Document Type
Article
Publication Date
6-21-2013
Abstract
We present a hardware-based method that can improve single molecule fluorophore observation time by up to 1500% and super-localization by 47% for the experimental conditions used. The excitation was modulated using an acousto-optic modulator (AOM) synchronized to the data acquisition and inherent data conversion time of the detector. The observation time and precision in super-localization of four commonly used fluorophores were compared under modulated and traditional continuous excitation, including direct total internal reflectance excitation of Alexa 555 and Cy3, non-radiative Förster resonance energy transfer (FRET) excited Cy5, and direct epi-fluorescence wide field excitation of Rhodamine 6G. The proposed amplitude-modulated excitation does not perturb the chemical makeup of the system or sacrifice signal and is compatible with multiple types of fluorophores. Amplitude-modulated excitation has practical applications for any fluorescent study utilizing an instrumental setup with time-delayed detectors.
Language
English
Publication Title
Methods and Applications in Fluorescence
Grant
GM94246-01A1
Rights
© 2013 IOP Publishing Ltd. This is the peer reviewed Accepted Manuscript version of an article accepted for publication. IOP Publishing Ltd is not responsible for any errors or omissions in this version of the manuscript or any version derived from it. This version is published under a Creative Commons CC-BY-NC-ND licence (https://creativecommons.org/licenses/BY-NC-ND/4.0/), which permits non-commercial copying and redistribution of the material in any medium or format, provided the original work is not changed in any way and is properly cited.
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Recommended Citation
Kisley, L., Chang, W. S., Cooper, D., Mansur, A. P., & Landes, C. F. (2013). Extending single molecule fluorescence observation time by amplitude-modulated excitation. Methods and applications in fluorescence, 1(3), 037001.
Manuscript Version
Accepted Manuscript