Low-cost and high-performance 3-D light-sheet fluorescence imaging on pre-owned conventional microscopes
Tingting Zhu, Xinlin Xie, Yao Yao, Dan Zhu2,* and Peng Fei1, 2, *
1. School of Optics and Electronic Information, Huazhong University of Science and Technology, Wuhan 430074, China
2. Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China
Abstract: light-sheet fluorescence microscopy (LSFM) is a promising imaging technique that enable imaging of samples in three dimensions at high speed and low phototoxicity1.LSFM is fundamentally characterized by its separate laser-sheet illumination path that provides optical sectioning of the samples and otherwise shows the same wide-field fluorescence detection with the conventional microscopes. Most of LSFM modalities are currently independent from either conventional microscopes, containing highly complicated setting with large form factors and high maintenance. Given the number of epifluorescence microscopes in service, a much easier way to access advanced LSFM imaging can be realized by creating a laser-sheet illumination and make use of existing conventional microscope for fluorescence readout. Here we present light-sheet imaging plugin (LIP) method that readily enables multi-dimensional, high spatiotemporal resolution imaging from single cells to whole organisms on an inverted microscope2.Furthermore, LIP can be also combined with microfluidics techniques to achieve high-throughput, high-resolution 3-D imaging/screening at a speeds up to 30 samples per seconds3.The ability to achieve high-speed, multi-dimensional imaging on a conventional microscope renders LIP device a valuable tool for many biomedical applications such as embryo development, tissue pathology and neuroscience. Furthermore, its compact add-on format allows the full use of pre-owned equipment and renovation at an affordable expense, which could substantially benefit smaller and less-funded departments / laboratories for not limiting the access to greater biological research in the absence of commercial light-sheet systems.
Keywords: Light-sheet microscopy, light-sheet imaging plugin, microfluidics, 3-D biomedical imaging
1. Power R M, Huisken J. A guide to light-sheet fluorescence microscopy for multiscale imaging.[J]. Nature Methods, 2017, 14(4):360.
2. Guan Z, Lee J, Jiang H, et al. Compact plane illumination plugin device to enable light sheet fluorescence imaging of multi-cellular organisms on an inverted wide-field microscope[J]. Biomedical Optics Express, 2015, 7(1):194.
3. Jiang H, Zhu T, Zhang H, et al. Droplet-based light-sheet fluorescence microscopy for high-throughput sample preparation, 3-D imaging and quantitative analysis on a chip[J]. Lab on A Chip, 2017.
Xinlin Xie, Yicong Yang, Yao Yao, Zhaoqiang Wang, Dan Zhu, and Peng Fei, "Low-cost and high-performance 3D light-sheet fluorescence imaging on pre-owned conventional microscopes (Conference Presentation)," Proc. SPIE 10493, Dynamics and Fluctuations in Biomedical Photonics XV, 104930S (Presented at SPIE BiOS: January 29, 2018; Published: 15 March 2018); https://doi.org/10.1117/12.2297157.5752212951001.
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