RIS ID
94197
Abstract
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 imagestability. 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.
Publication Details
Elenko, M. P., Szostak, J. W. & van Oijen, A. M. (2010). Single-molecule binding experiments on long time scales. Review of Scientific Instruments, 81 (8), 083705-1-083705-9.