Combined fluorescence and reflectance spectroscopy
First Claim
1. An apparatus for performing fluorescence and spatially resolved reflectance spectroscopy on a sample, comprising:
- a light source;
a monochromator in optical communication with said light source;
a reflectance illumination fiber in optical communication with said light source;
a fluorescence excitation fiber in optical communication with said monochromator;
an imaging spectrograph;
a fluorescence collection fiber in optical communication with said imaging spectrograph;
a reflectance collection fiber in optical communication with said imaging spectrograph and in spaced relation with said reflectance illumination fiber; and
a detector in optical communication with said imaging spectrograph;
wherein said reflectance illumination fiber, said fluorescence excitation fiber, said fluorescence collection fiber, and said reflectance collection fiber define a fiber optic probe, said probe comprising a center section and an outer section, said fluorescence excitation fiber and said fluorescence collection fiber being positioned in said center section, and said reflectance illumination fiber and said reflectance collection fiber being positioned in said outer section.
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Abstract
Methods and apparatus for performing fluorescence and spatially resolved reflectance spectroscopy on a sample. A sample is irradiated with a fluorescence excitation fiber and radiation is collected from the sample with a fluorescence collection fiber and detected to form fluorescence spectra. The sample is also illuminated with a reflectance illumination fiber and reflected light from the sample is collected at a plurality of collection positions and detected to form spatially resolved reflectance spectra. The fibers may form a probe arranged in concentric sections. The spectra are analyzed by pre-processing and reducing the dimensionality of the spectral data.
274 Citations
48 Claims
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1. An apparatus for performing fluorescence and spatially resolved reflectance spectroscopy on a sample, comprising:
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a light source;
a monochromator in optical communication with said light source;
a reflectance illumination fiber in optical communication with said light source;
a fluorescence excitation fiber in optical communication with said monochromator;
an imaging spectrograph;
a fluorescence collection fiber in optical communication with said imaging spectrograph;
a reflectance collection fiber in optical communication with said imaging spectrograph and in spaced relation with said reflectance illumination fiber; and
a detector in optical communication with said imaging spectrograph;
wherein said reflectance illumination fiber, said fluorescence excitation fiber, said fluorescence collection fiber, and said reflectance collection fiber define a fiber optic probe, said probe comprising a center section and an outer section, said fluorescence excitation fiber and said fluorescence collection fiber being positioned in said center section, and said reflectance illumination fiber and said reflectance collection fiber being positioned in said outer section. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)
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24. An apparatus for measuring fluorescence and spatially resolved reflectance spectra of a sample, comprising:
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a light source;
a monochromator in optical communication with said light source;
a fiber optic probe in optical communication with said light source and with said monochromator, said probe comprising a plurality of fluorescence excitation and collection fibers in spaced relation and surrounded at least partially by a plurality of reflectance collection fibers in spaced relation with a reflectance illumination fiber;
an imaging spectrograph in optical communication with said plurality of fluorescence collection fibers and with said plurality of reflectance collection fibers; and
a detector in optical communication with said imaging spectrograph. - View Dependent Claims (25, 26, 27, 28, 29, 30)
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31. A method for combined fluorescence and spatially resolved reflectance spectroscopy of a sample, comprising:
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providing a reflectance illumination fiber, a fluorescence excitation fiber, a fluorescence collection fiber, and a reflectance collection fiber that define a fiber optic probe comprising a center section and an outer section, said fluorescence excitation fiber and said fluorescence collection fiber being positioned in said center section, and said reflectance illumination fiber and said reflectance collection fiber being positioned in said outer section;
directing radiation to said sample with said fluorescence excitation fiber;
collecting radiation from said sample with said fluorescence collection fiber;
directing said radiation from said sample to an imaging spectrograph and a detector;
illuminating said sample with said reflectance illumination fiber;
collecting reflected light from said sample with said reflectance collection fiber; and
directing said reflected light from said sample to an imaging spectrograph and a detector. - View Dependent Claims (32, 33, 34, 35, 36, 37, 38, 39, 41, 42, 43)
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40. The method of claim 40, wherein said analyzing further comprises forming one or more composite algorithms.
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44. A method for combined fluorescence and spatially resolved reflectance spectroscopy of a sample, comprising:
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providing a probe comprising a plurality of fluorescence excitation and collection fibers in spaced relation surrounded at least partially by a plurality of reflectance collection fibers in spaced relation with a reflectance illumination fiber;
directing radiation to said sample with a fluorescence excitation fiber;
collecting radiation from said sample with a fluorescence collection fiber;
directing said radiation from said sample to an imaging spectrograph and a detector;
illuminating said sample with a reflectance illumination fiber;
collecting reflected light at a plurality of collection positions from said sample with said plurality of reflectance collection fibers arranged in spaced relation;
directing said reflected light from said sample to an imaging spectrograph and a detector to produce spectral data;
pre-processing said data; and
reducing a dimension of said data using principal component analysis. - View Dependent Claims (45, 46, 47, 48)
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Specification