Hi Jan IHC on plastic-embedded specimens is difficult, I think the antigens can get rather damaged from the embedding and/or deplastifying processes. In our experience, it's possible to do some markers, while others are irretrievable. One thing I notice is that our protocol for deplastification calls for 2x MEA 20 minutes, while yours does 3x. Perhaps cutting down to 2x would be sufficient (your sections are thinner than ours, too), and would potentially cause less damage. Good luck
__________________________________ Jean-Martin Lapointe, DMV, MS, dACVP AccelLAB Inc ------------------------------ Message: 4 Date: Fri, 21 Jan 2011 09:41:39 -0500 From: Jan Berry <jebe...@umich.edu> Subject: [Histonet] Immunohistochemistry in plastic sections To: histonet@lists.utsouthwestern.edu Message-ID: <a566bacb-4a03-4f7b-8702-98c6556d2...@umich.edu> Content-Type: text/plain; charset=us-ascii I'm wondering if anyone has experience with immunohistochemistry using plastic sections. I am currently working with 4um sections, and have tried doing staining with and without Dako antigen retrieval, but I am not getting very good results. Tissues (mostly mouse bones, but some soft tissues also) were fixed in PFA, cold embedded, and sections were deplastified using 3X 20 minutes methoxyethyl acetate and 2X 5minutes acetone, followed by 5 minutes in running dI water before beginning. I would appreciate any advice! Jan Berry, University of Michigan ************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
