How about phase contrast optics and identifying the cells by their shapes? Non-specific addition of colour to a section of unfixed tissue probably will not show the cells any more clearly. I have taken the liberty of including a colleague, Dr Paul Walton, in this reply. He does laser capture microdissection and may have something to suggest. John Kiernan Anatomy, UWO London, Canada = = = ----- Original Message ----- From: delphine eberle <eberledelph...@hotmail.com> Date: Wednesday, January 27, 2010 16:27 Subject: Stains for Macrophages for Laser Capture Microdissection purpose To: jkier...@uwo.ca, sharon.will...@bms.com Cc: histonet@lists.utsouthwestern.edu > Hi, > > I have another question following Macrophage staining. > I am setting up a Laser Capture Microdissection protocol to dissect > macrophages from atherosclerosis lesions (non fixed frozen sections) and > extract RNA for gene expression purpose. > I am looking at a quick staining (less than 30min otherwise RNA is > degradated) for macrophages in that context? Any suggestions? > > Thanks a lot, > Delphine > > Delphine Eberlé PhD > UCSF Department of Vascular Surgery > eberledelph...@hotmail.com > > > From: jkier...@uwo.ca > > To: sharon.will...@bms.com > > Date: Wed, 27 Jan 2010 00:22:53 -0500 > > Subject: Re: [Histonet] Histology Special Stains for Macrophages > > CC: histonet@lists.utsouthwestern.edu > > > > None of the methods mentioned in the enquiry are stains for macrophages. > > Research workers who never took Histology 101 often stain cells of the > > monocyte/macrophage lineage immunohistochemically (IHC), using very > > expensive primary antibodies and fairly expensive kits to amplify and > > detect the binding sites. IHC is necessary if you must find every > > macrophage, including a tissue's recent monocyte immigrants that haven't > > yet done any work. Macrophages that have been busily eating blood are full > > of brown granules that don't need any staining. > > > > Ordinary people recognize macrophages by their appearance in sections > > stained with a well done H&E or with one of the many Romanowsky-Giemsa > > blood stains. With the latter it's important to get the pH right - much > > lower for sections of formaldehyde-fixed objects (pH4 to 5) than for > > methanol-fixed films or smears (traditionally 6.8). It's all very well > > explained in RD Lillie & GM Fullmer's "Histopathologic Technic..." (4th ed > > 1976, pp.193-197; the 3rd edition, 1965, is OK too) and also in more recent > > textbooks including those by Bancroft & Gamble, Geoffrey Brown, Freida > > Carson and, of course > > Yours truly. > > > > John Kiernan > > Anatomy, UWO > > London, Canada > > http://publish.uwo.ca/~jkiernan/bookfind.htm > > = = = > > ----- Original Message ----- > > From: "Willman, Sharon" <sharon.will...@bms.com> > > Date: Tuesday, January 26, 2010 12:12 > > Subject: [Histonet] Histology Special Stains for Macrophages > > To: "histonet@lists.utsouthwestern.edu" <histonet@lists.utsouthwestern.edu> > > > > > Hi, > > > We are needing to do a special stain for macrophages. What > > > is the most common stain for that? Does anyone do a Sudan > > > Black, Alcian Blue or Van Gieson for macrophages? Any > > > information would be appreciated. > > > Thanks in advance. > > > Sharon > > > > > > > > > ________________________________ > > > This message (including any attachments) may contain > > > confidential, proprietary, privileged and/or private > > > information. The information is intended to be for the use of > > > the individual or entity designated above. If you are not the > > > intended recipient of this message, please notify the sender > > > immediately, and delete the message and any attachments. Any > > > disclosure, reproduction, distribution or other use of this > > > message or any attachments by an individual or entity other than > > > the intended recipient is prohibited. > > > _______________________________________________ > > > 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 >
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