CC BY
8DOI 10.24412/cl-37136-2023-1-148-149
HIGH THROUGHPUT 3D IMAGING AND ITS APPLICATION IN NEUROSCIENCE WEI GONG1, XIAO XIAO2, JIAJIA CHEN1 AND KE SI1,2
1MOE Frontier Science Center for Brain Science & Brain-Machine Integration, NHC and CAMS Key Laboratory of Medical Neurobiology, School of Brain Science and Brain Medicine, Zhejiang University,
Hangzhou, China
2College of Optical Science and Engineering, State Key Laboratory of Modern Optical Instrumentation,
Zhejiang University, Hangzhou, China
weigong@zj u.edu.cn
ABSTRACT
High-speed and high-resolution imaging for large volume biological samples is always an aspirational goal of microscopists. The unprecedented developments in optical imaging tools and fluorescence probes have provided numerous visual access to biological structure and function at cellular resolution, such as confocal laser scanning microscopy (CLSM), two photon-excited fluorescence laser-scanning microscopy (2PLSM), light field microscopy (LFM) and so on. Beyond these advanced techniques, however, perfect optical imaging complex larger volume samples also demands fast acquisition, wide field of view (FOV), protection against photobleaching and photoxicity and so on. One of promising approaches is light-sheet fluorescence microscopy (LSFM) combined with the optical clearing methods, which has been proved a powerful choice for large volumetric imaging in recent years.
Here, we provide two fast optical clearing methods for mouse and human brains, respectively. FOCM is a nontoxic ultrafast optical clearing method, which could clarify 300-^m-thick mouse brain slices within 2 min without morphology distortion [1]. The cheap reagents and easy operation make FOCM have great potential in the exploration of neuroscience. On this basis, we developed a multiscale aqueous method of rapid clearing and staining for human brain, which achieving high quality transparency of 1mm-thick human brain tissue within 1 day while maintaining deformation (Fig. 1a). Besides, this method can greatly increase the depth of staining and achieve exploration of the three-dimensional structure of the human brain (Fig. 1b). Overall, our method will provide great assistance to chart the cellular and molecular architecture of human brain and exploring the ultimate territory of humanity. In addition, we have developed imaging systems and image processing algorithms that are suitable for these two transparent methods [2-3], and established a high-throughput 3D imaging platform.
3. Before After
bi
2mm
Figure: (a) Transparency of lmm-thick human brain cleared by our method. (b) 3D imaging of 1mm-thick human brain with blood vessels labelled.
REFERENCES
[1] Zhu X, Huang L, Zheng Y, et al. Ultrafast optical clearing method for three-dimensional imaging with cellular resolution. Proc Natl Acad Sci U S A. 2019.
[2] Chen J, Du Z, Xu C, Xiao X, Gong W, Si K. Ultrafast 3D histological imaging based on a minutes-time scale tissue clearing and multidirectional selective plane illumination microscopy. Opt Lett. 2022.
[3] Hu L, Hu S, Gong W, Si K. Image enhancement for fluorescence microscopy based on deep learning with prior knowledge of aberration. Opt Lett. 2021.
