Re: BCIP/NBT insolubility

After our BCIP/NBT we go through a series of graded acetones... 3-5 dips in
each water- 30-60-90-60-30%acetones-back to water and check microscopically
for a nice royal blue.  Works for us.

          "            To:     "'Histonet'"                    
                    08/31/01 09:14        Subject:     BCIP/NBT insolubility      

 A research worker is finding that the BCIP/NBT end product she is
using on her insitu whole mount preparations (13.5dpc mouse
embryo's) is "leaching" out in the alcohols (despite only a few
minutes in each ethanol) on processing to paraffin wax. She is
looking at hair cells in the cochlea and although some colour is
retained, it is not strong enough for photography/publication.
Does anyone know of a method for stabilizing this end product
(which I thought was insoluble) or getting p/w sections without
alcohol dehydration (e.g. DMSO).Morphology is critical here so no
frozen sections and preferably not a water based resin!
I await your replies with the same optimism I have that England will
demolish the Germans (at football) this Saturday!
Many thanks,
Terry Hacker,
Head of Histopathology and Electron Microscopy
Medical Research Council,
Oxfordshire, OX11 ORD
01235 824521
Fax 01235 834776

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