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Label-free adaptive optics single-molecule localization microscopy for whole zebrafish
Sanghyeon Park1,2, Yonghyeon Jo1,2, Minsu Kang3
1Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science, Seoul, Republic of Korea.
Nature Communications
|July 13, 2023
Summary
This study introduces label-free adaptive optics for deep-tissue super-resolution imaging using single-molecule localization microscopy (SMLM). It overcomes specimen-induced aberrations, enabling clearer imaging deeper within biological tissues.
Area of Science:
- Biomedical Optics
- Microscopy
- Neuroscience
Background:
- Specimen-induced aberrations limit imaging depth in single-molecule localization microscopy (SMLM).
- Overcoming these aberrations is crucial for deep-tissue super-resolution imaging.
Purpose of the Study:
- To apply label-free wavefront sensing adaptive optics to SMLM for enhanced deep-tissue super-resolution imaging.
- To overcome the limitations of specimen-induced aberrations in SMLM.
Main Methods:
- Developed a label-free adaptive optics system for SMLM.
- Measured complex tissue aberrations using intrinsic reflectance, not fluorescence.
- Physically corrected wavefront distortions exceeding previous limits.
Main Results:
- Achieved super-resolution imaging up to 102 μm deep in whole zebrafish and mouse brain tissues.
- Resolved sub-diffraction morphologies of cilia, oligodendrocytes, and dendritic spines.
- Enhanced localization number by up to 37 times with precision comparable to aberration-free samples.
Conclusions:
- Label-free adaptive optics significantly expands SMLM's application range for deep-tissue imaging.
- The system effectively corrects severe tissue aberrations, preventing loss of localization number.
- Enables high-resolution imaging in previously inaccessible biological samples like whole zebrafish.
