Single Molecule Force Spectroscopy with a Micropipette
Abstract:To directly dissect molecular interactions between single macromolecules with dynamic force spectroscopy, measurement of small forces and precision in position are two of the critical factors for success. Here we described a relatively simple instrument incorporated on the platform of an inverted fluorescence microscope using a pulled micropipettes to detect sub-piconewton forces. The hollow micropipette allows convenient exchange of the molecules being studied with the same force sensor and, thus, is specifically suited for detecting small differences introduced by point mutation or mild external conditions. Moreover, such a system is directly compatible with single channel electrophysiology and single molecule fluorescence with only moderate complications in the instrumentation. The capabilities of this system are demonstrated with applications to the rupture of the biotin–streptavidin complex and the extraction of a biotinylated lipid molecule from a bilayer, which have produced results in excellent agreement with previous measurements.
Document Type: Research Article
Publication date: 2006-06-01
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- Scanning probe microscopy (SPM) typically covers atomic force microscopy (AFM), scanning tunneling microscopy (STM), near-field scanning optical microscopy (NSOM, or SNOM) and related technologies. Since its invention in early 1980s, SPM has now been regarded as one of the major driving forces for the rapid development of nanoscience and nanotechnology, and the tool of choice in many areas of research. Journal of Scanning Probe Microscopy (JSPM) provides a forum for rapid dissemination of important developments in SPM technology. JSPM offers scientists, engineers and developers timely, peer-reviewed research on SPM science and technology of the highest quality. JSPM publishes original rapid communications, full research papers and timely state-of-the-art reviews (with author's photo and biography) encompassing the fundamental and applied research on SPM in all fields of science, engineering, and medicine.
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