Fluorometer detection system
First Claim
1. A fluorometer for exciting a sample including a fluorophore and for detecting fluorescent emissions from the sample comprising:
- a laser diode for exciting the sample,a detector positioned to receive fluorescent emission, andmeans for determining the time of receipt of the fluorescent emission, said means including a ramp generator.
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Accused Products
Abstract
An improved fluorescence detection system is provided which utilizes a relatively high powered, relatively high repetition rate light source with high speed detection electronics to increase system sensitivity and accuracy. In the preferred embodiments, a laser diode is the light source. In one embodiment, the position of a time window is varied to compile a decay profile of a fluorophore. In another embodiment, the time to detection of a photon is used to compile the decay profile. In one aspect of this invention, a histogram of the fluorescence decay is generated by determining a preliminary histogram of the shape and multiplying it by the ratio of the total number of events divided by the number of events comprising the preliminary histogram. In another aspect of this invention, the time of detection after excitation of the photon is started from a random time, such as after a preceding event is detected and the data stored.
78 Citations
32 Claims
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1. A fluorometer for exciting a sample including a fluorophore and for detecting fluorescent emissions from the sample comprising:
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a laser diode for exciting the sample, a detector positioned to receive fluorescent emission, and means for determining the time of receipt of the fluorescent emission, said means including a ramp generator. - View Dependent Claims (2, 3, 4, 9)
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5. A method for generating a histogram of the intensity as a function of time for a transient state fluorescence determination comprising the steps of:
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detecting events of fluorescence, counting the total number of detected events, determining a preliminary histogram of shape of the histogram, counting the number of events comprising the preliminary histogram, and multiplying the preliminary histogram times the ratio of the count of the total number of events and the count of the number of events comprising the preliminary histogram. - View Dependent Claims (6)
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7. The method for generating a histogram of detected events from an excitation pulse in a transient state fluorometry system comprising the steps of:
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a) monitoring for a detected event, b) upon an event, determining the time of occurrence of the detected event, c) storing the time determined in step b and ignoring for purposes of step b other detected events while performing this step, d) upon completion of step c, resume monitoring at step a at a time after completion of step c not related to the time of the excitation pulse, and e) generating a histogram using the times stored in step c. - View Dependent Claims (8)
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10. A method for generating a histogram of detected events in the transient state fluorometry system comprising the steps of:
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a) monitoring for a detected event, b) upon an event, determining the time of occurrence of the detected event by sampling a ramp voltage, c) storing the time determined in step b and ignoring for purposes of step b other detected events while performing this step, d) upon completion of step c, resume monitoring at step a at a time after completion of step c and, e) generating a histogram using the times stored in step c. - View Dependent Claims (11)
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12. A fluorometer for exciting a sample including a fluorophore and for detecting fluorescent emissions from the sample comprising:
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a laser diode for exciting the sample, a detector positioned to receive fluorescent emission, and means for determining the time of receipt of the fluorescent emission, said means including a delay generator. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32)
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Specification