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FISH in biofilms

Ian Harper Ian.Harper at sci.monash.edu.au
Wed Sep 9 00:42:58 EST 1998

> Hello........I am looking for a technique for the simultaneous hyb'ing of
> 2 different bacterial species (occ. found in a biofilm), with 2 different
> fluorescently labeled oligo probes.......preferably using Texas Red and
> Fluroescein as the labels. I have gotten the Texas Red to work alone in a
> pH 6.0 hybridization environment, but am stumped as to how to get the
> Fluroescein probe to work since it needs a pH 9.0 environment.

I have no direct experience with FISH, but I would suggest you look dump the 
Fluorescein (which is notoriously pH sensitive - hence the slight 
modification of these dyes to make pH probes !!!!!), and move to 
either BODIPY-FL (not related to fluorescein) or better still Alexa
(not Greek, but is insensitive to pH bewteen 4 and 10). These dyes are
from Molecular Probes, and are insensitive to pH, more photostable and in
have narrower excitation/emission  spectra but with maxima similar to 
fluorescein. What more could you want ? 

Good luck,


[ I have no commercial interest in  Molecular Probes - just a very 
satisfied customer]. 
Their website, which is also searchable, is at http://www.probes.com 

>< >< >< >< >< >< >< >< >< >< >< >< ><
Dr Ian S. Harper
Confocal Microscopy Facility
Department of Biological Sciences
Monash University, Clayton, Vic 3168, Australia
Email: Ian.Harper at sci.monash.edu.au
Tel: +61 3 9905 5635;  Fax: +61 3 9905 5613
>< >< >< >< >< >< >< >< >< >< >< >< ><

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