Chen For mast cells you can fix in 10% NBF and be fine with toluidine blue. We stain with toluidine blue on formalin fixed samples all of the time. The trick is not to dehydrate the sections, let them air dry and then mount. I would stick with formalin fixation that's going to be your best bet for most specials and IHC stains. For eosinophils you can stain with giemsa, or we would run a modified dif-quik stain that worked well for eosinophils. For neutrophils I'm not sure but if you are staining for mouse neutrophils, serotec has a nice antibody. Macrophages would be IHC for ED-1 also from serotec, CD3 from Dako will work on rat tissue nicely. There are other markers that will work on cell lines.
Good Luck Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC PO Box 18592 Boulder, Colorado 80308 office (303) 682-3949 fax (303) 682-9060 www.premierlab.com Ship to Address: 1567 Skyway Drive, Unit E Longmont, Colorado 80504 -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Chen, Shu-Cheng Sent: Wednesday, February 02, 2011 11:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Rat lung histology for mast cells and eosinophils Hi, We are called to support an asthmatic rat lung project and need to stain for mast cells, eosinophils, neutrophils and possibly other inflammatory cells, such as Th2 cells etc. Any suggestions you can give us in terms of fixatives and special stains or IHC are very much appreciated. From my search, it seems that Carnoy's fixative with toluidine blue is the way to go for mast cells. But can one clearly differentiate eos from neuts with this fixative or a formalin fixed H&E stained lung section? If we need to do Trichrome, PAS, May Grunwald Giemsa and H&E etc. what would be the best fixative to accommodate these stains? BTW, I heard that formalin free Zn-fixative is good for IHC. Will it be good for all these stains above? Sorry for so many questions. It shows how little experience I have in this area. Thank you, Shu Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates Direct contact information for affiliates is available at http://www.merck.com/contact/contacts.html) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet