[Histonet] Stains for pneumocystis

[Gayle Callis]

We have one lab whose expertise is Pneuocystis research.  They use 
Dade-Behring Diff-Quick from VWR but extend time in Solution 2.

Protocol:

Fix cells with methanol
Solution 1 - according to package insert
Solution 2 - 30 to 40 minutes
Rinse per package insert, air dry and look under oil, a coverslip can be 
mounted over an air dry slide (fans help out with drying or forced air of 
some sort).

Any blood cells will be too dark but the Pneumocystis needs the extra time 
in Sol. 2 in order to stain the cytoplasm for viewing.

This is basically a Wright Giemsa stain, and if done like a blood smear, 
should probably work also.  Wright Giemsa is available under Harleco 
brand.  One might have to stain longer after buffering however.

They stain too numerous to count murine BAL samples, spun down with Cytospin.

GMS can be shortened by using a MW staining method and you may want to fix 
the smears/spins differently when using this protocol i.e. other than 
methanol.

Good luck

At 06:22 AM 1/10/2006, you wrote:
>My fellow Histonetters,
>
>I would like to know what stain you are using for pneumocystis.  We are
>presently using a "modified" GMS that takes approximately one hour and
>15 minutes to perform.  A few of our physicians would like something a
>bit faster.  (Of course, I would like to get the specimen sometimes
>before 3 p.m., but that is another story for another time!)  I have also
>found that a Thin Prep slide from a BAL is the best specimen.  We have
>tried direct smears and Cyto-spins with much less success.
>
>Thanks is advance,
>Lynne
>
>Lynne A. Bell, HT (ASCP)
>Laboratory
>Central Vermont Hospital
>P.O. Box 547
>Barre, VT 05641
>(802)371-4122
>
>
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)