Biomicrofluidics. 2010 Sep 30;4(3):32208. doi: 10.1063/1.3494287.
Recently, interest in single cell analysis has increased because of its potential for improving our understanding of cellular processes. Single cell operation and attachment is indispensable to realize this task. In this paper, we employed a simple and direct method for single-cell attachment and culture in a closed microchannel. The microchannel surface was modified by applying a nonbiofouling polymer, 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, and a nitrobenzyl photocleavable linker. Using ultraviolet (UV) light irradiation, the MPC polymer was selectively removed by a photochemical reaction that adjusted the cell adherence inside the microchannel. To obtain the desired single endothelial cell patterning in the microchannel, cell-adhesive regions were controlled by use of round photomasks with diameters of 10, 20, 30, or 50 μm. Single-cell adherence patterns were formed after 12 h of incubation, only when 20 and 30 μm photomasks were used, and the proportions of adherent and nonadherent cells among the entire UV-illuminated areas were 21.3%±0.3% and 7.9%±0.3%, respectively. The frequency of single-cell adherence in the case of the 20 μm photomask was 2.7 times greater than that in the case of the 30 μm photomask. We found that the 20 μm photomask was optimal for the formation of single-cell adherence patterns in the microchannel. This technique can be a powerful tool for analyzing environmental factors like cell-surface and cell-extracellular matrix contact.
最近，单细胞分析的兴趣增加了，因为它有可能提高我们对细胞过程的理解。单细胞操作和附着对于实现这一任务是必不可少的。在本文中，我们采用了一种简单直接的方法，在封闭的微通道中进行单细胞附着和培养。微通道表面通过应用非生物污染聚合物 2-甲基丙烯酰氧乙基磷酸胆碱（MPC）聚合物和硝基苄基光可裂解连接体进行修饰。通过紫外线（UV）光照射，通过光化学反应选择性地去除 MPC 聚合物，该光化学反应调整了微通道内的细胞附着。为了在微通道中获得所需的单个内皮细胞模式，通过使用直径为 10、20、30 或 50 μm 的圆形光掩模来控制细胞附着区域。孵育 12 小时后，仅当使用 20 和 30 μm 光掩模时，才形成单细胞附着模式，并且在整个 UV 照射区域中，附着细胞和非附着细胞的比例分别为 21.3%±0.3%和 7.9%±0.3%。在 20 μm 光掩模的情况下，单细胞附着的频率比在 30 μm 光掩模的情况下高 2.7 倍。我们发现 20 μm 光掩模是在微通道中形成单细胞附着模式的最佳选择。该技术可以成为分析细胞表面和细胞-细胞外基质接触等环境因素的有力工具。
