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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
