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Proceedings Article

Measurement of cell surface protein dynamics by two-photon image correlation spectroscopy and image cross-correlation spectroscopy

[+] Author Affiliations
Paul W. Wiseman

McGill Univ. (Canada)

Jeffrey A. Squier

Univ. of California/San Diego (USA)

Proc. SPIE 4633, Commercial and Biomedical Applications of Ultrafast and Free-Electron Lasers, 74 (April 5, 2002); doi:10.1117/12.461365
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From Conference Volume 4633

  • Commercial and Biomedical Applications of Ultrafast and Free-Electron Lasers
  • Glenn S. Edwards; Joseph Neev; Andreas Ostendorf; John C. Sutherland
  • San Jose, California, United States | January 20, 2002

abstract

Advances in laser-scanning microscopy and the advent of confocal microscopy permitted the development of image correlation spectroscopy (ICS). ICS is an imaging analog of fluorescence correlation spectroscopy (FCS) optimized for measuring the aggregation state of fluorescently labeled macromolecules on the surface of biological cells. The ICS method entails spatial autocorrelation analysis of fluorescence fluctuations within an image sampled from an area of the sample as well as temporal autocorrelation analysis of fluorescence fluctuations through a time series of images. Together, the spatial/temporal autocorrelation analysis enables measurement of fluorophore concentration, aggregation state and transport properties. ICS was first implemented on a confocal laser-scanning microscope (CLSM) using single photon excitation. More recently we have extended the method for two-photon ICS as well as image cross-correlation spectroscopy (ICCS). ICCS allows measurement of co-localization of non-identical molecules labeled with fluorophores of different emission wavelengths. We present a variety of applications of the ICS and ICCS methods in cellular systems. We will discuss the measurement of the transport and clustering properties of membrane receptors by single photon ICS and two-photon ICCS. As well, we will describe how spatial ICS may be used to quantify the distribution of fluorescently labeled dendritic spines in neurons.

© (2002) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.
Citation

Paul W. Wiseman and Jeffrey A. Squier
"Measurement of cell surface protein dynamics by two-photon image correlation spectroscopy and image cross-correlation spectroscopy", Proc. SPIE 4633, Commercial and Biomedical Applications of Ultrafast and Free-Electron Lasers, 74 (April 5, 2002); doi:10.1117/12.461365; http://dx.doi.org/10.1117/12.461365


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