Help on whole cell patch clamp

Ken Scholz kps2 at midway.uchicago.edu
Thu Jul 15 12:30:27 EST 1993


In article <jzempel.742701405 at morpheus>, jzempel at morpheus.wustl.edu (John
Zempel) wrote:
> 
> xiaoz at gandalf.rutgers.edu (Xiao Zhang) writes:
> 
> > Any suggestion on this matter would be very appreciated. 
>
> Two things:
> 
> Try different glass.
> 
> Try using Fl in your internal solution.  While this may not be desirable, it
> may get you started.
A few more suggestions:
Check the osmolarity of both your bath and pipette solutions, as well as
the
tissue culture media the cells are grown in.
Your difficulties may be an indication that the cells are not completely
healthy--check your cell culture media, incubator etc.  Be especially
watchful of pH changes (e.g. when the incubator door is left open or when
plates of cells are left outside the incubator in media that is intended
to be used in a 5% CO2 environment).
Look in the microscope at your pipette when suction is applied--does the
pipette move when you apply suction?
Try using younger (or older) cells.
How often are the cells fed?
Make sure neither pipette solution nor bath solution are, or have ever
been,
contaminated.  Once a solution is contaminated even sterile filtration will
leave behind bacterial toxins that can make cells very fragile.

--
Ken Scholz                         Univ. of Chicago
kps2 at midway.uchicago.edu           Dept. of Pharm. and Physiol. Sci.



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