[Histonet] warthin starry fading

[Gudrun Lang]

Hi all!

Today I stained Warthin Starry for Hp. This time I was lucky to have the
first time purchased control tissue (Spirochetes control from Sigma) for
comparison.

In the package there was also a ready stained slide with really nice black
spirochetes in yellow-brownish tissue/nuclei. This was stained with a Sigma
Steiner Kit, but I thought my Warthin Starry should look the same.

So, after staining, the patient tissue shows nice Hp in yellow/brownish
background, but nuclei didn't stain. The control tissue had black-sprinkled
nuclei and no spirochetes at all. I was a little bit disappointed, but at
least the patient tissue was ok for diagnosis.

A few hours later I looked at the control slide again and found, that the
whole black staining had faded till a few speckles. (Patient slide was out
of reach to look at)

 

Does anybody know why this fading did happen? I have no information about
the fixation and processing of the control slides, but thought it must fit
for FFPET. The only point is, that we received the slides several months
before. Is aging a factor? 

 

Staining protocol in brief:

Everything diluted in pH 4 water, made with citric acid.

30 min 1% AgNO3

5-7 min developer (2% AgNO3+gelatine+Hydrochinon)

Waterrinse

Dehydration, coverslipping

 

Gudrun