PROJECT SUMMARY – IMAGING SHARED RESOURCE The primary goal of the Imaging Shared Resource (ISR) is to provide Wistar Institute Cancer Center members with comprehensive and flexible imaging solutions, ranging from high-end microscopy and small animal imaging to complete image analysis services. In recent years, technological advances have made it possible to utilize quantitative, high resolution imaging approaches to dissect spatio-temporal aspects of cellular behavior related to malignant transformation, metastatic dissemination, and therapeutic response, and the ISR has amply positioned itself to support inquiries into these areas. During the last budget cycle, the ISR was further enhanced with new scientific leadership, additional personnel and a significant expansion of new instrumentation and facility renovations. The ISR continues to offer basic widefield, upright and inverted microscopy, as well as low magnification and specimen imaging for routine applications, but has also invested in a new Leica SP8X WLL white-light scanning confocal microscope to complement an existing Leica SP5 confocal. Combined, these systems respond to an increasing need for advanced microscopy usage within the Cancer Center. This new instrument is optimized for high resolution, rapid, live-cell imaging and has provided the instrumentation needed to successfully investigate dynamic, intracellular mechanisms in multiple dimensions. Additionally, a new Leica SP8 MP 2-Photon, intravital microscope has replaced an obsolete Prairie Instruments model and is capable of providing not only single session, deep imaging support in both live and fixed models, but also longitudinal studies of metastatic growth and tumorigenesis in live mouse models. A Nikon TiE inverted widefield system was also added to the facility and is especially useful for highly automated imaging including large area stitched acquisition of slides and plates. The TiE is fully configured for high-content, time-lapse recording and is set up for continuous, real-time image analysis. To further support longitudinal imaging studies in live mouse models designed to interrogate metastatic progression and response to targeted therapies, the ISR applied for, and was awarded an S10 shared instrument grant (S10 OD021669) for the purchase of a new Perkin Elmer IVIS SpectrumCT small animal imaging system. This new instrument replaced an older IVIS 200 and adds new functionality in the form of an integrated microCT unit as well as quantitative opportunities in 3D for more accurate localization and measurement of tumor burden. The highly experienced staff works directly with users to acquire their images, or trains users to operate equipment independently. The ISR specializes in supporting complex, multimodal imaging experiments including FRAP, FRET and 6D projects utilizing 3D tracking. The ISR stresses ethical practices in all aspects of image acquisition, post-process image manipulation and analysis. The overall miss...