Cryo-fluorescence micro-optical sectioning tomography for volumetric imaging of various whole organs with subcellular resolution

Lei Deng; Jianwei Chen; Yafeng Li; Yutong Han; Guoqing Fan; Jie Yang; Dongjian Cao; Bolin Lu; Kefu Ning; Shuo Nie; Zoutao Zhang; Dan Shen; Yunfei Zhang; Wenbin Fu; Wei Eric Wang; Ying Wan; Sha Li; Yu-Qi Feng; Qingming Luo; Jing Yuan

doi:10.1016/j.isci.2022.104805

Cryo-fluorescence micro-optical sectioning tomography for volumetric imaging of various whole organs with subcellular resolution

iScience. 2022 Jul 20;25(8):104805. doi: 10.1016/j.isci.2022.104805. eCollection 2022 Aug 19.

Authors

Lei Deng^{1

2}, Jianwei Chen^{1

2}, Yafeng Li^{1

2}, Yutong Han^{1

2}, Guoqing Fan^{1

2}, Jie Yang^{1

2}, Dongjian Cao^{1

2}, Bolin Lu^{1

2}, Kefu Ning^{1

2}, Shuo Nie^{1

2}, Zoutao Zhang^{1

2}, Dan Shen^{1

2}, Yunfei Zhang^{1

2}, Wenbin Fu³, Wei Eric Wang³, Ying Wan^{4

5}, Sha Li^{6

7}, Yu-Qi Feng^{6

7}, Qingming Luo^{1

2

8}, Jing Yuan^{1

2

9}

Affiliations

¹ Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics, MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Innovation Institute, Huazhong University of Science and Technology, Wuhan 430074, China.
² MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Innovation Institute, Huazhong University of Science and Technology, Wuhan 430074, China.
³ Department of Cardiology, Daping Hospital, Army Medical University, Chongqing 400038, China.
⁴ Biomedical Analysis Center, Army Medical University, Chongqing 400038, China.
⁵ Chongqing Key Laboratory of Cytomics, Chongqing 400038, China.
⁶ Department of Chemistry, Wuhan University, Wuhan 430072, China.
⁷ School of Public Health, Wuhan University, Wuhan 430071, China.
⁸ School of Biomedical Engineering, Hainan University, Haikou, 570228, China.
⁹ HUST-Suzhou Institute for Brainsmatics, JITRI Institute for Brainsmatics, Suzhou, 215123, China.

Abstract

Optical visualization of complex microstructures in the entire organ is essential for biomedical research. However, the existing methods fail to accurately acquire the detailed microstructures of whole organs with good morphological and biochemical preservation. This study proposes a cryo-fluorescence micro-optical sectioning tomography (cryo-fMOST) to image whole organs in three dimensions (3D) with submicron resolution. The system comprises a line-illumination microscope module, cryo-microtome, three-stage refrigeration module, and heat insulation device. To demonstrate the imaging capacity and wide applicability of the system, we imaged and reconstructed various organs of mice in 3D, including the healthy tongue, kidney, and brain, as well as the infarcted heart. More importantly, imaged brain slices were performed sugar phosphates determination and fluorescence in situ hybridization imaging to verify the compatibility of multi-omics measurements. The results demonstrated that cryo-fMOST is capable of acquiring high-resolution morphological details of various whole organs and may be potentially useful for spatial multi-omics.

Keywords: Biological sciences; Natural sciences; Neuroscience; Techniques in neuroscience.