[Histonet] manual processing help

Alfred Penzo Mendez alfredpe <@t> mail.med.upenn.edu
Fri Oct 9 08:51:15 CDT 2009


Dear Histoners,

I used to process mouse tissues using a vacuum oven and never ran into any
 problems. I recently moved to a new lab and there is no vacuum oves, but 
I've been told that vacuum is not an absolute requirement for paraffin
infiltration, and I only need to double the paraffin infiltration changes;
 however I've tried a coule of times and I can't get anything embedded. Liver
 and pancreas become vey hard and turn to dust when I try to section them. 
Gut cuts, but sections have lots of compressions and wrinkles. Does anyone
 have an idea of why is this happening? here is the full procedure:

-Tissues: adult mouse liver, pancreas and gut cut into small pieces (1mm3)
-Fix: zinc-formalin (polysciences) 1 h Room temp + o/n at 4oC. Tissues is fixed in 10 ml.
-Dehydration: 1 hour each 50%, 70%, 95%, 1005 ethanol + o/n in ethanol at 4oC.
-Clearing: 2x 30 minutes in xylene (also tried 2x 1 hour, same result, room temp
-Paraffin: 3x 1 hour in paraplast at 60oC

Many thanks,

Alfredo. 
  



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