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

  1. Wentao Yu
  2. Lei Kang
  3. Victor TC Tsang
  4. Yan Zhang
  5. Ivy HM Wong
  6. Terence TW Wong  Is a corresponding author
  1. Hong Kong University of Science and Technology, Hong Kong

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.

Data availability

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

Author details

  1. 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..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4712-3177
  2. 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..
  3. 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..
  4. 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..
  5. 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..
  6. 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..
    ORCID icon "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).

Reviewing Editor

  1. Kevin M Dean, University of Texas Southwestern Medical Center, United States

Publication history

  1. Preprint posted: November 13, 2021 (view preprint)
  2. Received: June 13, 2022
  3. Accepted: November 4, 2022
  4. Accepted Manuscript published: November 4, 2022 (version 1)
  5. Version of Record published: November 17, 2022 (version 2)

Copyright

© 2022, Yu et al.

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.

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  1. Wentao Yu
  2. Lei Kang
  3. Victor TC Tsang
  4. Yan Zhang
  5. Ivy HM Wong
  6. Terence TW Wong
(2022)
Translational rapid ultraviolet-excited sectioning tomography for whole-organ multicolor imaging with real-time molecular staining
eLife 11:e81015.
https://doi.org/10.7554/eLife.81015

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