Wokosin David L, Squirrell Jayne M, Eliceiri Kevin W, White John G
Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison, Madison, Wisconsin 53706.
Rev Sci Instrum. 2003 Jan;74(1):193-201. doi: 10.1063/1.1524716.
Experimental laser microbeam techniques have become established tools for studying living specimens. A steerable, focused laser beam may be used for a variety of experimental manipulations such as laser microsurgery, optical trapping, localized photolysis of caged bioactive probes, and patterned photobleaching. Typically, purpose-designed experimental systems have been constructed for each of these applications. In order to assess the consequences of such experimental optical interventions, long-term, microscopic observation of the specimen is often required. Multiphoton excitation, because of its ability to obtain high-contrast images from deep within a specimen with minimal phototoxic effects, is a preferred technique for in vivo imaging. An optical workstation is described that combines the functionality of an experimental optical microbeam apparatus with a sensitive multiphoton imaging system designed for use with living specimens. Design considerations are discussed and examples of ongoing biological applications are presented. The integrated optical workstation concept offers advantages in terms of flexibility and versatility relative to systems implemented with separate imaging and experimental components.
实验性激光微束技术已成为研究活体标本的既定工具。可操纵的聚焦激光束可用于多种实验操作,如激光显微手术、光镊、笼状生物活性探针的局部光解以及图案化光漂白。通常,针对这些应用中的每一种都构建了专门设计的实验系统。为了评估此类实验性光学干预的后果,通常需要对标本进行长期的显微镜观察。由于多光子激发能够以最小的光毒性效应从标本深处获得高对比度图像,因此它是体内成像的首选技术。本文描述了一种光学工作站,它将实验性光学微束装置的功能与专为活体标本设计的灵敏多光子成像系统相结合。讨论了设计考量,并展示了正在进行的生物学应用实例。相对于由单独的成像和实验组件实现的系统,集成光学工作站概念在灵活性和通用性方面具有优势。