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21 August 2020 Monitoring of PpIX-induced death dynamics in HeLa cells by digital holographic tomography
Daria A. Gorbenko, Andrey V. Belashov, Ilia K. Litvinov, Tatiana N. Belyaeva, Elena S. Kornilova, Irina V. Semenova, Oleg S. Vasyutinskii
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Proceedings Volume 11490, Interferometry XX; 1149013 (2020) https://doi.org/10.1117/12.2567498
Event: SPIE Optical Engineering + Applications, 2020, Online Only
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Abstract
We present the analysis of the response of human cervix epidermoid carcinoma HeLa cells to photodynamic treatment with protoporphyrin IX (PpIX) synthesized in living cells under accumulation of 5-aminolevulinic acid (5-ALA). PpIX synthesis was estimated by means of confocal uorescent microscopy basing on its uorescence intensity in the red wavelength range. The cell death dynamics and pathways caused by intracellular generation of reactive oxygen species under laser irradiation of photosensitized cells were examined using digital holographic tomography. These processes were studied both at early stages of cells death on living specimens and at later stages on paraformaldehyde-fixed specimens. Statistical analysis of changes in cellular morphology monitored using holographic tomography allowed us to conclude on cell death pathways, observed at different irradiation doses. Data obtained by holographic tomography have been validated by a standard AO/EB test for cell membrane integrity conducted using the confocal fluorescence microscope.
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Daria A. Gorbenko, Andrey V. Belashov, Ilia K. Litvinov, Tatiana N. Belyaeva, Elena S. Kornilova, Irina V. Semenova, and Oleg S. Vasyutinskii "Monitoring of PpIX-induced death dynamics in HeLa cells by digital holographic tomography", Proc. SPIE 11490, Interferometry XX, 1149013 (21 August 2020); https://doi.org/10.1117/12.2567498
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KEYWORDS
Digital holography
Holography
Tomography
Cell death
Confocal microscopy
Refractive index
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