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Proceedings Paper

Spatial fluorescence cross-correlation spectroscopy between core and ring pinholes
Author(s): Yoann Blancquaert; Antoine Delon; Jacques Derouard; Rodolphe Jaffiol
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Paper Abstract

Fluorescence Correlation Spectroscopy (FCS) is an attractive method to measure molecular concentration, mobility parameters and chemical kinetics. However its ability to descriminate different diffusing species needs to be improved. Recently, we have proposed a simplified spatial Fluorescence cross Correlation Spectroscopy (sFCCS) method, allowing, with only one focused laser beam to obtain two confocal volumes spatially shifted. Now, we present a new sFCCS optical geometry where the two pinholes, a ring and core, are encapsulated one in the other. In this approach all physical and chemical processes that occur in a single volume, like singlet-triplet dynamics and photobleaching, can be eliminated; moreover, this new optical geometry optimises the collection of fluorescence. The first cross Correlation curves for Rhodamine 6G (Rh6G) in Ethanol are presented, in addition to the effect of the size of fluorescent particules (nano-beads, diameters : 20, 100 and 200 nm). The relative simplicity of the method leads us to propose sFCCS as an appropriate method for the determination of diffusion parameters of fluorophores in solution or cells. Nevertheless, progresses in the ingeniering of the optical Molecular Detection Efficiency volumes are highly desirable, in order to improve the descrimination between the cross correlated volumes.

Paper Details

Date Published: 14 April 2006
PDF: 9 pages
Proc. SPIE 6191, Biophotonics and New Therapy Frontiers, 61910B (14 April 2006); doi: 10.1117/12.662244
Show Author Affiliations
Yoann Blancquaert, Lab. de Spectrométrie Physique, CNRS, Univ. Joseph Fourier (France)
Antoine Delon, Lab. de Spectrométrie Physique, CNRS, Univ. Joseph Fourier (France)
Jacques Derouard, Lab. de Spectrométrie Physique, CNRS, Univ. Joseph Fourier (France)
Rodolphe Jaffiol, Univ. de Technologie de Troyes (France)

Published in SPIE Proceedings Vol. 6191:
Biophotonics and New Therapy Frontiers
Romualda Grzymala; Olivier Haeberle, Editor(s)

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