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STORM imaging of mitochondrial dynamics using a vicinal-dithiol-proteins-targeted probe
Bingling Chen1, Wanjun Gong1, Zhigang Yang1
1Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen, 518060, China.
Biomaterials
|March 16, 2020
Summary
We enhanced Stochastic Optical Reconstruction Microscopy (STORM) for tracking mitochondrial dynamics. This new method achieves high spatial and temporal resolution, enabling visualization of ultra-fine structures and rapid cellular events.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Biophysics
Background:
- Stochastic Optical Reconstruction Microscopy (STORM) offers high resolution for visualizing cellular structures.
- Current STORM limitations include low temporal resolution, hindering the study of dynamic intracellular events.
- Mitochondrial dynamics are crucial for cellular function but challenging to observe in real-time.
Purpose of the Study:
- To develop an improved STORM strategy for high-resolution, high-temporal-resolution imaging of mitochondrial dynamics.
- To overcome the limitations of conventional STORM in capturing rapid cellular processes.
- To enable quantitative analysis of subcellular dynamics beyond the optical diffraction limit.
Main Methods:
- Implementation of a compressed sensing STORM algorithm.
- Pre-treatment of raw STORM data using noise-corrected principal component analysis.
- Application of K-factor image factorization for enhanced image quality.
- Utilized a vicinal-dithiol-proteins targeting probe for mitochondrial labeling.
Main Results:
- Achieved spatial resolution of 45 nm and temporal resolution of 0.8 s for mitochondrial dynamics.
- Successfully monitored dynamic mitochondrial tubulation retraction of approximately 746 nm within 1.2 s.
- Observed novel clusters (radii ~90 nm) of labeled conjugates on outer mitochondrial membranes.
Conclusions:
- The developed STORM strategy significantly enhances temporal resolution for observing mitochondrial dynamics.
- This approach allows for unprecedented visualization and quantitative analysis of subcellular behaviors.
- The findings open new avenues for studying rapid cellular processes with super-resolution microscopy.
