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Abstract

Flow cytometers are robust and ubiquitous tools of biomedical research, as they enable high-
throughput fluorescence-based multi-parametric analysis and sorting of single cells. How-
ever, analysis is often constrained by the availability of detection reagents or functional

changes of cells caused by fluorescent staining. Here, we introduce MAPS-FC (multi-angle
pulse shape flow cytometry), an approach that measures angle- and time-resolved scattered
light for high-throughput cell characterization to circumvent the constraints of conventional

flow cytometry. In order to derive cell-specific properties from the acquired pulse shapes, we
developed a data analysis procedure based on wavelet transform and k-means clustering. We

analyzed cell cycle stages of Jurkat and HEK293 cells by MAPS-FC and were able to assign

cells to the G1, S, and G2/M phases without the need for fluorescent labeling. The results
were validated by DNA staining and by sorting and re-analysis of isolated G1, S, and G2/M

populations. Our results demonstrate that MAPS-FC can be used to determine cell properties

that are otherwise only accessible by invasive labeling. This approach is technically com-

patible with conventional flow cytometers and paves the way for label-free cell sorting.