This application is for the purchase of the Nikon A1 confocal microscope, a laser line scanning confocal microscope system to support and augment the imaging capabilities of the user group within the Heart Institute. The new instrumentation will enable the user group to carry out state of the art imaging for all of their NIH-funded investigations at a level of resolution that is not currently possible with the existing instrument. It will also provide new capabilities to the user group;capabilities that have become critical to the group as they explore the phenotypes of their disease models of human cardiovascular disease. These new capabilities include: simultaneous and sequential 4 channel detection plus 1 transmitted light detection channel for DIC, FRAP/ photoactivation studies, a completely motorized confocal microscope with preprogrammed and custom programmed optical acquisition modes for ease of use, multipoint acquisition scanning that can be used in conjunction with Nikon's Perfect Focus System for long-term time lapse experiments, and large image stitching. The instrument will enable vital imaging, automatic focus during acquisition of dynamic processes such as aggresome formation and intracellular trafficking, imaging of calcium flux and the ability to use an increased range of fluorescent tags. Proposed use of this instrumentation will support and enhance existing NIH-funded projects and provide avenues of research support that will lead to new funding for future projects. Five NIH-supported users of this equipment will form the core research team and the value added to their research activities by this instrumentation is discussed. All of these individuals currently interact within the Heart Institute and are the principal users of the existing equipment. The identified projects are untenable using the equipment already in place. The group's research interests cover a wide spectrum but all deal with analyzing mouse and cell-based models of human cardiovascular disease in which cause-and-effect relationships are explored. All depend upon the ability to accurately image molecules at the subcellular level. The PI, who has been active in the field of heart research for ~23 years, will be responsible for the managerial oversight. He will report to an internal advisory board composed of scientists from various Cincinnati Children's Hospital departments and divisions. The equipment will be housed in the new research building at Cincinnati Children's Hospital Research Foundation. This 450,000 sq. ft. facility opened in 2008 and provides state of the art support and infrastructure for the instrument's operation. A dedicated research suite is already set aside for confocal analyses within the Heart Institute and will house this instrument: the space consists of a 450 sq ft room equipped with special airflow. The instrumentation will be operated and maintained by the PI and a Ph.D.-level facility staff scientist. Together these personnel have over 45 years of imaging experience, ensuring that the services will be provided in a timely manner.
| Meng, Qinghang; Bhandary, Bidur; Bhuiyan, Md Shenuarin et al. (2018) Myofibroblast-Specific TGF? Receptor II Signaling in the Fibrotic Response to Cardiac Myosin Binding Protein C-Induced Cardiomyopathy. Circ Res 123:1285-1297 |
| James, Jeanne; Robbins, Jeffrey (2016) Healing a Heart Through Genetic Intervention. Circ Res 118:920-2 |
| Gupta, Manish K; Gulick, James; James, Jeanne et al. (2013) Functional dissection of myosin binding protein C phosphorylation. J Mol Cell Cardiol 64:39-50 |
| Razzaque, Md Abdur; Gupta, Manish; Osinska, Hanna et al. (2013) An endogenously produced fragment of cardiac myosin-binding protein C is pathogenic and can lead to heart failure. Circ Res 113:553-61 |
| Bhuiyan, Md Shenuarin; Gulick, James; Osinska, Hanna et al. (2012) Determination of the critical residues responsible for cardiac myosin binding protein C's interactions. J Mol Cell Cardiol 53:838-47 |
