3/28/96

I am looking for available information about freezing a tissue(brain) for LM histochemistry and immunocytochemistry. Also, I need information how to estimate the optical density of a histochemically stained sections? Are there books, those could give me such information? Any information would be very much appreciated.

Igor_Polyakov@qmgate.arc.nasa.gov
Igor Polyakov

In response to your question for freezing brain tissue. We freeze brain for cryostat sectioning in our lab. After it is perfused, (generally in 4% paraformaldehyde) we run the brain through a sucrose series of 10%, 20% and 30% solution. This is for cryoprotection. We then make a cup with aluminum foil, place a small amount of Tissue Tek (freezing medium) in the bottom and place our 2mm slab of whole brain in the cup with the face we want to section, down. We then submerge the rest of the slab in Tissue tek and place the cup on dry ice with acetone poured over the dry ice to make it colder. It freezes pretty rapidly, and will last in the -20' freezer for a couple of weeks before sectioning. Let me know if you want more details...

Cheri Owen
cowen@cmb.biosci.wayne.edu

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