Direct method for the correction of pressure induced scrambling of polarized fluorescence intensities
OwnerLesley Davenport, Piotr Targowskiinventors
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Abstract
The present invention provides a simple and direct method for the simultaneous correction of steady-state polarized fluorescence intensities, depolarized (or scrambled) by the effects of applied hydrostatic pressure without having to first determine the scrambling factors from a separate experiment. The method involves direct recalculation of the measured polarized data obtained for the sample of interest at the time of data collection.
Claims16
A process for measuring and removing scrambling effects induced by an applied hydrostatic pressure (p), from fluorescence intensities while avoiding the need for a separate pressurized calibration experiment, comprising the acts of measuring polarized fluorescence intensities and then determining excitation and emission correction factors simultaneously.
A method for the correction of time dependent polarized fluorescence intensities obtained for a sample under an applied hydrostatic pressure (p), comprising the steps of:
- a) collecting four non-truncated polarized iVH, iHH, iHV) decay profiles;
- b) integrating said decay profiles;
- c) calculating emission and excitation correction factors X and Y, respectively, from integrals of said profiles;
- and
- d) using said emission and excitation factors, together with said iVV and iVH decay profiles, to perform a sum-difference analysis to obtain profiles for total corrected intensity (Scoff) and difference in polarized fluorescence intensity (Dcoff).
A computer-controlled instrument for measuring and removing scrambling effects, induced by an applied hydrostatic pressure (p), from fluorescence intensities while avoiding the need for a separate calibration experiment, comprising a computer/processor, a fluorescence spectrometer, and a computer readable storage medium comprising computer executable code for instructing the instrument to perform the acts of measuring polarized fluorescence intensities and then determining excitation and emission correction factors.
Dependencies are inferred from the claim text.