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。三维激光操纵：利用镜对激光进行二维扫描，利用物镜的垂直运动控制激光的三维聚焦。在此基础上，实现了对微观物体的高速三维激光操纵。在这里，CCD快门定时和镜头运动同步，成功地消除了模糊。微工具三维激光操纵：利用聚焦激光局部加热制成的热凝胶，成功地用凝胶珠操纵细胞。此外，利用激光捕获的微工具可以对有机纳米管进行三维操作。我们用光刻技术制造微型工具。这样我们就可以在不直接用激光照射细胞的情况下操纵细胞。片上封闭空间的制作：我们利用光交联树脂在微芯片上制作封闭空间（笼）。所述笼由可透膜制成。单个细胞在笼子里运输。我们还在微通道中制作了pH敏感表面。利用三维多尺度操作技术对芯片中的单个细胞进行测量：我们采用光刻技术制作了一种具有微通道和微笼的微芯片。细胞在笼子里运输。控制了细胞周围的环境条件，对细胞进行了长时间的稳定监测。

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.