Kim Youngho, Hong Su, Lee Sang Ho, Lee Kangsun, Yun Seok, Kang Yuri, Paek Kyeong-Kap, Ju Byeong-Kwon, Kim Byungkyu
Department of R&D, Cerno Instruments, Seoul 136-764, Korea.
Rev Sci Instrum. 2007 Jul;78(7):074301. doi: 10.1063/1.2751414.
To reduce the problem of cell loss due to adhesion, one of the basic phenomena in microchannel, we proposed the droplet-based magnetically activated cell separator (DMACS). Based on the platform of the DMACS-which consists of permanent magnets, a coverslip with a circle-shaped boundary, and an injection tube-we could collect magnetically (CD45)-labeled (positive) cells with high purity and minimize cell loss due to adhesion. To compare separation efficiency between the MACS and the DMACS, the total number of cells before and after separation with both the separators was counted by flow cytometry. We could find that the number (3241/59 940) of cells lost in the DMACS is much less than that (22 360/59 940) in the MACS while the efficiency of cell separation in the DMACS (96.07%) is almost the same as that in the MACS (96.72%). Practically, with fluorescent images, it was visually confirmed that the statistical data are reliable. From the viability test by using Hoechst 33 342, it was also demonstrated that there was no cell damage on a gas-liquid interface. Conclusively, DMACS will be a powerful tool to separate rare cells and applicable as a separator, key component of lab-on-a-chip.
为减少微通道中的基本现象之一——因黏附导致的细胞损失问题,我们提出了基于液滴的磁激活细胞分离器(DMACS)。基于由永久磁铁、具有圆形边界的盖玻片和注射管组成的DMACS平台,我们能够以高纯度收集磁性(CD45)标记(阳性)的细胞,并将因黏附导致的细胞损失降至最低。为比较MACS和DMACS之间的分离效率,使用流式细胞术对两种分离器分离前后的细胞总数进行了计数。我们发现,DMACS中损失的细胞数量(3241/59940)远少于MACS中的(22360/59940),而DMACS中的细胞分离效率(96.07%)与MACS中的(96.72%)几乎相同。实际上,通过荧光图像直观地证实了统计数据是可靠的。通过使用Hoechst 33342进行的活力测试还表明,气液界面上没有细胞损伤。总之,DMACS将成为分离稀有细胞的有力工具,并可作为芯片实验室的关键组件——分离器使用。