The current prevailing methods for identifying immune cell subsets exploit a group of differentiation markers (CDs) targeted by fluorochrome or metal conjugated antibodies. However, such labeling methods, requiring a staining process and specific reagents, prevent rapid and cost-effective identification of immune cell subsets. Therefore we developed a label-free imaging cytometry platform that synergistically used refractive index (RI) tomography and three-dimensional (3-D) deep learning. We constructed and trained a deep learning classifier that learns unique representations from the 3-D RI map of each cell obtained using RI tomography without labeling. In this study, we were able to classify human naïve, memory, and senescent T cells according to the expression of CD4, CD8, CD45RA, CCR7 and CD57 using the label-free classifier within milliseconds, with high precision (>95%) even though the morphological and biochemical characteristics extracted from the RI tomograms of the T cells are almost homogeneous. This cannot be achieved by conventional machine learning approaches that only exploit the set of manually extracted features. Our label-free cell sorting platform will facilitate rapid and cost-effective immunological and biomedical studies by eliminating the laborious labeling process.
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