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. ? ? ?
