A simple indirect immunofluorescence micromethod for cell typing

We describe a simple indirect immunofluorescence micromethod for cell typing. Cells are layered on 18-well immunofluorescence slides coated with high polymer poly-L-lysine. Expressed cell surface membrane markers are detected by indirect immunofluorescence using monoclonal antibodies and the cells are stained with Evans blue dye to permit easy morphological identification under fluorescence and light microscopy. We compared this method with a conventional method for lymphocyte typing in 16 blood samples and 10 bronchoalveolar lavage samples. The results of the two methods did not differ and correlated closely (r = 0.988 for blood samples; r = 0.995 for bronchoalveolar lavage samples). The principal advantages of this micromethod are: the small amounts of cells (10(4) cells) and reagents needed, the ease with which numerous antibodies can be tested and the convenience offered by fixation and staining of cell preparations for reading.