What are people using as a liquid cover glass for stained cells in 6 well culture plates to be view with inverted microscope with having that annoying refractile, funky look to the cells.
Cells are grown in 6 well culture plates and with previous cultures, been stained with Hema 3 (hematological stain) and Giemsa of which neither has been successful to see cell membranes and nuclei clearly. Viewing is done with inverted microscope and cells are counted. I know there are some stains that can be viewed left under water with inverted scope. H&E obviously cannot be left under water due to eosin washing out of cytoplasm. A possibility is letting H&E or T blue stained cells dry, then put a thin layer of permanent mounting media to flow over the cells to fill in spaces, let dry and view with inverted microscope. I think one could do an old bonehead trick we used on surface stained, thicker ground MMA embedded undecalcified bone, flood with immersion oil and put a cover glass on top. Very messy!!! However one could let H&E dry after last alcohol rinse, put a layer of immersion oil over the cells, and view with inverted microscope. Anything to fill in the cellular spaces on a confluent sheet of cells but let the oil be the cover glass. Growing cells on a cover glass or on slide with wells/gaskets is not an option. Any suggested stains with protocols would be appreciated too with thoughts here about using Toluidine blue. The recipe for making up the stain, staining method and preferred fixative would be appreciated. You can send information privately if you wish. A challenge to be sure Thanks Gayle Callis HTL/HT/MT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet