cell cultures


I recommend to all users of the microscopy core doing cell cultures for
immunofluorescence microscopy to grow their cells on 13 mm round glass
coverslips (1.5 thickness if you can find them) coated with whatever holds their
cell type to the surface (polylysine, collagen, etc). the coverslips are placed
in 24-well plates with growing media and then seeded with the cells. After they
grow to about 50-60% confluency, they are fixed in 4% em grade formaldehyde in
buffer (P04, PEM, etc) at room temp, washed and stained in the wells. The
coverslips are then picked up, carefully, (takes a little practiced) and placed
cell side down on a drop of Vectashield (can't be used with Cy2), Prolong,
Slowfade, or homemade, etc mountant on a slide.  Seal with clear nail varnish.
After a little practice you can place four coverslips on one slide.  You can
store them @ -20.C if needed.  Using the 24-well plates makes it very easy to
handle and economical to grow your cells.  As far as microscopy this method, I
believe, is superior optically, as opposed to using chamber slides or other
methods that grow cells attached to slides when imaging with oil immersion lens.

Hank Adams
Department of Molecular Genetics Microscopy Core
University of Texas MD Anderson Cancer Center
Houston, TX

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