期刊
REVIEW OF SCIENTIFIC INSTRUMENTS
卷 81, 期 8, 页码 -出版社
AMER INST PHYSICS
DOI: 10.1063/1.3473936
关键词
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资金
- Howard Hughes Medical Institute Funding Source: Medline
We describe an approach for performing single-molecule binding experiments on time scales from hours to days, allowing for the observation of slower kinetics than have been previously investigated by single-molecule techniques. Total internal reflection fluorescence microscopy is used to image the binding of labeled ligand to molecules specifically coupled to the surface of an optically transparent flow cell. Long-duration experiments are enabled by ensuring sufficient positional, chemical, thermal, and image stability. Principal components of this experimental stability include illumination timing, solution replacement, and chemical treatment of solution to reduce photodamage and photobleaching; and autofocusing to correct for spatial drift. (C) 2010 American Institute of Physics. [doi:10.1063/1.3473936]
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