Department of Microengineering, Kyoto University, Sakyo-ku, Kyoto, Japan.
Biomed Microdevices. 2012 Aug;14(4):791-7. doi: 10.1007/s10544-012-9656-5.
A microfluidic device integrated with a Total Internal Reflection (TIR)-based chip for cell observation and analysis was developed. This integrated device enables in situ Total Internal Reflection Fluorescence Microscopy (TIRFM) on adherent cells cultured under continuous medium perfusion. This TIR-based chip, allows TIRFM to be easily performed on cells without the assembly of complicated optical components and cell culture chambers. The integrated device was evaluated by tracking the movement of fluorescent beads and monitoring the location of insulin granules in mouse pancreatic β-cells. This system offers higher signal-to-noise (S/N) ratio than epi-fluorescence microscopy (EPIFM), and comparable image quality to commercial TIRFM systems when imaging insulin granules. We also detected repetitive changes in intracellular Ca(2+) concentration in MIN6-m9 cells stimulated with KCl, which demonstrates quick perfusion for cell analysis while maintaining high S/N ratio.
一种集成了全内反射（TIR）芯片的微流控设备用于细胞观察和分析。该集成设备可在连续介质灌注下培养的贴壁细胞上进行原位全内反射荧光显微镜（TIRFM）。该 TIR 芯片可轻松对细胞进行 TIRFM，无需组装复杂的光学组件和细胞培养室。通过跟踪荧光珠的运动和监测小鼠胰腺β细胞中胰岛素颗粒的位置来评估集成设备。与 epi-fluorescence microscopy (EPIFM) 相比，该系统具有更高的信噪比（S/N），并且在成像胰岛素颗粒时，图像质量与商业 TIRFM 系统相当。我们还检测到 KCl 刺激的 MIN6-m9 细胞内 Ca(2+)浓度的重复变化，这表明在保持高 S/N 比的同时，可快速进行细胞分析的灌注。
