Measurement of surface concentration of fluorophores by fluorescence fluctuation spectroscopy |
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Authors: | Delon A Derouard J Delapierre G Jaffiol R |
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Institution: | Laboratoire de Spectrométrie Physique (UMR CNRS-UJF 5588), Université Grenoble 1, Bo?te Postale 87, 38402 Saint Martin d'Hères, France. adelon@ujf-grenoble.fr |
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Abstract: | Fluorescence fluctuation spectroscopy is applied to study molecules passing through a small observation volume, usually subjected to diffusive or convective motion in a liquid phase. We suggest that such a technique could be used to measure the areal absolute concentration of fluorophores deposited on a substrate or embedded in a thin film, with a resolution of a few micrometers. The principle is to translate the solid substrate in front of a confocal fluorescence microscope objective and to record the subsequent fluctuations of the fluorescence intensity. The validity of this concept is investigated on model substrates (fluorescent microspheres) and DNA biochips. |
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