Oblique plane microscopy for 3D, multi-color, eight-day time-lapse imaging of genetically heterogeneous breast cancer spheroids in 384-well plates with quantitative 3D analysis

Leo Rowe-Brown; Nathan Curry; Sandesh Meyler; Nina Moderau; Chris Dunsby

doi:10.1117/12.2648952

15 March 2023 Oblique plane microscopy for 3D, multi-color, eight-day time-lapse imaging of genetically heterogeneous breast cancer spheroids in 384-well plates with quantitative 3D analysis

Leo Rowe-Brown, Nathan Curry, Sandesh Meyler, Nina Moderau, Chris Dunsby

Author Affiliations +

Proceedings Volume 12383, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XXI; 1238302 (2023) https://doi.org/10.1117/12.2648952
Event: SPIE BiOS, 2023, San Francisco, California, United States

Abstract

Oblique plane microscopy (OPM) is a light-sheet microscopy technique that illuminates a tilted plane of a specimen and collects the fluorescence with the same high NA objective. Here, we apply OPM with stage-scanned image volume acquisition to perform high-content, 3D time-lapse imaging of up to 180 breast cancer spheroids over eight days every eight hours. Each spheroid comprises a combination of one to three different cell populations, each co-expressing a unique oncogene-fluorescent protein pair. We trained the image segmentation tool Cellpose to segment nuclei in the resulting data in order to measure integrated fluorescence intensity, nuclear size, and nuclear count for each of the different oncogenes to quantify competition between mutations.

Conference Presentation

Citation Download Citation

Leo Rowe-Brown, Nathan Curry, Sandesh Meyler, Nina Moderau, and Chris Dunsby "Oblique plane microscopy for 3D, multi-color, eight-day time-lapse imaging of genetically heterogeneous breast cancer spheroids in 384-well plates with quantitative 3D analysis", Proc. SPIE 12383, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XXI, 1238302 (15 March 2023); https://doi.org/10.1117/12.2648952

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