Re: Brown-Hopps

From:Barbara Stancel

Dear Helen,

We have used the Twort Stain for 35 years. Our pathologists prefer it to the 
other Gram stains. The techs prefer it because it is simple and does not 
require picric acid. It is our most asked for stain, so we have it set up to 
run 25 slides at a time.

Our Twort stain is a "hybrid". It was developed by Charles West at our 
"mother" laboratory in Beltsville, MD, USA in the late 1950's or early 
1960's. The Gram+ bacteria stain a deep blue with the crystal 
violet/iodine/acetone. The Gram-bacteria are double stained with Safranin O 
and the Twort solution (neutral red and fast green). They stain a medium to 
dark red. The back ground (counterstain) is bright green from the fast 

I have been working on a copy of the procedure for publication, but have not 
had the time to finish. So, our procedure is not published. If you would 
like to give a try, I would be glad to send you a copy.

Histologically yours,

Barbara H. Stancel, HTL(ASCP)HT
USDA, FSIS, OPHS, Eastern Laboratory, Pathology
RRC, 950 College Station Road
Athens, Georgia  30604
phone: (706) 546-3556
fax: (706) 546-3589

>From: lam-helen 
>Subject: Brown-Hopps
>Date: Sun, 30 Jun 2002 22:55:26 +0800
>We have to stain for both gram positive and negative micro-organisms in the 
>same paraffin section.  However, we usually do conventional gram and 
>therefore not too familiar with the Twort's or the Brown Hopps method.  
>Which one is more reproducible and easier to master?  How do these methods 
>compare in yours hands?   As for the piciric acid-acetone employed in Brown 
>Hopps, do you think this counterstain is going to further decolorize the 
>gram positive organisms and nuclei?  As I have never seen a section stained 
>with Brown Hopps before, can anyone tell me what the expected results are?  
>I have tried both methods.  With the Brown Hopps, nuclei stain a brownish 
>(not bright at all) red and in the same colour and intensity as gram 
>negative organisms.  Is that OK?  It would be very nice if anyone can his 
>or her experience with me.  Thanks in advance.
>Helen Lam
>Tuen Mun Hospital,
>Hong Kong

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