Dr. Sadao Ota Profile

Dr. Sadao Ota

at Univ of Tokyo

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Proceedings Article | 15 March 2023 Presentation + Paper

High-throughput parallel 3D imaging flow cytometry

Masashi Ugawa, Sadao Ota

Proceedings Volume 12383, 1238309 (2023) https://doi.org/10.1117/12.2648669

KEYWORDS: Flow cytometry, 3D image processing, Imaging systems, Microscopy, Cameras, 3D acquisition, Particles, Structural analysis, Objectives

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Imaging flow cytometry has enabled high-throughput morphology-based analysis of large cell populations at the singlecell level. However, most of these systems were based on 2D imaging, and with the loss of the spatial information in the depth direction, its accuracy was limited, especially when performing analysis based on subcellular structures. Efforts to develop 3D imaging flow cytometry have been undertaken, but the throughput was limited because of the slow 3D imaging systems. Even by adopting light-sheet microscopy, the throughput was limited to less than 100 cells/s, due to the camera frame rate. To overcome this limitation, we combined oblique-plane microscopy with 1D acoustofluidic particle focusing to image multiple cells in parallel. With this, we were able to overcome the throughput limitation by the camera frame rate and achieve a detection throughput of 2,300 cells/s, the highest to our knowledge for 3D imaging flow cytometry. Furthermore, with such high throughput, we were able to capture 3D images of 4×10⁵ cells in an acquisition time of less than five min and perform subcellular-structure-based analysis of mitotic cells, demonstrating practical 3D imaging flow cytometry for the first time to our knowledge. Result show, that with 3D imaging flow cytometry, we can capture structural information that would be overlooked with its 2D imaging counterpart.

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