By applying the confocal fluorescence microscopy, we investigated the dynamics of the synthesis and accumulation of protoporphyrin IX induced by 5-aminolevulinic acid (5-ALA) in three cell lines: malignant - human epithelial carcinoma HeLa and human lung adenocarcinoma A549 - and fibroblast-like immortalized mouse embryonic 3T3 cells. The accumulation of protoporphyrin IX in cell cultures was evaluated based on the analysis of its fluorescence intensity. The dynamics of changes in fluorescence intensity was studied depending upon the duration of cells incubation with 5-ALA and its concentration.

The paper presents development of cell segmentation algorithms on two-dimensional phase images and three-dimensional refractive index distributions obtained, respectively, by means of digital holographic microscopy and tomography. The algorithms were optimized for measurements of cells morphological parameters: volume, projected area and membrane area. The developed segmentation algorithms were validated and volume calculation error was estimated by comparison with the values obtained using confocal fluorescence tomography.