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Recently we have run into a nasty problem with our NBT/BCIP in situ
detections. The in situ part seems to work fine and the alk phos
detections is sensitive.
However, under the microscope, the precipitate is completely granular
and seems to be located above the tissue, which is absolutely unsuitable for
photomicrography.
Has anyone encountered a similar problem, and how did you get rid of it?
Fri Apr 03, 2009 2:42 pm
Laoz
Rank: Member
Joined: Apr 13, 2007
Posts: 109
Post subject:
I don't really have this problem, but you can try vectorlab's mounting medium, which is claimed help for preventing NBT/BCIP in situ crack.
Laoz
Sat Apr 25, 2009 2:02 am
hrn
Rank: Site Admin
Joined: Sep 27, 2004
Posts: 246
Post subject:
If dehydrating and mounting in organic mountant, insufficient dehydration (water in xylen) can be the reason for this. I use to dip in aceton between alcohol and xylen. Looks like more intense stainings are prone to crystallize....
Mon Apr 27, 2009 7:35 am
hrn
Rank: Site Admin
Joined: Sep 27, 2004
Posts: 246
Post subject:
Digging deep in my archive, I found a horror example of NBT/BCIP crystals. See images
I also found in my notes, that these crystals are formed by formazan, a reaction product from NBT/BCIP.
Knock on wood...haven't had this problem in years ....
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