Light-field flow cytometry for high-resolution, volumetric and multiparametric 3D single-cell analysis

Xuanwen Hua, Keyi Han, Biagio Mandracchia, Afsane Radmand, Wenhao Liu, Hyejin Kim, Zhou Yuan, Samuel M. Ehrlich, Kaitao Li, Corey Zheng, Jeonghwan Son, Aaron D. Silva Trenkle, Gabriel A. Kwong, Cheng Zhu, James E. Dahlman and Shu Jia ()
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Xuanwen Hua: Georgia Institute of Technology and Emory University
Keyi Han: Georgia Institute of Technology and Emory University
Biagio Mandracchia: Georgia Institute of Technology and Emory University
Afsane Radmand: Parker H. Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology
Wenhao Liu: Georgia Institute of Technology and Emory University
Hyejin Kim: Georgia Institute of Technology and Emory University
Zhou Yuan: Parker H. Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology
Samuel M. Ehrlich: Parker H. Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology
Kaitao Li: Georgia Institute of Technology and Emory University
Corey Zheng: Georgia Institute of Technology and Emory University
Jeonghwan Son: Georgia Institute of Technology and Emory University
Aaron D. Silva Trenkle: Georgia Institute of Technology and Emory University
Gabriel A. Kwong: Georgia Institute of Technology and Emory University
Cheng Zhu: Georgia Institute of Technology and Emory University
James E. Dahlman: Georgia Institute of Technology and Emory University
Shu Jia: Georgia Institute of Technology and Emory University

Nature Communications, 2024, vol. 15, issue 1, 1-12

Abstract: Abstract Imaging flow cytometry (IFC) combines flow cytometry and fluorescence microscopy to enable high-throughput, multiparametric single-cell analysis with rich spatial details. However, current IFC techniques remain limited in their ability to reveal subcellular information with a high 3D resolution, throughput, sensitivity, and instrumental simplicity. In this study, we introduce a light-field flow cytometer (LFC), an IFC system capable of high-content, single-shot, and multi-color acquisition of up to 5,750 cells per second with a near-diffraction-limited resolution of 400-600 nm in all three dimensions. The LFC system integrates optical, microfluidic, and computational strategies to facilitate the volumetric visualization of various 3D subcellular characteristics through convenient access to commonly used epi-fluorescence platforms. We demonstrate the effectiveness of LFC in assaying, analyzing, and enumerating intricate subcellular morphology, function, and heterogeneity using various phantoms and biological specimens. The advancement offered by the LFC system presents a promising methodological pathway for broad cell biological and translational discoveries, with the potential for widespread adoption in biomedical research.

Date: 2024
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DOI: 10.1038/s41467-024-46250-7

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