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Macrophotography with Lighsheet Illumination Enables Whole Expanded Brain Imaging

DISCOVERIES (ISSN 2359-7232), 2021, July-September issue

CITATION: 

Lee CM, Tian X, Tsao C, Chen P, Huang TN, Hsueh YP, Chen BC. Macro photography with Lighsheet Illumination Enables Whole Expanded Brain Imaging with Single-cell Resolution. Discoveries 2021, 9(3): e133. DOI: 10.15190/d.2021.12


Submitted: May 21, 2021; Revised: June 26, 2021; Accepted: July 13, 2021; Published: August 04, 2021;

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Macro photography with Lighsheet Illumination Enables Whole Expanded Brain Imaging with Single-cell Resolution

Chia-Ming Lee1,#, Xuejiao Tian (1,2,#), Chieh Tsao (1,#), Peilin Chen (1), Tzyy-Nan Huang (3), Yi-Ping Hsueh (3), Bi-Chang Chen (1,2,4,*)

(1) Research Center for Applied Sciences, Academia Sinica, Taipei 11529, Taiwan

(2) Brain Research Center, National Tsing Hua University, Hsinchu 30013, Taiwan

(3) Institute of Molecular Biology, Academia Sinica, Taipei 11529, Taiwan

(4) Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 11529, Taiwan

# These authors contributed equally to this project.


*Corresponding authors: Bi-Chang Chen, Research Center for Applied Sciences, Academia Sinica, 128 Sec. 2, Academia Rd., Nankang, Taipei 11529, Taiwan; Phone: 886-2-27873133; Fax: 886-2-27873122; Email: chenb10@gate.sinica.edu.tw

Abstract

Macro photography allows direct visualization of the enlarged whole mouse brain by a combination of lightsheet illumination and expansion microscopy with single-cell resolution.  Taking advantage of the long working distance of a camera lens, we imaged a 3.7 cm thick, transparent, fluorescently-labeled expanded brain. In order to improve 3D sectioning capability, we used lightsheet excitation confined as the depth of field of the camera lens. Using 4x sample expansion and 5x optical magnification, macro photography enables imaging of expanded whole mouse brain with an effective resolution of 300 nm, which provides the subcellular structural information at the organ level

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References

1. Ueda HR, Dodt H-U, Osten P, Economo MN, et al.  Whole-brain profiling of cells and circuits in mammals by tissue clearing and light-sheet microscopy. Neuron. 2020 May 6; 106(3):369-387. doi: 10.1016/j.neuron.2020.03.004

2. Zhao Y, Bucur O, Irshad H, et al. Nanoscale imaging of clinical specimens using pathology-optimized expansion microscopy. Nat Biotechnol. 2017 Aug;35(8):757-764. doi: 10.1038/nbt.3892

3. Bucur O, Fu F, Calderon M, et al. Nanoscale imaging of clinical specimens using conventional and rapid-expansion pathology. Nat Protoc. 2020 May;15(5):1649-1672. doi:10.1038/s41596-020-0300-1

4. Gallagher B, Zhao Y. Nanoscale Imaging of Synaptic Connections with Expansion Microscopy. Discoveries. 2019 Sep 30;7(3):e101. doi: 10.15190/d.2019.14

5. Tillberg PW, Chen F, Piatkevich KD, et al. Protein-retention expansion microscopy of cells and tissues labeled using standard fluorescent proteins and antibodies. Nat Biotechnol. 2016 Sep;34(9):987-92. doi: 10.1038/nbt.3625

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7. Chung K, Wallace J, Kim SY, et al. Structural and molecular interrogation of intact biological systems. Nature. 2013 May 16; 497(7449):332-7. doi:10.1038/nature12107

8. Chen F, Tillberg PW, Boyden ES. Expansion Microscopy. Science. 2015 Jan 30; 347(6221):543-8. doi:10.1126/science.1260088

9. Ratzlaff EH, Grinvald A. A tandem-lens epifluorescence macroscope: Hundred-fold brightness advantage for wide-field imaging. J Neurosci Methods. 1991; 36(2–3):127–137. doi.10.1016/0165-0270(91)90038-2

10. Lu CH, Tang WC, Liu YT, et al. Lightsheet localization microscopy enables fast, large-scale, and three-dimensional super-resolution imaging. Commun Biol. 2019 May 9; 2, 177. doi:10.1038/s42003-019-0403-9

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