In this application, we propose to upgrade the existing Department of Biology Optical Sectioning Microscope (OSM) Facility. We are requesting several pieces of equipment that will facilitate the use of the instrument and provide new capabilities that will enhance the facility. The OSM uses the computational methods of out-of-focus light removal to provide high resolution 3-D images of both living and fixed specimens. Its main advantage over confocal microscopy is that is uses far lower levels of illumination to generate high quality images, thereby minimizing the damage to living specimens. Our request for upgrading the facility has four main components. First, we wish to add confocal and partially confocal optics to the OSM, so that depending on the sensitivity of the specimen, we may choose to apply computational methods to either non-confocal, partially confocal or fully confocal images. Second, we propose to add the capability for producing video output so that we have a means to store many movies of 3-D time-lapse sequences for our own analysis, and also a means to record representative movies for presentation to seminar audiences. Third, we request improvements that will facilitate the storage, retrieval and hard copy output of the massive data sets that we work with, so that the data are more easily accessible. Fourth, we seek to add the capability for microinjection to the OSM, so that several different sorts of in vivo injection experiments can be performed expeditiously, The main purpose of all of these enhancements is to make the instrument more accessible to the users and to remedy current shortcomings in the facility. With the addition of these components, our OSM facility will remain on the forefront of biological imaging and continue to facilitate our research for many years to come.
