Re: frozen sections

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From:"J. A. Kiernan" <>
To:Atoska Gentry <>
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

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