US 2003/0151000 A1Published application

Method for measuring flourescence, apparatus for measuring flourescence and apparatus for evaluating sample using it

AssigneeHamamatsu Photonics K.K.

InventorsMotoyuki Watanabe, Kazuya Iguchi

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Abstract

A sample S is irradiated with pulsed pumping light supplied from a pumping light source 1, fluorescence generated by the sample S is detected by a photodetector 5 by way of a condensing optical system 3 and a spectroscope 4, and the fluorescence time waveform is subjected to a data analysis in a data processing unit 62 in a controller 6, so as to compute waveform data and physical quantities such as fluorescence lifetime. Here, the pumping light time waveform is fixedly arranged with respect to a time axis used for the data analysis, a plurality of fitting calculations are carried out while moving the fluorescence time waveform and fitting range from an initial position earlier than a pumping light peak to a later end position, and optimal measurement waveform data is selected according to a predetermined selection criterion. This realizes fluorescence measuring method and apparatus which can compute waveform data and the like accurately and efficiently regardless of fluctuations in the fluorescence time waveform, and a sample evaluating apparatus using the same.

Claims13

Dependent claims

A fluorescence measuring method comprising:

a pumping step of irradiating a sample with pulsed pumping light;
a light-detecting step of detecting fluorescence released from said sample pumped with said pumping light;
and
a data processing step of subjecting a time waveform of fluorescence detected by said light-detecting step to a data analysis including a fitting calculation in a fitting range acting as a predetermined time range fixedly set for said time waveform of fluorescence so as to compute waveform data;
wherein, in said data processing step,
a time waveform of pumping light determined beforehand and said time waveform of fluorescence are arranged on a time axis used for said fitting calculation such that a fluorescence peak of said time waveform of fluorescence is placed at an initial position earlier or later by a predetermined time width than a time position substantially coinciding with a pumping light peak of said time waveform of pumping light, and
then, while moving said time waveform of fluorescence and fitting range or said time waveform of pumping light with respect to said time axis to an end position on said time axis on the opposite side of said time position where said fluorescence peak substantially coincides with said pumping light peak from said initial position, respective fitting calculations are carried out at a plurality of time positions different from each other with reference to said time waveform of pumping light, and
a waveform data item selected according to a predetermined selection criterion from a plurality of waveform data items respectively computed in said fitting calculations is employed as final measurement waveform data.

5 more elements

A fluorescence measuring apparatus comprising:

pumping means for irradiating a sample with pulsed pumping light;
light-detecting means for detecting fluorescence released from said sample pumped with said pumping light;
and
data processing means for subjecting a time waveform of fluorescence detected by said light-detecting means to a data analysis including a fitting calculation in a fitting range acting as a predetermined time range fixedly set for said time waveform of fluorescence so as to compute waveform data;
wherein said data processing means arranges a time waveform of pumping light determined beforehand and said time waveform of fluorescence on a time axis used for said fitting calculation such that a fluorescence peak of said time waveform of fluorescence is placed at an initial position earlier or later by a predetermined time width than a time position substantially coinciding with a pumping light peak of said time waveform of pumping light, and
then, while moving said time waveform of fluorescence and fitting range or said time waveform of pumping light with respect to said time axis to an end position on said time axis on the opposite side of said time position where said fluorescence peak substantially coincides with said pumping light peak from said initial position, carries out respective fitting calculations at a plurality of time positions different from each other with reference to said time waveform of pumping light, and
employs a waveform data item selected according to a predetermined selection criterion from a plurality of waveform data items respectively computed in said fitting calculations as final measurement waveform data.

4 more elements

Dependencies are inferred from the claim text.