Long-term super-resolution inner mitochondrial membrane imaging with a lipid probe

Shuai Zheng; Neville Dadina; Deepto Mozumdar; Lauren Lesiak; Kayli N Martinez; Evan W Miller; Alanna Schepartz

doi:10.1038/s41589-023-01450-y

Long-term super-resolution inner mitochondrial membrane imaging with a lipid probe

Nat Chem Biol. 2024 Jan;20(1):83-92. doi: 10.1038/s41589-023-01450-y. Epub 2023 Oct 19.

Authors

Shuai Zheng^#¹, Neville Dadina^#¹, Deepto Mozumdar², Lauren Lesiak¹, Kayli N Martinez¹, Evan W Miller^{1

3}, Alanna Schepartz^{4

5

6

7}

Affiliations

¹ Department of Chemistry, University of California, Berkeley, Berkeley, CA, USA.
² Department of Chemistry, Yale University, New Haven, CT, USA.
³ Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
⁴ Department of Chemistry, University of California, Berkeley, Berkeley, CA, USA. schepartz@berkeley.edu.
⁵ Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA. schepartz@berkeley.edu.
⁶ California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, CA, USA. schepartz@berkeley.edu.
⁷ Chan Zuckerberg Biohub, San Francisco, San Francisco, CA, USA. schepartz@berkeley.edu.

^# Contributed equally.

Abstract

The inner mitochondrial membrane (IMM) generates power to drive cell function, and its dynamics control mitochondrial health and cellular homeostasis. Here, we describe the cell-permeant, lipid-like small molecule MAO-N₃ and use it to assemble high-density environmentally sensitive (HIDE) probes that selectively label and image the IMM in live cells and multiple cell states. MAO-N₃ pairs with strain-promoted azide-alkyne click chemistry-reactive fluorophores to support HIDE imaging using confocal, structured illumination, single-molecule localization and stimulated emission depletion microscopy, all with significantly improved resistance to photobleaching. These probes generate images with excellent spatial and temporal resolution, require no genetic manipulations, are non-toxic in model cell lines and primary cardiomyocytes (even under conditions that amplify the effects of mitochondrial toxins) and can visualize mitochondrial dynamics for 12.5 h. This probe will enable comprehensive studies of IMM dynamics with high temporal and spatial resolution.

MeSH terms

Fluorescent Dyes*
HeLa Cells
Humans
Lipids
Microscopy, Fluorescence / methods
Mitochondrial Membranes*
Monoamine Oxidase

Substances

Fluorescent Dyes
Lipids
Monoamine Oxidase

Abstract

MeSH terms

Substances

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