This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).
This Major Research Instrumentation-Recovery and Reinvestment (MRI-R2) award funds the development of two versatile, modular fluorescence microscopy platforms for tracking and mechanically probing single molecules/particles at the University of Michigan (UM), and the installation of a third, user-friendly microscope system for user training and project development. The development and application of two open-access single molecule tracking and probing microscopes transforms the limited capability for high-content screening of biological samples. Benchmarks are set that make both instruments operational within two years and lay out plans for testing, further optimization and expansion over an additional two years, after which recharges will maintain them. The instruments are being utilized for research and education by the University of Michigan community and beyond, engaging physical and biological scientists as well as an engineer. This development activity helps establish a unique, nation-wide, educational Center of Single Molecule Analysis (CoSiMA) in a highly cross-disciplinary, emerging field of transformative power in the biosciences. CoSiMA is engaging in vigorous outreach efforts to experimentalists, theorists, undergraduates, and high school students with large, local, underrepresented groups, as well as the public at large, in close collaboration with UM's existing IDEA Institute. These efforts are facilitated by the creation of a central laboratory facility with access to multiple single molecule microscopes under guidance of a Steering Committee and a senior staff scientist, as well as by the broad inspirational appeal of the emerging single molecule biosciences. Results from the studies enabled by the new equipment are disseminated by student and faculty presentations at regional and national meetings, and through publication in peer-reviewed journals.
This project established the Single Molecule Analysis in Real-Time (SMART) Center at the University of Michigan, a unique suite of shared resources optimized for localization and characterization of single biomolecules. The Center is situated in ~1,000 sqft of specifically renovated space and currently hosts four single molecule microscopes: the Modular Super-Resolution Single-Particle Tracker (SPT), Single-Particle Nanomanipulator (SPN), time-resolved confocal ALBA, and a small atomic force microscope (AFM). The SMART lab also hosts a workstation and relevant software for post-processing and analysis. Additionally, the Center aims to provide support to users by building a local network of shared expertise in single molecule experimental design and analysis. At the conclusion of this funding period, the Center has successfully transitioned to a sustaining funding model based on user fees and college subsidy. Four specialized microscopes were assembled to establish this Center. The Single Particle Tracker offers localization of single fluorescent particles with nanoscale precision and allows users to track such particles’ motion in real-time. The time-resolved confocal microscope enables optical sectioning of samples and high-bandwidth detection of particle motions and intermolecular associations. The single particle nanomanipulator is a force microscope that uses light to hold and manipulate individual particles and measure forces down to the piconewton range, the range of forces that individual biomolecules exert. The atomic force microscope enables nanoscale detailed probing of sample topography. These tools provide researchers with the ability to probe the characteristics of and interactions between individual molecules such as proteins, RNA, or drugs. This type of investigation is critical in understanding the detailed mechanisms by which these complex macromolecules function. This leads to better understanding, for example, of how perturbation of these functions may either result in or alleviate disease conditions, or enable informed synthesis of novel macromolecules with designed functionality. The completion of this Center, and its successful transition to a sustained shared user facility, provides this unique suite of microscopy tools assembled specifically to analyze the nanoscale behavior of biomolecules to university researchers and to the larger research community. Individual investigators can now leverage the equipment and expertise of this pooled centralized resource to address questions relevant to basic science and nanotechnology that would be burdensome, both technically and financially, to pursue individually. Furthermore, the Center is committed to working with individual researchers to enable them to pursue their questions most effectively, implementing custom modifications and analysis support when appropriate. The modularity of the instrumentation and expertise of the center staff and collaborators facilitates such modification and enhancements to the facility in response to user needs. Similarly, we expect to successfully pursue resources to update instrumentation as the needs of the research community evolve. In addition to the resources provided specifically by the Center, we also provide a logical central location for university knowledge resources related to single molecule studies. We have contact with a significant portion of investigators at the university with expertise relevant to single molecule investigations. By providing a central location to aggregate these investigators’ knowledge, we can facilitate exchange of knowledge between specialties.
