Re: frozen sections
<< Previous Message | Next Message >>
From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> |
To: | Atoska Gentry <gentras@vetmed.auburn.edu> |
Reply-To: | |
Content-Type: | TEXT/PLAIN; charset=US-ASCII |
On Thu, 13 Apr 2000, Atoska Gentry wrote:
> >problem surfaces. After completing my H&E's the nuclei are very visible
> >but can any of you recommend a stain/stains that would reveal cell margins
> >(i.e. whereby astrocytes, and glial fibers are distinguishable)?
H&E is pretty uninformative for nervous tissue, for just the reason
you state. Use a basic dye instead of alum-haematoxylin, and this
will show the DNA and RNA (Nissl substance), thereby revealing the
shapes of the neurons. Normal astrocytes and oligodendrocytes can
usually be recognized by their nuclei. You need a special method
(immunohistochemical is easiest) to see the cytoplasmic processes
of neuroglial cells.
A blood stain such as Giemsa or Wright's will give you a nice
blue Nissl/nuclear stain with a pink background. You'll need to
use it at a more acid pH than the standard 6.8 that's usual for
blood smears. Try pH 4, 4.5, 5 and 5.5 and decide which gives
you the result you want. A combination of a basic and an acid
dye is much better than H&E (for nearly everything, IMHO, but
definitely for nervous tissue).
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
Phone: (519) 661-2111
FAX (Department): (519) 661-3936
E-mail: kiernan@uwo.ca
<< Previous Message | Next Message >>