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

Patent metadata

Field	Value
Publication number	US-2022276167-A1
Application number	US-202217578932-A
Country	US
Kind code	A1
Filing date	Jan 19, 2022
Priority date	Apr 1, 2016
Publication date	Sep 1, 2022
Grant date	—

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

1 .- 13 . (canceled)

14 . A system for classifying or characterizing a biological sample, the system comprising:
(a) a first transmission element configured to convey optical excitation signals; (b) a second transmission element configured to receive the optical excitation signals from the first transmission element and convey the optical excitation signals to the biological sample, wherein the biological sample generates responsive optical signals in response to the optical excitation signals and the responsive optical signals are received by the second transmission element; (c) a signal collection element coupled to the second transmission 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 second transmission element and temporally split the responsive optical signals into a plurality of temporally distinct spectral bands corresponding to the plurality of spectral filters; and (d) a processor coupled to the signal collection element and configured to 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.

15 . The system of claim 14 , wherein each of the temporally distinct spectral bands is time delayed with respect to another.

16 . The system of claim 14 , wherein the responsive optical signal comprises one or more of a fluorescence spectrum, a Raman spectrum, an ultraviolet-visible spectrum, or an infrared spectrum.

17 . The system of claim 14 , wherein the single pixel collection element comprises a photomultiplier tube.

18 . The system of claim 14 , wherein to characterize the biological sample of (d) comprises determining one or more of a concentration or a distribution of a molecule in the sample based on the plurality of temporally distinct spectral bands.

19 . The system of claim 18 , wherein the molecule comprises an exogenous fluorescent molecule.

20 . The system of claim 18 , wherein the molecule comprises an endogenous fluorescent molecule.

21 . The system of claim 18 , 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).

22 . The system of claim 14 , wherein the biological sample is characterized in about 100 ms or less.

23 . The system of claim 14 , 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.

24 . The system of claim 14 , wherein the first transmission element is configured to scan the optical excitation signals across a pre-determined portion of the biological sample.

25 . The system of claim 14 , 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.

26 . A method for assisting surgery, comprising:
(a) radiating a target tissue of a subject in an operating room during a surgery of the subject with at least one light pulse at a predetermined wavelength to cause the target tissue to produce a responsive optical signal; (b) collecting the responsive optical signal from the target tissue; (c) characterizing the target tissue based on the collected responsive optical signal using Time-Resolved Fluorescence Spectroscopy; and (d) displaying spectroscopy information of the characterized target tissue superimposed on an image of the target tissue.

27 . The method of claim 26 , wherein the tissue of the subject is radiated with the at least one light pulse prior to or after surgical resection of the tissue.

28 . The method of claim 26 , wherein characterizing the target tissue based on the collected optical signal using Time-Resolved Fluorescence Spectroscopy comprises splitting the responsive optical signal at pre-determined wavelength ranges with a plurality of spectral filters to obtain a plurality of distinct spectral bands and characterizing the target tissue based on decay information for the plurality of spectral bands.

29 . The method of claim 28 , wherein each distinct spectral bands of the plurality of distinct spectral bands is time delayed with respect to another.

30 . The method of claim 28 wherein the responsive optical signal is split into the plurality of distinct spectral bands with a filter wheel comprising the plurality of spectral filters.

31 . The method of claim 28 , wherein an identifier is encoded into one or more of the distinct spectral bands using one or more of the plurality of spectral filters.

32 . The method of claim 28 , wherein the plurality of distinct spectral bands is detected with a single pixel collection element.

33 . The method of claim 32 , wherein the single pixel collection element comprises a photomultiplier tube.

34 . The method of claim 26 , wherein the responsive optical signal comprises one or more of a fluorescence spectrum, a Raman spectrum, an ultraviolet-visible spectrum, or an infrared spectrum.

35 . The method of claim 26 , wherein the tissue is characterized by determining one or more of a concentration or a distribution of a molecule in the tissue based on the plurality of temporally distinct spectral bands.

36 . The method of claim 35 , wherein the molecule comprises an exogenous fluorescent molecule or an endogenous fluorescent molecule.

37 . The method of claim 35 , 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).

38 . The method of claim 26 , wherein the tissue is characterized in about 100 ms or less.

39 . The method of claim 26 , wherein characterizing the target tissue based on the collected responsive optical signal comprises passing the responsive optical signal through an optical assembly comprising one or more optical components having a numerical aperture of 0.22 or greater.

40 . The method of claim

Assignees

• Black Light Surgical Inc
• Cedars Sinai Medical Center

Inventors

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

Classifications

• G01J3/0218

  using optical fibers · CPC title

• G01N21/6402 Primary

  Atomic fluorescence; Laser induced fluorescence · CPC title

• G01J3/4406

  Fluorescence spectrometry · CPC title

• G01J3/32

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

• G01J3/10

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