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Illuminating Mitochondrial Dynamics: Ultrahigh Labeling Stability Probe for Long-Term SIM Super-Resolution Imaging of
Xiangpeng Lin1, Xuelei Pang1, Yue Huang1
1MOE Key Laboratory for Biomedical Photonics, Advanced Biomedical Imaging Facility-Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, Hubei 430070, China.
ACS Central Science
|September 29, 2025
Summary
Researchers developed new fluorescent probes, HZ Mito Red and HZ Mito Deep Red, for stable, long-term mitochondrial imaging. These probes enable detailed super-resolution microscopy of mitochondrial dynamics in diseases.
Area of Science:
- Cell Biology
- Mitochondrial Biology
- Biomedical Imaging
Background:
- Mitochondrial dynamics are crucial for understanding disease pathogenesis.
- Existing fluorescent probes lack sufficient labeling stability, photostability, and low cytotoxicity for long-term super-resolution imaging.
- Advanced imaging techniques require robust probes to visualize subtle mitochondrial changes.
Purpose of the Study:
- To develop novel covalent mitochondrial probes with enhanced stability, photostability, and low cytotoxicity.
- To enable high signal-to-noise, long-term dynamic super-resolution microscopy of mitochondria.
- To facilitate detailed analysis of mitochondrial morphology during cellular processes like ferroptosis, apoptosis, and autophagy.
Main Methods:
- Rational design and screening of novel covalent mitochondrial probes (HZ Mito Red and HZ Mito Deep Red).
- Comparative analysis of probe stability, photostability, and cytotoxicity against Mito Tracker Red (MTR).
- Application of probes in super-resolution microscopy (SIM) for dynamic imaging of mitochondrial morphology during cell death and autophagy.
- Assessment of probe compatibility with cell immunofluorescence staining.
Main Results:
- HZ Mito Red demonstrated 10-fold improved mitochondrial labeling stability and remarkable photostability (80% fluorescence retention after 300 SIM images).
- Negligible phototoxicity of HZ Mito Red preserved mitochondrial integrity during extended imaging (400 SIM images).
- HZ Mito Deep Red offered similar stability and near-phototoxicity-free dynamic tracking (60% fluorescence retention after 300 SIM images).
- Both probes enabled unprecedented detailed delineation of mitochondrial morphology during ferroptosis, apoptosis, and autophagy.
- Probes are compatible with cell immunofluorescence staining, enhancing their versatility.
Conclusions:
- HZ Mito Red and HZ Mito Deep Red are robust, versatile tools for advanced mitochondrial dynamics research.
- These probes overcome limitations of existing probes, enabling high-resolution, long-term imaging.
- The developed probes significantly advance the study of mitochondrial involvement in disease pathogenesis.
