Complete blood count and differentiation of leukocytes (DIFF) belong to the most frequently performed laboratory diagnostic tests. Here, a flow cytometry‐based method for label‐free DIFF 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 classification of nine leukocyte types, in addition to different types of leukemia and demonstrates 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.

中文翻译：

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通过全息显微镜进行无标记高通量白血病检测

全血细胞计数和白细胞分化 (DIFF) 属于最常进行的实验室诊断测试。在此，报道了一种基于流式细胞术的方法，使用高度受控的 2D 流体动力学聚焦条件，通过数字全息显微镜对外周白细胞图像的丰富相差进行未接触白细胞的无标记 DIFF。对重建图像的形态特征进行主成分分析，除了不同类型的白血病之外，还可以对九种白细胞类型进行分类，并证明缓解期急性髓系白血病细胞的消失。为了排除混杂效应，通过分析 20 个盲法临床样本来测试分类策略。此处，70% 的标本被正确分类，另外 20% 的分类接近正确诊断。总而言之，这些发现表明细胞计数方法在血液疾病的自动化和无试剂诊断方面具有广泛的临床适用性。