Fluorescent In situ

Bruce Appel appel at uoneuro.uoregon.edu
Thu Dec 22 15:41:24 EST 1994


In article <94Dec22.095229est.18900 at gpu.utcc.utoronto.ca>,
ilivne at UTCC.UTORONTO.CA ("I. Livne") wrote:

> Dear netters
> Has any one tried the Molecular Probes ELF substrate for detection of
Digoxigenin probes
> in whole mount in situ hybridization? On paper it looks as promising
alternative to the 
> NBT substrate and generates clean Fluorescet signal. Is it for real?
> 
> Livne Bar
> I.Livne at utoronto.ca

We've tried out the ELF substrate to detect several different
digoxygenin-labeled probes hybridized to whole-mount zebrafish embryos. We
find that it is less sensitive than NBT/BCIP and even slightly less
sensitive than Fast Red. In addition, the reaction product forms a
crytalline precipitate, which is not evenly distributed throughout the
cell, thus reducing cellular resolution. Finally, the substrate comes out
of solution after about 2 hr and sticks to the tissue, causing additional
problems.

Despite the problems, I can see it having some utility. For example, if
you want to mark a domain (as opposed to individual cells) with a
fluorescent signal, this may do it for you.

Hope this helps.
Bruce Appel

-- 
Bruce Appel
University of Oregon
appel at uoneuro.uoregon.edu



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