Analyte detection from steady-state luminescence lifetime

G - Physics – 01 – N

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G01N 21/64 (2006.01) A61B 5/00 (2006.01) A61B 6/00 (2006.01)

Patent

CA 2187735

Methods and apparatus determine analyte concentra- don in vivo and in vitro by the steady state determination of luminescence lifetime. A fluoro-phore that is quenched by the analyte is free to undergo Brownian rotation. The fluoro-phore (12) is irradiated with continuous linearly po- larized light (24). Emitted luminescence is resolved into vector components parallel and perpendicular to the plane of polarization of the excitation light (36, 38). A math- ematical function is employed which relates the lumines- cence anisotropy to quencher concentration. For analytes which do not quench excited staves, a known quantity of the analyte is conjugated to a quencher molecule or energy transfer acceptor, and a competition reaction is set up in which labelled and unlabelled analytes compete for sites on a labelled carrier molecule. An empirically determined mathematical function is employed which relates lumines- cence anisotropy at the carrier label emission band to ana- lyte concentration.

Procédés et appareil de détermination in vivo et in vitro de la concentration d'analytes, par détermination à l'état stationnaire de la durée de la luminescence. Un fluorophore éteint par l'analyte est libre de s'animer d'un mouvement de rotation brownien. On irradie le fluorophore (12) de lumière continue à polarisation linéaire (24). On décompose la luminescence émise en ses composantes vectorielles parallèles et perpendiculaires au plan de polarisation de la lumière d'excitation (36, 38). On utilise une fonction mathématique associant l'anisotropie de la luminescence à la concentration de l'extincteur. Pour les analytes qui n'éteignent pas les états excités, on conjugue une quantité connue de l'analyte avec une molécule extinctrice ou un accepteur de transfert d'énergie, et on établit une réaction de compétition dans laquelle les analytes marqués et non marqués se disputent des sites sur une molécule porteuse marquée. On emploie une fonction mathématique déterminée par voie empirique qui associe à la concentration de l'analyte l'anisotropie de la luminescence dans la bande d'émission du marqueur de la porteuse.

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