users of NovaRed - some questions

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From:Gayle Callis <>
Date:Thu, 8 Jul 1999 15:07:30 -0600

I have some questions for users of Vectors NovaRed

Are you using this chromogen on frozen sections, fixed after cryosectioning
with either Acetone or paraformaldehyde, and after immunostaining, using a
hematoxylin (Gill type) as a counterstain?

Do you notice any loss of cellular morphology (shown by the hematoxylin 

Have done both fixations, brilliant CD marker staining, but nuclei, other cells
are just not staining.  They tend to look washed out, and not distinct.
nuclei of immunostained cells are basically nonexistent, very frustrating.
I would asume, after chromogen color development and 5 min rinse, the 
pH of tissue should be adjusted for hematoxylin. 

Two buffering systems were tried, TBS and Dulbecco's PBS, a gentle
peroxidase block (Dako), everything that worked with either AEC or
DAB substrate, and after hematoxylin counterstain, cells always looked
like they were right out of Vogue with my oldies but goodies!

Would like feedback on what others are seeing.  I know the substrate has been
used for paraffin sections, probably NBF fixed, and they may stand up to
some rougher treatment. How does the hematoxylin staining look with these?

Just some groping around in the dark for input - love the substrate for
color and sensitivity.  Can't figure the poor hematoxylin/morphology loss,
isotonicity is off??? something is wrong here when it hasn't been with other

Gayle Callis


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