Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332.
Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Atlanta, GA 30322.
Proc Natl Acad Sci U S A. 2020 Jun 30;117(26):14779-14789. doi: 10.1073/pnas.2001404117. Epub 2020 Jun 19.
Hematological analysis, via a complete blood count (CBC) and microscopy, is critical for screening, diagnosing, and monitoring blood conditions and diseases but requires complex equipment, multiple chemical reagents, laborious system calibration and procedures, and highly trained personnel for operation. Here we introduce a hematological assay based on label-free molecular imaging with deep-ultraviolet microscopy that can provide fast quantitative information of key hematological parameters to facilitate and improve hematological analysis. We demonstrate that this label-free approach yields 1) a quantitative five-part white blood cell differential, 2) quantitative red blood cell and hemoglobin characterization, 3) clear identification of platelets, and 4) detailed subcellular morphology. Analysis of tens of thousands of live cells is achieved in minutes without any sample preparation. Finally, we introduce a pseudocolorization scheme that accurately recapitulates the appearance of cells under conventional staining protocols for microscopic analysis of blood smears and bone marrow aspirates. Diagnostic efficacy is evaluated by a panel of hematologists performing a blind analysis of blood smears from healthy donors and thrombocytopenic and sickle cell disease patients. This work has significant implications toward simplifying and improving CBC and blood smear analysis, which is currently performed manually via bright-field microscopy, and toward the development of a low-cost, easy-to-use, and fast hematological analyzer as a point-of-care device and for low-resource settings.
血液分析，通过全血细胞计数（CBC）和显微镜检查，对于筛选、诊断和监测血液状况和疾病至关重要，但需要复杂的设备、多种化学试剂、繁琐的系统校准和程序，以及经过高度培训的人员进行操作。在这里，我们介绍了一种基于免标记分子成像和深紫外显微镜的血液分析方法，该方法可以快速提供关键血液参数的定量信息，从而促进和改善血液分析。我们证明，这种免标记方法可以实现 1）定量的五分白细胞分类，2）定量的红细胞和血红蛋白特征，3）血小板的清晰识别，以及 4）详细的亚细胞形态。无需任何样品制备，在几分钟内即可对数万张活细胞进行分析。最后，我们引入了一种伪彩色化方案，该方案可以准确地再现传统染色方案下血涂片和骨髓抽吸物的细胞外观，用于显微镜分析。通过一组血液学家对来自健康供体和血小板减少症及镰状细胞病患者的血涂片进行盲法分析，评估了诊断效果。这项工作对于简化和改进目前通过明场显微镜手动进行的 CBC 和血涂片分析具有重要意义，并且对于开发低成本、易于使用和快速的血液分析仪作为即时护理设备和在资源有限的环境中具有重要意义。
