Method and device for referencing fluorescence intensity signals
First Claim
1. Process for optical determination of a biological, chemical or physical parameter of a sample, utilizing at least two different luminescent materials, the first of which being sensitive to the parameter at least with respect to luminescence intensity, and the second of which being insensitive to the parameter at least with respect to luminescence intensity and decay time, andthe luminescent materials exhibit different decay times;
- the process comprising measuring phase behavior of luminescence responses of both luminescent materials by a single detector and obtaining a reference value being independent of the total intensity of both luminescent materials by the phase behavior measured and determining the parameter utilizing the reference value;
wherein a phase shift (φ
m) of the phases of a sum signal of both luminescent materials is used as the reference value.
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Abstract
A method and device for fluorimetric determination of a biological, chemical or physical parameter of a sample utilize at least two different luminescent materials, the first of which is sensitive to the parameter, at least with respect to luminescence intensity, and the second of which is insensitive to the parameter, at least with respect to luminescence intensity and decay time. The luminescent materials have different decay times. The time- or phase behaviour of the resulting luminescence response is used to form a reference value for determination of a parameter.
61 Citations
14 Claims
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1. Process for optical determination of a biological, chemical or physical parameter of a sample, utilizing at least two different luminescent materials, the first of which being sensitive to the parameter at least with respect to luminescence intensity, and the second of which being insensitive to the parameter at least with respect to luminescence intensity and decay time, and
the luminescent materials exhibit different decay times; the process comprising measuring phase behavior of luminescence responses of both luminescent materials by a single detector and obtaining a reference value being independent of the total intensity of both luminescent materials by the phase behavior measured and determining the parameter utilizing the reference value;
wherein a phase shift (φ
m) of the phases of a sum signal of both luminescent materials is used as the reference value.- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 13, 14)
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11. Process for optical determination of a biological, chemical or physical parameter of a sample, utilizing at least two different luminescent materials, the first of which being sensitive to the parameter at least with respect to luminescence intensity, and the second of which being insensitive to the parameter at least with respect to luminescence intensity and decay time, and the luminescent materials exhibit different decay times;
- the process comprising measuring time behavior of a sum signal of luminescence responses of both luminescent materials during an excitation pulse and during the decay period by a single detector and obtaining a reference value being independent of total intensity of both luminescent materials by the time behavior measured and determining the parameter utilizing the reference value;
wherein the intensity ratio l1/l2 of the sum signal of both luminescent materials is used as the reference value and wherein l1 is the luminescence intensity during excitation and l2 is the luminescence intensity during decay.
- the process comprising measuring time behavior of a sum signal of luminescence responses of both luminescent materials during an excitation pulse and during the decay period by a single detector and obtaining a reference value being independent of total intensity of both luminescent materials by the time behavior measured and determining the parameter utilizing the reference value;
Specification