Abstract

This work considers the measurement of FAD fluorescence intensity as a method for the safety, simple and real-time detection of pathological cells and informative value of this approach. FAD participates in essential processes such as fatty acid oxidation, the Krebs cycle and other redox reactions. According to literature, cells in different physiological states have different levels of FAD intensity in green-blue spectrum. Hence, it is highly relevant to determine the physiological state of cells by the difference in FAD signal intensity.

The study was realized with skin fibroblasts as a model object. On the first stage of experiments 20-days cells cultured on a pre-marked coverslips were divided into two subgroups on the basis of the autofluorescence signal intensity. The first subgroup included cells with a high autofluorescence signal (presumably senescent or pathological), and the second - cells and low one. During subsequent experiments after 24 hours necrotic cells were analyzed in a culture using Hoechst 33342 and propidium iodide in two subgroups separately.

According to the results, over 50% of cells with high autofluorescence intensity were identified as necrotic, that can subsequently be used for early diagnosis of various pathologies states. Thus, this study, with its advantages such as non- invasiveness, high sensitivity and biosafety, shows the possibility of early diagnosis of various diseases by measuring the fluorescence signal of FAD and finding cells with high fluorescence levels, which are mostly necrotic.

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