Two‐photon image correlation spectroscopy and image cross‐correlation spectroscopy

  title={Two‐photon image correlation spectroscopy and image cross‐correlation spectroscopy},
  author={Paul W. Wiseman and Jeffrey Squier and Mark Ellisman and Kent R. Wilson},
  journal={Journal of Microscopy},
We introduce two‐photon image correlation spectroscopy (ICS) using a video rate capable multiphoton microscope. We demonstrate how video rate two‐photon microscopic imaging and image correlation analysis may be combined to measure molecular transport properties over ranges typical of biomolecules in membrane environments. Using two‐photon ICS, we measured diffusion coefficients as large as 10−8 cm2 s−1 that matched theoretical predictions for samples of fluorescent microspheres suspended in… 
k-Space image correlation spectroscopy: a method for accurate transport measurements independent of fluorophore photophysics.
We present the theory and application of reciprocal space image correlation spectroscopy (kICS). This technique measures the number density, diffusion coefficient, and velocity of fluorescently
Image correlation microscopy for uniform illumination
It is shown that image correlation can also be used to measure fluorescence dynamics in uniform illumination or wide‐field imaging systems and is called uniform illumination image correlation microscopy, and it is established that the temporal autocorrelation function depends strongly on particle size and not particle shape.
Fluorescence Correlation Spectroscopy and Fluorescence Lifetime Imaging Microscopy
FCS and FLIM allow information to be gathered beyond localization, including diffusional mobility, protein clustering and interactions, and molecular environment.
Arbitrary-Region Raster Image Correlation Spectroscopy
High Speed Two Photon Excitation Microscopy in Live Cell Imaging using Image Correlation Spectroscopy (ICS)
For live-cell imaging, two-photon excitation microscopy (TPEM) is proving to be a significant technological advancement. The unique features offered by TPEM are the ability to image thick sections,
Measuring fast dynamics in solutions and cells with a laser scanning microscope.
Novel extensions to ICS are developed that probe spatial correlations in previously inaccessible temporal windows and are used to determine for the first time spatially resolved diffusions of paxillin-EGFP stably expressed in CHOK1 cells.
Spatially resolved fluorescence correlation spectroscopy using a spinning disk confocal microscope.
We develop an extension of fluorescence correlation spectroscopy (FCS) using a spinning disk confocal microscope. This approach can spatially map diffusion coefficients or flow velocities at up to
Dynamic range and background filtering in raster image correlation spectroscopy
This study investigates methods to filter out immobile or slow moving background structures and their impact on RICS results, and the capacity of RICS analysis to separate two species is studied.
Brief introduction to fluorescence correlation spectroscopy.
  • E. Elson
  • Physics, Chemistry
    Methods in enzymology
  • 2013


Two-photon fluorescence correlation spectroscopy: method and application to the intracellular environment.
Image cross-correlation spectroscopy: A new experimental biophysical approach to measurement of slow diffusion of fluorescent molecules
The technique of image cross-correlation spectroscopy has been applied to measurements of diffusion of fluorescent beads in polymer solution. The technique is based on measurement and analysis of
Two-photon laser scanning fluorescence microscopy.
The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation.
Fluorescence correlation spectroscopy. III. Uniform translation and laminar flow
We extend fluorescence correlation spectroscopy to systems that undergo translation or laminar flow in a sample cell. We include theoretical and experimental results; we consider uniform and
Fluorescence correlation spectroscopy. II. An experimental realization
This paper describes the first experimental application of fluorescence correlation spectroscopy, a new method for determining chemical kinetic constants and diffusion coefficients. These quantities