[Histonet] Timms copper stain

[Sarah Glyn-Jones]

Hi,

 

I have recently begun trying to optimise Timms copper stain on kidney
tissue. I have frozen tissue sections and I leave them in 1% sodium sulphide
for 1hr before TCA treatment and then the Timms reagent.

 

The recipe I am using is:

 

Solution A
 
5% silver nitrate aqueous (5g in 100ml distilled H2O).
 
Solution B 
 
Hydroquinone                          2.0gms
Citric acid (monohydrate)          5.0gms
Distilled water                         100ml
 
Develop sections in freshly filtered solution of 1 part A and 5 parts B for
5min

 

There are a number of different recipes for the Timms reagent and the one I
am currently using is not working. I cannot seem to get the stain into the
tissue. I end up with lots of staining on top of the tissue despite heavy
washing. 

Does anyone have a different recipe that I could try? I have seen recipes
for Timms reagent using gum arabic and 10% hydroquinone, except hydroquinone
is only soluble in water to 7% (I tried making a 10% solution and it was a
disaster).

 

Any suggestions would be greatly appreciated.

 

Thanks!

 

Sarah

 

PhD Candidate

Proteomics and Biomedicine Group

School of Biological Sciences

Thomas Building

University of Auckland

Tel: (09) 373-7599 ext 85763