McConnell Gail
University of Strathclyde, Glasgow, Scotland.
Curr Protoc Cytom. 2006 Nov;Chapter 2:Unit2.13. doi: 10.1002/0471142956.cy0213s38.
Confocal laser scanning microscopy (CLSM) and multiphoton laser scanning microscopy (MPLSM) are methods both widely used by life-sciences researchers for imaging fluorescently labeled live cells and fixed tissue specimens. Key to the success of both CLSM and MPLSM is the application of a suitable laser source, namely one that provides sufficient average or peak power at the correct wavelength to excite fluorescence. High stability of the laser source output is required for three-dimensional imaging, time-lapse studies of live cells, and quantitative studies and inter-experiment comparisons. The laser technology associated with the design of such lasers is mature, yet is unfortunately rather complex. This complexity can be off-putting for the life-sciences researcher who needs to optimize the system for the best possible images, but this apprehension can be overcome by understanding the function of the system components. This unit summarizes the optimization of the most commonly used laser sources for CLSM and MPLSM, including power and wavelength tuning and methods for cleaning optical components.
共聚焦激光扫描显微镜（CLSM）和多光子激光扫描显微镜（MPLSM）都是生命科学研究人员广泛用于对荧光标记的活细胞和固定组织标本进行成像的方法。CLSM和MPLSM成功的关键在于应用合适的激光源，即在正确波长下提供足够平均功率或峰值功率以激发荧光的激光源。三维成像、活细胞的延时研究、定量研究以及实验间比较都需要激光源输出具有高稳定性。与这类激光器设计相关的激光技术已经成熟，但不幸的是相当复杂。这种复杂性可能会让需要优化系统以获得最佳图像的生命科学研究人员望而却步，但通过了解系统组件的功能可以克服这种担忧。本单元总结了CLSM和MPLSM最常用激光源的优化方法，包括功率和波长调谐以及光学组件清洁方法。
