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Making Coverslips. by Steve Smith, Cheng- Zhong Zhang and Amy Xu. Inspect coverslip for flatness. Interference fringes most visible in fluorescent light through a magnifying glass. Interference fringes. - PowerPoint PPT Presentation
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Inspect coverslip for flatness
Interference fringes most visible in fluorescent light through a magnifying glass.
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.
Focus Laser Beam
Position laser lens proper distance from coverslips by raising and lowering the table.
Thread Nescofilm through rollers
Suspend Nescofilm flat and taut, far away from any surface (including paper backing)!
Focus Laser Beam
Position laser lens proper distance from Nescofilm surface by raising and lowering the table.
Clamp Glass Tubing
Carefully clamp pipette at top and bottom, threading through the filament. Don’t touch glass tubing in the center. Fingerprints = bad.
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.
Flatten gaskets
Press gaskets against glass to make them stick. Do not get fingerprints in the center.
Pipette tip placement
Place glass tube over alignment holes. Place tip halfway through center channel. Press glass tube into gasket to make it stick.
Dispenser tube placement
2.5 mm long dispenser tubes placed at a diagonal to adjust length into channel. Too close to the walls = fills with melted Nescofilm. Too long = catches bubbles.
Place heated weight on glass slide
From bottom to top: hotplate, aluminum foil, microchamber, 1mm glass slide, aluminum foil, heated aluminum block (~ 250 grams), aluminum foil handles. Heat for 5 minutes.
Tighten microchamber on frame
Press down in the middle of the clamp while gently tightening the screws.
Epoxy enters tube by capillary action
Be careful epoxy does not run too far and clog back end of pipette!