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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics

| EXPLORING THE INTERFACE OF LIGHT AND BIOMEDICINE

  • Editors: Andrew Dunn and Anthony Durkin
  • Vol. 6, Iss. 1 — Jan. 3, 2011

EMCCD-based spectrally resolved fluorescence correlation spectroscopy

Felix Bestvater, Zahir Seghiri, Moon Sik Kang, Nadine Gröner, Ji Young Lee, Kang-Bin Im, and Malte Wachsmuth  »View Author Affiliations


Optics Express, Vol. 18, Issue 23, pp. 23818-23828 (2010)
http://dx.doi.org/10.1364/OE.18.023818


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Abstract

We present an implementation of fluorescence correlation spectroscopy with spectrally resolved detection based on a combined commercial confocal laser scanning/fluorescence correlation spectroscopy microscope. We have replaced the conventional detection scheme by a prism-based spectrometer and an electron-multiplying charge-coupled device camera used to record the photons. This allows us to read out more than 80,000 full spectra per second with a signal-to-noise ratio and a quantum efficiency high enough to allow single photon counting. We can identify up to four spectrally different quantum dots in vitro and demonstrate that spectrally resolved detection can be used to characterize photophysical properties of fluorophores by measuring the spectral dependence of quantum dot fluorescence emission intermittence. Moreover, we can confirm intracellular cross-correlation results as acquired with a conventional setup and show that spectral flexibility can help to optimize the choice of the detection windows.

© 2010 OSA

OCIS Codes
(040.1520) Detectors : CCD, charge-coupled device
(180.1790) Microscopy : Confocal microscopy
(300.6280) Spectroscopy : Spectroscopy, fluorescence and luminescence

ToC Category:
Spectroscopy

History
Original Manuscript: July 13, 2010
Revised Manuscript: September 29, 2010
Manuscript Accepted: October 1, 2010
Published: October 27, 2010

Virtual Issues
Vol. 6, Iss. 1 Virtual Journal for Biomedical Optics

Citation
Felix Bestvater, Zahir Seghiri, Moon Sik Kang, Nadine Gröner, Ji Young Lee, Kang-Bin Im, and Malte Wachsmuth, "EMCCD-based spectrally resolved fluorescence correlation spectroscopy," Opt. Express 18, 23818-23828 (2010)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=oe-18-23-23818


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