Characterization of the conformational changes of a tripartite molecular beacon

Cecile Couzon; Daniel Banks; James Jonkman; Aleksandra Dabkowska; Razvan Nutiu; Yingfu Li; Cecile Fradin

doi:10.1117/12.537423

21 June 2004 Characterization of the conformational changes of a tripartite molecular beacon

Cecile Couzon, Daniel Banks, James Jonkman, Aleksandra Dabkowska, Razvan Nutiu, Yingfu Li, Cecile Fradin

Proceedings Volume 5323, Multiphoton Microscopy in the Biomedical Sciences IV; (2004) https://doi.org/10.1117/12.537423
Event: Biomedical Optics 2004, 2004, San Jose, CA, United States

Abstract

We have investigated the kinetics of the conformational changes of a new type of molecular beacons called tripartite molecular beacons. The rate constants corresponding to the opening and closing of the beacons have been obtained from fluorescence correlation spectroscopy experiments. We found that both rate constants are larger for the tripartite molecular beacon than for the corresponding molecular beacon. This paper outlines the importance of using very low excitation intensities for this type of measurements, and of considering the fact that the beacon still emits a residual background fluorescence in its closed form. We also report on the exploration of several strategies to improve the precision of the measurements by increasing the characteristic time associated with the diffusion of the beacons so that it would not be confused with the relaxation time associated with the conformational changes.

Citation Download Citation

Cecile Couzon, Daniel Banks, James Jonkman, Aleksandra Dabkowska, Razvan Nutiu, Yingfu Li, and Cecile Fradin "Characterization of the conformational changes of a tripartite molecular beacon", Proc. SPIE 5323, Multiphoton Microscopy in the Biomedical Sciences IV, (21 June 2004); https://doi.org/10.1117/12.537423

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