Re: [Histonet] Grocott's Methenamine Silver Stain
Aha! I knew there was a way to do that, but I could not find it in the
archives. Thank you, I'll try it.
Jones, Sarah - RAS wrote:
>Since you don't know how long they were in the decal solution, you might
>also try putting the slides (following deparaffinization and hydration and
>before staining) into a saturated lithium carbonate solution for half an
>hour or longer. This helps restore nuclear detail in over-decalcified
>specimens, and might also help with your problem.
>From: Kathleen Roberts [mailto:email@example.com]
>Sent: Wednesday, May 05, 2004 3:16 PM
>To: Bartlett, Jeanine; 'firstname.lastname@example.org'
>Subject: Re: [Histonet] Grocott's Methenamine Silver Stain
>I am not sure how long they were in decal...but I supplied some of our
>Fisher Scientific Cal-EX, which has hydrochloric acid and EDTA in it, to
>the Rutgers vet who had this case of sneezing rats. I did ask him how
>long he decal'd them, but he doesn't remember either.
>The chromic acid was freshly ordered as a 10% solution from LabChem,
>Inc., which was then diluted down to 4% the morning I was to use it.
>Thanks for your help!
>Bartlett, Jeanine wrote:
>>How long were they in decal and what type of decal? And do you make
>>your chromic fresh?
>>Jeanine Bartlett, HT(ASCP)
>>Centers for Disease Control
>>Infectious Disease Pathology Activity
>>1600 Clifton Road, MS/G-32
>>Atlanta, GA 30333
>>[mailto:email@example.com] On Behalf Of Kathleen
>>Sent: Wednesday, May 05, 2004 1:54 PM
>>Subject: [Histonet] Grocott's Methenamine Silver Stain
>>I've been trying to stain some FFPE slides of decalcified rat sinuses
>>for fungi using the AFIP protocol for Grocott's stain, and despite using
>>fresh reagents, the positive control and unknowns do not stain at all,
>>except for some random deposition of silver that does not even begin to
>>look like fungi. The positive control is loaded with Candida, and can
>>be seen with H&E easily.
>>So far, we don't see any fungi in the rat sinuses and lungs (they were
>>sneezing, and the bedding came back positive for Aspergillus) with H&E
>>staining, which is why we decided to try the Grocott's-one way or
>>another we should be able to visualize it in the tissues.
>>Any ideas why the Candida-loaded positive control would not stain with
>>Grocott's? Right now I am thinking that perhaps they didn't sit long
>>enough in the 4% chromic acid, even though the protocol says to soak the
>>slides for an hour to oxidize them. Either that or a longer soak in the
>>methenamine-silver solution at 60 degrees C (also for one hour).
>>Thanks in advance for your help-
>>Principal Lab Technician
>>Dept of Pharmacology & Toxicology
>>Ernest Mario School of Pharmacy
>>41 B Gordon Rd
>>Piscataway, NJ 08854
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