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

Hua, Xuanwen; Han, Keyi; Mandracchia, Biagio; Radmand, Afsane; Liu, Wenhao; Kim, Hyejin; Yuan, Zhou; Ehrlich, Samuel M.; Li, Kaitao; Zheng, Corey; Son, Jeonghwan; Silva Trenkle, Aaron D.; Kwong, Gabriel A.; Zhu, Cheng; Dahlman, James E.; Jia, Shu

doi:10.1038/s41467-024-46250-7

Cited by 7 publications

References 72 publications

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Advances and Prospects in Liquid Biopsy Techniques for Malignant Tumor Diagnosis and Surveillance

Zhang,

Li,

Zhao

et al. 2024

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Liquid biopsy technology provides invaluable support for the early diagnosis of tumors and surveillance of disease course by detecting tumor‐related biomarkers in bodily fluids. Currently, liquid biopsy techniques are mainly divided into two categories: biomarker and label‐free. Biomarker liquid biopsy techniques utilize specific antibodies or probes to identify and isolate target cells, exosomes, or molecules, and these techniques are widely used in clinical practice. However, they have certain limitations including dependence on tumor markers, alterations in cell biological properties, and high cost. In contrast, label‐free liquid biopsy techniques directly utilize physical or chemical properties of cells, exosomes, or molecules for detection and isolation. These techniques have the advantage of not needing labeling, not impacting downstream analysis, and low detection cost. However, most are still in the research stage and not yet mature. This review first discusses recent advances in liquid biopsy techniques for early tumor diagnosis and disease surveillance. Several current techniques are described in detail. These techniques exploit differences in biomarkers, size, density, deformability, electrical properties, and chemical composition in tumor components to achieve highly sensitive tumor component identification and separation. Finally, the current research progress is summarized and the future research directions of the field are discussed.

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Advances and Prospects in Liquid Biopsy Techniques for Malignant Tumor Diagnosis and Surveillance

Zhang,

Li,

Zhao

et al. 2024

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Lensless On-Chip Chemiluminescence Imaging for High-Throughput Single-Cell Heterogeneity Analysis

Yang,

Fang,

Liu

et al. 2024

Nano Lett.

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High-throughput single-cell heterogeneity imaging and analysis is essential for understanding complex biological systems and for advancing personalized precision disease diagnosis and treatment. Here, we present a miniaturized lensless chemiluminescence chip for high-throughput single-cell functional imaging with subcellular resolution. With the sensitive chemiluminescence sensing and wide field of view of contact lensless imaging, we demonstrated the chemiluminescent imaging of over 1000 single cells, and their membrane glycoprotein and the highthroughput single-cell heterogeneity of membrane protein imaging were examined for precision analysis. Furthermore, the functional adhesion and heterogeneity of single live cells were imaged and explored. This miniaturized lensless on-chip CL-CMOS imaging platform enables high-throughput single-cell imaging and analysis with high sensitivity and subcellular resolution, providing new techniques for the cellular study of biological heterogeneity and has potential application in precision disease diagnosis and treatment at the point-of-care settings.

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Three-dimensional isotropic imaging of live suspension cells enabled by droplet microvortices

Cardenas-Benitez,

Hurtado,

Luo

et al. 2024

Proc. Natl. Acad. Sci. U.S.A.

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Fast, nondestructive three-dimensional (3D) imaging of live suspension cells remains challenging without substrate treatment or fixation, precluding scalable single-cell morphometry with minimal alterations. While optical sectioning techniques achieve 3D live cell imaging, lateral versus depth resolution differences further complicate analysis. We present a scalable microfluidic method capable of 3D fluorescent isotropic imaging of live, nonadherent cells suspended inside picoliter droplets with high-speed single-cell volumetric readout (800 to 1,200 slices in 5 to 8 s) and near-diffraction limit resolution (~216 nm). The platform features a droplet trap array that leverages flow-induced droplet interfacial shear to generate intradroplet microvortices, which rotate single cells on their axis to enable optical projection tomography (OPT)-based imaging. This allows gentle (~1 mPa shear stress) observation of cells encapsulated inside nontoxic isotonic buffer droplets, facilitating scalable OPT acquisition by simultaneous spinning of hundreds of cells. We demonstrate 3D imaging of live myeloid and lymphoid cells in suspension, including K562 cells, as well as naive and activated T cells—small cells prone to movement in their suspended phenotype. Our fully suspended, orientation-independent cell morphometry, driven by isotropic imaging and spherical harmonic analysis, enabled the study of primary T cells across various immunological activation states. This approach unveiled six distinct nuclear content distributions, contrasting with conventional 2D images that typically portray spheroid and bean-like nuclear shapes associated with lymphocytes. Our arrayed-droplet OPT technology is capable of isotropic, single live-cell 3D imaging, with the potential to perform large-scale morphometry of immune cell effector function states while providing compatibility with microfluidic droplet operations.

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

Cited by 7 publications

References 72 publications

Advances and Prospects in Liquid Biopsy Techniques for Malignant Tumor Diagnosis and Surveillance

Advances and Prospects in Liquid Biopsy Techniques for Malignant Tumor Diagnosis and Surveillance

Lensless On-Chip Chemiluminescence Imaging for High-Throughput Single-Cell Heterogeneity Analysis

Three-dimensional isotropic imaging of live suspension cells enabled by droplet microvortices

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