Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan.
Biotechnol Bioeng. 2012 Aug;109(8):2017-24. doi: 10.1002/bit.24471. Epub 2012 Feb 24.
Absolute counting of total leukocytes or specific subsets within small amounts of whole blood is difficult due to a lack of techniques that enable separation of all leukocytes from limited amounts of whole blood. In this study, a microfluidic device equipped with a size-controlled microcavity array for highly efficient separation of leukocytes from submicroliters of whole blood was developed. The microcavity array can separate leukocytes from whole blood based on differences in the size and deformability between leukocytes and other blood cells. Leukocytes recovered on aligned microcavities were continuously processed for image-based immunophenotypic analysis. Our device successfully recovered over 90% of leukocytes in 1 µL of whole blood without pretreatment such as density gradient centrifugation or erythrocyte lysis. In addition, the proposed system successfully performed absolute enumeration of human CD4(+) and CD8(+) leukocytes from 1 µL of whole blood, and the obtained data showed good correlation with conventional flow cytometric analysis. Our microfluidic device has great potential as a tool for a point-of-care leukocyte analysis system.
由于缺乏能够从有限量全血中分离所有白细胞的技术，因此难以对少量全血中的总白细胞或特定亚群进行绝对计数。在这项研究中，开发了一种配备尺寸可控微腔阵列的微流控装置，用于从亚微升全血中高效分离白细胞。微腔阵列可以根据白细胞与其他血细胞之间的大小和变形性差异来分离白细胞。在对齐的微腔上回收的白细胞被连续进行基于图像的免疫表型分析。我们的设备成功地从 1 µL 的全血中回收了超过 90%的白细胞，而无需进行密度梯度离心或红细胞裂解等预处理。此外，该系统成功地从 1 µL 的全血中对人 CD4(+)和 CD8(+)白细胞进行了绝对计数，并且获得的数据与传统的流式细胞术分析具有良好的相关性。我们的微流控装置非常有潜力成为即时白细胞分析系统的工具。
