[Histonet] Re: Double stain ?

[Johnson, Teri]

Rachel,

Are you using chromogenic or fluorescent visualization?  If chromogenic,
then do a sequential double stain using the Triton treatment and
antibody complex first, use a stable chromogen (like DAB), and then do
your enzyme digestion and second antibody complex.  If fluorescent, that
makes things a lot trickier.  Sometimes we just can't do fluorescent
doublestaining because the pretreatments are incompatible. The answer
then is to use serial sections and you can often do single staining on
each slide, and then compare the same area in adjacent sections.  Not
the best, but it's certainly a reasonable alternative if the chromogenic
techniques with DAB aren't suitable (as with expected co-expression).

Teri Johnson, HT(ASCP)QIHC
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, MO 64133