| 摘要 |
This invention relates to a method to correct for error in an anti-Stokes photoluminescence measurement in a Foerster-type resonance energy-transfer assay. The method comprises the steps of exciting anti-Stokes photoluminescent molecules, ions, phosphors, chelates, or particles, i.e. donors of the assay, with one wavelength or multiple wavelengths of light wherein the wavelength or wavelengths are greater than that of an emission wavelength of acceptor molecules in the assay; measuring emission light signal at a wavelength, which is the emission wavelength of the acceptor molecules in the Foerster-type resonance energy-transfer assay, and which differs from the emission wavelength of the anti-Stokes photoluminescent donor in at least two different time windows; a first time window within the time window defined by the excitation light pulse, i.e. within the time window opening when the excitation light pulse is switched on and closing when the excitation light pulse is switched off, and a second time window to follow the first time window not at all overlapping with the time window defined by the excitation pulse; and correcting the emission light signal measured, comprising signals originating from non-radiative and radiative energy transfer, within the first time window by estimating the ratio of the signals from non-radiative and radiative energy transfer or the signal originating from radiative energy transfer using at least one emission light signal measured in the second time window. This invention also relates to the use of a device according to the method of the invention, a system for carrying out the method and a software product for the system.
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