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Abstract
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.
Highlights
Various methods of interference microscopy has recently become rather popular in studies of cell morphology (e.g. (Barer, 1957; Byrne et al, 2008; Dunn, 1998; Marquet et al, 2015; Tychinsky & Tikhonov, 2010; Vishnyakov & Levin, 1998))
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
It was shown that there is no significant difference between the estimates of calculation of dead cells fraction obtained from the Laser Interference Microscopy (LIM) data and conventional technique
Summary
Various methods of interference microscopy has recently become rather popular in studies of cell morphology (e.g. (Barer, 1957; Byrne et al, 2008; Dunn, 1998; Marquet et al, 2015; Tychinsky & Tikhonov, 2010; Vishnyakov & Levin, 1998)). There are a lot of various type of cell were estimated using different methods of interference microscopy: erythrocytes It was shown that activation of macrophages is accompanied by an increase of cell area, a minor (less than 10%) decrease in macrophage dry mass, and pronounced decrease in cell mass in case of cell death (Zagubizhenko et al, 2012). This helped us to distinguish the populations of non-activated, dead and activated macrophages
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