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Article . 2011 . Peer-reviewed
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Multi-confocal fluorescence correlation spectroscopy

Authors: Galland, Rémi; Gao, Jie; Kloster, Meike; Herbomel, Gaetan; Destaing, Olivier; Balland, Martial; Souchier, Catherine; +4 Authors

Multi-confocal fluorescence correlation spectroscopy

Abstract

We report a multi-confocal Fluorescence Correlation Spectroscopy (mFCS) technique that combines a Spatial Light Modulator (SLM), with an Electron Multiplying-CCD camera (EM-CCD). The SLM is used to produce a series of laser spots, while the pixels of the EM-CCD play the roles of virtual pinholes. The phase map addressed to the SLM, calculated by using the spherical wave approximation, makes it possible to produce several diffraction limited laser spots. The fastest acquisition mode leads to a time resolution of 100 microseconds. By using solutions of sulforhodamine G we demonstrated that the observation volumes are similar to that of a standard confocal set-up. mFCS experiments have also been conducted on two stable cell lines: mouse embryonic fibroblasts expressing eGFP-actin and H1299 cells expressing the heat shock factor fusion protein HSF1-eGFP. In the first case we could recover the diffusion constant of G-actin within the cytoplasm, although we were also sensitive to interactions with F-actin. Concerning HSF1, we could clearly observe the modifications of the number of molecules and of the HSF1 dynamics during heat shock.

Country
France
Keywords

180.1790, [PHYS.PHYS.PHYS-BIO-PH]Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Green Fluorescent Proteins, MESH: Rhodamines, MESH: Actins, MESH: Spectrometry, Cell Line, Mice, MESH: Green Fluorescent Proteins, Computer-Assisted, correlations, MESH: Least-Squares Analysis, Image Processing, Computer-Assisted, Animals, MESH: Animals, Least-Squares Analysis, MESH: Mice, [SDV.IB] Life Sciences [q-bio]/Bioengineering, [PHYS.PHYS.PHYS-BIO-PH] Physics [physics]/Physics [physics]/Biological Physics [physics.bio-ph], Rhodamines, molecular transport, 170.6280, 540, molecular dynamics, Actins, MESH: Cell Line, MESH: Image Processing, Confocal microscopy, Spectrometry, Fluorescence, [SDV.IB]Life Sciences [q-bio]/Bioengineering, fluorescence

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
2
Average
Average
Average
Green
gold