[Histonet] Dark background with Gomori's reticulin impregnation

[Malika Benatti]

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Andre,

By looking at your method it does not look right from the start.
We use the Gordon and Sweet Method and it work wonder for demonstration
of reticulin fibers
Here it is Hope it helps

Malika

1.   Sections to water.
2.   Treat with 0.25% acidified Potassium permanganate.   	--- 5
mins
3.   Bleach in 1% oxalic acid.
4.   Rinse in distilled water.
5.   Treat with 5% iron alum.			--- 15 mins	   
6.   Rinse briefly in distilled water.
7.   Impregnate with ammoniacal silver solution. Filter Solution before
use  ---1 min
(Note:  Time in ammoniacal silver may need to be increased 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.

Results
	Reticulin	- Black
  	Collagen	- Grey
	Nuclei	- Red




> From: André Kougioumoutzakis <akoug <@t> hotmail.com>
> Date: 18 April 2010 23:08:35 GMT+01:00
> To: <histonet <@t> lists.utsouthwestern.edu>
> Subject: [Histonet] Dark background with Gomori's reticulin  
> impregnation
>

>
> Hi everyone,
>
>
>
> I've been performing Gomori's reticulin impregnation and never got  
> the results I see in textbooks. The reticulin fibers stain fine, but 

> I get a dark purple/gray background instead of a light pink  
> background. Does anyone know why ?
>
>
>
>
>
> Here is the technique I use :
>
>
>
> 1) Deparaffinize and rehydrate
>
> 2) 0,5% potassium permanganate           1 min
>
> 3) Running tapwater                             3 min
>
> 4) Distilled water
>
> 5) 5% Oxalic acid                                 2 min
>
> 6) Running tapwater                             3 min
>
> 7) Distilled water
>
> 8) 2% ammonium iron sulfate                  1 min
>
> 9) Running tapwater                             3 min
>
> 10) Distilled water
>
> 11) Ammoniacal silver                            1 min
>
> 12) Distilled water                                15 sec
>
> 13) 20% Formaldehyde                          3 min
>
> 14) Running tapwater                            3 min
>
> 15) Distilled water
>
> 16) 0,2% Gold chloride                           5 min
>
> 17) Distilled water
>
> 18) 3% potassium metabisuphite              5 min
>
> 19) Distilled water
>
> 20) Sodium thiosulfate                           1 min
>
> 21) Running tapwater                            3 min
>
> 22) Distilled water
>
> 23) 0,1% Nuclear fast red                     15 min
>
> 24) Distilled water
>
> 25) Dehydrate, clear and mount
>
>
>
> Thanks for your help!
>
> Andre
>
>
>
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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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