A quantitative method for measuring phototoxicity of a live cell imaging microscope

Methods in Enzymology
Jean-Yves TinevezSpencer L Shorte

Abstract

Fluorescence-based imaging regimes require exposure of living samples under study to high intensities of focused incident illumination. An often underestimated, overlooked, or simply ignored fact in the design of any experimental imaging protocol is that exposure of the specimen to these excitation light sources must itself always be considered a potential source of phototoxicity. This can be problematic, not just in terms of cell viability, but much more worrisome in its more subtle manifestation where phototoxicity causes anomalous behaviors that risk to be interpreted as significant, whereas they are mere artifacts. This is especially true in the case of microbial pathogenesis, where host-pathogen interactions can prove especially fragile to light exposure in a manner that can obscure the very processes we are trying to observe. For these reasons, it is important to be able to bring the parameter of phototoxicity into the equation that brings us to choose one fluorescent imaging modality, or setup, over another. Further, we need to be able to assess the risk that phototoxicity may occur during any specific imaging experiment. To achieve this, we describe here a methodological approach that allows meaningful measurement, and ...Continue Reading

Citations

Feb 26, 2016·Journal of the American Chemical Society·Marcus J C LongYimon Aye
Dec 9, 2014·Cell Adhesion & Migration·Julien CaronSpencer L Shorte
Dec 3, 2016·Wiley Interdisciplinary Reviews. Developmental Biology·Ellen T ArenaKevin W Eliceiri
Oct 8, 2016·Methods : a Companion to Methods in Enzymology·Jean-Yves TinevezKevin W Eliceiri
Sep 9, 2016·Journal of Cellular Physiology·Stephen Douthwright, Greenfield Sluder
Jan 24, 2017·The International Journal of Biochemistry & Cell Biology·Richard KasprowiczPeter O'Toole
Apr 1, 2017·Molecular Biology of the Cell·Kurt Thorn
Jul 1, 2017·Nature Methods·P Philippe LaissueHari Shroff
Jul 28, 2017·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Jaroslav IchaCaren Norden
Oct 19, 2017·Nature Methods·Ingo GregorJörg Enderlein
Jun 5, 2020·Current Protocols in Mouse Biology·Jeremy Sanderson
Apr 17, 2014·Biotechnology Journal·Régis TournebizeSpencer Shorte
Oct 14, 2017·FEMS Microbiology Reviews·Katrin RosenthalAndreas Schmid
Apr 14, 2017·Scientific Reports·John H StockleyRagnhildur T Káradóttir
Jun 23, 2016·PloS One·Bhavna RajasekaranAndrew C Oates
Aug 4, 2015·Physics in Medicine and Biology·A GeorgantzoglouR Jena
May 18, 2021·Journal of Physics D: Applied Physics·Kalina L ToshevaRicardo Henriques
Jun 8, 2021·Extreme Mechanics Letters·M A Bashar EmonM Taher A Saif

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