whole mount in situs

Bill BILL at Pharm.SOM.sunysb.edu
Fri Apr 30 07:41:05 EST 1993


Hi everyone,

I have been attempting to perform whole mount in situ
hybridization on mouse oocytes.  The hybe conditions I use seem
to work fine when I use an alk. phosphatase congugated antibody
to detect the digoxigenin UTP.  However, this detection system
does not have the resolution I hope to attain.  Therefore, I had
hoped to use a rhodamine congugated antibody, this is where the
problems begin.  Under the conditions I use the oocytes
autofluoresce.  This is where it gets a little strange.  Its
seems that three of the hybe components (formamide, CHAPS,
SSC) can cause autofluoresence on there own.  Since, this seems
pretty weird I wonder if the temp. (70C) could cause this.

Therefore my question has anyone tried this technique on oocytes
and if so did you encounter similar problems?  Any imput on
solutions is greatly appreciated.

Thanks,

Bill Richards
BHS T-8
Pharmacology
SUNY at Stony Brook
Bill at Pharm.som.sunysb.edu

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