beta gal staining in suspension cells

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From:Jih-tung Pai <>
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Hi All,

    I'm new in this group, so I don't know if this has been asked
before. I am trying to do a beta-gal staining on a lacZ transgenic
mouse. When I stain a piece of tissue or minced tissues, I will get very
strong staining with or without fixation. But, when I try to break the
tissue into single cell suspension by collagenase treatment or by
Medimachine (a machine design to break tissue into single cells), I can
no longer get staining in these single cell suspension. Although I know
for sure a portion of these cells should have strong beta-gal activity
which normally will turn whole staining solution into blue if they are
in intact tissue without fixation. What could be the reasons for that?
Is it necessary to stick cells to plate or slide before one can do
beta-gal staining? Beta-gal staining is desirable as I need to use this
procedure to verify my cell purification scheme, and I won't have enough
cells to do FACS analysis by FDG after the procedure. I use In Situ beta
Galactosidase staining kit from Strategene for staining. I would
appreciate any suggestions and comments.


Jih-tung Pai
The Burnham Institute
10901 N. Torrey Pines Rd.       Tel. 002-1-858-646-3100 x3229
La Jolla, CA 92037, USA         Fax  002-1-858-646-3199

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