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

[Prichett, Wendy]

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