Phototoxicity induced in living HeLa cells by focused femtosecond laser pulses: a data-driven approach

Nonlinear optical microscopy is a powerful label-free imaging technology, providing biochemical and structural information in living cells and tissues. A possible drawback is photodamage induced by high-power ultrashort laser pulses. Here we present an experimental study on thousands of HeLa cells,...

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Veröffentlicht in:Biomedical optics express 2021-12, Vol.12 (12), p.7886-7905
Hauptverfasser: Talone, B, Bazzarelli, M, Schirato, A, Dello Vicario, F, Viola, D, Jacchetti, E, Bregonzio, M, Raimondi, M T, Cerullo, G, Polli, D
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Sprache:eng
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Zusammenfassung:Nonlinear optical microscopy is a powerful label-free imaging technology, providing biochemical and structural information in living cells and tissues. A possible drawback is photodamage induced by high-power ultrashort laser pulses. Here we present an experimental study on thousands of HeLa cells, to characterize the damage induced by focused femtosecond near-infrared laser pulses as a function of laser power, scanning speed and exposure time, in both wide-field and point-scanning illumination configurations. Our data-driven approach offers an interpretation of the underlying damage mechanisms and provides a predictive model that estimates its probability and extension and a safety limit for the working conditions in nonlinear optical microscopy. In particular, we demonstrate that cells can withstand high temperatures for a short amount of time, while they die if exposed for longer times to mild temperatures. It is thus better to illuminate the samples with high irradiances: thanks to the nonlinear imaging mechanism, much stronger signals will be generated, enabling fast imaging and thus avoiding sample photodamage.
ISSN:2156-7085
2156-7085
DOI:10.1364/boe.441225