SLIM-assisted automatic cartography of cell death types and rates resulting from localized photodynamic treatment.

Abstract

We report a spatial light interference microscopy (SLIM)-based methodology aimed at automatic monitoring and analysis of changes in cellular morphology within extended fields of view in cytological samples. The experimental validation was performed on HeLa cells in vitro subjected to localized photodynamic treatment. The performed long-term noninvasive monitoring using the SLIM technique allowed us to estimate quantitative parameters characterizing the dynamics of average phase shift in individual cells and to reveal changes in their morphology specific for different mechanisms of cell death. The results obtained evidenced that the proposed SLIM-based methodology provides an opportunity for identification of cell death type and quantification of cell death rate in an automatic mode. The major sources of potential errors that can affect the results obtained are discussed. The developed methodology is promising for automatic monitoring of large ensembles of individual cells and for quantitative characterization of their response to various treatment modalities.