In response to PAR-09-028: Shared Instrumentation Grant Program (S10), this application requests support for a state-of-the-art BioScope Catalyst Atomic Force Microscope (AFM) system (Veeco Instruments, Plainview, NY) integrated with a new Olympus IX81 inverted microscope equipped with a Fluoview FV1000 confocal laser scanning system (LCSM, Olympus, Center Valley, PA) to be housed in the core facilities of the Department of Biomedical Engineering at Columbia University. Motivation for this request stems from the fact that there are currently no core facilities at Columbia University (or that we are aware of in New York City) that provide an AFM system with confocal capabilities for general life sciences applications. The AFM has become a staple instrument for biomedical engineering research, providing an important imaging and material testing tool spanning applications from single molecules to tissues. The integrated system will be used by multiple NIH- supported investigators for at least 85% of the time and will be made accessible for other researchers and academic purposes on an availability basis. The requested BioScope Catalyst system is user-friendly with integrated microscopy and analyses software (MIRO), as well as an open scan head design that permits simultaneous transmitted light applications, easy sample access and can be integrated with the FV1000 LSCM system. A microscope configuration combining these imaging sytems can be used to great advantage in cellular and tissue-level biomedical research where structures imaged with AFM can be cross-correlated to biomolecules identifiable via the LCSM. Similarly, using this system, AFM force measurements can permit mapping of sample mechanical stiffness to be correlated with underlying fluorescently labeled matrix structures visible by LSCM. Lastly, measurement of cellular responses using LCSM in response to AFM-induced mechanical stimulation provides unique opportunities for studies of mechanobiology, which investigates how cells perceive and respond to their physical environment. Further enhancing the capabilities of this integrated AFM system, the Fluoview FV1000 SIM (SIMultaneous) module provides photoactivation or bleaching and simultaneous image collection, enabling fluorescence recovery after photobleaching (FRAP) and molecular uncaging applications. Therefore, a research platform that delivers combined AFM and LSCM capabilities would offer a powerful research tool for biological applications. The BME department is home to the largest number of NIH-supported investigators at the downtown campus and is well-poised to lead this Shared Instrumentation effort, serving to further bridge the two campuses (downtown Morningside Heights to the uptown Medical Campus) and foster interdisciplinary research collaborations at Columbia.
