On-chip cell measurement in closed space using multi-scale manipulation

我们研究了基于封闭空间中单个细胞的刺激来理解细胞未知功能的基本方法.三维激光操控：利用检流镜对激光进行二维扫描，并通过物镜透镜的垂直运动，实现激光在三维空间的聚焦控制。实现了激光对微观物体的三维高速操纵。在这里，CCD快门定时和透镜运动同步，并成功地消除了模糊.三维激光微工具操纵：利用聚焦激光局部加热制备的热凝胶，成功地实现了对细胞的三维激光微工具操纵。此外，通过使用激光捕获的微工具，有机纳米管的3D操作是可能的。我们用光刻法制造微型工具。这样我们就可以在没有激光直接照射细胞的情况下操纵细胞。芯片上封闭空间的制造：我们通过使用光交联树脂在微芯片中制造封闭空间（笼）。笼子是由渗透膜制成的。单个细胞在笼中运输。我们还在微通道中制作了pH敏感表面.使用3D多尺度操作的芯片中的单个细胞测量：我们通过光刻法制作了具有微通道和笼的微芯片。细胞在笼中运输。控制了细胞周围的环境条件，使细胞长时间稳定生长。

We studied on fundamental methods for understanding of unknown function of cells based on stimulation of individual cells in a closed space.1. Three-dimensional laser manipulation: By using two-dimensional scanning of laser with Galvano mirrors and vertical motion of the objective lens, we controlled focus of laser in 3D. Then, we realized three-dimensional laser manipulation of microscopic objects in high speed. Here, CCD shutter timing and lens motion was synchronized and the blur was eliminated successfully.2. Three-dimensional laser manipulation with microtools: By using thermal gel which can be made by local heating of focused laser, we succeeded in manipulating cells with the gel-bead. Moreover, 3D manipulation of an organic nanotube was possible by suing laser trapped microtools. We made microtools by photolithography. Then we can manipulate cells without direct laser irradiation to the cells.3. Fabrication of on-chip closed space: We made a closed space (cage) in the microchip by using photo crosslinkable resin. The cage is made is made of permeable membrane. Individual cells are transported in the cage. We also made a pH sensitive surface in the microchannel.4. Individual cell measurement in the chip using 3D multi-scale manipulation: We made a microchip having microchannels and cages by photolithography. The cell was transported in the cage. Environmental condition around the cell was controlled and the cell was rnonitored stably for long time.