Year: 2010
Type: article
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 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.
Source: Review of Scientific Instruments
DOI: 10.1063/1.3473936
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: hybrid
Funder: Howard Hughes Medical Institute
OpenAlex ID: W2012555900