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Publications

Journal Article Ppublish 7 citations

PubMed: 39449993 DOI: 10.1016/j.mitoco.2024.03.001

Photobleaching and phototoxicity of mitochondria in live cell fluorescent super-resolution microscopy.

Mitochondrial Commun 2:38-47

Authors: Chia-Hung Lee, Douglas C Wallace, Peter J Burke

Affiliations

  • Department of Biomedical Engineering, University of California, Irvine, CA, 92697, United States.
  • Center for Mitochondrial and Epigenomic Medicine, Children's Hospital of Philadelphia and Department of Pediatrics, Division of Human Genetics, University of Pennsylvania, Philadelphia, PA, 19104, United States.
  • Department of Electrical Engineering and Computer Science, United States.

Abstract

Photobleaching and phototoxicity can induce detrimental effects on cell viability and compromise the integrity of collected data, particularly in studies utilizing super-resolution microscopes. Given the involvement of multiple factors, it is currently challenging to propose a single set of standards for assessing the potential of phototoxicity. The objective of this paper is to present empirical data on the effects of photobleaching and phototoxicity on mitochondria during super-resolution imaging of mitochondrial structure and function using Airyscan and the fluorescent structure dyes Mitotracker green (MTG), 10-N-nonyl acridine orange (NAO), and voltage dye Tetramethylrhodamine, Ethyl Ester (TMRE). We discern two related phenomena. First, phototoxicity causes a transformation of mitochondria from tubular to spherical shape, accompanied by a reduction in the number of cristae. Second, phototoxicity impacts the mitochondrial membrane potential. Through these parameters, we discovered that upon illumination, NAO is much more phototoxic to mitochondria compared to MTG or TMRE and that these parameters can be used to evaluate the relative phototoxicity of various mitochondrial dye-illumination combinations during mitochondrial imaging.

MeSH

Cells, Mitochondria, Microscopy, Lighting

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