[Histonet] Dark background with Gomori's reticulin impregnation

[André Kougioumoutzakis]

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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