CN1397794A - Method and instrument for detecting infrared spectrum of biological tissue - Google Patents

Abstract

Translated fromChinese

本发明提供了一种采用傅立叶变换中红外光谱检测装置检测生物体组织红外光谱的方法及其专用装置。检测装置是傅立叶变换中红外光谱装置，包括电器系统、干涉仪、检测器、试样装置、红外光源和转镜，红外光源的红外光辐射波长范围为2.5μm～25μm；干涉仪为折射扫描干涉仪；检测器由检测波段为2.5μm～25μm的DTGS或MCT检测器构成；试样装置有三个可互换的装置：水平ATR平台、光纤ATR平台、发射光纤探头装置。采用本发明的方法和装置，3～5分钟即可获得生物体组织分子水平的信息，通过与正常组织的红外光谱信息进行比较，为鉴定组织是否发生变异、变异程度提供重要的参考信息。

The invention provides a method for detecting the infrared spectrum of biological tissue by using a Fourier transform mid-infrared spectrum detection device and a special device thereof. The detection device is a Fourier transform mid-infrared spectroscopy device, including an electrical system, an interferometer, a detector, a sample device, an infrared light source and a rotating mirror. The infrared light radiation wavelength range of the infrared light source is 2.5 μm to 25 μm; The detector is composed of a DTGS or MCT detector with a detection band of 2.5 μm to 25 μm; the sample device has three interchangeable devices: a horizontal ATR platform, an optical fiber ATR platform, and a launching optical fiber probe device. By adopting the method and device of the present invention, information at the molecular level of biological tissue can be obtained within 3 to 5 minutes, and by comparing with the infrared spectrum information of normal tissue, important reference information is provided for identifying whether the tissue has mutated and the degree of mutated.

Description

Translated fromChinese

一种检测生物体组织红外光谱的方法及其专用装置A method and special device for detecting infrared spectrum of biological tissue

技术领域：Technical field:

本发明涉及一种检测生物体组织的红外光谱的方法及其专用装置，属于红外光谱测定及装置技术领域。The invention relates to a method for detecting infrared spectrum of biological tissue and a special device thereof, belonging to the technical field of infrared spectrum measurement and devices.

背景技术：Background technique:

红外光谱法是研究分子结构变化的重要手段之一，红外光谱是研究分子结构变化的灵敏探针，因此，检测生物体组织的红外光谱，可以在早期发现生物体组织的变化信息，发现其在分子水平的变化情况。从理论上讲，这种研究生物体组织分子水平的变化情况的方法，可发展为一种早期诊断的方法。Infrared spectroscopy is one of the important means to study molecular structure changes. Infrared spectroscopy is a sensitive probe for studying molecular structure changes. Therefore, detecting the infrared spectrum of biological tissues can detect the changes in biological tissues at an early stage, and find out the changes in biological tissues. Changes at the molecular level. Theoretically, this method of studying the changes at the molecular level of organism tissues can be developed into a method of early diagnosis.

譬如，癌症是威胁人类健康的一大疾病，而癌症的治愈很大程度决定于早期诊断。癌症的发展过程可概括为：在细胞癌变前，首先是相应的生物分子如核酸、蛋白、磷脂、脂肪、糖及粘蛋白等，其组成、结构、构型和构象等发生变化，进而发展成为癌细胞和肿瘤组织等。一般癌症检测方法，如X-光，CT和MRI都需要在癌细胞发展成肿瘤实体，具有可观测的尺寸大小才能以影像方式看到；而一般常用的病理活检也需要将癌变组织冰冻切片，染色后在显微镜下观察。这一过程至少需要半小时，在手术过程中往往需要等待活检结果出来以后，才能确定具体的后继手术方案。For example, cancer is a major disease that threatens human health, and the cure of cancer largely depends on early diagnosis. The development process of cancer can be summarized as follows: before the cells become cancerous, firstly, corresponding biomolecules such as nucleic acid, protein, phospholipid, fat, sugar and mucin change in composition, structure, configuration and conformation, and then develop into Cancer cells and tumor tissues, etc. General cancer detection methods, such as X-ray, CT and MRI, need to develop into a tumor entity after the cancer cells have an observable size before they can be seen in images; and the commonly used pathological biopsy also requires frozen sections of cancerous tissues. Observe under a microscope after staining. This process takes at least half an hour. During the operation, it is often necessary to wait for the results of the biopsy before determining the specific follow-up operation plan.

用红外光谱检测癌细胞，国际上最早是1986年Benedeth的报道(AppliedSpectroscopy，1986，40，39)，以后陆续有很多报道。一般测定需经过匀浆提取等过程之后进行测定，这不仅需要较长时间，而且破坏样品；同时，现有的光谱装置体积较大，价格昂贵，移动后调试费时费力，不易普及应用。The detection of cancer cells by infrared spectroscopy was first reported by Benedeth in 1986 in the world (Applied Spectroscopy, 1986, 40, 39), and there have been many reports since then. Generally, the measurement needs to be carried out after homogenization and extraction, which not only takes a long time, but also destroys the sample; at the same time, the existing spectroscopic devices are large in size, expensive, time-consuming and laborious to adjust after moving, and are not easy to popularize and apply.

发明内容：Invention content:

本发明的目的是提供一种检测生物体组织红外光谱的方法。The purpose of the present invention is to provide a method for detecting the infrared spectrum of biological tissue.

本发明的另一目的是提供实现所述方法的测定装置，该装置体积小、稳定性好、光通量高、测定精确，并可经长途搬运、稍加调试即可正常使用。Another object of the present invention is to provide a measurement device for realizing the method, which is small in size, good in stability, high in luminous flux, accurate in measurement, and can be used normally after long-distance transportation and a little adjustment.

本发明的检测生物体组织红外光谱的方法是：采用傅立叶变换中红外光谱检测装置进行检测。The method for detecting the infrared spectrum of the biological tissue of the present invention is: adopting a Fourier transform mid-infrared spectrum detection device for detection.

按照离体与活体的生物组织分为两种方式：According to the biological tissue in vitro and in vivo, it can be divided into two ways:

一、离体生物组织的红外光谱检测：1. Infrared spectrum detection of isolated biological tissues:

1.通过外科手术、组织活检获得待测的离体生物组织样品；1. Obtain the isolated biological tissue sample to be tested through surgical operation or tissue biopsy;

2.以标记或描述的方式指定待测的区域、部位；2. Designate the area and part to be tested by marking or describing;

3.将样品置于傅立叶变换中红外光谱检测装置的试样装置上；3. Place the sample on the sample device of the Fourier transform mid-infrared spectrum detection device;

4.控制条件，检测器采集红外光谱信息；4. Control conditions, the detector collects infrared spectrum information;

5.显示或打印该生物组织样品的红外光谱检测结果。5. Display or print the infrared spectrum detection result of the biological tissue sample.

二、活体(在体)生物组织的红外光谱检测：2. Infrared spectrum detection of living (in vivo) biological tissues:

1.借助各种内窥镜(胃镜、十二指肠镜、结肠镜、鼻咽镜、支气管镜、膀胱镜、宫腔镜)、腔镜(腹腔镜、胸腔镜、关节腔镜)或在手术过程中暴露术野，选择待测生物组织部位；1. With the help of various endoscopes (gastroscope, duodenoscope, colonoscope, nasopharyngoscope, bronchoscope, cystoscope, hysteroscope), laparoscopy (laparoscopy, thoracoscopy, arthroscopy) or in Expose the surgical field during the operation and select the biological tissue site to be tested;

2.将傅立叶变换中红外光谱检测装置中的试样装置的ATR光纤探头或发射光纤探头置于待检测部位；2. Place the ATR optical fiber probe or the emission optical fiber probe of the sample device in the Fourier transform mid-infrared spectrum detection device on the part to be detected;

3.控制条件，检测器采集红外光谱信息；3. Control conditions, the detector collects infrared spectrum information;

4.显示或打印该生物组织样品的红外光谱检测结果。4. Display or print the infrared spectrum detection result of the biological tissue sample.

实现上述方法的红外光谱装置，是一种傅立叶变换中红外光谱检测装置，其组成如图1所示，包括：电器系统1，干涉仪2，检测器3，试样装置4，红外光源和转镜5。红外光源的红外光幅射波长范围为2.5μm～25μm，检测器的波段为2.5μm～25μm。其中的电器系统包括电源和各种器件的控制电路和FFT电路是常规的系统，而所述的：The infrared spectrum device realizing the above method is a Fourier transform mid-infrared spectrum detection device, and its composition is as shown in Figure 1, including:electrical system 1,interferometer 2,detector 3,sample device 4, infrared light source and rotary Mirror 5. The infrared radiation wavelength range of the infrared light source is 2.5 μm to 25 μm, and the wavelength band of the detector is 2.5 μm to 25 μm. The electrical system including the power supply and the control circuit and FFT circuit of various devices is a conventional system, and the above:

A.干涉仪：采用折射扫描干涉仪，其中反射镜均为空心直角反射镜，分束镜是楔形镜，所有光学件均固定在同一个五面体铸件中，从而提高仪器的稳定性；同时干涉仪又是密封的，使干涉仪内部与外界不产生空气交换，进一步保证稳定性和避免空气中各种物质对光谱的干扰影响。具有高稳定性、高光通量、小体积的优点。A. Interferometer: A refraction scanning interferometer is used, in which the mirrors are hollow right-angle mirrors, the beam splitter is a wedge mirror, and all optical parts are fixed in the same pentahedron casting, thereby improving the stability of the instrument; simultaneous interference The instrument is also sealed, so that there is no air exchange between the interior of the interferometer and the outside world, further ensuring stability and avoiding the interference of various substances in the air on the spectrum. It has the advantages of high stability, high luminous flux and small volume.

B.检测器由检测波段2.5μm～25μm的DTGS(氘代硫酸三甘肽)或MCT(碲镉汞)检测器构成。B. The detector is composed of a DTGS (deuterated triglyceride sulfate) or MCT (mercury cadmium telluride) detector with a detection band of 2.5 μm to 25 μm.

C.试样装置：有下述三个可互换的装置C. Sample device: There are three interchangeable devices as follows

1.水平ATR平台：如图2所示，为衰减全反射(ATR)组织试样信息采集平台，所用的采样晶体7由(20～40)×20×2mm³的高折射率的Ge、ZnSe或KRS-5晶体构成。传送光幅射的反射镜9由平面镜、球面镜、抛物镜或椭球镜构成；进、出光反射镜为非球面镜，其光斑直径在2mm以下。试样可直接放在水平ATR平台上；该平台上还可以备有给试样加压的装置如压力板11，以提高检测光谱信噪比。1. Horizontal ATR platform: as shown in Figure 2, it is an attenuated total reflection (ATR) tissue sample information acquisition platform, and^the used sampling crystal 7 is made of (20～40) × 20 × 2mm High refractive index Ge, ZnSe Or KRS-5 crystal composition. The reflecting mirror 9 that transmits light radiation is made of plane mirror, spherical mirror, parabolic mirror or ellipsoidal mirror; the incoming and outgoing light reflecting mirrors are aspherical mirrors, and the diameter of its spot is below 2mm. The sample can be directly placed on the horizontal ATR platform; the platform can also be equipped with a device for pressurizing the sample, such as a pressure plate 11, to improve the signal-to-noise ratio of the detection spectrum.

2.光纤ATR平台：如图3所示，采用中红外光纤ATR组织试样采集探头，晶体探头12为由30～75°的Ge、ZnSe或KRS-5材料构成的圆锥体或角体，有两个取样点，以提高信号强度；光纤13为中空的中红外光纤，内径为1mm～5mm，光纤内壁涂有金、金属卤化物(如AgCl)、金属硫化物(如金属硫化镧)等物质，光纤的通光范围为2.5μm～25μm；光纤接口14为用高透过率中红外材料制成的球形聚光镜和扩束镜或平面镜；光纤与光学系统间的耦合件由BaF₂、CaF₂、ZnSe或KRS-5等材料制成。2. Optical fiber ATR platform: as shown in Figure 3, the mid-infrared optical fiber ATR tissue sample collection probe is adopted, and thecrystal probe 12 is a cone or horn made of Ge, ZnSe or KRS-5 materials at 30 to 75°. Two sampling points to increase signal strength;optical fiber 13 is a hollow mid-infrared optical fiber with an inner diameter of 1 mm to 5 mm, and the inner wall of the optical fiber is coated with gold, metal halides (such as AgCl), metal sulfides (such as metal lanthanum sulfide) and other substances , the light transmission range of the optical fiber is 2.5 μm ~ 25 μm; theoptical fiber interface 14 is a spherical condenser lens and a beam expander mirror or a plane mirror made of high transmittance mid-infrared material; the coupling between the optical fiber and the optical system is made of BaF₂ , CaF₂ , ZnSe or KRS-5 and other materials.

3.发射光纤探头装置：如图4所示，采集组织试样发射信息的探头由光纤探头和黑体辐射源组成。中红外光纤18为中空的，直径1～5mm，内壁镀有金、金属卤化物、金属硫化物等物质，通光范围为2.5μm～25μm；光纤探头17和光纤接口19分别为用中红外高透过率的材料BaF2、CaF2或ZnSe制成的聚光镜和扩束镜或平面镜。黑体辐射源22为用石墨等黑体材料制成，用恒定电流控制其温度的装置。3. Emitting optical fiber probe device: as shown in Figure 4, the probe for collecting emission information of tissue samples consists of an optical fiber probe and a black body radiation source. The mid-infraredoptical fiber 18 is hollow, with a diameter of 1 to 5 mm, and the inner wall is plated with gold, metal halide, metal sulfide and other substances, and the light transmission range is 2.5 μm to 25 μm; theoptical fiber probe 17 and the optical fiber interface 19 are used for mid-infrared high Condenser and beam expander or plane mirror made of BaF2, CaF2 or ZnSe materials with high transmittance. The blackbody radiation source 22 is made of blackbody materials such as graphite, and its temperature is controlled by a constant current.

采用本发明的检测生物体组织红外光谱的方法和装置，对于离体组织如手术取下的组织，可以直接测定，3～5分钟即可获得生物体组织分子水平的信息，通过与正常组织的红外光谱信息进行比较，为鉴定生物体组织是否发生变异、变异程度提供重要的参考信息。如对人体的正常组织，癌变组织，以及癌旁组织分别在波段2.5μm～25μm范围内进行扫描检测并作比较，可以为确定组织是否异常或其变异程度提供重要的参考信息。Using the method and device for detecting the infrared spectrum of biological tissue of the present invention, it can be directly measured for isolated tissues such as surgically removed tissues, and information at the molecular level of biological tissues can be obtained in 3 to 5 minutes. Infrared spectral information is compared to provide important reference information for identifying whether biological tissues have mutated and the degree of variation. For example, the normal tissue, cancerous tissue, and paracancerous tissue of the human body are scanned and compared in the range of 2.5 μm to 25 μm, which can provide important reference information for determining whether the tissue is abnormal or its degree of variation.

采用中红外光纤ATR组织试样采集探头时，还可用于生物体组织的原位测定，可能发展成在手术过程中对组织进行直接原位测定的方法。这种直接测定组织红外光谱的方法，既节省测量时间，也免除一切破坏组织的切片染色等步骤，在临床上可减轻病人痛苦，缩短检测时间。When the mid-infrared optical fiber ATR tissue sample collection probe is used, it can also be used for in-situ determination of biological tissues, and may be developed into a method for direct in-situ determination of tissues during surgery. This method of directly measuring the infrared spectrum of the tissue not only saves the measurement time, but also avoids all steps such as section staining that destroys the tissue, which can relieve the pain of the patient and shorten the detection time in clinical practice.

利用本发明的检测方法和检测装置，可以快速获得生物体组织分子水平的变异与否和变异程度，这在生物医学中有广泛的用途，特别是在医疗临床手术中，能快速提供可靠的参考信息。任何离体组织皆可在该本发明的装置的信息采集平台上或利用该装置中的信息采集探头与组织接触进行扫描检测，扫描范围400-4000cm^-1，记录下所测得的红外光谱。如需原位检测，可将中红外光纤的ATR探头置于待测组织的部位，然后进行扫描测定，记录下所测得的红外光谱，将数据储存备用。Utilizing the detection method and detection device of the present invention, the variation and degree of variation at the molecular level of biological tissues can be quickly obtained, which is widely used in biomedicine, especially in medical clinical operations, and can quickly provide reliable reference information. Any isolated tissue can be scanned and detected on the information collection platform of the device of the present invention or by contacting the tissue with the information collection probe in the device. The scanning range is 400-4000cm^-1 , and the measured infrared spectrum can be recorded. If in-situ detection is required, the ATR probe of the mid-infrared optical fiber can be placed on the tissue to be measured, and then the scanning measurement is performed, the measured infrared spectrum is recorded, and the data is stored for future use.

本发明的进一步发展：可以将所获得的患者组织的红外光谱信息(包括峰位、强度、峰宽、谱峰强度等)，与患者病历中的基本数据包括年龄、组织部位、诊断结果、发展阶段等，编成数据库；将所检测得到的各种肿瘤和正常组织的光谱特征结合统计分析结果写成专用软件，在测定新样品组织光谱后输入到软件系统，根据统计分析的结果，进行光谱特征比较，可以为确定组织正常或癌变提供重要的参考。采用本发明的方法和装置，发明人已经对人的口腔、食道、胃、肠、胆囊、肺、肝、肾、甲状腺、卵巢、淋巴结和皮肤等的离体肿瘤组织进行了检测并记录了其光谱图，建立了相应的数据库。Further development of the present invention: the obtained infrared spectral information (including peak position, intensity, peak width, spectral peak intensity, etc.) Stages, etc., compiled into a database; the spectral characteristics of various tumors and normal tissues detected were combined with statistical analysis results to write special software, and after measuring the tissue spectra of new samples, input them into the software system, and perform spectral characteristics analysis according to the results of statistical analysis. The comparison can provide an important reference for determining whether the tissue is normal or cancerous. Using the method and device of the present invention, the inventor has detected and recorded the isolated tumor tissues of human oral cavity, esophagus, stomach, intestine, gallbladder, lung, liver, kidney, thyroid, ovary, lymph node and skin, etc. The corresponding database was established for the spectrogram.

本发明的优点及积极效果：Advantage of the present invention and positive effect:

1.本发明的从分子水平检测生物体光谱的方法，可以得到生物体组织的准确度较高的信息。根据所检测的红外光谱，比较正常组织和变异组织的光谱区别，可以为生物体组织的病变提供早期诊断参考。根据发明人对数百个样品的检测所得的上千张光谱的统计分析，早期诊断准确度可达到85％以上。对于一些疑难病变，采用活体切片镜检难以作出结论的，可以根据所检测到的光谱变化获得重要的参靠信息。1. The method for detecting the spectrum of a living body at the molecular level according to the present invention can obtain high-accuracy information of living body tissues. According to the detected infrared spectrum, comparing the spectral difference between normal tissue and mutant tissue can provide an early diagnosis reference for biological tissue lesions. According to the inventor's statistical analysis of thousands of spectra obtained from the detection of hundreds of samples, the accuracy of early diagnosis can reach more than 85%. For some difficult lesions, it is difficult to draw a conclusion by microscopic examination of biopsy, and important reference information can be obtained according to the detected spectral changes.

2.本发明的红外光谱装置，体积小、稳定性好、光通量高、测定精确，可以放置在可移动的工作台上移动到手术台旁进行检测。2. The infrared spectroscopy device of the present invention is small in size, good in stability, high in luminous flux, and accurate in measurement, and can be placed on a movable workbench and moved to the side of the operating table for detection.

3.利用本发明的红外光谱装置，可以进行原位检测或进行离体组织的检测。3. Using the infrared spectroscopy device of the present invention, in-situ detection or detection of isolated tissues can be performed.

4.利用本发明的方法和装置，检测速度快，只需3～5分钟即可获得生物体组织的光谱信息，比起活检切片法节省很多时间。4. Using the method and device of the present invention, the detection speed is fast, and the spectral information of the biological tissue can be obtained in only 3 to 5 minutes, which saves a lot of time compared with the biopsy section method.

5.本发明的方法和装置，与内窥镜结合，可以发展成为癌症早期诊断方法。5. The method and device of the present invention, combined with an endoscope, can be developed into a method for early diagnosis of cancer.

附图说明：Description of drawings:

图1为本发明的红外光谱装置的方框图，图中：Fig. 1 is the block diagram of infrared spectrum device of the present invention, among the figure:

1-电器系统，2-干涉仪，3-检测器，4-水平ATR平台，5-转镜，6-光纤ATR平台。1-electrical system, 2-interferometer, 3-detector, 4-horizontal ATR platform, 5-rotating mirror, 6-optical fiber ATR platform.

图2(a)为水平ATR平台结构示意图，图2(b)为水平ATR平台沿图2(a)中A-A’方向俯视图，图中：Figure 2(a) is a schematic diagram of the structure of the horizontal ATR platform, and Figure 2(b) is a top view of the horizontal ATR platform along the A-A' direction in Figure 2(a), in the figure:

7-采样晶体，8-晶体托板，9-反射镜，10-支架，11-压力板。7-sampling crystal, 8-crystal support plate, 9-mirror, 10-support, 11-pressure plate.

图3(a)为光纤ATR平台结构示意图，图3(b)为光纤ATR平台中光纤探头结构示意图，图中：Fig. 3 (a) is a schematic structural diagram of an optical fiber ATR platform, and Fig. 3 (b) is a schematic structural diagram of an optical fiber probe in an optical fiber ATR platform. In the figure:

12-晶体探头，13-光纤，14-光纤接口，15-ATR平台，16-光纤探台12-Crystal Probe, 13-Fiber, 14-Fiber Interface, 15-ATR Platform, 16-Fiber Probe

图4为发射光纤探头装置结构示意图，(a)为发射光纤探头结构示意图，(b)为发射光纤探头接口转接和黑体辐射源示意图，图中：Figure 4 is a schematic diagram of the structure of the launching fiber optic probe device, (a) is a schematic diagram of the structure of the launching fiber optic probe, (b) is a schematic diagram of the interface transfer of the launching fiber optic probe and a blackbody radiation source, in the figure:

17-光纤探头，18-光纤，19-光纤接口，20-光纤接口孔，21-光源转镜，22-黑体辐射源，23-光纤探头插孔。17-optical fiber probe, 18-optical fiber, 19-fiber optic interface, 20-fiber optic interface hole, 21-light source rotating mirror, 22-blackbody radiation source, 23-fiber optic probe jack.

图5为良性乳腺肿瘤的红外光谱。Figure 5 is the infrared spectrum of a benign breast tumor.

图6为口腔组织的红外光谱，a为正常组织，b为恶性肿瘤组织。Figure 6 is the infrared spectrum of oral tissue, a is normal tissue, b is malignant tumor tissue.

图7(a)为直肠正常组织的红外光谱，图7(b)为直肠癌组织的红外光谱。Figure 7(a) is the infrared spectrum of normal rectal tissue, and Figure 7(b) is the infrared spectrum of rectal cancer tissue.

图8为甲状腺组织的红外光谱，a为正常组织，b为甲状腺肿瘤组织。Figure 8 is the infrared spectrum of thyroid tissue, a is normal tissue, b is thyroid tumor tissue.

图9为食道组织的红外光谱，a为正常组织，b为食道癌组织。Figure 9 is the infrared spectrum of esophageal tissue, a is normal tissue, b is esophageal cancer tissue.

图10为肺组织的红外光谱，a为正常组织，b为肺癌组织。Figure 10 is the infrared spectrum of lung tissue, a is normal tissue, b is lung cancer tissue.

具体实施方式：Detailed ways:

实例1Example 1

如图1和图2所示，本发明的红外光谱装置，是一种傅立叶变换中红外光谱检测装置，包括：电器系统1，干涉仪2，检测器3，试样装置4，红外光源和转镜5。红外光源的红外光幅射波长范围为2.5μm～25μm，检测器的波段为2.5μm～25μm。其中的电器系统包括电源和各种器件的控制电路和FFT电路是常规的系统。干涉仪2采用折射扫描干涉仪，其中反射镜均为空心直角反射镜，分束镜是楔形镜，所有光学件均固定在同一个五面体铸件中，从而提高仪器的稳定性；同时干涉仪又是密封的，使干涉仪内部与外界不产生空气交换，进一步保证稳定性和避免空气中各种物质对光谱的干扰影响。检测器由检测波段2.5μm～25μm的DTGS(氘代硫酸三甘肽)或MCT(碲镉汞)检测器构成。如图2所示，试样装置为水平ATR平台，所用的采样晶体7由30×20×2mm³的高折射率的Ge晶体构成。传送光幅射的反射镜9由平面镜构成；进、出光反射镜为非球面镜，其光斑直径在2以下。试样组织可直接放在水平ATR平台上；该平台上还可以备有给试样加压的压力板11，以提高检测光谱信噪比。As shown in Figures 1 and 2, the infrared spectroscopy device of the present invention is a Fourier transform mid-infrared spectroscopy detection device, comprising: anelectrical system 1, aninterferometer 2, adetector 3, asample device 4, an infrared light source and a rotary Mirror 5. The infrared radiation wavelength range of the infrared light source is 2.5 μm to 25 μm, and the wavelength band of the detector is 2.5 μm to 25 μm. Among them, the electrical system including power supply, control circuit and FFT circuit of various devices is a conventional system.Interferometer 2 uses a refraction scanning interferometer, in which the mirrors are hollow right-angle mirrors, the beam splitter is a wedge mirror, and all optical parts are fixed in the same pentahedron casting, thereby improving the stability of the instrument; at the same time, the interferometer is also It is sealed, so that there is no air exchange between the inside of the interferometer and the outside world, further ensuring stability and avoiding the interference of various substances in the air on the spectrum. The detector consists of a DTGS (deuterated triglyceride sulfate) or MCT (mercury cadmium telluride) detector with a detection band of 2.5 μm to 25 μm. As shown in Figure 2, the sample device is a horizontal ATR platform, and the sampling crystal 7 used is composed of a 30×20×2 mm³ Ge crystal with a high refractive index. The reflecting mirror 9 that transmits light radiation is made of plane mirror; The sample tissue can be directly placed on the horizontal ATR platform; the platform can also be equipped with a pressure plate 11 for pressurizing the sample to improve the signal-to-noise ratio of the detection spectrum.

采用上述装置，将在医院手术后的离体乳腺肿块样品置于ATR平台，样品的组织外观颇似癌变，在做冰冻切片活检之前，该组织进行中红外光谱检测，所得的光谱如图5所示，测定全过程约需要3分钟。将该光谱与正常组织的红外光谱进行对比，其光谱特征相似，初步判断为良性肿瘤，与活检结果一致。冰冻切片病理检查全过程约30分钟。Using the above-mentioned device, the isolated breast mass sample after surgery in the hospital was placed on the ATR platform. The tissue appearance of the sample was quite cancerous. Before the frozen section biopsy, the tissue was detected by mid-infrared spectrum, and the obtained spectrum was shown in Figure 5. The whole measurement process takes about 3 minutes. Comparing the spectrum with the infrared spectrum of normal tissue, the spectral features are similar, and it is initially judged as a benign tumor, which is consistent with the biopsy results. The whole process of frozen section pathological examination takes about 30 minutes.

实例2Example 2

同实施例1，区别在于红外光谱检测装置采用的试样装置为光纤ATR平台，如图3所示，晶体探头12为由30～75°的Ge、ZnSe或KRS-5材料构成的圆锥体或角体，有两个取样点，以提高信号强度；光纤13为中空的中红外光纤，内径为1mm～5mm，光纤内壁涂有金、金属卤化物(如AgCl)、金属硫化物(如金属硫化镧)等物质，光纤的通光范围为2.5μm～25μm；光纤接口14为用高透过率中红外材料制成的球形聚光镜和扩束镜或平面镜；光纤与光学系统间的耦合件由BaF₂、CaF₂、ZnSe或KRS-5等材料制成。Withembodiment 1, the difference is that the sample device adopted by the infrared spectrum detection device is an optical fiber ATR platform, as shown in Figure 3, thecrystal probe 12 is a cone or There are two sampling points to improve the signal strength; theoptical fiber 13 is a hollow mid-infrared optical fiber with an inner diameter of 1 mm to 5 mm, and the inner wall of the optical fiber is coated with gold, metal halide (such as AgCl), metal sulfide (such as metal sulfide Lanthanum) and other substances, the light-passing range of the optical fiber is 2.5 μm to 25 μm; theoptical fiber interface 14 is a spherical condenser lens and a beam expander mirror or a plane mirror made of a high-transmittance mid-infrared material; the coupling between the optical fiber and the optical system is made of BaF_2. Made of materials such as CaF₂ , ZnSe or KRS-5.

采用上述装置，获得口腔腮腺正常组织和癌变组织的中红外光纤光谱，可观察到明显的的差别，如图6所示。正常组织光谱图a可观察到明显的CH伸缩振动谱带(2800-3000cm^-1)和1746cm^-1峰强度都很高；而恶性肿瘤组织的光谱图b相应谱带的峰强度都很低。Using the above-mentioned device, the mid-infrared optical fiber spectra of the normal tissue and the cancerous tissue of the oral parotid gland can be obtained, and obvious differences can be observed, as shown in FIG. 6 . In normal tissue spectrogram a, obvious CH stretching vibration bands (2800-3000cm^-1 ) and 1746cm^-1 peak intensities are both very high; in malignant tumor tissue spectrogram b, the peak intensities of corresponding bands are very low.

实例3Example 3

同实施例1的方法和装置，手术后离体肠道组织样品，分别进行中红外光谱测定，得到正常直肠和直肠癌的光谱图7(a)和图7(b)。离体的直肠癌组织光谱与正常直肠组织光谱的比较：Using the same method and device as in Example 1, mid-infrared spectrum measurements were performed on isolated intestinal tissue samples after surgery, and the spectra of normal rectum and rectal cancer were obtained in Figure 7(a) and Figure 7(b). Comparison of the spectrum of isolated rectal cancer tissue with that of normal rectal tissue:

从图7(b)可以看到与核酸有关的1082cm^-1处相对峰强度比图7(b)中1086cm^-1明显增多，与基于生物学认识的癌组织中细胞增殖活跃、核酸增多观点相吻合。From Figure 7(b), it can be seen that the relative peak intensity at 1082cm^-1 related to nucleic acid is significantly higher than that at 1086cm^-1 in Figure 7(b), which is consistent with the view of active cell proliferation and increased nucleic acid in cancer tissue based on biological understanding match.

实例4Example 4

同实施例1，区别在于红外光谱检测装置采用的试样装置为发射光纤探头装置，如图4所示，采集组织试样发射信息的探头由光纤探头和黑体辐射源组成。中红外光纤18为中空的，直径1～5mm，内壁镀有金、金属卤化物、金属硫化物等物质，通光范围为2.5μm～25μm；光纤探头17和光纤接口19分别为用中红外高透过率的材料BaF2、CaF2或ZnSe制成的聚光镜和扩束镜或平面镜。黑体辐射源22为用石墨等黑体材料制成，用恒定电流控制其温度的装置。Same asEmbodiment 1, the difference is that the sample device used by the infrared spectrum detection device is an emission fiber optic probe device, as shown in Figure 4, the probe for collecting emission information of tissue samples consists of a fiber optic probe and a blackbody radiation source. The mid-infraredoptical fiber 18 is hollow, with a diameter of 1 to 5 mm, and the inner wall is plated with gold, metal halide, metal sulfide and other substances, and the light transmission range is 2.5 μm to 25 μm; theoptical fiber probe 17 and the optical fiber interface 19 are for mid-infrared high Condenser and beam expander or plane mirror made of BaF2, CaF2 or ZnSe materials with high transmittance. The blackbody radiation source 22 is made of blackbody materials such as graphite, and its temperature is controlled by a constant current.

如图8所示，采用上述装置获得甲状腺正常组织光谱a和肿瘤组织光谱b，可以发现：在1000～1300cm^-1谱带与1400～1460cm^-1谱带相对峰强度比在正常组织光谱图a中接近于1，而在光谱图b中的吸收峰较弱，相对强度比小于1。As shown in Figure 8, using the above device to obtain the spectrum a of normal thyroid tissue and the spectrum b of tumor tissue, it can be found that the relative peak intensity ratio between the 1000-1300 cm^-1 band and the 1400-1460 cm^-1 band is in the normal tissue spectrum a is close to 1, while the absorption peak in spectrogram b is weak, and the relative intensity ratio is less than 1.

实例5Example 5

同实施例1的方法和装置，获得正常食道组织和食道癌组织的光谱。如图9所示，正常食道组织光谱图a与食道癌组织光谱图b比较：在CH伸缩振动谱带(2800～3000cm^-1)，图a可见吸收峰，而图b看不到；图a的1240cm^-1处磷酸二酯键的对称伸缩振动明显，而图b此处较弱。The spectra of normal esophageal tissue and esophageal cancer tissue were obtained with the method and device in Example 1. As shown in Figure 9, the normal esophageal tissue spectrogram a is compared with the esophageal cancer tissue spectrogram b: in the CH stretching vibration band (2800～3000cm^-1 ), the absorption peak can be seen in figure a, but not in figure b; in figure a The symmetrical stretching vibration of the phosphodiester bond at 1240cm^-1 is obvious, while it is weaker in Figure b.

实例6Example 6

同实施例1的方法和装置，获得离体肺组织的光谱，图10列出了正常肺组织和肺癌组织的红外光谱。正常肺组织光谱图a与肺癌组织光谱图b比较：在图a中酰胺II带1549cm^-1较强，而在图b中明显减弱；在正常组织中酰胺I带峰位1644cm^-1，而在癌组织中为1640cm^-1。The spectrum of the isolated lung tissue was obtained with the method and device in Example 1, and FIG. 10 lists the infrared spectra of normal lung tissue and lung cancer tissue. Comparison of normal lung tissue spectrogram a and lung cancer tissue spectrogram b: in picture a, the amide II band is stronger at 1549 cm^-1 , but in picture b it is obviously weaker; in normal tissue, the peak of amide I band is 1644 cm^-1 It is 1640cm^-1 in cancer tissue.

Claims (5)

1. the method for a detection of biological soma infrared spectrum is characterized in that it being to adopt Fourier transform mid-infrared light spectrum detection device that the bio-tissue of stripped or live body is detected.

2. realizing the infrared spectrum device of the described method of claim 1, is a kind of Fourier transform mid-infrared light spectrum detection device, comprises electric system, interferometer, and detecting device, the sample device, infrared light supply and tilting mirror is characterized in that:

The infrared radiat-ion wavelength coverage of described infrared light supply is 2.5 μ m～25 μ m;

Described interferometer is the refraction scanning interferometer, and wherein catoptron is hollow corner cube mirror, and beam splitter is the wedge shape mirror, and all optical elements all are fixed in the same pentahedron foundry goods, and interferometer seals simultaneously;

Described detecting device is made of the DTGS or the MCT detecting device that detect wave band 2.5 μ m～25 μ m;

Described sample device is horizontal ATR platform, and used sampling crystal is by (20～40) * 20 * 2mm³Ge, the ZnSe of high index of refraction or KRS-5 crystal constitute; The catoptron that the transmission light width of cloth is penetrated is made of level crossing, spherical mirror, parabolic lens or ellipsoidal mirror; Into and out of light reflection mirror is aspheric mirror, and its spot diameter is below 2mm.

3. infrared spectrum device as claimed in claim 2 is characterized in that being equipped with on the described sample device device to the test samples pressurization.

4. realizing the infrared spectrum device of the described method of claim 1, is a kind of Fourier transform mid-infrared light spectrum detection device, comprises electric system, interferometer, and detecting device, the sample device, infrared light supply and tilting mirror is characterized in that:

The infrared radiat-ion wavelength coverage of described infrared light supply is 2.5 μ m～25 μ m;

Described interferometer is the refraction scanning interferometer, and wherein catoptron is hollow corner cube mirror, and beam splitter is the wedge shape mirror, and all optical elements all are fixed in the same pentahedron foundry goods, and interferometer seals simultaneously;

Described detecting device is made of the DTGS or the MCT detecting device that detect wave band 2.5 μ m～25 μ m;

Described sample device is an optical fiber ATR platform: adopt middle infrared optical fiber ATR tissue sample acquisition probe, cone or the angle body of crystal probe for being made of 30～75 ° Ge, ZnSe or KRS-5 material has two sampling spots; Optical fiber is the middle infrared optical fiber of hollow, and internal diameter is 1mm～5mm, and the optical fiber inwall scribbles gold or metal halide or metal sulfide, and the logical optical range of optical fiber is 2.5 μ m～25 μ m; Spherical condenser and beam expanding lens or the level crossing of optical fiber interface for making with the high permeability middle infrared material; Male part between optical fiber and optical system is by BaF₂, CaF₂, ZnSe or KRS-5 material make.

5. realizing the infrared spectrum device of the described method of claim 1, is a kind of Fourier transform mid-infrared light spectrum detection device, comprises electric system, interferometer, and detecting device, the sample device, infrared light supply and tilting mirror is characterized in that:

The infrared radiat-ion wavelength coverage of described infrared light supply is 2.5 μ m～25 μ m;

Described interferometer is the refraction scanning interferometer, and wherein catoptron is hollow corner cube mirror, and beam splitter is the wedge shape mirror, and all optical elements all are fixed in the same pentahedron foundry goods, and interferometer seals simultaneously;

Described detecting device is made of the DTGS or the MCT detecting device that detect wave band 2.5 μ m～25 μ m;

Described sample device is the launching fiber probe apparatus: the probe of gathering tissue sample emission information is made up of fibre-optical probe and blackbody radiation source, optical fiber is the middle infrared optical fiber of the hollow of diameter 1mm～5mm, inwall is coated with gold or metal halide or metal sulfide, and the logical optical range of optical fiber is 2.5 μ m～25 μ m; Material B aF2, the CaF2 of infrared high permeability or condenser and beam expanding lens or the level crossing that ZnSe makes during fibre-optical probe and optical fiber interface are respectively and use; Blackbody radiation source is controlled the device of its temperature for to make with the black matrix material with steady current.

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