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Single-molecule detection using continuous wave excitation of two-photon fluorescence |
Optics Letters, Vol. 36, Issue 16, pp. 3185-3187 (2011)
http://dx.doi.org/10.1364/OL.36.003185
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Abstract
Two-photon fluorescence (TPF) is one of the most important discoveries for biological imaging. Although a cw laser is known to excite TPF, its application in TPF imaging has been very limited due to the perceived low efficiency of excitation. Here we directly excited fluorophores with an IR cw laser used for optical trapping and achieved single-molecule fluorescence sensitivity: discrete stepwise photobleaching of enhanced green fluorescent proteins was observed. The single-molecule fluorescence intensity analysis and on-time distribution strongly indicate that a cw laser can generate TPF detectable at the single-molecule level, and thus opens the door to single-molecule TPF imaging using cw lasers.
© 2011 Optical Society of America
OCIS Codes
(140.7010) Lasers and laser optics : Laser trapping
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(180.2520) Microscopy : Fluorescence microscopy
(190.4180) Nonlinear optics : Multiphoton processes
(350.4855) Other areas of optics : Optical tweezers or optical manipulation
ToC Category:
Nonlinear Optics
History
Original Manuscript: May 27, 2011
Revised Manuscript: July 17, 2011
Manuscript Accepted: July 18, 2011
Published: August 11, 2011
Virtual Issues
Vol. 6, Iss. 9 Virtual Journal for Biomedical Optics
Citation
Ximiao Hou and Wei Cheng, "Single-molecule detection using continuous wave excitation of two-photon fluorescence," Opt. Lett. 36, 3185-3187 (2011)
http://www.opticsinfobase.org/ol/abstract.cfm?URI=ol-36-16-3185
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