FFPE tissue sections of various murine tissues including the heart and liver, when stained using Polysciences' PSR kit (Cat# 24901), display 'blotches' or regions where there is a clear difference in the background (non-collagen) stain. Typically, PSR staining gives a straw or golden-colored background in the cytoplasm (sometimes pale pink), while collagen is very intense pink/red. I've attached an example low magnification image of a liver section we stained displaying the phenomenon I describe - non-uniformity in the background. I would love to hear if anyone else has experienced such an issue and what can be done to eliminate it. Although the collagen is stained well regardless of the background, it is visually unappealing. Thanks!
I have stained nearly 2000 cardiac sections with PSR and can't recall ever having a similar issue. We do get some batch variation but not variation within individual slides. I would be thinking of things along the lines of tissue sections drying, incomplete immersion or rinse in a reagent, reagents not being mixed or a tissue based phenomenon such as pooling of blood/regional disease. Are you able to post a higher magnification image of the transition?
This picture could be met in case of variation of the level of the rack on which the slides are stained. Air bubbles or dryness gives another picture. Please check about the level of the rack and also don't induce too much pressure on the cover slip.
Thanks, Joseph and Mamdouh. I'm also thinking it had to be an issue with tissue drying or incomplete immersion during the staining process.
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