CN101975769A - Human tissue autofluorescence detection system based on excitation of light sources with different wavelength - Google Patents
Human tissue autofluorescence detection system based on excitation of light sources with different wavelength Download PDFInfo
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Abstract
The invention relates to an autofluorescence detection system for exciting specific endogenous fluorescent substances in a human tissue by light sources with different wavelengths and detecting information of fluorescence with specific wavelength corresponding to the fluorescent substances in endoscopy for the human early tumor tissue. The autofluorescence detection system mainly consists of an excitation light source part, an excitation-detection-acquisition part, an optical collection part, a signal preprocessing part and a computer part. The autofluorescence detection system is characterized in that a tunable laser (1) can adjust the needed pulse laser wavelengths according to the specific endogenous fluorescent substances in the human tissue, and a filter wheel (10) is equipped with narrow band pass filters with different center wavelengths. The system can excite the specific endogenous fluorescent substances in the human tissue by a plurality of optimal excitation wavelengths sequentially, thus avoiding the problem of no benefit for analysis owing to insufficient fluorescence information or information overlap and the like caused by excitation of single wavelength or broad spectrum light sources, and improving sensitivity and specificity of early tumor detection.
Description
Technical field
The invention belongs to biomedical quasi-instrument, relate to a kind of tissue autofluorescence detection system, especially a kind of endoscopy that can be used for human body infantile tumour tissue, utilize endogenous fluorescent material specific in the light sources with different wavelengths exciting human tissue, and the autofluorescence detection system of detection and the corresponding specific wavelength fluorescence information of these fluorescent materials.
Technical background
Existing tissue autofluorescence detection system is by utilizing conventional endoscope, and the endogenous fluorescent material of the light source activation tissue of employing single wavelength or wide spectrum, utilization autofluorescence SPECTRAL DIVERSITY normal and tumor tissues is carried out fluoroscopic examination to tissue system.The autofluorescence of tissue is mainly derived from materials such as amino acid, structural proteins, enzyme, coenzyme, fat, vitamin and porphyrin, they have different best fluorescence exciting wavelengths and corresponding with it fluorescent emission wavelength separately, wherein common endogenous fluorescent material such as pairing maximum excitation wavelengths such as tryptophane, collagen, nicotinamide adenine dinucleotide reduced, porphyrin and flavin adenine dinucleotide (FAD) are respectively 280,325,340,400 and 450nm, and corresponding optimal fluorescence emission wavelength is respectively 335,390,460,635 and 535nm.Under the exciting of a certain specific wavelength excitation source, tissue autofluorescence spectrum is the stack of multiple endogenous fluorescent material spectrum.So far, obtained the autofluorescence detection system that clinical principium uses (1) Xillix-LIFE lung/GI/ENT (Xillix Technologies Corp, Richmond, BC, Canada) (excitation wavelength 442nm) have been arranged; (2) D-light System (Karl Storz, Tuttlingen, Germany) (excitation wavelength 442nm); (3) WavSTAT, Optical Biopsy System (SpectraScience, San Diego, USA) (excitation wavelength 410nm); (4) SAFE-3000 (Pentax, Tokyo, Japan) (excitation wavelength 408nm); (5) Evis LuceraSpectrum (Olympus, Japan) (excitation wavelength 395-475nm).These autofluorescence endoscopy examining systems adopt the single wavelength (408,410 and 442nm) or the wide excitation of spectra light source (395-475nm) of visible light wave ranges, these excitation wavelengths not only can not excite the fluorescence of tryptophane, collagen and nicotinamide adenine dinucleotide reduced, and can not excite the fluorescence of porphyrin effectively.Therefore, the detected fluorescence signal of system is mainly derived from part porphyrin and flavin adenine dinucleotide (FAD).Excitation wavelength is too single or too broadly all fail to excite the fluorescence information with diagnostic significance effectively, still can't realize the high sensitivity and the specific detection of human body infantile tumour tissue in clinical practice.
Summary of the invention
Adopt single wavelength or broad spectrum light source as excitation source at existing tissue autofluorescence detection system, fail to excite effectively the fluorescence information of endogenous fluorescent material with diagnostic significance, in clinical practice, there is the not high deficiency of sensitivity and specificity, the invention provides a kind of tissue autofluorescence detection system based on the multi-wavelength light source excitation, this system can adopt specific endogenous fluorescent material in a plurality of maximum excitation wavelength exciting human tissues, detect corresponding fluorescence information, avoid adopting single wavelength or broad spectrum light source to excite causing the abundant inadequately or information overlap of fluorescence information to be unfavorable for problems such as analysiss, thus the detection sensitivity and the specificity of raising human body infantile tumour tissue.
For realizing that the technical scheme that purpose of the present invention adopts is: the autofluorescence detection system is by the excitation source part, excite-detect-collecting part, the optical collection part, Signal Pretreatment part and computing machine partly constitute, wherein the Signal Pretreatment part is connected by signal wire with the computing machine part, excitation source part by exciting light with excite-detect-optical fiber of exciting light input end in the collecting part links to each other, excite-detect-optical fiber of fluorescence output terminal in the collecting part partly links to each other with optical collection by fluorescent signal path, fluorescent signal path enters the Signal Pretreatment part after partly transmitting through optical collection, become electric signal after photomultiplier photoelectricity transforms in the Signal Pretreatment part, enter the computing machine part.
Described excitation source part is made up of Wavelength tunable laser, optical attenuator and coupled lens, constitutes the exciting light path.Described tunable laser can be regulated required pulse laser wavelength according to endogenous fluorescent material specific in the tissue, the wavelength coverage that can regulate successively is from 210nm to 2200nm, having overcome existing tissue autofluorescence detection system adopts single wavelength or broad spectrum light source as excitation source, fail to excite effectively the fluorescence information of endogenous fluorescent material, in clinical practice, have the not high deficiency of sensitivity and specificity with diagnostic significance; The pulsed laser power of tunable laser output can be regulated by optical attenuator, and the exciting light after the adjusting is coupled into through coupled lens in the optical fiber of exciting light input end of y-type optical fiber.
Described exciting-detect-collecting part is made up of optical fiber and endoscope that two-way is the Y type, and wherein a route six roots of sensation of y-type optical fiber is transmitted the optical fiber of exciting light, and another road is the fluorescence output terminal of single transmission fluorescence optical fiber.Optical fiber one end of transmission exciting light is the exciting light input end, and the other end is an incoming end, and in the biopsy channel access endoscope by endoscope, the exciting light that excitation source partly sends is by this fiber-optic illuminated tissue to be measured; Optical fiber one end of transmission fluorescence is the fluorescence output terminal, and the other end also is an incoming end, in the biopsy channel access endoscope by endoscope.The incoming end of the optical fiber incoming end of transmission exciting light and transmission fluorescence optical fiber merges the afterbody that constitutes Y-shaped, and the output terminal of the optic fibre input end of transmission exciting light and transmission fluorescence optical fiber separately constitutes two heads of Y-shaped.The autofluorescence that tissue to be measured sends partly transmits fluorescence by the fluorescence output terminal of this optical fiber to optical collection.Constitute " exciting light-optical fiber-endoscope-irradiation-fluorescence excitation " and " collection fluorescence-optical fiber-spread out of fluorescence " light-path like this.
Described optical collection part is made up of filter wheel and two coupled lens, the fluorescence of the optical fiber fluorescence output terminal output of transmission fluorescence, by the coupling of preceding coupled lens after filter in the halo and the corresponding optical filter of excitation wavelength, again by back coupled lens coupling input signal preprocessing part.Filter wheel adopts the universal porous filter wheel that can buy on the market, and the narrow band pass filter of different centre wavelengths is installed according to the wavelength of correspondence in the hole.When on the filter wheel 6 narrow band pass filters being installed, centre wavelength can be selected 335nm respectively, 390nm, 460nm, 520nm, 535nm, and 635nm, bandwidth is 20nm, be used for extracting respectively the fluorescence signal that the 280nm excitation produces 335nm, the fluorescence signal of generation 390 of 340nm excitation and 460nm, the 460nm excitation produces the fluorescence signal of 520nm, and the 450nm excitation produces the fluorescence signal of 535nm and the fluorescence signal that the 400nm excitation produces 635nm.Wherein, Wavelength tunable laser and filter wheel be by computing machine part synchro control, realizes optical filter corresponding one by one on excitation wavelength and the filter wheel.
Described Signal Pretreatment part is made up of photomultiplier, prime amplifier, photon counter and counting integrated circuit board, the light signal that enters photomultiplier passes through and becomes electric signal after photoelectricity transforms, pass through prime amplifier, photon counter and counting integrated circuit board successively, enter computing machine and partly carry out data acquisition and processing (DAP), constitute the Signal Pretreatment part.
The beneficial effect of patent of the present invention is, the laser instrument that adopts tunable wave length obtains the maximum excitation wavelength at specific endogenous fluorescent material as excitation source, detects corresponding emitting fluorescence simultaneously, and the best that has realized fluorescence information excites and detects; Simultaneously, adopt optical filter from continuous fluorescence spectrum, to decompose and obtain discrete wavelength of fluorescence, this on hardware, realize from continuous spectrum decomposite can clear and definite characterizing tissues fluorescent characteristic fluorescence signal, the process that reduced data is handled, the high sensitivity and the specific detection of realization tissue autofluorescence.
Description of drawings
Fig. 1 is a system architecture synoptic diagram of the present invention.
Among Fig. 1,1 is tunable laser, 2 is optical attenuator, and 3 is coupled lens, and 4 is the exciting light input end of optical fiber one end of transmission exciting light, 5 the other ends for transmission exciting light optical fiber access endoscope, 6 is endoscope, and 7 is tissue to be measured, the 8 fluorescence output terminals for transmission fluorescence, 9 is preceding coupled lens, 10 is filter wheel, and 11 is the back coupled lens, and 12 are the cooling voltage controller, 13 is refrigeratory, 14 is photomultiplier, and 15 is prime amplifier, and 16 is single photon counter, 17 are the counting integrated circuit board, and 18 is computing machine.
Embodiment
Below in conjunction with accompanying drawing invention is further described.
As shown in Figure 1, the autofluorescence detection system figure that excites based on different wave length.Choose a routine human colon tissue and carry out the autofluorescence Research of Spectrum Characteristics.Before experimentizing, by cooling off voltage controller (12) with refrigeratory (13) (C9144, Hamamatsu Corp., Japan) open and cooled off 30 minutes, make photomultiplier (14) (R928, Hamamatsu Corp., Hamamatsu Japan) reaches steady-working state.According to requirement of experiment, open computing machine (18), and stored corresponding three threshold value I in advance with colon
T1And I
T2Its numerical value is respectively 30000 and 3, open tunable laser (1) (PIV OPO 355, OPOTEK, USA), the output wavelength of regulating tunable laser (1) by the Control Software on the computing machine (18) is 400nm, and filter wheel (10) (FW102B is selected in control simultaneously, Thorlabs Inc., USA) going up centre wavelength is the optical filter of 635nm.The exciting light that tunable laser (1) is sent carries out power adjustments through optical attenuator (2), passes through the optical fiber that coupled lens (3) is coupled and enters the six roots of sensation transmission exciting light on the exciting light input end (4) that transmits exciting light optical fiber afterwards.The optical fiber incoming end (5) of transmission exciting light is by endoscope (6) (CF-1T140I/L, Olympus Corp., Japan) exciting light is passed through the biopsy channel uniform irradiation on tissue to be measured (7), the fluorescence that sends after tissue to be measured (7) is excited is collected by the optical fiber incoming end of the transmission fluorescence in the y-type optical fiber again, exports from the fluorescence output terminal (8) of y-type optical fiber at last.The fluorescence of output is coupled by preceding coupled lens (9) after the last centre wavelength of filtration halo (10) is the optical filter of 635nm, after carrying out narrow-band filtering, be coupled to photomultiplier (14) by back coupled lens (11) again, by being converted to electric signal after the conversion of photomultiplier (14) photoelectricity, through prime amplifier (15) (C6438, Hamamatsu Corp., Hamamatsu, Japan) carrying out electric signal amplifies, electric signal after the amplification is again by single photon counter (16) (C9744, Hamamatsu Corp., Hamamatsu Japan) differentiates that the back is by counting integrated circuit board (17) (M8784, Hamamatsu Corp., Hamamatsu Japan) counts and by the fluorescence signal I of computing machine (18) record 635nm
635, its numerical value is 10000, less than threshold value I
T1, failing to judge types of organization, must further detect.Output wavelength by the adjusting tunable laser of the Control Software on the computing machine (18) (1) is 280nm again, it is the optical filter of 335nm that centre wavelength is selected in control simultaneously, is counted and write down by computing machine (18) the fluorescence signal I of 335nm by counting integrated circuit board (17)
335, its numerical value is 96000; Output wavelength by the adjusting tunable laser of the Control Software on the computing machine (18) (1) is 340nm again, it is the optical filter of 390nm that centre wavelength is selected in control simultaneously, is counted and write down by computing machine (18) the fluorescence signal I of 390nm by counting integrated circuit board (17)
390, its numerical value is 50000; Selecting centre wavelength again is the optical filter of 460nm, the fluorescence signal I of record 460nm
460, its numerical value is 82000.Selecting tunable laser (1) output wavelength again is 450nm, and selecting centre wavelength simultaneously is the fluorescence signal I of the optical filter detection 535nm of 535nm
535, its numerical value is 15000.At last, selecting tunable laser (1) output wavelength again is 460nm, and selecting centre wavelength simultaneously is the fluorescence signal I of the optical filter detection 520nm of 520nm
520, its numerical value is 20000.Handle at the enterprising line data of computing machine (18), calculate I
r=(I
460+ I
520+ I
535) * I
335/ (I
390)
2The result be 4.49, greater than threshold value I
T2, showing nicotinamide adenine dinucleotide reduced, flavin adenine dinucleotide (FAD) and tryptophane are higher with respect to the relative content of collagen, judge the colon tumor tissue that is organized as to be measured.
Choose another routine human colon tissue and carry out the autofluorescence Research of Spectrum Characteristics.According to the process of embodiment 1, at first select the 400nm exciting light, detect the fluorescence signal I of 635nm
635, the numerical value of its record is 9000, less than threshold value I
T1, failing to judge types of organization, must further detect.Then, select the 280nm excitation, detect the fluorescence signal I of 335nm
335, its numerical value is 75000.Select the 340nm exciting light then, detect the fluorescence signal I of 390nm and 460nm
390And I
460, its numerical value is respectively 70000 and 81000.Select the 450nm excitation again, detect the fluorescence signal I of 535nm
535, its numerical value is 10000.At last, select the 460nm exciting light, detect 520 fluorescence signal I
520, its numerical value is 18000.Calculate I
r=(I
460+ I
520+ I
535) * I
335/ (I
390)
2The result be 1.67, less than threshold value I
T2, showing nicotinamide adenine dinucleotide reduced, flavin adenine dinucleotide (FAD) and tryptophane are lower with respect to the relative content of collagen, judge the colon normal structure that is organized as to be measured.
Choose another routine human colon tissue and carry out the autofluorescence Research of Spectrum Characteristics.According to the process of embodiment 1, at first select the 400nm exciting light, detect the fluorescence signal I of 635nm
635, the numerical value of its record is 60000, is higher than threshold value I
T1, show that porphyrin content is higher, judge the colon tumor tissue that is organized as to be measured.
Claims (4)
1. tissue autofluorescence detection system based on the different wavelengths of light source excitation, mainly by the excitation source part, excite-detect-collecting part, the optical collection part, Signal Pretreatment part and computing machine partly constitute, wherein the Signal Pretreatment part is connected by signal wire with the computing machine part, excitation source part by exciting light with excite-detect-optical fiber of exciting light input end in the collecting part links to each other, excite-detect-optical fiber of fluorescence output terminal in the collecting part partly links to each other with optical collection by fluorescent signal path, fluorescent signal path enters the Signal Pretreatment part after partly transmitting through optical collection, become electric signal after photomultiplier photoelectricity transforms in the Signal Pretreatment part, enter the computing machine part, it is characterized in that tunable laser (1) can be equipped with the narrow band pass filter of different centre wavelengths according to the required pulse laser wavelength of endogenous fluorescent material adjusting specific in the tissue on the filter wheel (10).
2. a kind of tissue autofluorescence detection system based on the different wavelengths of light source excitation according to claim 1 is characterized in that wavelength coverage that described tunable laser can regulate successively is from 210nm to 2200nm.
3. a kind of tissue autofluorescence detection system based on the different wavelengths of light source excitation according to claim 1 is characterized in that being equipped with on the described filter wheel narrow band pass filter of different centre wavelengths.
4. a kind of tissue autofluorescence detection system according to claim 1 based on the different wavelengths of light source excitation, it is characterized in that can be by the synchro control software on the computing machine, the synchronous conversion of corresponding narrow band pass filter on the excitation wavelength of realization tunable laser output and the filter wheel.
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| CN102607608A (en) * | 2012-03-08 | 2012-07-25 | 天津奇谱光电技术有限公司 | Extensible electro-optical sensing system |
| CN103608662A (en) * | 2011-06-29 | 2014-02-26 | 京都府公立大学法人 | Tumor site identification device and method |
| CN103781253A (en) * | 2014-01-23 | 2014-05-07 | 安徽理工大学 | Method and device for controlling light intensity of broadband light source to be stable |
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| CN103917155A (en) * | 2011-11-07 | 2014-07-09 | 皇家飞利浦有限公司 | Detection apparatus for determining state of tissue |
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| CN104066385A (en) * | 2012-01-30 | 2014-09-24 | Ccs株式会社 | Illumination device for tumor detection and illumination device for examinations |
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| CN103781253A (en) * | 2014-01-23 | 2014-05-07 | 安徽理工大学 | Method and device for controlling light intensity of broadband light source to be stable |
| CN104807792A (en) * | 2015-04-27 | 2015-07-29 | 电子科技大学 | Negative refractive index photonic crystal bioluminescence detector |
| CN104807792B (en) * | 2015-04-27 | 2017-06-30 | 电子科技大学 | A kind of negative index photonic crystal type bioluminescence detector |
| CN106092996A (en) * | 2016-08-03 | 2016-11-09 | 复旦大学 | A kind of cancer diagnosis system based on the autofluorescence life-span |
| CN106092996B (en) * | 2016-08-03 | 2024-03-15 | 复旦大学 | Cancer diagnosis system based on autofluorescence life |
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