RE: H&E, (and how to get advice)

From:"Tarpley, John" <jtarpley@amgen.com>

Circle,
If you want to try immunohistochemistry you might check with the various
antibody vendors to see if they have samples sizes of antibodies available.
Sometimes these sizes are available at lower cost or even free. This might
give you the opportunity to demonstrate the value of IHC to your supervisor
in order to obtain more funds to continue the project. I agree with John
Kiernan's point that while an H&E may be useful for normal vessels, you will
probably find it difficult to distinguish between endothelial cells and
smooth muscle cells in various pathological conditions.

John E. Tarpley 5-1-A
Associate Scientist
Amgen Inc.
One Amgen Center Drive
Thousand Oaks, CA 91320 
These Opinions are my own and not necessarily those of my employer.

-----Original Message-----
From: Circle :> [mailto:s991002@mailserv.cuhk.edu.hk]
Sent: Monday, November 06, 2000 1:03 AM
To: J. A. Kiernan
Cc: Histonet
Subject: Re: H&E, (and how to get advice)


Dear John,

Oh, sorry for any inconvenience. My name is Circle. Sounds strange to
Westerners, but it's a friendly name in my place. You may called my
surname Chan.

The reason why I asked this question is that I borrow a CD-rom from
library, (called Histology image review, by Frank J. Wilson) which
contains lots of histology and IHC photos. And I noticed
that the stain they used is H&E. However, I found the colour is different
among different slides. I used H&E to counter stain some cells before. The
technicans told me that H gives the colour of purple/blue (on the
nucleus) and E gives orange/red (on the cytoplasm). What I found in the
CD-rom is that I can identify different structures on the slides. For
example, I can distinguish endothelial cells and the arterial muscle
cells. That is, they have differnt tone on differnet parts of the tissue!

Other reason I would like to try on H&E rather than IHC is the price of
the antibody. I know factor VIII can identify endothelial
cells. Unfortuantely, my supervisor has already rejected as the antibody
cost much. 

I have the alpha-actin to stain the vascular smooth muscle cells only. 

I am not just a beginner of histotechnology, but also an
outsider. However, as a student, I hope I can learn more and practice
more.

Yours,
Circle Chan





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