SGER - Atomic Force Microscopy System with Single Molecule Fluorescence Capabilites

点击翻译按钮获取中文摘要

Single molecule studies can provide unique information on the structure and dynamics ofbiological molecules and cellular complexes that is impossible to obtain using traditionalstructural techniques. These studies, to be performed at conditions close to physiologicalenvironments, open the way for looking directly at the function of individual biologicalmolecules and their complexes. Scanning probe microscopy and atomic force microscope(AFM) in particular has proven to be a very useful technique permitting static and dynamicstudies of molecules at nanometer range resolution. However, the requirement for the sample tobe bound to the surface restricts considerably the dynamical studies. A breakthrough hasrecently been made in the real-time observation of dynamics of molecular complexes at thesingle-molecule level using single-photon sensitive fluorescence microscopes. However, theresolution of this technique is limited by the wavelength of the light and does not revealstructural details of molecules. The combination of AFM and a single molecule fluorescence(SMF) microscope into one instrument would enable us to identify structural details ofbiological systems at the molecular level by AFM and to use the SMF microscope to observedirectly the dynamics of an experimental system at a the single molecule level. Importantly, thedynamics can be performed on molecular systems structurally characterized by AFM andlocalized on a comparatively large surface area. Therefore, the major objective of this proposalis to build AFM/SMF system based on the currently available Bioscope AFM/fluorescencemicroscope instrument. The sensitivity of the fluorescent system will be tested using speciallydesigned samples. In addition, potential pitfalls of integrated AFM will be identified andanalyzed for solutions.