CN103335992A - Fluorescence type glucose capillary biosensor - Google Patents
Fluorescence type glucose capillary biosensor Download PDFInfo
- Publication number
- CN103335992A CN103335992A CN2013102500939A CN201310250093A CN103335992A CN 103335992 A CN103335992 A CN 103335992A CN 2013102500939 A CN2013102500939 A CN 2013102500939A CN 201310250093 A CN201310250093 A CN 201310250093A CN 103335992 A CN103335992 A CN 103335992A
- Authority
- CN
- China
- Prior art keywords
- fluorescence
- kapillary
- sensor
- nano
- probe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Abstract
The invention relates to a fluorescence type glucose capillary biosensor. The biosensor comprises nano sensors, a capillary coupling device, capillaries, a light source, a fiber optic spectrometer and a probe, wherein the nanometer sensors are distributed on the inner walls of the capillaries; the capillary coupler is connected with optical fibers and hoses; the optical fibers can be used for receiving an exciting light generated by the light source; the hoses can be used for receiving liquid to be detected; and a new fluorescence spectrum generated by the liquid to be detected and the nano sensors under the excitation of an evanescent field produced by the exciting light is received by the probe, and the probe transmits the received fluorescence spectrum to the fiber optic spectrometer to be analyzed. According to the fluorescence type glucose capillary biosensor, the capillaries are adopted for sampling, thus the dosage of samples to be detected can be obviously decreased, and reagents are saved. A fluorescence signal is excited through the evanescent field, therefore, the background fluorescence in a biological sample can be reduced efficiently, and the lower detection limit can be elevated.
Description
Technical field
The present invention relates to biochemical analysis and quantitative technique field, particularly relate to a kind of fluorescence formula glucose kapillary biology sensor, the concentration that can be applied to micro-glucose in the micro-example detects.
Background technology
When light entered optically thinner medium from optically denser medium, the refraction angle was greater than incident angle.When incident angle increased to a certain angle, the refraction angle equaled 90 °, at this moment, the refract light complete obiteration, incident light all returns in the original medium, and this phenomenon is called total reflection.
Experiment shows, when total reflection light wave be not utterly on the interface by whole reflected back optically denser mediums, but penetrate the very thin layer of surface of optically thinner medium (an about wavelength) and propagate some distances (wavelength magnitude) along the interface, return optically denser medium at last.Because electromagnetic field can not interrupt on the interphase of two media, it should satisfy boundary relation, thereby will there be transmitted wave in optically thinner medium.This transmitted wave is evanescent wave or evanscent field.The amplitude of evanscent field is exponential damping with the increase with the perpendicular degree of depth of interphase.
Some material is subjected to can launch the fluorescence different with optical maser wavelength behind ultraviolet light or the radiation of visible light.The excitation spectrum of material and fluorescence spectrum can be as the qualitative analyses of this material.When conditions such as excitating light strength, wavelength, agents useful for same and temperature fixedly the time, material is in the finite concentration scope, and its emitted luminescence intensity becomes correlationship with the concentration of this material in the solution, can be used as quantitative test.Utilize fluorescence that unknown sample is carried out chemico-analytic method and be called fluorometry.
Conventional optical fiber biosensor is as probe with optical fiber, fix corresponding fluorescent material as probe on the surface of fibre-optical probe, fibre-optical probe is put into solution to be measured, test substance interacts and sends fluorescence in fibre-optical probe surface fluorescence material and the solution, by surveying the concentration determination that fluorescence intensity realizes micro substance to be measured in the solution.The subject matter that exists in the prior art has:
1, the conventional fluoroscopic examination based on the evanscent field mode that adopts the detection optical fiber probe to separate with solution to be measured, effective sample size that solution usage to be measured detects much larger than actual participation concentration, so reagent a large amount of in the conventional sense are wasted, the experimental cost height is difficult to realize the trace detection analysis of sample.
2, operating process is simple inadequately in the Chang Gui detection.
3, the large usage quantity of conventional sense solution so the influence of other disturbing factors is bigger, causes detection sensitivity.
Summary of the invention
At above the deficiencies in the prior art, the invention provides a kind of fluorescence formula glucose kapillary biology sensor.
Purpose of the present invention is achieved through the following technical solutions:
A kind of fluorescence formula glucose kapillary biology sensor, this biology sensor comprises nano-sensor, the kapillary coupling device, kapillary, light source, fiber spectrometer and probe, described nano-sensor is distributed in capillary tube inner wall, described kapillary coupling device links to each other with flexible pipe with optical fiber, receive the exciting light that described light source sends by optical fiber, receive liquid to be measured by flexible pipe, described liquid to be measured and nano-sensor produce new fluorescence spectrum and are received by described probe under the evanscent field of described excitation light generation excites, this probe is transferred to the fiber spectrometer analysis with the fluorescence spectrum that receives.
Described probe is arranged on extracapillary and presses close to kapillary.
Described nano-sensor is evenly distributed on capillary tube inner wall.
Described nano-sensor is glucose oxidase fluorescence nano sensor, inside nanoparticles embedding fluorescence reference molecule and oxygen probe molecule in the sensor, the outside glucose oxidase of modifying.
Described kapillary is quartz capillary.
Measuring principle of the present invention:
1. evanscent field
After laser incides the kapillary end face at a certain angle, in capillary tube inner wall generation total reflection, as shown in Figure 3; Simultaneously perpendicular to axis of bore to direction produce evanscent field, the amplitude of evanscent field is exponential damping with the increase with the perpendicular degree of depth of interphase, as shown in Figure 4.
When light incides optically thinner medium from optically denser medium, establish optically denser medium and the optically thinner medium refractive index is respectively n
1, n
2By total reflection formula
Then
Namely
θ
cCritical angle during for total reflection.
For given refractive index parameter, improve that evanscent field excites and the zone that absorbs must make incident angle at critical angle θ
cNear, when incident angle surpasses critical angle θ
cThe time, the intensity of evanscent field will descend fast.
The present invention uses quartz capillary and a kind of aqueous solution, and quartzy refractive index is n
1=1.46, the refractive index of solution to be measured is approximately the refractive index of water, is taken as n
2=1.33, by the cirtical angle of total reflection
Calculate θ
c=65.638 °.Exciting light is with θ among the present invention
c=65.638 ° incident angle incides capillary wall, can make the intensity of evanscent field reach maximum.
2. fluoroscopic examination principle
Detection of the present invention is based on the reaction of glucose and oxygen:
Glucose oxidase fluorescence nano sensor is inner embedding fluorescence oxygen probe molecule and reference molecule and the outside nano particle of modifying glucose oxidase, and what of dissolved oxygen amount in the solution are its fluorescence intensity depend on.Nano-sensor inside is by PFO, C
6Obtain with the certain proportion mixing with PtOEP, under exciting, the 405nm evanscent field sends fluorescence, in the fluorescent line of nano particle, can see three emission peaks, be respectively 440nm, 466nm, the 650nm place, the fluorescence of 650nm is sent by PtOEP, and the fluorescence intensity of PtOEP can strengthen along with the decline of oxygen concentration in the environment; Nano-sensor is outside modifies glucose oxidase, but oxygen reaction reduces nano-sensor oxygen concentration on every side in glucose oxidase catalysis glucose and the solution.Glucose oxidase fluorescence nano sensor can be fixed on by the mode of self assembly and adapt on the capillary tube inner wall.When the exciting light of wavelength 405nm incides capillary wall, perpendicular to axis of bore to direction form evanscent field, the PtOEP in the evanscent field excitation nano sensor sends the fluorescence that wavelength is 650nm; After containing the unknown concentration glucose solution and entering kapillary, glucose under the catalysis of nanoparticle surface glucose oxidase with oxygen reaction, nano-sensor oxygen concentration is on every side reduced, be reflected in the fluorescence of PtOEP in the nano particle, then be weakening of red fluorescence intensity, spectrum by spectrometer measurement kapillary sensor, calculate ruddiness and green intensity ratio in the fluorescence spectrum and obtain the relation in fluorescence intensity ratio and reaction time, and then draw the concentration of glucose in the solution to be measured.
Put it briefly, under the exciting of 405nm light, the intensity that is fixed on the ratio fluorescent of the glucose oxidase nano-sensor generation on the capillary tube inner wall is directly proportional with glucose content in the solution, utilize fiber spectrometer to collect fluorescence signal and analyze, can draw the concentration of glucose in the solution to be measured.
The invention has the advantages that:
The present invention has replaced the fibre-optical probe and the solution to be measured that separate in the popular response with kapillary, has realized that the microanalysis of test substance is measured.
1, the present invention adopts kapillary to carry out the measurement of concetration of material, has significantly reduced and has detected the reagent that uses, and has reduced experimentation cost, realizes the microanalysis of micro-example.
2, the capillary portion among the present invention can be produced in batches, can simplify the step of sample detection, shortens the time of detecting.
3, fluorescence signal of the present invention excites by evanscent field, can effectively reduce the background fluorescence in the biological sample, improves and detects lower limit.
Description of drawings
Fig. 1: a kind of fluorescence formula of the present invention glucose kapillary biosensor structure schematic diagram;
Fig. 2: the oxygen nano-sensor structural representation that the surface is not modified;
Fig. 3: laser is at the total reflection phenomenon of capillary wall;
Fig. 4: the evanscent field that laser produces in the capillary wall total reflection.
Embodiment
Be illustrated in figure 1 as a kind of fluorescence formula glucose kapillary biosensor structure schematic diagram, this fluorescence formula glucose kapillary biology sensor comprises nano-sensor 1, kapillary coupling device 2, kapillary 3, light source 4, fiber spectrometer 5 and pops one's head in 6.Described nano-sensor 1 is distributed in the inwall of kapillary 3, described kapillary coupling device 2 links to each other with flexible pipe with an optical fiber, can receive the exciting light that described light source 4 sends by optical fiber, can receive liquid to be measured by flexible pipe, described kapillary coupling device can allow exciting light to incide capillary wall simultaneously and solution enters in the capillary cavity.Described liquid to be measured and nano-sensor excite the new fluorescence spectrum of generation down to be received by described probe 6 at the evanscent field of described excitation light generation, and this probe 6 is transferred to fiber spectrometer 5 with the fluorescence spectrum that receives and analyzes.
Described probe is arranged on extracapillary and presses close to kapillary, and described nano-sensor is evenly distributed on capillary tube inner wall.Described kapillary is quartz capillary.Capillary wall can play the effect of optical fiber, exciting light is in capillary wall total reflection and propagation vertically, simultaneously perpendicular to axis of bore to direction produce evanscent field, evanscent field excites glucose oxidase fluorescence nano sensor to produce ratio fluorescent, the intensity of ratio fluorescent is directly proportional with concentration of glucose in the solution, obtain fluorescence signal by fiber spectrometer, the analysis of fluorescence signal intensity can reach the purpose that detects concentration of glucose in the solution.Capillary tube inner wall is modified with the 3-aminopropyl triethoxysilane in the present embodiment, so by with the crosslinked nano-sensor of realizing of nano-sensor in the self assembly of capillary tube inner wall.
Described nano-sensor is glucose oxidase fluorescence nano sensor, the inner embedding fluorescence reference molecule of nano-sensor wherein and oxygen probe molecule, the outside glucose oxidase of modifying.Under the 405nm optical excitation, the ratio fluorescent intensity of glucose oxidase nano-sensor is directly proportional with the glucose content in the solution.
The preparation of glucose oxidase fluorescence nano sensor: at first utilize reprecipitation method to prepare the oxygen nano-sensor that poly-D-lysine coats, pass through the crosslinked realization glucose oxidase of glutaraldehyde then in the finishing of nano-sensor.1, the preparation of oxygen nano-sensor: with PFO, C
6Be dissolved in the tetrahydrofuran with certain mass ratio with PtOEP, getting this solution of 200 microlitres is injected under the ultrasound wave condition in 8 ml waters (PPL that contains 0.001mg/ml), after consequent suspending liquid leaves standstill 2 hours, be that the filtering membrane of 400nm filters with the aperture, can obtain the aqueous solution of nano particle.Fig. 2 is the not modified oxygen nano-sensor that has prepared.
2, the finishing of nano particle:
The nano particle that has prepared is carried out emulsion with poly-D-lysine mix, utilize the electrostatic interaction of nano particle and poly-D-lysine to carry out the oxygen nano-sensor that polymerization forms the poly-D-lysine coating.
Oxygen nano-sensor and glucose oxidase that poly-D-lysine is coated carry out cross-linking reaction by glutaraldehyde, realize that glucose oxidase is in the finishing of nano-sensor.
Glucose oxidase fluorescence nano sensor is in the self assembly mode of capillary tube inner wall: the kapillary material among the present invention is that quartz is silicon dioxide, therefore capillary tube inner wall can be carried out APTS and modify, nano-sensor and capillary tube inner wall carry out self assembly by cross-linking reaction then.
1. the acetone soln that will be dissolved with suitable concentration 3-aminopropyl three TMOSs (APTS) injects quartz capillary, leaves standstill amido fully to be attached on the capillary tube inner wall in 40 minutes.
2. clean kapillary and dry down at 70 ℃ with acetone.
3. the suspending liquid of the nano-sensor that the process that will prepare is modified injects quartz capillary, finishes nano-sensor in the self assembly of capillary tube inner wall by nano particle and the cross-linking reaction of the capillary tube inner wall of process APTS modification.
Among the present invention in the fluorescence formula kapillary biology sensor fluoroscopic examination principle of nano-sensor be that exciting light is producing evanscent field perpendicular to the kapillary axial direction when propagating along capillary wall, excite the glucose oxidase fluorescence nano sensor of capillary tube inner wall to produce ratio fluorescent by evanscent field, utilize fiber spectrometer to collect fluorescence signal and carry out glucose detection.
During use, the kapillary of handling well is fixed on the experiment table, by regulating the angle of experiment table and surface level, makes kapillary and exciting light be optimal incident angle in fact θ
c, after kapillary is regulated the fibre-optical probe of spectrometer pressed close to the position and will pop one's head in and fix of windowing of kapillary outer wall, fiber spectrometer is connected with computing machine.After the detection light path regulates, open laser instrument, open fiber spectrometer and computing machine polishing wax testing software simultaneously, record fluorescence spectrum image at this moment.The solution that will contain glucose enters in the kapillary it by peristaltic pump, records first order fluorescence spectrum at regular intervals, no longer changes up to the fluorescence spectrum image.Drawn the concentration of glucose in the solution to be measured by ratio fluorescent intensity and reaction time relation.
Should be appreciated that the above detailed description of technical scheme of the present invention being carried out by preferred embodiment is illustrative and not restrictive.Those of ordinary skill in the art is reading on the basis of instructions of the present invention and can make amendment to the technical scheme that each embodiment puts down in writing, and perhaps part technical characterictic wherein is equal to replacement; And these modifications or replacement do not make the essence of appropriate technical solution break away from the spirit and scope of various embodiments of the present invention technical scheme.
Claims (5)
1. fluorescence formula glucose kapillary biology sensor, it is characterized in that, this biology sensor comprises nano-sensor, the kapillary coupling device, kapillary, light source, fiber spectrometer and probe, described nano-sensor is distributed in capillary tube inner wall, described kapillary coupling device links to each other with flexible pipe with optical fiber, receive the exciting light that described light source sends by optical fiber, receive liquid to be measured by flexible pipe, described liquid to be measured and nano-sensor produce new fluorescence spectrum and are received by described probe under the evanscent field of described excitation light generation excites, this probe is transferred to the fiber spectrometer analysis with the fluorescence spectrum that receives.
2. a kind of fluorescence formula glucose kapillary biology sensor according to claim 1 is characterized in that described probe is arranged on extracapillary and presses close to kapillary.
3. a kind of fluorescence formula glucose kapillary biology sensor according to claim 1 is characterized in that described nano-sensor is evenly distributed on capillary tube inner wall.
4. a kind of fluorescence formula glucose kapillary biology sensor according to claim 1, it is characterized in that, described nano-sensor is glucose oxidase fluorescence nano sensor, inside nanoparticles embedding fluorescence reference molecule and oxygen probe molecule in the sensor, the outside glucose oxidase of modifying.
5. a kind of fluorescence formula glucose kapillary biology sensor according to claim 1 is characterized in that described kapillary is quartz capillary.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310250093.9A CN103335992B (en) | 2013-06-21 | 2013-06-21 | A kind of fluorescence type glucose capillary biosensor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310250093.9A CN103335992B (en) | 2013-06-21 | 2013-06-21 | A kind of fluorescence type glucose capillary biosensor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103335992A true CN103335992A (en) | 2013-10-02 |
| CN103335992B CN103335992B (en) | 2016-02-10 |
Family
ID=49244196
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310250093.9A Expired - Fee Related CN103335992B (en) | 2013-06-21 | 2013-06-21 | A kind of fluorescence type glucose capillary biosensor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103335992B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103616361A (en) * | 2013-12-04 | 2014-03-05 | 北京交通大学 | Fluorescent glucose nano biosensor and preparation method thereof |
| CN106308787A (en) * | 2016-08-23 | 2017-01-11 | 北京大学 | Fluorescence-visible electrophysiologic micro electrode structure and production method thereof |
| CN106568754A (en) * | 2016-11-06 | 2017-04-19 | 浙江大学 | Optical system used for measuring liquid sample multiphoton fluorescence spectrum |
| CN108896539A (en) * | 2018-05-15 | 2018-11-27 | 武汉大学 | Measure the optofluidic detector of phosphorus content in seawater |
| CN109580563A (en) * | 2018-11-23 | 2019-04-05 | 南方科技大学 | A kind of nano-sensor and its preparation method and application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4981779A (en) * | 1986-06-26 | 1991-01-01 | Becton, Dickinson And Company | Apparatus for monitoring glucose |
| US20030152308A1 (en) * | 2002-02-08 | 2003-08-14 | The Research Foundation Of State University Of New York | Capillary waveguide fluorescence sensor |
| CN1873450A (en) * | 2006-06-30 | 2006-12-06 | 清华大学 | Biosensor of full fiber optic evanescent wave |
| US7871573B2 (en) * | 2004-07-26 | 2011-01-18 | University Of Louisville Research Foundation, Inc. | Enhancement of sensitivity of fluorophore mediated biosensing and bioimaging |
-
2013
- 2013-06-21 CN CN201310250093.9A patent/CN103335992B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4981779A (en) * | 1986-06-26 | 1991-01-01 | Becton, Dickinson And Company | Apparatus for monitoring glucose |
| US20030152308A1 (en) * | 2002-02-08 | 2003-08-14 | The Research Foundation Of State University Of New York | Capillary waveguide fluorescence sensor |
| US7871573B2 (en) * | 2004-07-26 | 2011-01-18 | University Of Louisville Research Foundation, Inc. | Enhancement of sensitivity of fluorophore mediated biosensing and bioimaging |
| CN1873450A (en) * | 2006-06-30 | 2006-12-06 | 清华大学 | Biosensor of full fiber optic evanescent wave |
Non-Patent Citations (2)
| Title |
|---|
| HAO XU ET AL.: "Fluorescent nano-PEBBLE sensors designed for intracellular glucose imaging", 《ANALYST》, vol. 127, no. 11, 27 September 2002 (2002-09-27), pages 1472 - 1474 * |
| PATRICK S.GRANT ET AL.: "Nanostructured Fluorescent Particles for Glucose Sensing", 《PROCEEDINGS OF SPIE, OPTICAL DIAGNOSTICS AND SENSING OF BIOLOGICAL FLUIDS AND GLUCOSE AND CHOLESTEROL MONITORING II》, vol. 4624, 31 December 2002 (2002-12-31), XP009065823, DOI: doi:10.1117/12.468326 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103616361A (en) * | 2013-12-04 | 2014-03-05 | 北京交通大学 | Fluorescent glucose nano biosensor and preparation method thereof |
| CN106308787A (en) * | 2016-08-23 | 2017-01-11 | 北京大学 | Fluorescence-visible electrophysiologic micro electrode structure and production method thereof |
| CN106308787B (en) * | 2016-08-23 | 2019-07-30 | 北京大学 | Visible electro physiology microelectrode structure of fluorescence and preparation method thereof |
| CN106568754A (en) * | 2016-11-06 | 2017-04-19 | 浙江大学 | Optical system used for measuring liquid sample multiphoton fluorescence spectrum |
| CN108896539A (en) * | 2018-05-15 | 2018-11-27 | 武汉大学 | Measure the optofluidic detector of phosphorus content in seawater |
| CN109580563A (en) * | 2018-11-23 | 2019-04-05 | 南方科技大学 | A kind of nano-sensor and its preparation method and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103335992B (en) | 2016-02-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gong et al. | Highly sensitive SERS detection and quantification of sialic acid on single cell using photonic-crystal fiber with gold nanoparticles | |
| EP1903330B1 (en) | Surface plasmon enhanced fluorescence sensor and fluorescence detecting method | |
| US20090304551A1 (en) | Ultra Sensitive Tapered Fiber Optic Biosensor For Pathogens, Proteins, and DNA | |
| CN103335992B (en) | A kind of fluorescence type glucose capillary biosensor | |
| CN106959370B (en) | A kind of biological sensor and detection method based on coupling grating | |
| CN103245641B (en) | Multi-channel planar waveguide evanescent wave biosensor | |
| CN106290279B (en) | A kind of single cell protein detection system and its application | |
| TW201305548A (en) | System and method for cell analysis | |
| CN103398974B (en) | A kind of Fibre Optical Sensor, preparation method and the system of measurement | |
| CA2638060A1 (en) | Method for using a photonic crystal fiber as a raman biosensor | |
| CN103063645B (en) | High-efficiency fluorescence detection based on novel Microstructure optical fiber | |
| US20090283700A1 (en) | Fluorescence detecting method | |
| CN109520994A (en) | A kind of micro-fluidic biological detection system and method | |
| JP2007114026A (en) | Standard substance for particle analyzer | |
| JP6098523B2 (en) | SPFS measurement sensor chip, SPFS measurement method using SPFS measurement sensor chip, and SPFS measurement apparatus equipped with SPFS measurement sensor chip | |
| US9134201B2 (en) | Fluid providing apparatus | |
| EP2542900B1 (en) | Apparatus and method for detecting and measuring biomolecular interactions | |
| JP4480130B2 (en) | Optical analyzer | |
| CN102859345B (en) | Automatic analyzer | |
| Lucas et al. | Cellular point-of-care diagnostics using an inexpensive layer-stack microfluidic device | |
| CN105793706B (en) | Disposable photometric measurement end | |
| US8716008B2 (en) | Detection method and detection system | |
| JP2009192259A (en) | Sensing device | |
| CN103558183A (en) | MZ interference type optical biochemistry sensor chip embedded with FP cavity | |
| CN107219192A (en) | A kind of biomolecule based on photonic crystal fiber is in fine detecting system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160210 Termination date: 20160621 |