Please advise.
I was sent a bunch of sample by an investigator who is not too sharp. The samples were prepared thus: Pancreases were removed, washed placed in a modified Zamboni fixative (2% formaldehyde, 15%Picric acid in 0.1M PBS, pH 7.5). Sample portions were cut from the head, body and tail of the pancreas in 3 mm by 3 mm sections. The pancreatic tissue samples from the various regions of the pancreas were fixed overnight; samples were equilibrated in 50% sucrose in 0.01M PBS for 12 hours at 4°C, and mounted in Tissue Tek OCT Compound (Miles Inc.); 0.015 mm (15 μm) thick sections were obtained by cryostat sectioning. SAMPLES WERE FROZEN IN 50% Tissue Tek 50% sucrose , then shipped on wet ice, not dry ice. These frozen tissue samples were in small embedding molds with a tiny dab of the OCT/sucrose gel on top of them, it looked like none was under them. They shipped these supposedly previous frozen samples to me on wet ice, not dry ice (dumb move), the molds were open on top in a box, the sample thawed and the gel and sample was leaking out of their molds, it was a mess. The investigator who sent the samples to me said this "the samples arrived within 18hrs as signed for, so they should have still been frozen", how this person got to be the CEO of a Biotech company I cannot imagine. They wanted me to just refreeze them and cryosection them to test some abs they sent with them, I said that I would not do that, I did agree to take 2 samples as a pilot study and fix them in formalin overnight and then process and embed them in paraffin, which I have done. If I can get them to section (they are still soft and squishy ) I was planning on doing an H&E stain and look at the samples to see if they look preserved at all. My question to you all is this: if the tissue looks viable by H&E what should I try to test immuno reactivity viability? I was thinking of running an antibody like vimentin or cytokeratin or something, but these are ms pancreas so maybe I should use insulin ab or something else. Thank you for your valuable advise, Regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. Ste.215 Aurora, CO 80045 720-859-4060 fax 720-859-4110 www.ihctech.net www.ihcrg.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet