[Histonet] Rat lung histology for mast cells and eosinophils

[Liz Chlipala]

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 
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Chen,
Shu-Cheng
Sent: Wednesday, February 02, 2011 11:29 AM
To: histonet <@t> 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

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