Fluorescence enhanced photothermal infrared spectroscopy and confocal fluorescence imaging
AssigneePhotothermal Spectroscopy Corp.
InventorCraig Prater
Life of this patent
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Abstract
Embodiments disclosed include methods and apparatus for Fluorescent Enhanced Photothermal Infrared (FE-PTIR) spectroscopy and chemical imaging, which enables high sensitivity and high spatial resolution measurements of IR absorption with simultaneous confocal fluorescence imaging. In various embodiments, the FE-PTIR technique utilizes combined/simultaneous OPTIR and fluorescence imaging that provides significant improvements and benefits compared to previous work by simultaneous detection of both IR absorption and confocal fluorescence using the same optical detector at the same time.
Claims24
A method of performing infrared imaging of a sample, the method comprising:
- illuminating the sample with a beam of excitation radiation to excite autofluorescent emission from the sample;
- capturing at least a portion of the autofluorescent emission from the sample with a camera-based detector to produce a frame autofluorescent image of the sample;
- illuminating at least a portion of the sample corresponding to the frame autofluorescent image with a beam of infrared radiation;
- capturing at least a portion of the autofluorescent emission from the sample with the camera-based detector while the portion of the autofluorescent emission from the sample is perturbed by the infrared light absorbed by the sample to produce an IR-absorbed frame autofluorescent image;
- and
- constructing a signal indicative of infrared absorption by the sample using the frame autofluorescent image and the IR-absorbed frame autofluorescent image.
A microscope system of performing infrared imaging of a sample, the microscope system comprising:
- an excitation light source configured to illuminate the sample with a beam of excitation radiation to excite autofluorescent emission from the sample;
- a camera-based detector arranged to capture at least a portion of the autofluorescent emission from the sample to produce a frame autofluorescent image of the sample;
- an infrared light source configured to illuminate at least a portion of the sample corresponding to the frame autofluorescent image with a beam of infrared radiation;
- a camera-based detector configured to capture at least a portion of the autofluorescent emission from the sample while the portion of the autofluorescent emission from the sample is perturbed by the infrared light absorbed by the sample to produce the IR-absorbed frame autofluorescent image;
- and
- a processor configured to construct a signal indicative of infrared absorption by the sample using the autofluorescent frame and the autofluorescent IR-absorbed frame.
Dependencies are inferred from the claim text.