Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Koganei, Tokyo 184-8588, Japan.
Biosens Bioelectron. 2013 Feb 15;40(1):110-4. doi: 10.1016/j.bios.2012.06.043. Epub 2012 Jun 28.
Monitoring of hematotoxicity, which requires serial blood collection, is difficult to carry out in small animals due to a lack of non-invasive, individual animal-appropriate techniques that enable enumeration of leukocyte subsets from limited amounts of whole blood. In this study, a microfluidic device equipped with a microcavity array that enables highly efficient separation of leukocytes from submicroliters of whole blood was applied for hematotoxicity monitoring in mice. The microcavity array can specifically separate leukocytes from whole blood based on differences in the size and deformability between leukocytes and other blood cells. Mouse leukocytes recovered on aligned microcavities were continuously processed for image-based immunophenotypic analysis. Our device successfully recovered almost 100% of mouse leukocytes in 0.1 μL of whole blood without the effect of serial blood collection such as changes in body weight and total leukocyte count. We assessed benzene-associated hematotoxicity in mice using this system. Mice were administered with benzene once daily and the depression of leukocyte numbers induced in individual mice was successfully monitored from tail vein blood collected every other day for 2 weeks. Serial monitoring of the leukocyte number in individual mice will contribute to the understanding of hematotoxicity and reduction of the number of animal experiment trials.
由于缺乏非侵入性、适用于个体动物的技术，无法从小动物中有限量的全血中计数白细胞亚群，因此难以对血液毒性进行监测。在这项研究中，应用了一种配备微腔阵列的微流控装置，该装置能够从亚微升全血中高效分离白细胞，用于监测小鼠的血液毒性。微腔阵列可以根据白细胞和其他血细胞在大小和变形性上的差异，特异性地从全血中分离白细胞。在对齐的微腔上回收的小鼠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 0.1 μL 的全血中回收了近 100%的小鼠白细胞，而不会像连续采血那样引起体重和总白细胞计数的变化。我们使用该系统评估了苯诱导的小鼠血液毒性。每天给小鼠注射苯一次，并成功地从尾静脉每隔一天采集的血液中监测到 2 周内个体小鼠白细胞数量的减少。对个体小鼠白细胞数量的连续监测将有助于了解血液毒性并减少动物实验的次数。
