LeGO-3D: 3D imaging of lung metastases and vascularisation using light sheet fluorescence microscopy

Lewis, Sabrina M; Berthelet, Jean; Whitehead, Lachlan W; Rajasekhar, Pradeep; El-Saafin, Farrah; Bell, Caroline; Naik, Shalin; Merino, Delphine; Wimmer, Verena C; Rogers, Kelly L

doi:10.1038/s44303-025-00111-0

LeGO-3D: 3D imaging of lung metastases and vascularisation using light sheet fluorescence microscopy

https://doi.org/10.1038/s44303-025-00111-0
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journal contribution

posted on 2025-12-08, 02:55 authored by Sabrina M Lewis, Jean BertheletJean Berthelet, Lachlan W Whitehead, Pradeep Rajasekhar, Farrah El-SaafinFarrah El-Saafin, Caroline BellCaroline Bell, Shalin Naik, Delphine MerinoDelphine Merino, Verena C Wimmer, Kelly L Rogers

Cancer metastasis involves a complex cascade of events, where cancer cells migrate from their site of origin to secondary sites via the lymphatic and circulatory system. During this process, some cancer subclones will successfully ‘seed’ at distant organs to generate lethal metastases. Here, we optimised a method for tracking cancer cells in metastatic breast cancer tumours and investigated their complex interplay with the lung vasculature using lentiviral-based optical barcoding (LeGO). Given the regional heterogeneity in lung tissue microenvironments as well as lobar asymmetry, we used light sheet microscopy to perform three-dimensional (3D) imaging of wholemount lung lobes. The results revealed that polychromatic metastases occurred less frequently than monochromatic metastases and were more likely to be located nearer to blood vessels in both spontaneous (i.e. mammary fat pad injections) and experimental (i.e. tail vein injections) mouse assays of metastasis. This 3D imaging and analytic pipeline can provide unique insights about metastatic heterogeneity and dynamics, and represents a new avenue for studying therapeutic response across large volumes of lung tissue.<p></p>

Funding

S.L. was supported by a Melbourne Research Scholarship from the University of Melbourne, a WEHI top up scholarship and Avis MacPhee PhD Award. J.B. is supported by the NHMRC (GNT2036844). D.M. is supported by the Victorian Cancer agency (MCRF21011), the NBCF (Investigator Initiated Research Grant IIRS0049), NHMRC (GNT2027459) and Love Your Sister. This research is supported by the Victorian Government through the Victorian Cancer Agency and the Operational Infrastructure Support Program at the Olivia Newton-John Cancer Research Institute and the Walter and Eliza Hall Institute. The authors and Olivia Newton-John Cancer Research Institute gratefully acknowledge the generous support of the Love Your Sister Foundation. We also thank the Austin BRF. This publication was supported through state funds approved by the State Parliament of Baden-Württemberg for the Innovation Campus Health + Life Science Alliance Heidelberg Mannheim.

History

Publication Date

2025-11-07

Journal

npj Imaging

Volume

3

Article Number

58

Pagination

11p.

Publisher

Springer Nature

ISSN

2948-197X

Rights Statement

© The Author(s) 2025 This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.