[Histonet] Grocott's Methenamine Silver Stain

[Kathleen Roberts]

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.

-Kathleen

Jones, Sarah - RAS wrote:

>Kathleen,
>
>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.  
>
>Sarah
>
>-----Original Message-----
>From: Kathleen Roberts [mailto:kgrobert <@t> rci.rutgers.edu] 
>Sent: Wednesday, May 05, 2004 3:16 PM
>To: Bartlett, Jeanine; 'histonet <@t> lists.utsouthwestern.edu'
>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!
>Kathleen
>
>Bartlett, Jeanine wrote:
>
>  
>
>>Kathleen:
>>
>>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
>>
>>
>>-----Original Message-----
>>From: histonet-bounces <@t> lists.utsouthwestern.edu
>>[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kathleen
>>Roberts
>>Sent: Wednesday, May 05, 2004 1:54 PM
>>To: 'histonet <@t> lists.utsouthwestern.edu'
>>Subject: [Histonet] Grocott's Methenamine Silver Stain
>>
>>
>>Hey, all-
>>
>>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-
>>Kathleen Roberts
>>Principal Lab Technician
>>Neurotoxicology Labs
>>Dept of Pharmacology & Toxicology
>>Ernest Mario School of Pharmacy
>>Rutgers University
>>41 B Gordon Rd
>>Piscataway, NJ 08854
>>
>>
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>>    
>>
>
>
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