Intravital confocal imaging often requires multiple fluorescent markers with similar spectral properties, the ability to capture highly dynamic events, and stringent specimen environmental needs. Specialized instrumentation optimized for acquisition speed, spatial resolution, or optical flexibility has evolved to address the individual needs of these applications, but no single system has provided the optimal combination of these performance indices. University of Cincinnati researchers have a strong history of quality microscopic imaging and a number of confocal systems at their disposal, but none of the currently available systems offer a suitable combination of speed, quantum efficiency, optical flexibility, and environmental control for optimal intravital biomedical research. This application requests an advanced dual-scanner confocal microscope, the Zeiss LSM 7 Duo, for the Live Microscopy Core located at the UC College of Medicine, Department of Molecular and Cellular Physiology. This advanced confocal microscope utilizes a pair of confocal scan heads optimized to provide coordinated high-resolution and high speed imaging capacity for live cell, tissue, and small animal imaging. The system will serve the legitimate scientific needs of our 4 major research groups, the established core facility client base, and the regional scientific community. The groups require this instrumentation to: 1) permit ultra-high speed imaging for dynamic in vivo processes, 2) provide spectral imaging capability for enhanced fluorophore separation and detection, and 3) accommodate complex photobleaching and photoactivation experiments that require concurrent manipulation and imaging. All major users have demonstrated a significant need for this instrumentation in support of their ongoing NIH-funded research projects. The needs of our users cannot be met by the equipment currently at our institution as no other available system provides the necessary combination of speed and resolution. Combining a fully-automated microscope platform with a 5Live high-speed confocal scanner, a high-resolution LSM710 scanner, and a computer-controlled environmental chamber, this workstation will ensure an optimal imaging environment for our demanding intravital biomedical research projects. The Zeiss LSM 7 Duo confocal will be installed within the current imaging core facility near to the Core facility manager, next to the image processing and analysis suite, and central to all of the investigators. This application is supported by substantial interest, need and financial support at the University, and will serve a strongly NIH-funded scientific community. We have secured funding from the University ($296,000) to assist with the initial instrumentation purchase and departmental funds for ongoing maintenance costs, service contracts, and personnel to run the microscope on a day-to-day basis.
