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Several methods are available for measuring viable attached cells.
In general, each method begins with the coating of plastic tissue culture surfaces, such as 96-well plates, with different amounts of a given attachment factor. A suspension containing a known number of cells is then added to the wells and incubated at 37 C for about one hour to allow cell attachment. The unattached cells are washed away and the number of attached cells determined. Probably the most commonly used attachment assay method is to fix the attached cells to the plastic surface with paraformaldehyde and then stain the cells with Toluidine blue. After several washing steps, the number of cells can be counted using a microscope.