Fluorescent imaging fluorescence correlation spectroscopy

Finally, in Chapter 11 some advanced techniques are briefly described fluorescence up-conversion, fluorescence microscopy (confocal excitation, two-photon excitation, near-field optics, fluorescence lifetime imaging), fluorescence correlation spectroscopy, and single-molecule fluorescence spectroscopy. 

Z. P. Huang and N. L. Thompson, Imaging fluorescence correlation spectroscopy Nonuniform IgE distributions on planar membranes, Biophys. J. 70, 2001-2007 (1996). 

Fluorescence Correlation Spectroscopy and Fluorescence Burst Analysis. Several nanoscopic chemical imaging approaches work very well for measurements of chemical kinetics, interactions, and mobility in solution. Fluorescence correlation spectroscopy (FCS) measures the temporal fluctuations of fluorescent markers as molecules diffuse or flow in solution through a femtoliter focal volume.54 Their average diffusive dwell times reveal their diffusion coefficients, and additional faster fluctuations can reveal chemical reactions and their kinetics if the reaction provides fluorescence modulation. Cross-correlation of the fluorescence of two distinguishable fluorophore types can very effectively reveal chemical binding kinetics and equilibria at nanomolar concentrations. 

FCS FT fMRI FTIR Fluorescence Correlation Spectroscopy Fourier Transform Functional Magnetic Resonance Imaging Fourier Transform Infrared Spectroscopy... 

Whereas FCS measures fluorescence fluctuations over time, a related technique, ICS, measures fluorescence fluctuations over space, in particular from images collected using a laser-scanning microscope (56). ICS analysis of pixels within a single image provides information about protein clustering and density. A variation of ICS known as image cross-correlation spectroscopy evaluates the interactions of molecules labeled with different fluorescent probes. ICS can also be performed on stacks of... 

Thompson NL, Steele BL. Total internal reflection with fluorescence correlation spectroscopy. Nat. Protoc. 2007 2 878-890. Sund SE, Axelrod D. Actin dynamics at the living cell submembrane imaged by total internal reflection fluorescence photobleach-ing. Biophys. J. 2000 79 1655-1669. 

Q. Ruan, Y. Chen, E. Gratton, M. Glaser, W.M. Mantulin, Cellular characterization of adenylate kinase and its isoform two-photon excitation fluorescence imaging and fluorescence correlation spectroscopy, Biophys. J. 83, 3177-3187(2002)... 

W.R. Zipfel, W.W.W. Webb, In vivo Diffusion Measurements Using Multiphoton Excitation Fluorescence Photobleaching Recovery and Fluorescence Correlation Spectroscopy, in A. Periasamy (ed.), Methods in Cellular Imaging, Oxford University Press, 216 (2001)... 

M. Doroshenko, et al., Monitoring the dynamics of phase separation in a polymer blend by confocal imaging and fluorescence correlation spectroscopy, Macromol. Rapid Commun. 33 (18) (2012) 1568—1573. 

Considering potential pharmaceutical applications, encapsulation of different substrates at the same time is demanded but is also a scientific challenge [267]. The polymersome system presented here offers the possibility for encapsulation of both hydrophilic and hydrophobic substances at the very same time in the very same polymersome. Potential loading systems are typically limited by the lack of suitable characterization methods. We utilized fluorescence correlation spectroscopy (PCS) in combination with cryogenic TEM imaging and DLS to characterize hydrophobic loading. 

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