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Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
Published on: June 30, 2018
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Long-term super-resolution inner mitochondrial membrane imaging with a lipid probe
Shuai Zheng1, Neville Dadina1, Deepto Mozumdar2
1Department of Chemistry, University of California, Berkeley, Berkeley, CA, USA.
Nature Chemical Biology
|October 19, 2023
Summary
Researchers developed new probes to visualize the inner mitochondrial membrane (IMM) in live cells. These high-density environmentally sensitive (HIDE) probes offer improved imaging resolution and photobleaching resistance without genetic modification.
Area of Science:
- Cell Biology
- Mitochondrial Biology
- Biochemistry
Background:
- The inner mitochondrial membrane (IMM) is crucial for cellular energy production and homeostasis.
- Understanding IMM dynamics is key to comprehending mitochondrial health and cellular function.
- Current imaging techniques for the IMM face limitations in resolution, photostability, and cellular compatibility.
Purpose of the Study:
- To develop a novel, cell-permeant probe for high-resolution imaging of the IMM in live cells.
- To create high-density environmentally sensitive (HIDE) probes using a new small molecule, MAO-N3.
- To assess the probe's utility across various microscopy techniques and cell types, including primary cells.
Main Methods:
- Synthesis and characterization of the cell-permeant small molecule MAO-N3.
- Assembly of high-density environmentally sensitive (HIDE) probes by conjugating MAO-N3 with fluorophores.
- Application of HIDE probes in live-cell imaging using confocal, structured illumination, single-molecule localization, and stimulated emission depletion microscopy.
- Evaluation of probe toxicity in model cell lines and primary cardiomyocytes.
- Long-term imaging of mitochondrial dynamics (up to 12.5 hours).
Main Results:
- MAO-N3 enables the assembly of HIDE probes that selectively label the IMM in live cells.
- HIDE probes demonstrate significantly improved resistance to photobleaching compared to conventional probes.
- Imaging achieved excellent spatial and temporal resolution across multiple advanced microscopy techniques.
- Probes were non-toxic to model cell lines and primary cardiomyocytes, even under stress conditions.
- Mitochondrial dynamics were visualized for extended periods (12.5 hours) without genetic manipulation.
Conclusions:
- The MAO-N3-based HIDE probes provide a powerful, non-invasive tool for studying IMM dynamics.
- These probes overcome limitations of existing methods, enabling high-resolution, long-term imaging of mitochondrial function.
- This technology facilitates comprehensive investigations into mitochondrial health and cellular homeostasis.
