WO2017206201A1 - 一种基于纤维内窥镜的拉曼光谱检测装置及其实现方法 - Google Patents

一种基于纤维内窥镜的拉曼光谱检测装置及其实现方法 Download PDF

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WO2017206201A1
WO2017206201A1 PCT/CN2016/085988 CN2016085988W WO2017206201A1 WO 2017206201 A1 WO2017206201 A1 WO 2017206201A1 CN 2016085988 W CN2016085988 W CN 2016085988W WO 2017206201 A1 WO2017206201 A1 WO 2017206201A1
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fiber
raman
endoscope
optic probe
excitation
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PCT/CN2016/085988
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English (en)
French (fr)
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陈荣
黄伟
冯尚源
陈冠楠
李永增
黄组芳
曾海山
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福建师范大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
    • A61B5/0075—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by spectroscopy, i.e. measuring spectra, e.g. Raman spectroscopy, infrared absorption spectroscopy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B1/00—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor
    • A61B1/00163—Optical arrangements
    • A61B1/00165—Optical arrangements with light-conductive means, e.g. fibre optics
    • A61B1/00167—Details of optical fibre bundles, e.g. shape or fibre distribution
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B1/00—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor
    • A61B1/06—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor with illuminating arrangements
    • A61B1/07—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor with illuminating arrangements using light-conductive means, e.g. optical fibres
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B1/00—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor
    • A61B1/233—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor for the nose, i.e. nasoscopes, e.g. testing of patency of Eustachian tubes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B1/00—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor
    • A61B1/307—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor for the urinary organs, e.g. urethroscopes, cystoscopes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B1/00—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor
    • A61B1/313—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor for introducing through surgical openings, e.g. laparoscopes
    • A61B1/317—Instruments for performing medical examinations of the interior of cavities or tubes of the body by visual or photographical inspection, e.g. endoscopes; Illuminating arrangements therefor for introducing through surgical openings, e.g. laparoscopes for bones or joints, e.g. osteoscopes, arthroscopes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/48—Other medical applications
    • A61B5/4887—Locating particular structures in or on the body

Definitions

  • the present invention relates to the field of fiber endoscopes, and in particular to a Raman spectrum detecting device based on a fiber endoscope and an implementation method thereof.
  • Cancer is a serious threat to human health and has become the leading cause of human disease-related death.
  • the China Cancer Statistics Annual Report shows that there are 3.5 million new cancer cases in China and 2.5 million deaths due to cancer. In the cure rate of cancer, the developed countries have reached 65%, while China has only about 25%.
  • Fiberscope is a commonly used medical device. It enters the human cavity through the cavity of the human body or through a small incision made by surgery. Observing the lesions in the cavity, determining its location and extent, it can be operated and videotaped. A reliable tool for diagnosis and treatment is widely used in clinical practice.
  • the existing medical fiber endoscope detection systems mostly rely on traditional white light reflecting fiber endoscopes (such as fiberscopes, electronic fiber endoscopes, etc.) to observe the morphological lesions of cancer, and the diagnosis is based on the doctor's Visual observation, combined with personal experience, to judge and identify the structure and shape of abnormal parts of the tissue, microscopic tissue lesions may be difficult to observe, from However, the diagnosis rate is greatly reduced, resulting in missed diagnosis or misdiagnosis, and there are certain difficulties in the clinical diagnosis of early cancer.
  • Raman spectroscopy is an inelastic scatter spectrum that can obtain fingerprint information of molecular structure, vibration mode, functional groups, etc.
  • the biochemical composition is very sensitive and can be widely used in the analysis of biomolecular structures. It is a non-destructive, fast and sensitive optical detection technology.
  • the current research shows that not only the fingerprint region of Raman spectroscopy (200-2000 cm -1 , fingerprint) has diagnostic significance for human tissue diseases, but also the Raman spectrum of high-wavenumber region (2600-3500 cm -1 , high wavenumber). Provide important biochemical information. Therefore, it is necessary to simultaneously acquire the Raman spectrum of the fingerprint region and the high wavenumber region when performing human tissue Raman spectroscopy.
  • a common method is to use a larger area detector to receive a Raman spectrum signal of a wider range of wave numbers. The disadvantage of this method is that it increases the volume of the Raman detection system, and also increases the cost of the system, which is not conducive to the miniaturization of the system and the widespread application.
  • the device that enters the human cavity through the fiber endoscope biopsy channel if it touches the mucous membrane due to improper operation, or even causes the mucous membrane to break and bleed, according to the classification rules of the Medical Device Classification Rules, "pass all or part of A medical device that invades the human body and contacts the tissues of the body, the blood circulation system, the central nervous system, etc. is an invasive device, that is, it is invasive, and it belongs to a device with significant risks. Therefore, it is important to ensure that the observation instruments entering the human body cavity do not cause damage to the human body.
  • a feasible way is to limit the length of the instrument to enter the body cavity, that is, increase the limit organization to avoid the operation of personnel Improperly touched the organization.
  • an object of the present invention is to provide a fiber endoscope-based Raman spectroscopy detecting device and an implementation method thereof, which are suitable for a living body, real-time Raman spectroscopy detection and diagnosis device for human body cavity tissue.
  • a Raman spectrum detecting device based on fiber endoscope comprising a dual wavelength laser, a Raman fiber probe, a fiber endoscope, a white light cold light source, a camera device, a Raman spectrometer, and data processing and a display device;
  • the white light source is connected to an optical interface of the fiberscope, and the camera is disposed on an upper portion of the fiberscope for acquiring an image in the fiberscope, the camera device
  • An output is coupled to the data processing and display device for displaying an image within the fiberscope;
  • an output of the dual wavelength laser is coupled to an input of the Raman fiber optic probe;
  • the Raman The output of the fiber optic probe is coupled to the Raman spectrometer and its detector.
  • the Raman fiber optic probe is Y-shaped, including a first branch and a second branch, the first branch includes an excitation fiber, and the second branch includes a plurality of collection fibers, the middle of the laser fiber The middle portion of the plurality of collection fibers is combined into a bundled fiber.
  • the combined optical fiber has a plurality of collecting optical fibers circumferentially arranged around the excitation optical fiber at an end surface of the detecting end, and the detecting end portion is fixed by a metal sleeve.
  • the combined fiber of the Raman fiber optic probe is disposed near the branch An adjustable limiting device, wherein the limiting device is configured to adjust a length of the fiber optic probe into the fiber endoscope biopsy channel, wherein the limiting device comprises a fixing sleeve and a fixing screw, and the fixing sleeve is sleeved On the surface of the Raman fiber optic probe, the fixing screw is vertically disposed on the fixing sleeve for fixing the Raman fiber optic probe.
  • excitation fiber and the collection fiber are both wrapped with a polymer material.
  • a fiber end surface of the detecting end portion of the Raman fiber optic probe is provided with a plating film, and an end surface of the excitation fiber is plated with a low-pass film that allows two wavelengths of excitation light to pass therethrough, and the end surface of the collecting fiber is plated with useful A high-pass film that passes through the excitation light and allows a larger wavelength of Raman scattered light to pass.
  • the output fiber of the dual-wavelength laser is connected to the excitation fiber of the Raman fiber probe through a filter assembly for alternately outputting excitation light of two different wavelengths;
  • the collection fiber of the Raman fiber probe is The filter assembly is connected, and the output light is connected to the Raman spectrometer and its detector via the collecting fiber.
  • the dual-wavelength laser alternately outputs two different wavelengths of excitation light, including 785 nm excitation light and 690 nm excitation light; the 785 nm excitation light is used to complete detection of a fingerprint region Raman spectrum, and the 690 nm excitation light is used to complete Detection of Raman spectra in high wavenumber regions.
  • the fiberscope is a fiberscope that meets different parts of a human body cavity, including a nasopharyngoscope, a cystoscope, a hysteroscope, a vocal cord arthroscope, and a ureter-nephroscope.
  • the white light cold light source is a 300W short arc xenon lamp white light cold light source.
  • the data processing and display device is a PC.
  • the invention is also implemented by the following method: a method for implementing a Raman spectrum detecting device based on a fiber endoscope, comprising: the following steps:
  • Step S1 extending the Raman fiber optic probe into the biopsy channel of the fiber endoscope, using a limiting device to adjust the length of the fiber optic probe into the biopsy channel, and extending the fiber endoscope into the human body cavity Detection
  • Step S2 turning on a white light cold light source, disposing the image capturing device on an upper portion of the fiber endoscope, acquiring an image in the fiber endoscope, and connecting an output end of the image capturing device to the data processing and display device, Used to display an image within the fiberscope;
  • Step S4 Turn on the dual-wavelength laser and emit excitation light of two different wavelengths, the excitation light is incident through a laser fiber, and the collection fiber collects the Raman scattered light and transmits it to the Raman spectrometer and the detector thereof. The detection of the Raman spectrum in the fingerprint region and the detection of the Raman spectrum in the high-wavenumber region are completed.
  • normal and tumor tissue Raman spectra have obvious Raman peaks at 851, 943, 1004, 1096, 1124, 1265, 1316, 1450, 1621 and 1660 cm -1 ; compared with normal tissue, nasopharynx
  • the Raman spectral characteristics of the fingerprint region of cancerous tissues also changed significantly, that is, the peaks at 851, 943, 1096, and 1124 decreased, while the peaks at 1004, 1265, 1316, 1450, 1621, and 1660 cm -1 Elevation occurred; the shape of the spectrum of normal tissues and tumor tissues was also significantly different between 1120-1360 cm -1 and 1560-1680 cm -1 .
  • Raman spectra of normal and nasopharyngeal carcinoma tissues have obvious Raman peaks at 2854, 2940 and 3009 cm -1 and 3067 cm -1 ; compared with normal tissues, high wavenumber regions of nasopharyngeal carcinoma tissues The spectral characteristics of the man also changed significantly, that is, the peak of the nasopharyngeal carcinoma tissue at 2854 and 2940 cm -1 increased compared with the normal tissue.
  • the fiber endoscope-based Raman spectroscopy detecting device established by the invention adopts a specially designed fiber optic probe, and can conveniently enter the human body cavity through the biopsy channel of the fiber endoscope. Inside, the Raman spectroscopy of the human body cavity is performed; the excitation of two different wavelengths is alternately outputted by the dual-wavelength laser as the excitation light, and the filter component and the control software are used to complete the spectrum detector with the same small area. Simultaneous detection of the Raman spectrum of the fingerprint region and the high wavenumber region.
  • the invention can realize the non-destructive, real-time and high-efficiency Raman spectroscopy detection of the human body cavity; effectively avoid the damage of the optical fiber probe to the human mucosa tissue during the clinical examination process, and the Raman spectroscopy detection system has a wide wave number coverage and a small volume.
  • Such advantages thus providing effective clinical testing tools for the non-destructive, rapid analysis and diagnosis of living tissue, have important application value.
  • using the fiber endoscope-based Raman spectroscopy detection device and the detection method established by the invention the fingerprint regions of the human living nasopharyngeal carcinoma tissue and the normal nasopharyngeal tissue, and the Raman spectral features of the high wavenumber region are obtained. The difference between the two.
  • 1 is a Raman spectrum detecting device based on a fiber endoscope.
  • FIG. 2 is a structural view of the fiber optic probe of FIG. 1.
  • FIG. 3 is a schematic view showing the use of the structure of the limiting device in the fiber optic probe of FIG. 2.
  • FIG. 4 is a schematic view showing the coating of the fiber end surface of the detecting end in the fiber optic probe of FIG. 2.
  • Figure 5 is a comparison of the mean Raman spectra of the nasopharyngeal tissue at normal sites and the low-wavenumber mean Raman spectra of nasopharyngeal carcinoma tissue obtained under excitation of 785 nm excitation light.
  • Figure 6 is a comparison of the mean Raman spectra of the nasopharyngeal tissue at normal sites and the high-wavenumber mean Raman spectra of nasopharyngeal carcinoma tissues obtained by excitation with 690 nm excitation light.
  • the embodiment provides a Raman spectrum detecting device based on a fiber endoscope, as shown in FIG. 1 , comprising a dual wavelength laser, a Raman fiber probe, a fiber endoscope, a white light cold light source, a camera device, a Raman spectrometer, and the like.
  • the white light cold light source is connected to an optical interface of the fiber endoscope, and the image capturing device is disposed on an upper portion of the fiber endoscope for collecting an image in the fiber endoscope,
  • An output end of the camera device is coupled to the data processing and display device for displaying an image in the fiberscope;
  • an output end of the dual-wavelength laser is coupled to an input end of the Raman fiber optic probe;
  • the output of the Raman fiber optic probe is coupled to the Raman spectrometer and its detector.
  • the Raman fiber optic probe is Y-shaped, including a first branch and a second branch, the first branch includes an excitation fiber, and the second branch includes a plurality of collection fibers, the laser The middle portion of the optical fiber and the middle portion of the plurality of collecting optical fibers are combined into one combined optical fiber.
  • the fiber optic probe can be conveniently inserted into the human body cavity through the biopsy channel of the fiberscope to perform Raman spectroscopy measurement of the tissue in the human body cavity.
  • the combined optical fiber has a plurality of collecting optical fibers arranged circumferentially around the excitation fiber at the end surface of the detecting end, and the detecting end is fixed by a metal sleeve to ensure the flatness and firmness of the end surface of the detecting portion.
  • the combined fiber of the Raman fiber optic probe is provided with an adjustable limiting device near the branch, and the limiting device is used for adjusting the length of the fiber probe into the fiber endoscope biopsy channel for limiting.
  • the limiting device includes a fixing sleeve and a fixing screw, and the fixing sleeve is sleeved on a surface of the Raman fiber optic probe, and the fixing screw is vertically disposed on the fixing sleeve for The Raman fiber optic probe is fixed.
  • the limit device can adjust the length of the fiber probe into the endoscope biopsy channel according to the detection requirement, and ensure that the fiber probe does not contact the human body cavity when the fiber probe passes through the biopsy channel to enter the cavity for detection.
  • the mucosal tissue avoids medical risks such as bleeding and infection caused by damage to the mucosal tissue in the cavity; and the fiber probe can be fixed by a tapered rubber sleeve to prevent the fiber probe from tilting and moving during the measurement process. Measuring the deviation of the point, and improving the positioning accuracy of the fiber probe to the lesion.
  • the excitation fiber and the collection fiber are both wrapped with a polymer material.
  • FIG. 4 is a schematic diagram of a coating end of a fiber probe detecting end, wherein a fiber end surface of the detecting end portion of the Raman fiber optic probe is provided with a plating film, and an end surface of the excitation fiber is plated with two wavelengths of excitation light.
  • Low-pass film to reduce the interference of the excitation light on the tissue caused by the non-excitation light generated by the optical component such as the excitation fiber; the end face of the collection fiber is plated to cut off the excitation light and allow the wavelength to be larger.
  • Man scattering light The high-pass film is passed to reduce the interference of the excitation light through the tissue into the collecting fiber to interfere with the tissue Raman signal.
  • the output fiber of the dual-wavelength laser is connected to the excitation fiber of the Raman fiber probe through a filter assembly for alternately outputting excitation light of two different wavelengths;
  • the collection fiber is connected to the filter assembly, and the output light is connected to the Raman spectrometer and its detector via the collection fiber.
  • the dual-wavelength laser alternately outputs two different wavelengths of excitation light, including 785 nm excitation light and 690 nm excitation light; and the 785 nm excitation light is used to complete detection of a fingerprint region Raman spectrum, the 690 nm excitation light. Used to complete the detection of high-wavenumber region Raman spectroscopy.
  • the two-wavelength laser optical components and complex control software to achieve the same small area of a spectral detector for fingerprint Raman spectrum region (200 ⁇ 2000cm -1, fingerprint), and the high wavenumber region (2600 ⁇ 3500cm -1 , high wavenumber) simultaneous detection of Raman spectroscopy, effectively reducing the size of the system and reducing the design cost.
  • the fiber endoscope is a fiber endoscope that satisfies different parts of the human body cavity, including a nasopharyngoscope, a cystoscope, a hysteroscope, a vocal cord arthroscope, and a ureter-nephroscope.
  • the white light cold light source is a 300W short arc xenon lamp white light cold light source.
  • the data processing and display device is a PC.
  • the device can realize the non-destructive, real-time and high-efficiency Raman spectroscopy detection of the human body cavity; effectively avoiding damage to the human mucosa tissue during the clinical examination of the fiber optic probe, and the Raman spectroscopy detection system has wave number coverage Wide, small size, etc., from It provides an effective clinical testing tool for the non-destructive and rapid diagnosis of living tissue, and has important application value.
  • the fiber endoscope may adopt a fiber nasopharyngoscope with an inner diameter of 2.2 mm of the biopsy channel and a 1.6 mm outer diameter of the fiber optic probe.
  • the combined portion of the fiber optic probe passes through the nasopharynx.
  • the mirror biopsy channel enters the nasal cavity and reaches the vicinity of the nasopharynx tissue.
  • An adjustable limiting device is arranged near the branch of the fiber optic probe, and the binding portion of the fiber optic probe is fixed by the fastening screw.
  • the limiting device follows The fiber optic probe is approached to the upper end opening of the biopsy channel.
  • the limiting device can be embedded in the opening of the biopsy channel and restrict the fiber probe from moving into the body cavity.
  • FIG. 2 is a structural diagram of the limiting device in the optical fiber probe.
  • FIG. 3 is a structural diagram of the limiting device in the optical fiber probe. Use the schematic.
  • the dotted line frame at the upper left of Figure 3 is a partial schematic view of the positional relationship between the limiting device on the fiber optic probe and the endoscope biopsy channel inlet 3.
  • the limiting device of the Raman fiber optic probe 2 comprises a fixing sleeve 5 and a fixing screw 4 of the fixing sleeve;
  • the fixing sleeve 5 is made of rubber material and has a tapered shape to ensure a firm connection to the Raman fiber optic probe 2 Fixing to prevent measurement error caused by tilting and moving of the fiber probe during measurement;
  • the fixed sleeve can be freely moved on the surface of the Raman fiber optic probe 2, and matched with the fixing screw 4 to the Raman fiber optic probe 2
  • the length of the scope 1 is precisely adjusted and adjusted according to specific test conditions to meet various test requirements.
  • the Raman spectroscopy of the normal and nasopharyngeal carcinoma tissues is performed by using the intraluminal tissue Raman spectroscopy apparatus of the fiberscope described above, and the test spectra are as shown in FIG. 5 and FIG. .
  • Fig. 5 is an average Raman spectrum of nasopharyngeal tissue and a mean Raman spectrum of nasopharyngeal carcinoma in the normal portion of the fingerprint region measured by 785 nm laser excitation. To the best of our knowledge, this is the first Raman spectral signal to measure the low wavenumber of living tissue in nasopharyngeal carcinoma.
  • the Raman peaks are attributed to specific biochemical substances, the intensity changes of the Raman peak indicate that certain biochemical components in the tissue have undergone specific changes with the development of nasopharyngeal carcinoma.
  • the contents of proteins such as tryptophan, phenylalanine, and tyrosine have changed, and the structure of some proteins has also changed.
  • the contents of nucleic acids, lipids, and saccharides have also changed accordingly.
  • the significant changes in these spectra indicate that the nasopharyngeal carcinoma Raman system we have constructed can detect specific changes in nasopharyngeal carcinoma tissue and is expected to achieve non-invasive detection of nasopharyngeal carcinoma.
  • Figure 6 is an average Raman spectrum of normal tissue and nasopharyngeal carcinoma tissue measured in a high wavenumber region as measured by 690 nm laser excitation. This is also the first Raman spectroscopy signal to measure the high wavenumber of nasopharyngeal carcinoma in vivo. It can be observed that normal and nasopharyngeal carcinoma tissues can obtain 2854, 2940 and 3009cm -1 and 3067cm -1 in the high wavenumber range. Peak, and the difference between the two: the peak of nasopharyngeal carcinoma tissue at 2854, 2940 cm -1 increased compared to normal tissue.

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Abstract

一种基于纤维内窥镜的拉曼光谱检测装置及其实现方法，包括双波长激光器、拉曼光纤探头（2）、纤维内窥镜（1）、白光冷光源、摄像装置、拉曼光谱仪以及显示装置；所述白光冷光源与所述纤维内窥镜（1）的光学接口相连，所述摄像装置设置于纤维内窥镜（1）上部，用以采集所述纤维内窥镜（1）内的图像，所述摄像装置的输出端与所述显示装置相连，用以显示所述纤维内窥镜（1）内的图像；所述双波长激光器的输出端与所述拉曼光纤探头（2）的输入端相连接；所述拉曼光纤探头（2）的输出端与所述拉曼光谱仪及其检测器相连接。该拉曼光谱检测装置及其实现方法适用于人体腔内组织的活体、实时的拉曼光谱检测与分析。

Description

一种基于纤维内窥镜的拉曼光谱检测装置及其实现方法 技术领域
本发明涉及纤维内窥镜领域，特别是涉及一种基于纤维内窥镜的拉曼光谱检测装置及其实现方法。
背景技术
癌症是一种严重威胁人类健康的疾病，已成为人类疾病相关死亡首要原因。《中国肿瘤统计年报》显示：我国每年新发癌症病例为350万，因癌症死亡的有250万。在癌症的治愈率上，目前发达国家已达65％，而我国仅有25％左右。
在癌症的治愈率上，目前发达国家已达65％，而我国仅有25％左右，癌症的治愈率与癌症发现的阶段密切相关，若在早期发现其治愈率就会极大提高。然而许多癌症患者早期并无明显症状，甚难发现。如鼻咽癌。纤维内窥镜是一种常用的医疗器械，经人体的腔道，或者是经手术做的小切口进入人体腔内，观察其腔内病变，确定其部位、范围，可进行手术和摄像,是诊疗的可靠工具，在临床上得到广泛应用。
然而，现有的医用纤维内窥镜检测系统大多依赖于传统白光反射纤维内窥镜(如纤维内窥镜、电子纤维内窥镜等)观察癌症的形态学病变，诊断时，单凭医生的肉眼观察，结合个人经验，对组织异常部位结构和形状进行判断和识别，微小的组织病变可能难以观察到，从 而大大降低了诊出率，造成漏诊或误诊，在早期癌症的临床诊断上存在一定的困难。拉曼光谱是一种非弹性散射光谱，可以获得物质丰富的分子结构、振动模式、官能团等指纹信息不需要复杂的样品准备过程，生物组织中水分干扰小，对蛋白质、核酸、磷脂和糖分的生化成分变化非常敏感等优势，可广泛应用于生物分子结构的分析，是一种无损、快速、高灵敏度的光学检测技术。
目前的研究表明，不仅拉曼光谱的指纹区(200～2000cm-1,fingerprint)对人体组织的疾病具有诊断意义，其高波数区(2600～3500cm-1,high wavenumber)的拉曼光谱也有能够提供十分重要的生化信息。因此，在进行人体组织拉曼光谱检查时，同时获取指纹区和高波数区的拉曼光谱十分必要。然而，为获取较大光谱范围的拉曼光谱，常用的方法是采用较大面积的检测器，以接收更宽波数范围的拉曼光谱信号。这种办法的缺点是，增加了拉曼检测系统的体积，同时也增加了系统的成本，不利于系统的小型化和大范围推广应用。
另一方面，经纤维内窥镜活检通道进入人体腔内的器械，如因操作不当而触碰到粘膜，甚至使粘膜破损而出血，根据《医疗器械分类规则》的分类规则“全部或者部分通过体表侵入人体，接触体内组织、血液循环系统、中枢神经系统等部位的医疗器械”为侵入器械，即是有创的，则属于有重大风险性的器械。因此，确保进入人体腔内的观察器械不对人体造成损伤十分重要。一种可行的办法是对要进入人体腔内的器械的伸入长度加以限制，即增加限位组织，避免因人员的操 作不当而触碰到组织。
发明内容
有鉴于此，本发明的目的是提供一种基于纤维内窥镜的拉曼光谱检测装置及其实现方法，旨在适用于人体腔内组织的活体、实时的拉曼光谱检测和诊断的装置。
本发明采用以下方案实现：一种基于纤维内窥镜的拉曼光谱检测装置，包括双波长激光器、拉曼光纤探头、纤维内窥镜、白光冷光源、摄像装置、拉曼光谱仪以及数据处理与显示装置；所述白光冷光源与所述纤维内窥镜的光学接口相连，所述摄像装置设置于纤维内窥镜上部，用以采集所述纤维内窥镜内的图像，所述摄像装置的输出端与所述数据处理与显示装置相连，用以显示所述纤维内窥镜内的图像；所述双波长激光器的输出端与所述拉曼光纤探头的输入端相连接；所述拉曼光纤探头的输出端与所述拉曼光谱仪及其检测器相连接。
进一步地，所述拉曼光纤探头为Y字形，包括第一分支与第二分支，所述第一分支包括一根激发光纤，所述第二分支包括若干根收集光纤，所述激光光纤的中部与所述若干根收集光纤的中部合束为一根合束光纤。
进一步地，所述合束光纤在探测端部的端面将若干根收集光纤围绕所述激发光纤作圆周排列，探测端部采用金属套筒固定。
进一步地，所述拉曼光纤探头的合束光纤靠近分支处设置有一可 调节的限位装置，所述限位装置用以调节光纤探头进入纤维内窥镜活检通道的长度进行限位，所述限位装置包括包括固定套筒与固定螺丝，所述固定套筒套置在所述拉曼光纤探头的表面，所述固定螺丝垂直设置于所述固定套筒上，用以对所述拉曼光纤探头进行固定。
进一步地，所述激发光纤与所述收集光纤均采用聚合物材料包裹。
进一步地，所述拉曼光纤探头的探测端部的光纤端面设置有镀膜，所述激发光纤的端面上镀有允许两个波长激发光通过的低通膜，所述收集光纤的端面上镀有用以截止激发光并允许波长更大的拉曼散射光通过的高通膜。
进一步地，所述双波长激光器的输出光纤通过滤光组件与所述拉曼光纤探头的激发光纤相连接，用以交替输出两种不同波长的激发光；所述拉曼光纤探头的收集光纤与滤光组件连接，其输出光经收集光纤与拉曼光谱仪及其检测器相连接。
进一步地，所述双波长激光器交替输出两种不同波长的激发光包括785nm激发光与690nm激发光；所述785nm激发光用以完成指纹区拉曼光谱的检测，所述690nm激发光用以完成高波数区拉曼光谱的检测。
进一步地，所述纤维内窥镜为满足人体腔内不同部位的纤维内窥镜，包括鼻咽镜、膀胱镜、子宫镜、声带关节镜以及输尿管-肾镜。
进一步地，所述白光冷光源为300W短弧氙灯白光冷光源。
进一步地，所述数据处理与显示装置为一PC机。
本发明还采用以下方法实现：一种基于纤维内窥镜的拉曼光谱检测装置的实现方法，其特征在于：包括以下步骤:
步骤S1：将所述拉曼光纤探头伸入所述纤维内窥镜的活检通道，采用限位装置调节光纤探头进入所述活检通道的长度，将所述纤维内窥镜伸入人体腔内进行检测；
步骤S2：开启白光冷光源，将所述摄像装置设置于纤维内窥镜上部，采集所述纤维内窥镜内的图像；将所述摄像装置的输出端与所述数据处理与显示装置相连，用以显示所述纤维内窥镜内的图像；
步骤S4：开启所述双波长激光器，并发出两种不同波长的激发光，所述激发光通过激光光纤入射，收集光纤采集所述拉曼散射光传输至所述拉曼光谱仪及其检测器，完成指纹区拉曼光谱的检测与高波数区拉曼光谱的检测。
进一步地，所获得的人活体鼻咽癌组织和正常鼻咽组织的拉曼光谱特征中，
在指纹区：正常与肿瘤组织拉曼光谱均在851、943、1004、1096、1124、1265、1316、1450、1621和1660cm-1处有明显的拉曼峰；相较于正常组织，鼻咽癌组织的指纹区拉曼光谱特性也发生了明显的变化，即在851、943、1096、1124处的峰值发生了下降，而在1004、1265、1316、1450、1621和1660cm-1处的峰值发生了升高；正常组织和肿瘤组织的光谱的形状在1120-1360cm-1以及1560-1680cm-1区 间也存在着明显的差异。
在高波数区：正常与鼻咽癌组织拉曼光谱均在2854、2940和3009cm-1和3067cm-1处有明显的拉曼峰；相较于正常组织，鼻咽癌组织的高波数区拉曼光谱特性也发生了明显的变化，即相较于正常组织，鼻咽癌组织光谱在2854、2940cm-1处的峰值发生了上升。
与现有技术相比，本发明具有如下优点：本发明建立的基于纤维内窥镜的拉曼光谱检测装置，采用特殊设计的光纤探头，可通过纤维内窥镜的活检通道便捷地进入人体腔内，进行人体腔内组织的拉曼光谱测量；采用双波长激光器交替输出两种不同波长的激发作为激发光，配合滤光组件和控制软件，实现了以同一个较小面积的光谱检测器完成指纹区和高波数区拉曼光谱的同时检测。另外，本发明可实现人体腔内组织无损、实时、高效的拉曼光谱检测；有效避免光纤探头在临床检查过程中对人体粘膜组织的损伤，拉曼光谱检测系统具有波数覆盖范围广，体积小等优点，从而为活体组织的无损、快速分析诊断提供有效的临床检测工具，具有重要的应用价值。同时，利用本发明建立的基于纤维内窥镜的拉曼光谱检测装置和检测方法，获得了人活体鼻咽癌组织和正常鼻咽组织的指纹区、及高波数区的拉曼光谱特征及其二者间的差异。
附图说明
图1为基于纤维内窥镜的拉曼光谱检测装置。
图2为图1中光纤探头的结构图。
图3为图2光纤探头中限位装置的结构图使用示意图。
图4为图2光纤探头中探测端光纤端面的镀膜示意图。
图5为785nm激发光激发下获得的正常部位鼻咽部组织平均拉曼光谱与鼻咽癌组织低波数平均拉曼光谱对比图。
图6为690nm激发光激发下获得的正常部位鼻咽部组织平均拉曼光谱与鼻咽癌组织高波数平均拉曼光谱对比图
具体实施方式
下面结合附图及实施例对本发明做进一步说明。
本实施例提供一种基于纤维内窥镜的拉曼光谱检测装置，如图1所示，包括双波长激光器、拉曼光纤探头、纤维内窥镜、白光冷光源、摄像装置、拉曼光谱仪以及数据处理与显示装置；所述白光冷光源与所述纤维内窥镜的光学接口相连，所述摄像装置设置于纤维内窥镜上部，用以采集所述纤维内窥镜内的图像，所述摄像装置的输出端与所述数据处理与显示装置相连，用以显示所述纤维内窥镜内的图像；所述双波长激光器的输出端与所述拉曼光纤探头的输入端相连接；所述拉曼光纤探头的输出端与所述拉曼光谱仪及其检测器相连接。
在本实施例中，所述拉曼光纤探头为Y字形，包括第一分支与第二分支，所述第一分支包括一根激发光纤，所述第二分支包括若干根收集光纤，所述激光光纤的中部与所述若干根收集光纤的中部合束为一根合束光纤。该光纤探头可通过纤维内窥镜的活检通道便捷地进入人体腔内，进行人体腔内组织的拉曼光谱测量。
在本实施例中，所述合束光纤在探测端部的端面将若干根收集光纤围绕所述激发光纤作圆周排列，探测端部采用金属套筒固定，以确保探测部的端面的平整与牢固。
在本实施例中，所述拉曼光纤探头的合束光纤靠近分支处设置有一可调节的限位装置，所述限位装置用以调节光纤探头进入纤维内窥镜活检通道的长度进行限位，所述限位装置包括包括固定套筒与固定螺丝，所述固定套筒套置在所述拉曼光纤探头的表面，所述固定螺丝垂直设置于所述固定套筒上，用以对所述拉曼光纤探头进行固定。通过所述限位装置可根据检测需要，调节光纤探头进入内窥镜活检通道的长度进行限位，确保光纤探头在穿过活检通道进入腔内进行检测时，其探测端的端面不接触人体腔内的粘膜组织，避免了对腔内粘膜组织的损伤而导致的出血、感染等医疗风险；并可通过锥形橡胶套筒对光纤探头进行固定，防止测量过程中由光纤探头侧倾、移动等造成测量点位的偏差,提高光纤探头对病灶的定位精度。
在本实施例中，所述激发光纤与所述收集光纤均采用聚合物材料包裹。
在本实施例中，图4中为光纤探头探测端镀膜示意图，所述拉曼光纤探头的探测端部的光纤端面设置有镀膜，所述激发光纤的端面上镀有允许两个波长激发光通过的低通膜，以减少激发光经激发光纤等光学元件产生的非激发光照射到组织上对测量造成的干扰；所述收集光纤的端面上镀有用以截止激发光并允许波长更大的拉曼散射光通 过的高通膜，以减少激发光经组织的反射进入收集光纤对组织拉曼信号的干扰。
在本实施例中，所述双波长激光器的输出光纤通过滤光组件与所述拉曼光纤探头的激发光纤相连接，用以交替输出两种不同波长的激发光；所述拉曼光纤探头的收集光纤与滤光组件连接，其输出光经收集光纤与拉曼光谱仪及其检测器相连接。
在本实施例中，所述双波长激光器交替输出两种不同波长的激发光包括785nm激发光与690nm激发光；所述785nm激发光用以完成指纹区拉曼光谱的检测，所述690nm激发光用以完成高波数区拉曼光谱的检测。所述双波长激光器配合滤光组件和控制软件，实现了以同一个较小面积的光谱检测器完成指纹区拉曼光谱(200～2000cm-1,fingerprint)和高波数区(2600～3500cm-1,high wavenumber)拉曼光谱的同时检测，有效减小了系统的体积并降低了设计成本。
在本实施例中，所述纤维内窥镜为满足人体腔内不同部位的纤维内窥镜，包括鼻咽镜、膀胱镜、子宫镜、声带关节镜以及输尿管-肾镜。
在本实施例中，所述白光冷光源为300W短弧氙灯白光冷光源。
在本实施例中，所述数据处理与显示装置为一PC机。
在本实施例中，该装置可实现人体腔内组织无损、实时、高效的拉曼光谱检测；有效避免光纤探头在临床检查过程中对人体粘膜组织的损伤，拉曼光谱检测系统具有波数覆盖范围广，体积小等优点，从 而为活体组织的无损、快速诊断提供有效的临床检测工具，具有重要的应用价值。
在本实施例中，所述纤维内窥镜可采用纤维鼻咽镜，其活检通道的内径为2.2mm，光纤探头合束部分外径1.6mm，使用时，光纤探头的合束部分通过鼻咽镜活检通道进入鼻腔，并到达鼻咽组织附近。在光纤探头的合束部分靠近分支处设有可调节的限位装置，其与光纤探头合束部分通过紧固螺丝相固定，当光纤探头在伸入内窥镜的过程中，限位装置随光纤探头往活检通道的上端开口处靠近，当限位装置到达活检通道开口处时，可将限位装置内嵌于活检通道的开口处，并限制光纤探头继续往人体腔内移动。
在本实施例中，“Y”形光纤探头及其与双波长激光器、检测器、限位装置之间的连接关系如图2所示，在图3中为光纤探头中限位装置的结构图使用示意图。图3左上方的虚线框为光纤探头上的限位装置与内窥镜活检通道入口3之间的位置关系的局部示意图。所述拉曼光纤探头2的限位装置包括固定套筒5和所述固定套筒的固定螺丝4；所述固定套筒5为橡胶材质并成锥形状，以确保牢固地对拉曼光纤探头2进行固定，防止测量过程中由于光纤探头侧倾、移动造成的测量误差；所述固定套筒可在拉曼光纤探头2表面进行自由移动，并配合固定螺丝4对拉曼光纤探头2进入内窥镜1的长度进行精确调节，并根据具体检测条件进行合理调节以满足各种测试要求。
在本实施例中，采用所述的纤维内窥镜的活体腔内组织拉曼光谱 检测装置，分别对正常和鼻咽癌活体组织进行拉曼光谱测试，测试光谱如图5和图6所示。图5是利用785nm激光激发所测到的指纹区正常部位鼻咽部组织平均拉曼光谱与鼻咽癌组织平均拉曼光谱。据我们所知，这是首次测得鼻咽癌活体组织低波数的拉曼光谱信号。通过对比，虽然正常与肿瘤组织拉曼光谱存在相似之处，都出现了851、943、1004、1096、1124、1265、1316、1450、1621和1660cm-1的拉曼峰；但同时也发现，相较于正常组织，鼻咽癌组织的某些光谱特性也发生了明显的变化，例如在851、943、1096、1124处的峰值发生了下降；而在1004、1265、1316、1450、1621和1660cm-1处的峰值发生了升高。除此之外，正常组织和肿瘤组织的光谱的形状在1120-1360cm-1以及1560-1680cm-1区间也存在着明显的差异。由于拉曼峰分别归属于特定的生化物质，因此拉曼峰位的强度变化表明随着鼻咽癌的发展，组织中的某些生化成分发生了特定的变化。例如色氨酸、苯丙氨酸、酪氨酸等蛋白的含量发生了变化，此外某些蛋白的结构也发生了改变。另外，核酸、脂类、糖类物质的含量也发生了相应的变化。这些光谱的显著性变化说明，我们构建的鼻咽癌活体拉曼系统可检测出鼻咽癌组织的特异性变化，有望实现鼻咽癌无损活体检测。图6是利用690nm激光激发所测到的高波数区正常组织和鼻咽癌组织的平均拉曼光谱。这也是首次测得鼻咽癌活体组织高波数的拉曼光谱信号，从中可观察到正常与鼻咽癌组织在高波数区间的都可以获得2854、2940和3009cm-1和3067cm-1等拉曼峰，以及二者间的差异差异：相较于正常组织，鼻咽癌组织光谱在2854、2940cm-1处 的峰值发生了上升。
以上所述仅为本发明的较佳实施例，凡依本发明申请专利范围所做的均等变化与修饰，皆应属本发明的涵盖范围。

Claims (10)

  1. 一种基于纤维内窥镜的拉曼光谱检测装置，其特征在于：包括双波长激光器、拉曼光纤探头、纤维内窥镜、白光冷光源、摄像装置、拉曼光谱仪以及数据处理与显示装置；所述白光冷光源与所述纤维内窥镜的光学接口相连，所述摄像装置设置于纤维内窥镜上部，用以采集所述纤维内窥镜内的图像，所述摄像装置的输出端与所述数据处理与显示装置相连，用以显示所述纤维内窥镜内的图像；所述双波长激光器的输出端与所述拉曼光纤探头的输入端相连接；所述拉曼光纤探头的输出端与所述拉曼光谱仪及其检测器相连接。
  2. 根据权利要求1所述的一种基于纤维内窥镜的拉曼光谱检测装置，其特征在于：所述拉曼光纤探头为Y字形，包括第一分支与第二分支，所述第一分支包括一根激发光纤，所述第二分支包括若干根收集光纤，所述激光光纤的中部与所述若干根收集光纤的中部合束为一根合束光纤；所述合束光纤在探测端部的端面将若干根收集光纤围绕所述激发光纤作圆周排列，探测端部采用金属套筒固定；所述激发光纤与所述收集光纤均采用聚合物材料包裹。
  3. 根据权利要求2所述的一种基于纤维内窥镜的拉曼光谱检测装置，其特征在于：所述拉曼光纤探头的合束光纤靠近分支处设置有一可调节的限位装置，所述限位装置用以调节光纤探头进入纤维内窥镜活检通道的长度进行限位，所述限位装置包括包括固定套筒与固定螺丝，所述固定套筒套置在所述拉曼光纤探头的表面，所述固定螺丝 垂直设置于所述固定套筒上，用以对所述拉曼光纤探头进行固定。
  4. 根据权利要求1所述的一种基于纤维内窥镜的拉曼光谱检测装置，其特征在于：所述拉曼光纤探头的探测端部的光纤端面设置有镀膜，所述激发光纤的端面上镀有允许两个波长激发光通过的低通膜，所述收集光纤的端面上镀有用以截止激发光并允许波长更大的拉曼散射光通过的高通膜。
  5. 根据权利要求1所述的一种基于纤维内窥镜的拉曼光谱检测装置，其特征在于：所述双波长激光器的输出光纤通过滤光组件与所述拉曼光纤探头的激发光纤相连接，用以交替输出两种不同波长的激发光；所述拉曼光纤探头的收集光纤与滤光组件连接，其输出光经收集光纤与拉曼光谱仪及其检测器相连接。
  6. 根据权利要求1所述的一种基于纤维内窥镜的拉曼光谱检测装置，其特征在于：所述双波长激光器交替输出两种不同波长的激发光包括785nm激发光与690nm激发光；所述785nm激发光用以完成指纹区拉曼光谱的检测，所述690nm激发光用以完成高波数区拉曼光谱的检测。
  7. 根据权利要求1所述的一种基于纤维内窥镜的拉曼光谱检测装置，其特征在于：所述纤维内窥镜为满足人体腔内不同部位的纤维内窥镜，包括鼻咽镜、膀胱镜、子宫镜、声带关节镜以及输尿管-肾镜。
  8. 根据权利要求1所述的一种基于纤维内窥镜的拉曼光谱检测装 置，其特征在于：所述白光冷光源为300W短弧氙灯白光冷光源。
  9. 一种根据权利要求1所述的基于纤维内窥镜的拉曼光谱检测装置的实现方法，其特征在于：包括以下步骤:
    步骤S1：将所述拉曼光纤探头伸入所述纤维内窥镜的活检通道，采用限位装置调节光纤探头进入所述活检通道的长度，将所述纤维内窥镜伸入人体腔内进行检测；
    步骤S2：开启白光冷光源，将所述摄像装置设置于纤维内窥镜上部，采集所述纤维内窥镜内的图像；将所述摄像装置的输出端与所述显示装置相连，用以显示所述纤维内窥镜内的图像；
    步骤S3：开启所述双波长激光器，并发出两种不同波长的激发光，所述激发光通过激光光纤入射，收集光纤采集所述拉曼散射光传输至所述拉曼光谱仪及其检测器，完成指纹区拉曼光谱的检测与高波数区拉曼光谱的检测。
  10. 根据权利要求1所述的一种基于纤维内窥镜的拉曼光谱检测装置的实现方法，其特征在于：采用所述纤维内窥镜为鼻咽镜时，所述拉曼光谱仪及其检测器获取到人活体鼻咽癌组织和正常鼻咽组织的拉曼光谱特征中，
    在指纹区，正常与肿瘤组织拉曼光谱均在851、943、1004、1096、1124、1265、1316、1450、1621和1660cm-1处有明显的拉曼峰；相较于正常组织，鼻咽癌组织的指纹区拉曼光谱特性在851、943、1096、1124处的峰值发生了下降，而在1004、1265、1316、1450、1621和 1660cm-1处的峰值发生了升高；正常组织和肿瘤组织的光谱的形状在1120-1360cm-1以及1560-1680cm-1区间也存在着明显的差异；
    在高波数区，正常与鼻咽癌组织拉曼光谱均在2854、2940和3009cm-1和3067cm-1处有明显的拉曼峰；相较于正常组织，鼻咽癌组织的高波数区拉曼光谱特性在2854、2940cm-1处的峰值发生了上升。
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