Abstract

Today’s imaging flow cytometer (IFC) systems are limited by the projection problem: collapsing three-dimensional (3D) information onto a two-dimensional (2D) image causes a lack of tomographic 3D resolution and reduced information content, limiting the reliability of spot counting or co-localization crucial to cell phenotyping. We present 3D imaging flow cytometry as a solution to the problem. Our high-throughput 3D cell imager based on optical sectioning microscopy combines orthogonal light-sheet scanning illumination with our previous spatiotemporal transformation detection to produce 3D cell image reconstruction from a cameraless single-pixel photodetector readout. We demonstrate this capability by cocapturing 3D fluorescence and label-free side-scattering images of single cells in flow with a throughput of approximately 500 cells per second.

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