By combining digital imaging (recording the structure within a sample) and flow cytometer (interrogating samples that continuously flow), imaging flow cytometry (IFC) can record the internal structure or the distribution of fluorophores within individual cells continuously flowing in a small channel. Existing IFC systems can acquire only one 2D cross-section image for each cell, failing to capture the abundant biological information available in the cell’s 3-D volume. Three-dimensional (3D) imaging flow cytometry is challenging, because acquiring the volumetric information usually requires a mechanism to scan the objective lens focus or to change the illumination direction. Snapshot 3D microscopy techniques have been developed to address this challenge, but they have insufficient spatial resolution for single-cell imaging and suffer from long data processing time. We overcome these limitations by combining two novel snapshot techniques developed by the PI with the most rigorous optical imaging theories and cutting-edge component technologies.
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Funding sources: National Institutes of Health (National Institute of General Medical Sciences, 1R21GM135848-01); UWM Research Growth Initiative; Lynde and Harry Bradley Foundation.
Collaborators: Dr. Sridhar Rao (Blood Research Institute), Dr. Woo Jin Chang (UWM).