FW: Cell Culture for IHC


FW: Cell Culture for IHC

Dear Laura,
at the our dept, cell cultures are used many times mostly as a positive control for different antigens (not viral), you have at least 2 options - you can grow your cells as usually, prepare cell suspension from them and make cytospins or you grow the cells directly on the slide, it is advisable to use commercially prepare special slides for this which can be plastic or glass (I prefer glass slide).

How to prepare cell suspension from cell culture is depend on how your cells grow, if you have cells which adhere on dish you can use police rubber to detach them or enzymes, as a cell medium for cell suspension being cytocentrifugated we use home made cell medium, but you can also try using PBS.

hope this help

Irena Kirbis
Institute of Oncology
dept of Cytopathology

-----Original Message-----
From: bliven.laura@marshfieldclinic.org
Sent: Thursday, February 13, 2003 5:12 PM
To: histonet@pathology.swmed.edu
Subject: Cell Culture for IHC

I have been asked to do an immunohistochemistry stain (West Nile Virus) on cell culture.  I usually on worked with paraffin blocks and smears.  I'm not even sure of the questions to ask, but here it goes....

In growing the cells, can I used glass slides (I don't have any plastic)?
Are the glass slides plain or charged?
How do I fix the cells?  (I know if they're not fixed in formalin there will be no antigen retrieval.)
Anything special I need to know before the jump such as potential problems or background staining?

Thanks in advance,
Laura Bliven
Marshfield Laboratories

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