QPI, FLIM AND AI IN ANALYSIS OF CELLS RESPONSE TO PHOTODYNAMIC TREATMENT

Abstract

A multimodal approach is developed which combines fluorescence lifetime imaging microscopy (FLIM) and quantitative phase imaging (QPI) in the realization of spatial light interference microscopy (SLIM) in a single complex. This combination allowed for a concurrent analysis of cell samples in vitro, where the SLIM channel provided data on cellular morphology and its variations, while the FLIM channel enabled monitoring fluorescence parameters of intracellular fluorophores, both endogenous and exogenous. The developed methodology was used for analysis of the response of cells of the two established lines, HeLa and A549, to photodynamic treatment with Radachlorin photosensitizer. The dynamics of changes in the phase shift introduced by cells gave information on the type and rate of cell death. Variations of fluorescence lifetimes of endogenous coenzymes NADH and FAD provided data on changes in cellular energy metabolism, while those of Radachlorin gave insight on its transportation and photobleaching. SLIM technique allowed for noninvasive monitoring of the response of both individual cells and their population in a sample. Processing of the data obtained from the analysis of entire populations of cells was performed using the developed machine-learning classification algorithm, which demonstrated successful evaluation of the relative amounts of live, necrotic and apoptotic cells and changes in these amounts as a function of treatment dose. The financial support of this work from Russian Science Foundation under the grant # 26-75-31002 is gratefully acknowledged.

Speaker

Irina Semenova
Ioffe Institute
Russia

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