Artifacts from photobleaching and dye-dissociation in fluorescence correlation spectroscopy

Rudra P. Kafle; Molly R. Liebeskind; Julia T. Bourg; Elizabeth White; Jens-Christian Meiners

doi:10.1117/12.2191402

25 August 2015 Artifacts from photobleaching and dye-dissociation in fluorescence correlation spectroscopy

Rudra P. Kafle, Molly R. Liebeskind, Julia T. Bourg, Elizabeth White, Jens-Christian Meiners

Proceedings Volume 9548, Optical Trapping and Optical Micromanipulation XII; 95481M (2015) https://doi.org/10.1117/12.2191402
Event: SPIE Nanoscience + Engineering, 2015, San Diego, California, United States

Abstract

Fluorescence correlation spectroscopy (FCS) is an optical technique in which the fluctuations in fluorescence intensity are quantified. The time correlation function gives insight into the dynamics of the molecule in its environment: typically the diffusion coefficient in a dilute solution is measured, but the technique has been expanded to uses in more complex environments, including living cells. In these environments photobleaching and dye-dissociation can substantially introduce artifacts in the FCS data. We present a technique to correct for the artifacts introduced by photobleaching and study dye dissociation in DNA solutions.

Citation Download Citation

Rudra P. Kafle, Molly R. Liebeskind, Julia T. Bourg, Elizabeth White, and Jens-Christian Meiners "Artifacts from photobleaching and dye-dissociation in fluorescence correlation spectroscopy", Proc. SPIE 9548, Optical Trapping and Optical Micromanipulation XII, 95481M (25 August 2015); https://doi.org/10.1117/12.2191402

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