Dual-View Transport of Intensity Phase Imaging-Based Flow Cytometry for Label-Free Cell Analysis and Classification

We introduce a quantitative phase imaging-based flow cytometer that integrates dual-view transport of intensity phase imaging with microfluidics into a commercial microscope, enabling label-free cell analysis and classification. By capturing under-focus and over-focus images simultaneously, the phase distributions of flowing cells are reconstructed to extract morphological parameters for subsequent classification. This system achieves high-accuracy phase imaging, as demonstrated by tests on a standard phase plate sample, and successfully recognizes and classifies cells, validated using mixtures of RAW264.7 cells and MC3T3-E1 cells in varying proportions. Given its simple configuration, precise phase retrieval, and robust classification capabilities, we believe this quantitative phase imaging-based flow cytometer holds great promise as an efficient tool for cell analysis in microfluidics, with potential applications in both fundamental research and clinical studies.