Upgrade of Femtosecond Laser Sources for Optical Workstation

描述(申请人提供)：实验性激光微束技术已成为研究活标本的既定工具。可定向聚焦的激光光束可用于各种实验操作，如激光显微手术、光学捕获、笼式生物活性探针的局部光解和图案化光漂白。通常，为这些应用中的每一种都构建了专门设计的实验系统。为了评估这种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。多光子激发，由于其能够在光毒效应最小的情况下从标本内部获得高对比度图像，是体内成像的首选技术。我们开发了一种光学工作站(OWS)，它结合了实验光学微束设备的功能和专为活体标本设计的灵敏多光子成像系统。该系统是由威斯康星大学麦迪逊分校光学和计算仪器实验室(LOCI)开发的。与使用独立成像和实验组件实施的系统相比，集成光学工作站概念在灵活性和通用性方面具有优势。此应用程序用于设备以增强OWS的能力，并替换已废弃的组件。需要两个飞秒激光光源。这两种资源都将取代/升级现有的激光器：一种是老式的钛宝石激光器，另一种是现已废弃的1047激光器。所要求的激光器是：一台麦台计算机可调谐的钛宝石激光器，调谐范围为700-1000 nm，以及一台1030 nm的锁定t-Pulse 20激光器。激光器将与目前在OWS上运行的激光消融和双扫描控制系统一起运行。这将既增加新的功能，又恢复失去的功能，例如双荧光激发以及同时的光学捕获和去胶合。升级后的OWS将通过改进功能更好地服务于当前项目，并通过培育能够利用其扩展的波长和操作能力的新项目，在华盛顿大学园区发挥更大的作用。与公共卫生相关：对活细胞和组织进行非侵入性成像的能力对公共卫生具有巨大的潜在益处，因为它可以为癌症进展等关键疾病过程提供新的见解。所要求的激光光源将使几个与健康相关的成像项目受益，范围从研究细胞分裂到了解信号在乳腺癌中的作用。

DESCRIPTION (provided by applicant): 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. We have developed an Optical Workstation (OWS) that combines the functionality of an experimental optical microbeam apparatus with a sensitive multiphoton imaging system designed for use with living specimens. The system was developed at the Laboratory for Optical and Computational Instrumentation (LOCI) at the University of Wisconsin, Madison. The integrated optical workstation concept offers advantages in terms of flexibility and versatility relative to systems implemented with separate imaging and experimental components. This application is for equipment to enhance the capabilities of the OWS and to replace a defunct component. Two femtosecond laser sources are requested. Both sources will replace/upgrade existing lasers: an older Ti:Sapphire laser and a now defunct 1047 laser. The requested lasers are: a Mai Tai computer tunable Ti:Sapphire laser with a tuning range of 700-1000nm and a 1030nm modelocked t-Pulse 20 laser. The lasers will operate with the laser ablation and dual scanning control systems currently functioning on the OWS. This will both add new features and restore lost functionality, such as dual fluorophore excitation together with simultaneous optical trapping and uncaging. The upgraded OWS will have an expanded role on the UW campus by better serving current projects through improved functionality, and by fostering new projects that can take advantage of its expanded wavelength and manipulation capabilities. Relevance to Public Health: The ability to image non-invasively into live cells and tissue is of great potential benefit to public health as it can provide new insight into key disease processes such as cancer progression. The requested laser sources will benefit several health related imaging projects ranging from studying cell division to understanding the role of signaling in breast cancer.