Making Blanks

• Use a pencil to write numbers on a piece of paper and cut them out so that they will fit nicely into each spot in the silicone mold.
• Use disposable pipette to inject resin halfway into the mold, and then place the numbered paper inside with forceps.
• Fill the rest of the mold with resin, making sure to fill to the top so the rectangle is solid, and that the numbered paper is in the middle of the resin.
• Place mold into 60°C oven for 24 hours or longer so the resin can polymerize, then pop out the blanks (use the better molds without cracks so the blanks pop out easier).

Mounting

• Take resin strips in labeled scintillation vial and record number in chart (pg. 13)
• Use microscope to determine optimal places within the resin strip where the cells are located, and have not peeled off (“good”, flat side, with cells, facing up).
• Use small ticks with the Sharpie on the “good” side of the strip to create coordinates, then flip the strip over to the “bad” side and circle the spot to indicate where the cells are.
• Continue searching through each resin strip until all ideal cell locations are marked.
• Take Ploy/Bed out of -20°C freezer so it has a chance to thaw before it is needed.
• Place resin strip in vice (with silicone strips on the grip parts) and use the saw to cut out the circled areas. If the strip is particularly thin and brittle, use a razor blade to score and snap the circled areas out. Use sandpaper, if necessary, to carefully wear down the resin, making the sample small enough to fit on top of the blank with little overhang.

*Put on a mask when cutting the resin so it is not inhaled*

• Move over to the fume hood, while making sure you remember which side is the “good” side (you can tell by looking at which side the marker is on, or which side is flat and which is curved)! Sand the top of the blank and the “bad” side of the resin sample, then place a small drop of resin on the top of the blank. Try to make the sample sit as horizontally as possible, to make it easier for sectioning.
• Stand the blanks up in the molds so it is easy to stick on the samples (see diagram on pg. 34). Stick the resin sample, “good” side with the cells facing up, onto the blank, so the two sanded sides are glued together. Once this has been done for all the samples, stick the entire mold into the 60°C oven for 24 hours or more, until polymerized.
• Record the numbers on the blanks and the corresponding number on the scintillation vial into the table on pg. 35.

Last modified by Whitney Moore, August 2007