Slide 44
  • Thread PE10 tubing onto pipette Tighten bottom clamp with pipette slot facing down.
  • Slide 45
  • Dispense small amount of epoxy Norland UV cure optical adhesive
  • Slide 46
  • Touch epoxy to end of tubing
  • Slide 47
  • Epoxy enters tube by capillary action Be careful epoxy does not run too far and clog back end of pipette!
  • Slide 48
  • UV crosslink epoxy 2-5 minutes in crosslinking chamber oven
  • Slide 49
  • Screw in nylon tube fittings Rinse out fittings first
  • Slide 50
  • Completed microchamber Congratulations!