[Histonet] Re: Stains for Macrophages for Laser Capture Microdissection purpose

[John Kiernan]

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 <eberledelphine <@t> hotmail.com>
Date: Wednesday, January 27, 2010 16:27
Subject: Stains for Macrophages for Laser Capture Microdissection purpose
To: jkiernan <@t> uwo.ca, sharon.willman <@t> bms.com
Cc: histonet <@t> 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
> eberledelphine <@t> hotmail.com 
> 
> > From: jkiernan <@t> uwo.ca
> > To: sharon.willman <@t> bms.com
> > Date: Wed, 27 Jan 2010 00:22:53 -0500
> > Subject: Re: [Histonet] Histology Special Stains for Macrophages
> > CC: histonet <@t> 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.willman <@t> bms.com>
> > Date: Tuesday, January 26, 2010 12:12
> > Subject: [Histonet] Histology Special Stains for Macrophages
> > To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> 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
> > > 
> > > 
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