Translational rapid ultraviolet-excited sectioning tomography for whole-organ multicolor imaging with real-time molecular staining

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Abstract

Rapid multicolor three-dimensional (3D) imaging for centimeter-scale specimens with subcellular resolution remains a challenging but captivating scientific pursuit. Here, we present a fast, cost-effective, and robust multicolor whole-organ 3D imaging method assisted with ultraviolet (UV) surface excitation and vibratomy-assisted sectioning, termed translational rapid ultraviolet-excited sectioning tomography (TRUST). With an inexpensive UV light-emitting diode (UV-LED) and a color camera, TRUST achieves widefield exogenous molecular-specific fluorescence and endogenous content-rich autofluorescence imaging simultaneously while preserving low system complexity and system cost. Formalin-fixed specimens are stained layer by layer along with serial mechanical sectioning to achieve automated 3D imaging with high staining uniformity and time efficiency. 3D models of all vital organs in wild-type C57BL/6 mice with the 3D structure of their internal components (e.g., vessel network, glomeruli, and nerve tracts) can be reconstructed after imaging with TRUST to demonstrate its fast, robust, and high-content multicolor 3D imaging capability. Moreover, its potential for developmental biology has also been validated by imaging entire mouse embryos (~2 days for the embryo at the embryonic day of 15). TRUST offers a fast and cost-effective approach for high-resolution whole-organ multicolor 3D imaging while relieving researchers from the heavy sample preparation workload.

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Data availability. The authors declare that all data supporting the findings of this study are available within the paper and its Supplementary Information.

Article and author information

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Wentao Yu

Department of Chemical and Biological Engineering, Hong Kong University of Science and Technology, Hong Kong, Hong Kong

Competing interests
Wentao Yu, has applied for a patent (US Provisional Patent Application No.: 63/254,546) related to the work reported in this manuscript..

"This ORCID iD identifies the author of this article:" 0000-0002-4712-3177
Lei Kang

Department of Chemical and Biological Engineering, Hong Kong University of Science and Technology, Hong Kong, Hong Kong

Competing interests
Lei Kang, has applied for a patent (US Provisional Patent Application No.: 63/254,546) related to the work reported in this manuscript..
Victor TC Tsang

Department of Chemical and Biological Engineering, Hong Kong University of Science and Technology, Hong Kong, Hong Kong

Competing interests
Victor TC Tsang, has a financial interest in PhoMedics Limited, which, however, did not support this work..
Yan Zhang

Department of Chemical and Biological Engineering, Hong Kong University of Science and Technology, Hong Kong, Hong Kong

Competing interests
Yan Zhang, has applied for a patent (US Provisional Patent Application No.: 63/254,546) related to the work reported in this manuscript..
Ivy HM Wong

Department of Chemical and Biological Engineering, Hong Kong University of Science and Technology, Hong Kong, Hong Kong

Competing interests
Ivy HM Wong, has a financial interest in V Path Limited, which, however, did not support this work..
Terence TW Wong

Department of Chemical and Biological Engineering, Hong Kong University of Science and Technology, Hong Kong, Hong Kong

For correspondence
ttwwong@ust.hk

Competing interests
Terence TW Wong, has a financial interest in PhoMedics Limited, which, however, did not support this work. Has applied for a patent (US Provisional Patent Application No.: 63/254,546) related to the work reported in this manuscript..

"This ORCID iD identifies the author of this article:" 0000-0001-6399-758X

Funding

Research Grants Council, University Grants Committee (16208620)

Terence TW Wong

Research Grants Council, University Grants Committee (26203619)

Terence TW Wong

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal experiments were conducted in conformity with a laboratory animal protocol approved by the Health, Safety and Environment Office of the Hong Kong University of Science and Technology (HKUST) (license number: AEP16212921).

Copyright

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.