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US 7,733,470 B2Granted
Method and apparatus for real-time measurement and calculation of a fluorescent lifetime
AssigneePhoton Technology International, Inc.
InventorsAleksander Siemiarczuk, Jens Wulf
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Abstract
The invention comprises a real-time stroboscopic acquisition protocol for a measurement of the fluorescence decay and a method and apparatus for real-time calculation of the fluorescence lifetime from that measurement.
Claims12
A method for determining the fluorescent lifetime of a sample in real-time using a stroboscopic instrument, the method comprising the steps of:
- measuring the instrument response function;
- pre-scanning the decay curve;
- identifying a peak intensity (Ip) of the decay curve and a peak time (Tp) at which the peak intensity occurs;
- determining a first delay time (Ta) and a second delay time (Tb);
- measuring a first intensity (Ia) at the first delay time;
- measuring a second intensity (Ib) at the second delay time;
- and
- calculating the fluorescent lifetime (T) of the sample in real-time based on the first and second delay times and the corresponding intensities,
- wherein the step of calculating the fluorescent lifetime (T) comprise the steps of:
- setting a comparison time parameter;
- determining whether the difference between the peak time and the first delay time is greater than the comparison time parameter;
- and
- if the difference between the peak time and the first delay time is greater than the comparison time parameter, calculating the fluorescent lifetime (T) according to the equation
- T=(Ta−Tb)/ln(Ib)/(Ia).
11 more elements
A computer-readable storage media storing computer executable instructions for a method of determining the fluorescent lifetime of a sample in real-time using a stroboscopic instrument, the method comprising the steps of:
- measuring the instrument response function;
- pre-scanning the decay curve;
- identifying a peak intensity (Ip) of the decay curve and a peak time (Tp) at which the peak intensity occurs;
- determining a first delay time (Ta) and a second delay time (Tb);
- measuring a first intensity (Ia) at the first delay time;
- measuring a second intensity (Ib) at the second delay time;
- and
- calculating the fluorescent lifetime (T) of the sample in real-time,
- wherein the step of calculating the fluorescent lifetime (T) further comprises the steps of:
- setting a comparison time parameter;
- determining whether difference between the peak time and the first delay time is greater than the comparison time parameter;
- and
- if the difference between the peak time and the first delay time is greater than the comparison time parameter, calculating the fluorescent lifetime (T) according to the equation
- T=(Ta−Tb)/ln(Ib/(Ia).
11 more elements
An apparatus for determining the fluorescent lifetime of a sample in real-time using a stroboscopic instrument, said apparatus comprising:
- means for measuring the instrument response function;
- means for pre-scanning the decay curve;
- means for identifying a peak intensity (Ip) of the decay curve and a peak time (Tp) at which the peak intensity occurs;
- means for determining a first delay time (Ta) and a second delay time (Tb);
- means for measuring a first intensity (Ia) at the first delay time;
- means for measuring a second intensity (Ib) at the second delay time;
- and
- means for calculating the fluorescent lifetime (T) of the sample in real-time sample in real-time based on the first and second delay times and the corresponding intensities,
- wherein said means for calculating the fluorescent lifetime (T) further comprises:
- means for setting a comparison time parameter;
- means for determining whether the difference between the peak time and the first delay time is greater than the comparison time parameter;
- and
- means for, if the difference between the peak time and the first delay time is greater than the comparison time parameter, calculating the fluorescent lifetime (T) according to the equation
- T=(Ta−Tb)/ln(Ib/(Ia).
11 more elements
A method for determining the fluorescent lifetime of a sample in real-time using a stroboscopic instrument, the method comprising the steps of:
- measuring the instrument response function;
- pre-scanning the decay curve;
- identifying a peak intensity (Ip) of the decay curve and a peak time (Tp) at which the peak intensity occurs;
- determining a first time delay (Ta) and a second time delay (Tb);
- measuring a first intensity (Ia) at the first delay time;
- measuring a second intensity (Ib) at the second delay time;
- and
- calculating the fluorescent lifetime (T) of the sample in real-time based on the first (Ta) and second (Tb) delay times and the corresponding intensities,
- wherein the first delay time (Ta) is determined by scanning the decay curve starting at the peak time (Tp) until the intensity is less than (0.95*Ip), and the second delay time (Tb) is determined by scanning the decay curve starting at the peak time (Tp) until the intensity is less than (0.10*Ip).
6 more elements
A computer-readable storage media storing computer executable instructions for a method of determining the fluorescent lifetime of a sample in real-time using a stroboscopic instrument, the method comprising the steps of:
- measuring the instrument response function;
- pre-scanning the decay curve;
- identifying a peak intensity (Ip) of the decay curve and a peak time (Tp) at which the peak intensity occurs;
- determining a first time delay (Ta) and a second time delay (Tb);
- measuring a first intensity (Ia) at the first delay time;
- measuring a second intensity (Ib) at the second delay time;
- and
- calculating the fluorescent lifetime (T) of the sample in real-time based on the first (Ta) and second (Tb) delay times and the corresponding intensities,
- wherein the first delay time (Ta) is determined by scanning the decay curve starting at the peak time (Tp) until the intensity is less than (0.95*Ip), and the second delay time (Tb) is determined by scanning the decay curve starting at the peak time (Tp) until the intensity is less than (0.10*Ip).
6 more elements
An apparatus for determining the fluorescent lifetime of a sample in real-time using a stroboscopic instrument, said apparatus comprising:
- means for measuring the instrument response function;
- means for prerscanning the decay curve;
- means for identifying a peak intensity (Ip) of the decay curve and a peak time (Tp) at which the peak intensity occurs;
- means for determining a first delay time (Ta) and a second delay time (Tb);
- means for measuring a first intensity (Ia) at the first delay time;
- means for measuring a second intensity (Ib) at the second delay time;
- and
- means for calculating the fluorescent lifetime (T) of the sample in real-time based on the first and second delay times and the corresponding intensities;
- wherein the first delay time is determined by scanning the decay curve starting at the peak time (Tp) until the intensity is less than (0.95*Ip) and the second delay time (Tb) is determined by scanning the decay curve starting at the peak time (Tp) until the intensity is less than (0.10*Ip).
6 more elements
Dependencies are inferred from the claim text.