30 March 2015 Performance of computer vision in vivo flow cytometry with low fluorescence contrast
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Detection and enumeration of circulating cells in the bloodstream of small animals are important in many areas of preclinical biomedical research, including cancer metastasis, immunology, and reproductive medicine. Optical in vivo flow cytometry (IVFC) represents a class of technologies that allow noninvasive and continuous enumeration of circulating cells without drawing blood samples. We recently developed a technique termed computer vision in vivo flow cytometry (CV-IVFC) that uses a high-sensitivity fluorescence camera and an automated computer vision algorithm to interrogate relatively large circulating blood volumes in the ear of a mouse. We detected circulating cells at concentrations as low as 20  cells/mL. In the present work, we characterized the performance of CV-IVFC with low-contrast imaging conditions with (1) weak cell fluorescent labeling using cell-simulating fluorescent microspheres with varying brightness and (2) high background tissue autofluorescence by varying autofluorescence properties of optical phantoms. Our analysis indicates that CV-IVFC can robustly track and enumerate circulating cells with at least 50% sensitivity even in conditions with two orders of magnitude degraded contrast than our previous in vivo work. These results support the significant potential utility of CV-IVFC in a wide range of in vivo biological models.
© 2015 Society of Photo-Optical Instrumentation Engineers (SPIE)
Stacey Markovic, Stacey Markovic, Siyuan Li, Siyuan Li, Mark J. Niedre, Mark J. Niedre, } "Performance of computer vision in vivo flow cytometry with low fluorescence contrast," Journal of Biomedical Optics 20(3), 035005 (30 March 2015). https://doi.org/10.1117/1.JBO.20.3.035005 . Submission:


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