08 June, 2009

Obviously Not Quite Right

DAB stain, mouse stomach, 40X.

This particular staining protocol had diaminobenzidine, hemotoxylin, and McGill's modified EA solution as stains. The diaminobenzidine, from what I understand, was supposed to color neutrophils and monocytes varying shades of grey-black intracellularly. Light blue is supposed to correspond to basophils. Either I've discovered some new property of basophils in which they invade and form sheets in the connective tissues and basement membrane of the stomach (which'd be even weirder considering that the system in question skews to TH1/TH17-driven inflammation), or I've screwed up the staining protocol somehow. The latter is much more likely, and it also looks like I left the slides in the hemotoxylin for way too long.

I just wish I could remember where I'd put the Coplin jars, because this time around I was staining out of Petri dishes (Toaster does not recommend putting xylenes in a Petri dish, unless it is a fancy glass Petri dish).
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Ambivalent Academic said...

Ah Toaster, histology is an ART not a science. You're way over-analyzing this shit. Also, try white-balancing your image in photoshop - it's totally kosher (go to image: levels: pick the white eyedropper and click it in the blank field in the center of your image). It will correct the weird hues you're seeing here, and maybe then it won't look so wonky.

Also, I've never used DAB alone, only as a colorometric indicator for immhunohistochemistry. I don't recall it giving any background staining in non-immunodetected tissue layers. Not sure if that means anything to you.

Anonymous said...

Not quite right? It's lovely! I love the corals and peachy tones. The scale bar kinda ruins it for me hanging on the wall in the living room.