Measurement of surface concentration of fluorophores using fluorescence fluctuation spectroscopy

Antoine Delon (1) , Jacques Derouard (1) , Guillaume Delapierre (2) , Rodolphe Jaffiol (1)

1 LSP - Laboratoire de Spectrométrie Physique
2 DTBS - Département Microtechnologies pour la Biologie et la Santé

Fluorescence fluctuation spectroscopy is applied to study molecules, passing through a small observation volume, usually subjected to diffusive or convective motion in liquid phase. We suggest that such a technique could be used to measure the areal absolute concentration of fluorophores deposited on a substrate or imbedded 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.

Mots clés: Spectroscopy; fluorescence laser induced; confocal microscopy; optics at surfaces

Domaines: Optique [physics.optics]

Antoine Delon, Jacques Derouard, Guillaume Delapierre, Rodolphe Jaffiol. Measurement of surface concentration of fluorophores using fluorescence fluctuation spectroscopy. Optics Letters, 2006, 31 (8), pp.1142. ⟨hal-00196035⟩