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Principles & Protocols

Protocols - Prepare Pioloform Grids

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1. Clean a Premium microscope slide (Fisher Scientific cat# 12-544-2) by dipping in 2 coplin staining jars (Ted Pella cat# 428-3) that containing 100% Ethanol and wipe it out with Kimwipes.
2. Place the clean, dry slide into cylinder of Film Casting Device(Electron Microscopy Sciences cat # 71305-01) that contains 0.5 to 0.7% Pioloform (Ted Pella cat # 19244) in chloroform in its 500ml flash and slowly squeeze the Atomizer bulb to pump up Pioloform staining solution to close to frosted label slide and hold for 20 seconds by turn off both air-in and air-out valve.
3. Turn on the air-out valve (black) to drain the solution down to flash by venting the pressure via the air-out valve and wait for 30 seconds.
4. Take the coated slide out and score 4 sides with a razor blade.
5. Breath slowly to the scored side of coated slide to create a frosting layer.
6. Quickly lower the coated slide held at an angle in Smith Grid Coating Trough (Ladd Research Industries Inc., cat #6-10840) that contains distilled water at the top Trough so that the film is detached from the slide. The color of the film should be gold. Disregard the film if it is silver.
7. Place a number of clean grids on a film.
8. Cut out piece of parafilm, place on top and pull it out the water.
9. Let a film with number of grids dries out on the petri dish o/n before using. Carbon coating if necessary in the same day making grids.

-- KdDerr - 02 Oct 2009