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Optical Clearing and Imaging of Immunolabeled Kidney Tissue
Published on: July 22, 2019
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Continuous imaging of large-volume tissues with a machinable optical clearing method at subcellular resolution
Can Zhou1,2, Ting Zheng1,2, Ting Luo1
1Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Huazhong University of Science and Technology, Wuhan 430074, China.
Biomedical Optics Express
|January 7, 2021
Summary
We developed M-CUBIC, a novel optical clearing method using poly-N-acryloyl glycinamide hydrogel embedding. This technique enhances tissue transparency and sectioning properties for high-resolution 3D imaging of whole organs.
Area of Science:
- Biological Imaging
- Tissue Clearing
- Biotechnology
Background:
- Optical clearing is crucial for whole-organ 3D imaging.
- Challenges include non-uniform clearing and poor sectioning properties of cleared tissues.
- Existing methods struggle to balance transparency with mechanical integrity.
Purpose of the Study:
- To develop an improved optical clearing method that enhances both tissue transparency and sectioning capabilities.
- To enable high-resolution, isotropic 3D imaging of intact biological organs.
- To facilitate detailed microstructural analysis of various organs.
Main Methods:
- Developed M-CUBIC (machinable CUBIC), combining a modified CUBIC clearing protocol with poly-N-acryloyl glycinamide (PNAGA) hydrogel embedding.
- Utilized high-throughput light-sheet tomography platform (HLTP) and fluorescent micro-optical sectioning tomography (fMOST) for imaging.
- Applied the method to intact mouse brains and kidneys.
Main Results:
- M-CUBIC successfully rendered tissues transparent while significantly improving their sectioning properties.
- Acquired continuous, subcellular-resolution datasets of mouse brains for single neuron tracing.
- Visualized the fine vascular structures of kidneys with high fidelity.
- Demonstrated the method's applicability to multiple organ types.
Conclusions:
- M-CUBIC offers a robust solution for whole-organ 3D imaging by overcoming limitations of traditional optical clearing.
- The enhanced sectioning property allows for detailed microstructural analysis at subcellular resolution.
- This technique holds significant potential for advancing biological research requiring high-resolution organ imaging.
