Re: [Histonet] staining cell cultures with Giemsa

From:koellingr@comcast.net



Helen,
What "kind" of cell cultures?  Primary bone marrow cultures? Immortal cell culture? Other primary lines?  Why Giemsa? And grown in what?  T-25's, T-150's?  Or on chambered glass slides? Chambered glass slides?  Have done Giemsa and many other histologic stains on "cell cultures" but the protocols (H&E, Giemsa, Oil Red O, etc) didn't vary much from standard histology lab other than I just used some common sense in adapting the particular situation to the stain.  A primary bone marrow culture in a T150 flask, I would wash and "fix" the cells (methanol) for Giemsa. Other fixative for other stain.  If I didn't mind viewing through the 2 flat sides, just do the stain in flask, emptying and washing.  Or if I wanted to get a "close" view with higher power, after fixation saw through the "top" "flat" half, do stain in what was an open container.  Or if I wanted really high power, I'd put sterile coverslips in the flask before seeding cells, grow to a monolayer, remove coverslips, fix and s
tain them.  Depends on the pickiness of the cell line. Or grow culture in chambered glass slides, remove chamber material, fix, stain.  I guess my point is you might not need to look for a specific Giemsa modification (or histologic stain) for "cell cultures".  You need to follow basic staining protocols and use common sense to adapt the stain to your very particular siutuation with type of cell and culture environment taken into consideration and not do something unwise (take culture flask through cyclic hydrocarbons like xylene to clear).  This is all time consuming and sounds like a lot of work but common sense for your particular project unique to you will take you further than someone elses directions.

Ray Koelling
PhenoPath Labs
Seattle, WA

-------------- Original message -------------- 
From: Helen E Johnson  

> 
> Hi Histonetters, 
> Does anyone have a protocol staining cell cultures with Giemsa? 
> Thanks. Helen Johnson 
> (hej01@health.state.ny.us) 
> 
> 
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