Rapid and sensitive photometric quantification of the proliferation of adherent vascular endothelial and smooth muscle cells in 96-multiwell plates after may-grünwald giemsa staining.

A simple, sensitive, and rapid photometric method has been developed to measure the proliferation of adherent cells in 96-multiwell plates quantitatively. The cultured cells were fixed with methanol and stained with May-Grünwald Giemsa. The multiwell plate was scanned at two wavelengths (lambda max 540 nm and lambda reference 405 nm or 690 nm). The correlation coefficient (r) between cell densities measured by direct microscopic cell counting or photometric light absorbance was greater than 0.9, in the range 400 to 66,000 cells/cm2. The method is applicable to adherent cells such as vascular endothelial cells, vascular smooth muscle cells, fibroblasts, or the breast adenocarcinoma cell line MCF-7. In addition, the morphology of the fixed and stained cells was examined and 3H-thymidine incorporation in cells was studied by radioactive counting of solubilized cells.