Systems, devices, and methods for time-resolved fluorescent spectroscopy

Patent metadata

Field	Value
Publication number	US-12025557-B2
Application number	US-202217578932-A
Country	US
Kind code	B2
Filing date	Jan 19, 2022
Priority date	Apr 1, 2016
Publication date	Jul 2, 2024
Grant date	Jul 2, 2024

Abstract

Provided herein are devices, systems, and methods for characterizing a biological sample in vivo or ex vivo in real-time using time-resolved spectroscopy. A light source generates a light pulse or continuous light wave and excites the biological sample, inducing a responsive fluorescent signal. A demultiplexer splits the signal into spectral bands and a time delay is applied to the spectral bands so as to capture data with a detector from multiple spectral bands from a single excitation pulse. The biological sample is characterized by analyzing the fluorescence intensity magnitude and/or decay of the spectral bands. The sample may comprise one or more exogenous or endogenous fluorophore. The device may be a two-piece probe with a detachable, disposable distal end. The systems may combine fluorescence spectroscopy with other optical spectroscopy or imaging modalities. The light pulse may be focused at a single focal point or scanned or patterned across an area.

First claim

What is claimed is:

1. A system for classifying or characterizing a biological sample, the system comprising:
(a) a light source configured to irradiate a biological sample with at least one light pulse, wherein the biological sample generates responsive optical signals in response to the at least one light pulse;
(b) a signal collection element, wherein the signal collection element comprises a single pixel detector and a filter wheel comprising a plurality of spectral filters, wherein the signal collection element is configured to receive the responsive optical signals from the biological sample and temporally split the responsive optical signals into a plurality of temporally distinct spectral bands corresponding to the plurality of spectral filters; and
(c) a processor coupled to the signal collection element and configured to: (i) characterize the biological sample using time-resolved fluorescence spectroscopy in response to the plurality of temporally distinct spectral bands in near real-time or real-time, and (ii) direct a display in communication with the processor to show spectroscopic information of the plurality of temporally distinct spectral bands alongside or overlaid on an image of the biological sample.

2. The system of claim 1 , wherein each of the temporally distinct spectral bands is time delayed with respect to another.

3. The system of claim 1 , wherein the responsive optical signal comprises one or more of a fluorescence spectrum, a Raman spectrum, an ultraviolet-visible spectrum, or an infrared spectrum.

4. The system of claim 1 , wherein the single pixel detector comprises a photomultiplier tube.

5. The system of claim 1 , wherein the biological sample is characterized by determining one or more of a concentration or a distribution of a molecule in the biological sample based on the plurality of temporally distinct spectral bands.

6. The system of claim 5 , wherein the molecule comprises an exogenous fluorescent molecule.

7. The system of claim 5 wherein the molecule comprises an endogenous fluorescent molecule.

8. The system of claim 5 , wherein the molecule comprises one or more of Flavin mononucleotide (FMN) riboflavin, Flavin adenine dinucleotide (FAD) riboflavin, lipopigments, endogenous porphyrin, free nicotinamide adenosine dinucleotide (NADH), bound NADH, lipofuscin, PpIX, purodaxine, retinol, riboflavin, chlorotoxin (CTX), 5-aminolevulinic acid (5-ALA), sodium fluorescein, ICG-labeled CTX, ICG-labeled knottin, Cy5-labeled knottin, Cy7-labeled knottin, a fluorescently-conjugated tumor-targeting antibody, a fluorescently-labeled tumor-targeting moiety, a fluorescently labeled antibody, fluorescently-labeled toxin, fluorescently-labeled endotoxin, fluorescently-labeled exotoxins, fluorescently-labeled tumor markers, or pyridoxal phosphate-glutamate decarboxylase (PLP-GAD).

9. The system of claim 1 , wherein the biological sample is characterized in about 100 ms or less.

10. The system of claim 1 , wherein an identifier is encoded into one or more of the temporally distinct spectral bands using one or more of the plurality of spectral filters.

11. The system of claim 1 , wherein the light source is scanned across a pre-determined portion of the biological sample.

12. The system of claim 11 , wherein the light source is scanned with a raster scanning pattern.

13. The system of claim 1 , wherein the signal collection element is configured to pass the responsive optical signal through an optical assembly comprising one or more optical components having a numerical aperture of 0.22 or greater.

14. The system of claim 13 , wherein the numerical aperture is of one or more fibers of the optical assembly, and wherein the total cross-sectional area is based on a number of the one or more fibers and a diameter of the one or more fibers.

15. The system of claim 1 , wherein a gain of the single pixel detector is automatically adjusted.

16. The system of claim 1 , wherein the at least one light pulse comprises a wavelength in a range of about 300 nm to about 1100 nm.

17. The system of claim 1 , wherein the biological sample is obtained from a surgical procedure of a patient.

18. The system of claim 1 , wherein the filter wheel comprises a rotating filter wheel.

19. The system of claim 1 , wherein the biological sample is characterized as normal, benign, malignant, scar tissue, necrotic, hypoxic, viable, non-viable, or inflamed.

20. The system of claim 1 , further comprising an optical delay element configured to apply an optical delay to the responsive optical signals and optically couple the responsive optical signal to the signal collection element.

21. The system of claim 1 , wherein the image of the biological sample comprises a brightfield image.

22. The system of claim 1 , wherein the image of the biological sample comprises a pre-operative image.

23. The system of claim 1 , wherein the image of the biological sample comprises an intra-operative image.

24. The system of claim 1 , wherein the image of the biological sample comprises a post-operative image.

Assignees

• Black Light Surgical Inc
• Cedars Sinai Medical Center

Inventors

• Butte Pramod
• Black Keith
• Kavanaugh Jack
• BORTNIK Bartosz
• Nie Zhaojun

Classifications

• G01J2003/1265

  the wavelengths being separated in time, e.g. through optical fibre array · CPC title

• G01J3/10

  Arrangements of light sources specially adapted for spectrometry or colorimetry · CPC title

• G01J3/2889

  Rapid scan spectrometers; Time resolved spectrometry · CPC title

• G01J3/32

  Investigating bands of a spectrum in sequence by a single detector · CPC title

• G01J2003/1213

  Filters in general, e.g. dichroic, band · CPC title