Coating on the slides depends a lot on the cell type you're working with.
The best bet is to just seed some cells on an a sterile slide in a dish and
see if they attach. Poly-L-lysine seems to work almost universally as a
coating cells will attach to, but it's often unnecessary.
As for antigen retrieval, this has a large element of trial-and-error as
well, and depends on the antigen. The first thing to try is to vary
fixation time and formalin concentration to achieve optimal results. If you
still don't get consistent results, detergent, boiling, acid, and/or
protease treatments may be necessary. I would recommend trying them in that
order, starting with some relatively mild, nonionic detergents.
We really have found that this process is as much an art as science. I hope
this helps. If you have some specific problems, post those and we can
KAREN TILMANT wrote:
> Hi, can anyone tell me how to prepare a cell smear so that it's possible
> to localisate some proteins with immunocytochemistry. Do I have to use
> coated glass slides? When I'm using formalin to fixate, do I've to do
> some antigen-retrieval?
> Thanks for your reply!
> Karen Tilmant
Eli V. Hestermann
ehestermann at whoi.eduhttp://www.mit.edu/people/octavian/eli.html
"Vita brevis est, ars longa" - Seneca