The advantages and possibilities of fluorescent methods for visualization of apoptosis and autophagy in human tumor cells in vitro

The possibilities of using fluorescent research methods and their advantages for visualization and determination of the type of programmed cell death of human tumor cells under the action of flavonoids in experiments in vitro were investigated. The object of the study were the tumor cells of cervical cancer HeLa and A498 kidney carcinoma, the flavonoid containing extract of hedge hyssop (Gratiola officinalis L.) was used for testing in the experiments. The following fluorescent methods were used: the “living and dead” test with double staining – iodide propidium and acridine orange, the method of double staining with annexin and iodide propidium. The confirmation of autophagy induction was performed using Muse cell analyzer with fluorescent reagents MuseAutophagy LC3-Antibody Based Kit. The application of fluorescent staining methods using double staining with acridine orange and iodide propidium in the “living and dead” test compared to phase microscopy allows to visualize the formation of apoptotic cells and autophagosomes in cells and, therefore, can serve as one of the methods of evaluation screening of the effectiveness of various chemotherapy drugs.