IMR Press / FBE / Volume 3 / Issue 2 / DOI: 10.2741/E263

Frontiers in Bioscience-Elite (FBE) is published by IMR Press from Volume 13 Issue 2 (2021). Previous articles were published by another publisher on a subscription basis, and they are hosted by IMR Press on imrpress.com as a courtesy and upon agreement with Frontiers in Bioscience.

Open Access Article
Multi-confocal fluorescence correlation spectroscopy
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1 Universite de Grenoble 1, CNRS, Laboratoire de Spectrometrie Physique UMR 5588, BP 87, 38402 Saint Martin d'Heres, France
2 Universite de Grenoble 1/ INSERM, Institut Albert Bonniot, U823, equipe 10, Stress et DyOGen, La Tronche, BP 170, 38042 Grenoble Cedex 9, France
3 Universite de Grenoble 1/ INSERM, Institut Albert Bonniot, U823, equipe DySAD ERL CNRS 3148, La Tronche, BP 170, 38042 Grenoble Cedex 09, France
4 Universite de Grenoble 1, Institut d'Ingenierie et de l'Information de Sante, Laboratoire TIMC, La Tronche, 38706 La Tronche cedex, France

*Author to whom correspondence should be addressed,

Academic Editor: Vladana Vukojevic

Front. Biosci. (Elite Ed) 2011, 3(2), 476–488; https://doi.org/10.2741/E263
Published: 1 January 2011
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 µs. 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.

Keywords
Fluorescence Correlation Spectroscopy
microscopy
spatial light modulator
diffusion
single molecules
multi-confocal
actin
cytoskeleton
heat shock
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