This is a proposal to acquire an atomic force microscope (AFM) on an inverted optical microscope and to develop two AFM-related non-imaging instruments: one for measuring single-molecule force spectroscopy and inter-molecular forces; the other, for measuring elasticities of soft samples under physiological conditions at the nano-scale. Over the past 10 years, atomic force microscopy (AFM) has become an increasingly important tool in biological research. It has gained popularity in biological applications because, unlike electron microscopy, it can image samples under physiological conditions, including live cells undergoing biological processes. The AFM acquires a topographical image of the sample surface by raster scanning an atomically sharp probe over the sample. In addition to its different imaging modes, the AFM is a versatile instrument that can be applied as a nano-indenter and as a molecular force apparatus to probe the mechanical properties of the sample. As a nano-indenter, the AFM has provided direct measurements of the local viscoelastic properties of samples on the nanometer scale. As a molecular force apparatus, the AFM has been used to measure the unbinding force of individual ligand-receptor complexes and the unfolding of individual proteins. Another attractive feature of the AFM is that it can be readily combined with optical microscopy techniques such as FRET, FRAP, TIRF and confocal microscopy. By integrating optical microscopy and AFM into a single experimental platform, the optical image can be directly correlated with the AFM data, providing a powerful tool for studying biological process in situ and in real time.
The acquisition and development of these three instruments is the first step toward establishing an ultramicroscopy center at the university. The two instruments to be developed can be constructed very economically, based on the designs of existing AFMs from the principal investigator's laboratory; this will permit the commercial AFM to be dedicated to imaging applications. The commercial AFM will be the first imaging AFM in the South Florida area and will provide a much needed resource for the local research community. These instruments will provide valuable research opportunities for undergraduates and students from underrepresented groups as well as researchers from different disciplines within the university.
