Estrogen fluoroimmunoassay with a fluorimeter designed for low-intensity light detection |
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Authors: | Jamila El Jabri Solange De Lauzon Nicole Cittanova Patrice Gervais Jacques Mugnier Jacques Pouget Bernard Valeur |
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Affiliation: | UFR Biomédicale des Saints-Pères, 75270 Paris Cedex 06 France;Laboratoire de Chimie Générale (CNRS UA 1103), Conservatoire National des Arts et Métiers, 292 rue Saint-Martin, 75141 Paris Cedex 03 France |
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Abstract: | A small, highly sensitive, inexpensive fluorimeter for the detection of very low fluorescence intensities for analytical and clinical purposes is described. High sensitivity is achieved by use of an inexpensive continuous wave laser with a power of a few milliwatts. The optical arrangement of the instrument has been thoroughly designed so that the effects of Rayleigh and Raman scattering are minimized. Fluorescence is dected through optical filters with a side-on photomultiplier. Concentrations of rhodamine B in ethanol as low as 10?11 M can be measured. To use this instrument for a fluoroimmunoassay of urinary estrogens (10?9?10?7 M), a purified monoclonal IgG, raised against estradiol, was labelled with RITC (rhodamine isothiocyanate) in a 1:1 molar ratio. It was possible to measure IgG-RITC concentrations between 10?9 and 10?10 M with good accuracy and reproducibility. |
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