Re: [Histonet] Stains for pneumocystis

From:Rene J Buesa

Hi Lynne:
  I also used Grocott's Methenamine silver but it took about 25-30 minutes using the following protocol:
  a- after dewaxing, hydrating → dist. water
  b- to chromic acid heated x30 secs. in my 480W microwave oven (MWO)[in my MWO this was equivalent to 3.6 kcal  of energy; you should adjust the time to the energy output of your MWO in order to not overheat the sections]
  c- to a water bath at 60ºC x 5 min.
  d-wash with dist water
  e- to the methenamine working solution → MWO x 45 secs. (= 5.4 kcal energy)
  f- to 60ºC water bath until sections are dark brown (approx. 5-7 min.)
  g- wash dist. water
  h- gold chloride sol x 5 min.
  i- wash with dist water
  j- sodium thiosulfate sol. x 30 secs.
  k- wash
  l- counterstain with light green sol. just a few seconds
  m-dehydrate, clear, cover slip.
  This procedure takes less than 30 min. total
  Hope this will help you.
  René J.`

"Bell, Lynne"  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 A. Bell, HT (ASCP)
Central Vermont Hospital
P.O. Box 547
Barre, VT 05641

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