Capability enhancements for confocal laser scanning microscope

共聚焦激光扫描显微镜（CLSM）是一种功能强大的工具，使生物学家能够通过固体组织或细胞获得光学激光“切片”，而无需切割物理切片。整个细胞/组织的三维重建可以从这些光学切片组装。组织或单细胞中的目标分子可以使用抗体和荧光标记来识别，当被特定波长的激光照射时，荧光标记会发光。因此，CLSM是全世界许多细胞生物学家和生理学家的核心，强大的工具。约克生物系CLSM设施于2002年安装。自那时以来，CLSM设备和技术有了很大的改进。大约10000小时/年是登录在我们的CLSM由部门的很大一部分，它是在迫切需要一些新的组件和某些旧的更换。它只有红色和绿色激光。重要的是，我们获得了405 nm（蓝色）激光。这将使三种不同的分子同时可视化，并允许分析几种细胞结构彼此之间的关系。它还将使活细胞染色（如DAPI）的使用，允许亚细胞结构的运动的延时研究，最小的激光光漂白。目前，许多荧光染色程序需要几天才能完成;我们希望获得一种新型的微波系统，该系统可以加速分子通过膜的渗透，而无需加热它们，这将使处理时间减少到几个小时，并允许研究人员在同一时间内实现更多。最后，2002年CLSM的几个原始部件已经达到使用寿命，如果CLSM要继续运行，就需要更换。即使是8岁的计算机，也无法运行CLSM制造商要求的当前软件。约克大学在细胞生物学和生理学方面都有杰出的研究记录，但只有在我们的部门CLSM中安装了所需的增强功能时，才能保持这一点。

Confocal Laser Scanning Microscopy (CLSM) is a powerful tool that enables biologists to obtain optical laser 'slices' through solid tissue or cells without the need for cutting physical sections. Three-dimensional reconstructions of whole cells/tissues can be assembled from these optical sections. Molecules of interest in tissues or single cells can be identified using antibodies to them and fluorescent tags that glow when illuminated by particular wavelengths of laser light. CLSM is thus a central, powerful tool for very many cell biologists and physiologists worldwide. The York Biology Dept CLSM facility was installed in 2002. CLSM equipment and techniques have improved greatly since then. About 10000hr/year are logged on our CLSM by a large segment of the department, and it is in desperate need of some new components and replacement of certain old ones. It has only red and green lasers. It is important that we obtain a 405nm (blue) laser. This will enable visualization of three different molecules simultaneously and permit analysis of the relationships of several cellular structures with each other. It will also enable use of stains on living cells (eg DAPI), permitting time-lapse studies of movements of subcellular structures, with minimal photobleaching by the laser. Many fluorescent staining procedures require several days at present to complete; we wish to acquire a novel microwave system that accelerates penetration of molecules through membranes without heating them, which will reduce processing time to a few hours and allow researchers to achieve much more in the same time. Finally, several original components of the 2002 CLSM have reached the end of their useful life and require replacement if the CLSM is to continue to operate. Even the computer is 8 years old and cannot run the current software required by the CLSM manufacturer. York University has a distinguished research record in both cell biology and physiology, but this can be maintained only if the requested enhancements are installed in our departmental CLSM.