Complete blood count and differentiation of leukocytes belong to the most frequently performed laboratory diagnostic tests. Here, a flow cytometry-based method for label-free differentiation of untouched leukocytes by digital holographic microscopy on the rich phase contrast of peripheral leukocyte images, using highly controlled 2D hydrodynamic focusing conditions is reported. Principal component analysis of morphological characteristics of the reconstructed images allows to classify not only nine leukocyte types, but also different types of leukemia and demonstrate disappearance of acute myeloid leukemia cells in remission. To exclude confounding effects, the classification strategy is tested by the analysis of 20 blinded clinical samples. Here, 70% of the specimens are correctly classified with further 20% classifications close to a correct diagnosis. Taken together, the findings indicate a broad clinical applicability of the cytometry method for automated and reagent-free diagnosis of hematological disorders.
Oliver Hayden, "Hematology analysis with holographic imaging cytometry (Conference Presentation)," Proc. SPIE 10887, Quantitative Phase Imaging V, 108871I (Presented at SPIE BiOS: February 05, 2019; Published: 4 March 2019); https://doi.org/10.1117/12.2513270.6008593371001.
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