Application of Phase Images for Estimation of Peritoneal Macrophages State

Procedure based on phase images obtained by the method of Laser Interference Microscopy (LIM) for detection of number non-activated, activated and dead isolated peritoneal macrophages in a sample in vitro was provided. The phase images provide quantitative estimates of the optical path difference (OPD) in each point of object. The OPD varies with cell thickness and concentration of cellular substances. Therefore, it provides useful information on cell structure, localization of subcellular structures, local concentrations of substances and dry mass of cell. In these work we propose a new assay for estimation a number of non-activated, activated and dead isolated peritoneal macrophages in a sample in vitro based on cell phase images. The results were compared with phagocytic index, which was evaluated using latex beads, and the percentage of cells with damaged membranes (dead cells). It was shown that with increasing time after the isolation, the percentage of dead macrophages estimated by LIM was similar to that of the cells with damaged membranes. However, the percentage of activated cells calculated using latex beads exceeded the percentage calculated using the phase images significantly.