Smith Zachary J, Gao Tingjuan, Chu Kaiqin, Lane Stephen M, Matthews Dennis L, Dwyre Denis M, Hood James, Tatsukawa Keith, Heifetz Laurence, Wachsmann-Hogiu Sebastian
Center for Biophotonics Science and Technology, University of California, Davis, 2700 Stockton Blvd. Suite 1400, Sacramento, CA 95817, USA.
Lab Chip. 2014 Aug 21;14(16):3029-36. doi: 10.1039/c4lc00567h. Epub 2014 Jun 23.
Current flow-based blood counting devices require significant medical infrastructure and are not appropriate for field use. In this article we report on the development of a sample preparation, measurement, and analysis method that permits automated and accurate counting of red blood cells (RBCs), white blood cells (WBCs), and platelets, as well as allowing a 3-part differential of the WBCs to be performed on extremely small volumes of whole blood. This method is compatible with portable instrumentation that can be deployed in the field. The method consists of serially diluting blood samples first with sodium dodecyl sulfate dissolved in phosphate buffered saline, then in acridine orange dissolved in phosphate buffered saline, followed by fluorescence and dark field imaging with low magnification objectives. Image analysis is performed to extract cell counts and differentials. We performed a paired analysis of 20 volunteers with complete blood count values both within and beyond the normal reference range using a commercial automated hematology analyzer and the image-based method, with the new method achieving accuracies comparable to that of the commercial system. Because the sample preparation and imaging are simple and inexpensive to implement, this method has applications for pediatrics, clinician offices, and global health in regions that do not have access to central hematology laboratories.
基于电流的血液计数设备需要大量的医疗基础设施，不适合现场使用。在本文中，我们报告了一种样品制备、测量和分析方法的开发，该方法能够自动且准确地计数红细胞（RBC）、白细胞（WBC）和血小板，还能对极少量全血中的白细胞进行三分群分类。这种方法与可部署在现场的便携式仪器兼容。该方法包括先用溶解在磷酸盐缓冲盐水中的十二烷基硫酸钠对血样进行系列稀释，然后用溶解在磷酸盐缓冲盐水中的吖啶橙进行稀释，接着用低倍物镜进行荧光和暗场成像。通过图像分析来提取细胞计数和分类结果。我们使用商用自动血液分析仪和基于图像的方法，对20名全血细胞计数结果在正常参考范围内和超出正常参考范围的志愿者进行了配对分析，新方法的准确性与商用系统相当。由于样品制备和成像操作简单且成本低廉，该方法适用于儿科、临床医生办公室以及无法使用中央血液学实验室的全球卫生领域。
