High-definition imaging using line-illumination modulation microscopy
- 1 March 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Methods
- Vol. 18 (3), 309-315
- https://doi.org/10.1038/s41592-021-01074-x
Abstract
The microscopic visualization of large-scale three-dimensional (3D) samples by optical microscopy requires overcoming challenges in imaging quality and speed and in big data acquisition and management. We report a line-illumination modulation (LiMo) technique for imaging thick tissues with high throughput and low background. Combining LiMo with thin tissue sectioning, we further develop a high-definition fluorescent micro-optical sectioning tomography (HD-fMOST) method that features an average signal-to-noise ratio of 110, leading to substantial improvement in neuronal morphology reconstruction. We achieve a >30-fold lossless data compression at a voxel resolution of 0.32 × 0.32 × 1.00 μm3, enabling online data storage to a USB drive or in the cloud, and high-precision (95% accuracy) brain-wide 3D cell counting in real time. These results highlight the potential of HD-fMOST to facilitate large-scale acquisition and analysis of whole-brain high-resolution datasets.Keywords
Funding Information
- Science Fund for Creative Research Groups (61721092)
- National Natural Science Foundation of China (81827901, 91749209, 81827901, 61890950, 61890954, 61890950,61890953)
- Fundamental Reserach Funds for the Central Universities
- Fundamental Research Funds for the Central Universities
- National Key Research and Development Program of China
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