Method and apparatus for identifying background fluorescence using spread spectrum excitation-source broadening in Raman spectroscopy
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Publication Number
US-11193827-B2
Publication Date
2021-12-07
Expiration Date
Abstract
A method and apparatus for determining a level of background fluorescent light produced during photometric interrogation of a sample is provided. The method includes applying an excitation light to a sample using a laser at a plurality linewidths different from one another, the excitation light at each of the plurality of different linewidths applied at an excitation wavelength operable to cause emission of light from the sample, the light emitted from the sample including Raman scattered light and background fluorescent light; detecting light emitted from the tissue sample at each of the plurality of linewidths using a detector and producing light signals representative of the detected light; and determining a level of the background fluorescent using the light signals representative of the detected light for each of the plurality of different linewidths.
Core Innovation
The invention relates to determining and mitigating background fluorescent light during photometric interrogation and Raman spectroscopy of a sample by using a laser at different linewidths. The excitation light causes emission from the sample that includes Raman scattered light and background fluorescent light, and the emitted light is detected at each applied linewidth. By using a broader linewidth in addition to a narrower linewidth, the background fluorescent level is determined from the resulting detected light signals.
In one aspect, excitation light is applied to the sample using a laser at a first linewidth and separately at a second linewidth, where the second linewidth is broader than the first. Light emitted from the tissue sample is detected with a spectrometer and a detector for each linewidth, producing first and second light signals. The level of the background fluorescent light is determined using the first light signals and the second light signals, to separate and quantify fluorescence relative to the Raman scattered light.
In another aspect, a method and apparatus analyze Raman spectroscopy data by applying excitation light at a plurality of different linewidths produced by modulating the laser. The emitted light is detected at each plurality linewidth using a spectrometer and detector to produce light signals representative of the detected light, and a level of the background fluorescent is determined for each linewidth. Raman scattered light is then analyzed while accounting for the determined level of background fluorescence.
Claims Coverage
The independent claims cover three inventive features: determining background fluorescent light using first and second excitation linewidths; analyzing a sample in Raman spectroscopy while accounting for determined background fluorescence across multiple linewidths; and an apparatus that performs linewidth-based background fluorescence determination during Raman spectroscopic analysis using a laser, spectrometer, detector, and analyzer with executable instructions.
Linewidth-dependent determination of background fluorescent light using two laser linewidths
Applying excitation light to a sample using a laser at a first linewidth and separately at a second linewidth, the second linewidth broader than the first linewidth; detecting first and second emitted light signals using a spectrometer and detector for each linewidth; determining a level of the background fluorescent using the first light signals and the second light signals.
Multi-linewidth Raman analysis accounting for background fluorescence determined from modulated laser linewidths
Applying an excitation light to a sample using a laser at a plurality linewidths different from one another, the plurality of different linewidths produced by modulating the laser; detecting light emitted from the tissue sample at each of the plurality of linewidths using a spectrometer and a detector to produce light signals; determining a level of the background fluorescent using the light signals for each of the plurality of different linewidths; analyzing the sample based on the Raman scattered light, including accounting for the determined level of background fluorescence.
Apparatus for determining background fluorescence during Raman spectroscopic analysis using a plurality of linewidths controlled by executable instructions
A laser; a spectrometer; a detector; and an analyzer having a processor and a memory device storing executable instructions that cause the processor to control the laser to apply excitation light to a sample at a plurality linewidths different from one another; control the spectrometer and the detector to detect light emitted from the tissue sample at each plurality of linewidths to produce light signals representative of detected light; and determine a level of the background fluorescent using the light signals representative of the detected light for each of the plurality of different linewidths.
All independent claims implement the same core approach: excitation light is applied at one or more different laser linewidths, emitted light including Raman scattered light and background fluorescent light is detected, and the level of background fluorescence is determined from the detected light signals at those linewidths. In claims directed to Raman analysis, the determined background fluorescence is used to account for background during analysis of Raman scattered light.
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