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

Method and instrument for detecting infrared spectrum of biological tissue Download PDF

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Publication number
CN1397794A
CN1397794A CN 02129410 CN02129410A CN1397794A CN 1397794 A CN1397794 A CN 1397794A CN 02129410 CN02129410 CN 02129410 CN 02129410 A CN02129410 A CN 02129410A CN 1397794 A CN1397794 A CN 1397794A
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infrared
optical fiber
interferometer
mirror
spectrum
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CN1217180C (en
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吴瑾光
翁诗甫
徐怡庄
杨丽敏
张莉
杨展澜
李维红
徐端夫
赵莹
武惠忠
石景森
任予
周孝思
张元福
徐智
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Peking University
Beijing Second Optical Instrument Factory
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Peking University
Beijing Second Optical Instrument Factory
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Abstract

A method and equipment for detecting the infrared spectrum of biological tissue is disclosed. The said equipment is an infrared spectrometer using fourier transform, which is composed of electric system, refraction-scan interferometer, DTGS or MCT detector, specimen unit, infrared light source and rotary mirror. The said specimen unit has three exchangeable parts: horizontal ATR platform, optical fibre ATR platform and optical-fibre emitting probe. The molecular information about biological tissue can be obtained within 3-5 min.

Description

The method and the isolated plant thereof of a kind of detection of biological soma infrared spectrum
Technical field:
The present invention relates to the method and the isolated plant thereof of the infrared spectrum of a kind of detection of biological soma, belong to infrared spectrum measurement and device technique field.
Background technology:
Infra-red sepectrometry is one of important means of research molecule structure change, infrared spectrum is the sensitive probe of research molecule structure change, therefore, and the infrared spectrum of detection of biological soma, can find the change information of bio-tissue in early days, find its situation of change at molecular level.Theoretically, the method for the situation of change of this research bio-tissue molecular level can develop into a kind of method of early diagnosis.
For example, cancer is a big disease that threatens human health, and the healing of cancer largely is decided by early diagnosis.The evolution of cancer can be summarized as: before cell carcinogenesis, at first be corresponding biomolecule such as nucleic acid, albumen, phosphatide, fat, sugar and mucin etc., its composition, structure, configuration and conformation etc. change, and then develop into cancer cell and tumor tissues etc.General method for detecting cancer, as X-light, CT and MRI need develop into the tumour entity at cancer cell, and having observable size could see in the image mode; And general pathological biopsy commonly used also needs the cancerous issue frozen section, examines under a microscope after the dyeing.This process needs half an hour at least, often need in surgical procedure to wait for that the biopsy result comes out after, could determine concrete follow-up operation plan.
Detecting cancer cell with infrared spectrum, is the report (AppliedSpectroscopy, 1986,40,39) of Benedeth in 1986 in the world the earliest, and a lot of reports are arranged later on successively.General mensuration needs through measuring after the processes such as homogenate extraction, and this not only needs the long period, and destroys sample; Simultaneously, existing spectral device volume is bigger, costs an arm and a leg, and mobile post debugging is wasted time and energy, and is difficult for popularization and application.
Summary of the invention:
The method that the purpose of this invention is to provide a kind of detection of biological soma infrared spectrum.
Another object of the present invention provides the determinator of realizing described method, and this device volume is little, good stability, luminous flux height, measure accurately, and can be through long-distance carrying, debugging can normally be used a little.
The method of detection of biological of the present invention soma infrared spectrum is: adopt Fourier transform mid-infrared light spectrum detection device to detect.
Be divided into dual mode according to the biological tissue that exsomatizes with live body:
One, the infrared spectrum of stripped biological tissue detects:
1. obtain stripped biological tissue samples to be measured by operation, biopsy;
2. specify zone to be measured, position in the mode of mark or description;
3. sample is placed on the sample device of Fourier transform mid-infrared light spectrum detection device;
4. controlled condition, detecting device is gathered infrared spectrum information;
5. show or print the infrared spectrum testing result of this biological tissue samples.
Two, the infrared spectrum of live body (at body) biological tissue detects:
1. expose the art open country by various endoscopes (gastroscope, duodenoscope, Sigmoidoscope, masopharyngeal mirror, bronchoscope, cystoscope, hysteroscope), chamber mirror (laparoscope, thoracoscope, articular cavity mirror) or in surgical procedure, select biological tissue to be measured position;
2. ATR fibre-optical probe or the launching fiber probe with the sample device in the Fourier transform mid-infrared light spectrum detection device places position to be detected;
3. controlled condition, detecting device is gathered infrared spectrum information;
4. show or print the infrared spectrum testing result of this biological tissue samples.
Realizing the infrared spectrum device of said method, is a kind of Fourier transform mid-infrared light spectrum detection device, and it is formed as shown in Figure 1, comprising: electric system 1, interferometer 2, detecting device 3, sample device 4, infrared light supply and tilting mirror 5.The infrared radiat-ion wavelength coverage of infrared light supply is 2.5 μ m~25 μ m, and the wave band of detecting device is 2.5 μ m~25 μ m.Electric system wherein comprises that the control circuit of power supply and various devices and fft circuit are conventional systems, and described:
A. interferometer: adopt the refraction scanning interferometer, 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, thereby improves the stability of instrument; Simultaneously interferometer seal again, makes that interferometer is inner not to produce air exchange with the external world, further guarantees stability and avoids in the air various materials to the disturbing effect of spectrum.Advantage with high stability, high light flux, small size.
B. detecting device is made of DTGS (deuterium thiosulfuric acid triglycine) that detects wave band 2.5 μ m~25 μ m or MCT (mercury cadmium telluride) detecting device.
C. sample device: following three interchangeable devices are arranged
1. horizontal ATR platform: as shown in Figure 2, be attenuated total reflection (ATR) tissue sample information collecting platform, used sampling crystal 7 is by (20~40) * 20 * 2mm 3Ge, the ZnSe of high index of refraction or KRS-5 crystal constitute.The catoptron 9 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.Sample can be placed directly on the horizontal ATR platform; Can also have device such as pressure plare 11 on this platform, detect spectral signal-noise ratio to improve to the sample pressurization.
2. optical fiber ATR platform: as shown in Figure 3, adopt middle infrared optical fiber ATR tissue sample acquisition probe, cone or the angle body of crystal probe 12 for being made of 30~75 ° Ge, ZnSe or KRS-5 material has two sampling spots, to improve signal intensity; Optical fiber 13 is the middle infrared optical fiber of hollow, and internal diameter is 1mm~5mm, and the optical fiber inwall scribbles gold, metal halide (as AgCl), metal sulfide materials such as (as the metal lanthanum sulfides), 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 14 for making with the high permeability middle infrared material; Male part between optical fiber and optical system is by BaF 2, CaF 2, material such as ZnSe or KRS-5 makes.
3. launching fiber probe apparatus: as shown in Figure 4, gather the probe of tissue sample emission information and form by fibre-optical probe and blackbody radiation source.Middle infrared optical fiber 18 is a hollow, and diameter 1~5mm, inwall are coated with materials such as gold, metal halide, metal sulfide, and logical optical range 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 17 and optical fiber interface 19 are respectively and use.Blackbody radiation source 22 is controlled the device of its temperature for to make with black matrix materials such as graphite with steady current.
Adopt the method and apparatus of detection of biological of the present invention soma infrared spectrum, the tissue that takes off in vitro tissue such as operation, can directly measure, can obtain the information of bio-tissue molecular level in 3~5 minutes, compare by infrared spectrum information, for whether identification of organism soma morphs, degree of variation provides important reference information with normal structure.As the normal structure to human body, whether cancerous issue, and cancer beside organism scans in wave band 2.5 μ m~25 mu m ranges respectively and detect and make comparisons can be for determining to organize unusual or its degree of variation provides important reference information.
When adopting middle infrared optical fiber ATR tissue sample acquisition probe, also can be used for the in-site detecting of bio-tissue, may develop in surgical procedure tissue is carried out the direct in-situ method for measuring.This direct mensuration is organized the method for infrared spectrum, has both saved Measuring Time, also exempts steps such as all histoclastic section statinings, can alleviate patient's misery clinically, shortens detection time.
Utilize detection method of the present invention and pick-up unit, the variation that can obtain the bio-tissue molecular level fast whether and degree of variation, this has been widely used in biomedicine, particularly in medical clinical operation, can provide reliable reference information fast.Any in vitro tissue all can be on the information collecting platform of this device of the present invention or is utilized the information acquisition probe in this device to scan detection, sweep limit 400-4000cm with organizing to contact -1, note measured infrared spectrum.As the need in situ detection, the ATR probe of middle infrared optical fiber can be placed the position of tissue to be measured, carry out sweep measuring then, note measured infrared spectrum, data storing is standby.
Of the present invention further developing: can be with the infrared spectrum information (comprising peak position, intensity, peak width, spectral strength etc.) of the patient tissue that obtained, comprise age, tissue site, diagnostic result, developing stage etc. with the master data in the patient medical record, weave into database; With detect the various tumours that obtain and the spectral signature of normal structure is write as special software in conjunction with statistic analysis result; after measuring the fresh sample tissue spectrum, be input to software systems; according to The result of statistics; carry out spectral signature relatively, can provide important reference for determining the normal or canceration of tissue.Adopt method and apparatus of the present invention, the inventor has carried out detecting and having write down its spectrogram to the tumor tissue in vitro of people's oral cavity, esophagus, stomach, intestines, gall-bladder, lung, liver, kidney, thyroid gland, ovary, lymph node and skin etc., has set up corresponding database.
Advantage of the present invention and good effect:
1. the method from molecular level detection of biological body spectrum of the present invention can obtain the higher information of accuracy of bio-tissue.According to the infrared spectrum that is detected, relatively the spectrum difference of normal structure and variation tissue can provide the early diagnosis reference for the pathology of bio-tissue.According to the statistical study of inventor to the last thousand sheets spectrum of the detection gained of hundreds of samples, the early diagnosis accuracy can reach more than 85%.For some difficult pathologies, adopt the vivisection microscopy to be difficult to draw a conclusion, can obtain important ginseng by information according to detected spectrum change.
2. infrared spectrum device of the present invention, volume is little, good stability, luminous flux height, measure accurately, can be placed on movably to move on the worktable that operating table is other to be detected.
3. utilize infrared spectrum device of the present invention, can carry out in situ detection or carry out the detection of in vitro tissue.
4. utilize method and apparatus of the present invention, detection speed is fast, only needs can obtain in 3~5 minutes the spectral information of bio-tissue, saves a lot of times compared with the biopsy slice method.
5. method and apparatus of the present invention combines with endoscope, can develop into the early diagnosis of cancer method.
Description of drawings:
Fig. 1 is the block scheme of infrared spectrum device of the present invention, among the figure:
The 1-electric system, 2-interferometer, 3-detecting device, the horizontal ATR platform of 4-, 5-tilting mirror, 6-optical fiber ATR platform.
Fig. 2 (a) is horizontal ATR platform structure synoptic diagram, and Fig. 2 (b) is horizontal ATR platform along A-A ' direction vertical view among Fig. 2 (a), among the figure:
The 7-crystal of sampling, 8-crystal supporting plate, 9-catoptron, 10-support, 11-pressure plare.
Fig. 3 (a) is an optical fiber ATR platform structure synoptic diagram, and Fig. 3 (b) is a fibre-optical probe structural representation in the optical fiber ATR platform, among the figure:
The 12-crystal probe, 13-optical fiber, the 14-optical fiber interface, the 15-ATR platform, 16-optical fiber is visited platform
Fig. 4 is a launching fiber probe apparatus structural representation, (a) is launching fiber sonde configuration synoptic diagram, (b) is switching of launching fiber probe interface and blackbody radiation source synoptic diagram, among the figure:
The 17-fibre-optical probe, 18-optical fiber, 19-optical fiber interface, 20-fiber interface hole, 21-light source tilting mirror, 22-blackbody radiation source, 23-fibre-optical probe jack.
Fig. 5 is the infrared spectrum of optimum tumor of breast.
Fig. 6 is the infrared spectrum of oral cavity tissue, and a is a normal structure, and b is a malignant tumor tissue.
Fig. 7 (a) is the infrared spectrum of rectum normal structure, and Fig. 7 (b) is the infrared spectrum of rectum cancer tissue.
Fig. 8 is the infrared spectrum of parathyroid tissue, and a is a normal structure, and b is a thyroid neoplasm tissue.
Fig. 9 is the infrared spectrum of esophageal tissue, and a is a normal structure, and b is the cancer of the esophagus tissue.
Figure 10 is the infrared spectrum of lung tissue, and a is a normal structure, and b is a cancerous lung tissue.
Embodiment:
Example 1
As depicted in figs. 1 and 2, infrared spectrum device of the present invention is a kind of Fourier transform mid-infrared light spectrum detection device, comprising: electric system 1, interferometer 2, detecting device 3, sample device 4, infrared light supply and tilting mirror 5.The infrared radiat-ion wavelength coverage of infrared light supply is 2.5 μ m~25 μ m, and the wave band of detecting device is 2.5 μ m~25 μ m.Electric system wherein comprises that the control circuit of power supply and various devices and fft circuit are conventional systems.Interferometer 2 adopts 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, thereby improves the stability of instrument; Simultaneously interferometer seal again, makes that interferometer is inner not to produce air exchange with the external world, further guarantees stability and avoids in the air various materials to the disturbing effect of spectrum.Detecting device is made of DTGS (deuterium thiosulfuric acid triglycine) that detects wave band 2.5 μ m~25 μ m or MCT (mercury cadmium telluride) detecting device.As shown in Figure 2, the sample device is horizontal ATR platform, and used sampling crystal 7 is by 30 * 20 * 2mm 3The Ge crystal of high index of refraction constitute.Transmitting catoptron that the light width of cloth penetrates 9 is made of level crossing; Into and out of light reflection mirror is aspheric mirror, and its spot diameter is below 2.The sample tissue can be placed directly on the horizontal ATR platform; Can also have pressure plare 11 on this platform, detect spectral signal-noise ratio to improve to the sample pressurization.
Adopt said apparatus, the stripped breast lump sample behind hospital surgical is placed the ATR platform, the tissue appearance of sample is quite like canceration, before cooking the frozen section biopsy, this tissue carries out middle infrared spectrum and detects, the spectrum of gained as shown in Figure 5, measuring overall process needs 3 minutes approximately.The infrared spectrum of this spectrum and normal structure is compared, and its spectral signature is similar, tentatively is judged as benign tumour, and the result is consistent with biopsy.About 30 minutes of frozen section pathologic finding overall process.
Example 2
With embodiment 1, difference is that the sample device that the infrared spectrum pick-up unit adopts is an optical fiber ATR platform, as shown in Figure 3, and cone or the angle body of crystal probe 12 for constituting by 30~75 ° Ge, ZnSe or KRS-5 material, two sampling spots are arranged, to improve signal intensity; Optical fiber 13 is the middle infrared optical fiber of hollow, and internal diameter is 1mm~5mm, and the optical fiber inwall scribbles gold, metal halide (as AgCl), metal sulfide materials such as (as the metal lanthanum sulfides), 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 14 for making with the high permeability middle infrared material; Male part between optical fiber and optical system is by BaF 2, CaF 2, material such as ZnSe or KRS-5 makes.
Adopt said apparatus, obtain the middle infrared optical fiber spectrum of oral cavity parotid gland normal structure and cancerous issue, can be observed tangible difference, as shown in Figure 6.Normal tissue spectrum figure a can be observed tangible CH stretching vibration bands of a spectrum (2800-3000cm -1) and 1746cm -1Peak intensity is all very high; And the peak intensity of the corresponding bands of a spectrum of spectrogram b of malignant tumor tissue is all very low.
Example 3
With the method and apparatus of embodiment 1, the stripped intestinal tissue sample in operation back carries out middle infrared spectrum respectively and measures, and obtains spectrogram 7 (a) and Fig. 7 (b) of the normal rectum and the carcinoma of the rectum.Rectum cancer tissue's spectrum that exsomatizes and the normally comparison of rectal tissue spectrum:
Can see the 1082cm relevant from Fig. 7 (b) with nucleic acid -1Locate relative peak intensity than the middle 1086cm of Fig. 7 (b) -1Showed increased increases viewpoint and matches with, nucleic acid active based on cell proliferation in the cancerous tissue of biology understanding.
Example 4
With embodiment 1, difference is that the sample device that the infrared spectrum pick-up unit adopts is the launching fiber probe apparatus, as shown in Figure 4, gathers the probe of tissue sample emission information and is made up of fibre-optical probe and blackbody radiation source.Middle infrared optical fiber 18 is a hollow, and diameter 1~5mm, inwall are coated with materials such as gold, metal halide, metal sulfide, and logical optical range 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 17 and optical fiber interface 19 are respectively and use.Blackbody radiation source 22 is controlled the device of its temperature for to make with black matrix materials such as graphite with steady current.
As shown in Figure 8, adopt said apparatus to obtain thyroid gland normal tissue spectrum a and tumor tissues spectrum b, can find: at 1000~1300cm -1Bands of a spectrum and 1400~1460cm -1The relative peak intensity of bands of a spectrum approaches 1 than in normal tissue spectrum figure a, and the absorption peak in spectrogram b a little less than, relative intensity ratio is less than 1.
Example 5
With the method and apparatus of embodiment 1, obtain the spectrum of normal esophageal tissue and cancer of the esophagus tissue.As shown in Figure 9, normal spectrogram a of esophageal tissue and cancer of the esophagus tissue spectrum figure b are relatively: at CH stretching vibration bands of a spectrum (2800~3000cm -1), figure a visible absorbance peak, and figure b can't see; The 1240cm of figure a -1The place phosphodiester bond symmetrical stretching vibration obvious, and figure b herein a little less than.
Example 6
With the method and apparatus of embodiment 1, obtain the spectrum of stripped lung tissue, Figure 10 has listed the infrared spectrum of normal lung tissue and cancerous lung tissue.Spectrogram a of normal lung tissue and cancerous lung tissue spectrogram b compare: acid amides II band 1549cm in figure a -1Stronger, and in figure b, obviously weaken; Acid amides I band peak position 1644cm in normal structure -1, and be 1640cm in cancerous tissue -1

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 * 2mm 3Ge, 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 BaF 2, CaF 2, 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.
CN 02129410 2002-08-21 2002-08-21 Method and instrument for detecting infrared spectrum of biological tissue Expired - Fee Related CN1217180C (en)

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WO2005072599A1 (en) * 2004-01-19 2005-08-11 Peking University Method and apparatus for noninvasively detecting the organism tissue
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WO2012075698A1 (en) * 2010-12-10 2012-06-14 广州宝胆医疗器械科技有限公司 Hysteroscope system with infrared thermal scanning function
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CN108603832A (en) * 2015-12-30 2018-09-28 蓝海新星股份有限公司 Equipment for analyzing the article to be analyzed being located in product chambers
CN105699323A (en) * 2016-03-08 2016-06-22 中国科学院合肥物质科学研究院 Drug detection system based on Fourier transform NIRSA (near infrared spectroscopy analysis) technology
CN112384786A (en) * 2018-05-09 2021-02-19 国立研究开发法人量子科学技术研究开发机构 Tissue recognition device, tissue recognition system and method, examination method, endoscope system, tissue recognition program, and recording medium
CN111631681A (en) * 2020-04-21 2020-09-08 上海市杨浦区市东医院 Diagnosis system for detecting oral cancer based on infrared Fourier spectrum
CN112881330A (en) * 2021-03-23 2021-06-01 颜旭东 Intelligent tumor infrared spectrum detection system and detection method based on 5G and block chain
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