Re: intestine cryosection problem

From:Patti Loykasek

I have an acetone fixation method for frozen sections that has always worked
well for me. It results in excellent morphology, and no loss of sections.
100% Acetone
70% Acetone - made with tap water!!
50% Acetone - made with tap water!!
Chill all solutions 4 degrees C.

1. Cut frozen sections, air dry for 10-15 minutes.
2. Fix in cold 100% Acetone for 10 minutes.
3. Air dry for 10-20 minutes.
4. Fix in cold 100% acetone for 10 minutes.
5. Place in cold 70% acetone for 30 seconds.
6. place in cold 50% acetone for 30 seconds.
7. Rinse in tap water.
8. Place in buffer.
You must use tap water - the ionic concentration is one of the key elements.
This procedure is from the old Biotek company. I mourn their demise.
I have great success with this fixation when using lymphoma markers,
cytokeratins, and most other antibodies. The only antibody that has not
worked with this fixation is TTF.

Hope this helps. 

Patti Loykasek
Phenopath Laboratories
Seattle, WA

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