[Histonet] Gordon & Sweet Reticulin Stain problem on BMT Sections

[Rene J Buesa]

Usually an excess of ammonium in the amoniacal-silver solution is the culprit. Check this solution.
René J.

--- On Tue, 6/8/10, Malika Benatti <BenatM <@t> gosh.nhs.uk> wrote:


From: Malika Benatti <BenatM <@t> gosh.nhs.uk>
Subject: [Histonet] Gordon & Sweet Reticulin Stain problem on BMT Sections
To: histonet <@t> lists.utsouthwestern.edu
Date: Tuesday, June 8, 2010, 10:33 AM


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Dear all,

Lately we have been experiencing problem with our Retic Stains, who keep lifting off slides after the gold chloride step.

We have changed all solutions one by one, trying to see which one was causing problem without success.
BMT Bx are cut at 3 micron on X-tra adhesive slides.
BMT BX are decal in 10 % Formic Formaldehyde.

Here is the protocol that we are currently using.

1.   Sections to water.
2.   Treat with 0.25% acidified Potassium permanganate.       --- 5 mins
3.   Bleach in oxalic acid.
4.   Rinse in distilled water.
5.   Treat with 5% iron alum.                            --- 15 mins       
6.   Rinse briefly in distilled water.
7.   Impregnate with filtered ammoniacal silver solution          ---1 min
NB: Time in ammoniacal silver may need to be increased up to 5 min for bone marrow trephines
8.   Rinse well in distilled water.
9.   Reduce in 10% formalin (in tap water).                    --- 1min
10.  Rinse in distilled water.
11.  Tone in 0.2% Gold Chloride                         ---1 min
12.  Rinse in distilled water.
13.  Fix in 5%  sodium thiosulphate (hypo)                    --- 30 sec
14.  Wash in water.
15.  Counterstain with neutral red.
16.  Dehydrate, clear and mount. 


Any suggestions would be  appreciated.

Malika


Malika Benatti

Specialist BMS 
Camelia Botnar Laboratories
Histopathology Department
Great Ormond Street Hospital
London WC1N 3JH


Tel:  +44 20 7405 9200 ext 5475
Fax:  +44 20 7829 7875

benatm <@t> gosh.nhs.uk


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