[Histonet] Growing PC12 cells onto coverslips, their differentiation and subsequent immunohistochemistry

[Gayle Callis]

Look into buying chamber slides from NUNC designed for this use.  They are 
mini culture chambers and you do not have to manipulate coverslips, 
sometimes a tedious, delicate proposition. They come in all sizes, large or 
multichambered.  After doing cell cultures, you can do all l the staining 
right on the slide from fixation to final product.  When finished, you 
mount coverslip and view.

Are you using an inverted microscope by any chance to see fluorescent 
staining?    If so there are petri dishes designed for confocal use with 
inverted where the bottom of dish IS a coverslip.   They come sterile and 
in different coverslip thicknesses and sizes, and are made of glass.  Very 
tidy little items as you can coat them or whatever, grow cells, fix, stain 
and do inverted fluorescent scope or confocal laster scanning with inverted 
setup.

At 12:41 PM 5/6/2005, you wrote:
>Hi,
>
>I have mainly been doing immunohistocchemistry on tissues for years but
>our lab is branching out to manipulate cell cultures.  We want to start
>immunohistochemistry on the PC12 cell line.  I have done tissue culture
>in the past however I need some advice on these cells.  We could like to
>seed them onto glass coverslips if possible and then get them to
>differentiate into neurons with NGF.  I have read some literature on
>this but need some advice along with clarification.  If anyone can give
>me their protocols for seeding and differentiation on coverslips (either
>matrix coated or using poly-L-lysine? Not sure which to use?) it would
>be much appreciated.  These neurons are going to be stained with
>immunofluorescence with a regular fixing, permeabilization, and staining
>protocol, which I have already optimized.
>
>Thanks in advance for any help.
>
>Wendy Pejic
>Research Technician
>OHRI, Loeb Building
>Ottawa, Ontario
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)