Capability enhancements for confocal laser scanning microscope

共聚焦激光扫描显微镜（CLSM）是一种强大的工具，使生物学家能够通过实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。组织或单细胞中感兴趣的分子可以通过抗体和荧光标签来识别，当被特定波长的激光照射时，荧光标签会发光。因此，CLSM是世界上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。从那时起，CLSM设备和技术有了很大的改进。在我们的CLSM上，每年大约有10000个小时是由很大一部分部门登录的，它迫切需要一些新的组件和替换某些旧的组件。它只有红色和绿色的激光。我们获得405nm（蓝色）激光是很重要的。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之间的关系。它还可以在活细胞上使用染色剂（如DAPI），允许对亚细胞结构的运动进行延时研究，而激光的光漂白作用最小。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的微波系统，它可以在不加热的情况下加速分子穿过膜的渗透，这将把处理时间缩短到几个小时，并使研究人员能够在相同的时间内完成更多的工作。最后，2002 CLSM的几个原始组件已达到其使用寿命的终点，如果CLSM要继续运行，则需要更换。即使计算机已经使用了8年，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，但只有在我们部门的CLSM中安装了所要求的增强功能，才能保持这一记录。

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.