CN108645827A - Based on the hypersensitive NO sensors for simplifying microstructured optical fibers - Google Patents
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Abstract
The invention discloses a kind of sensor fibres for detecting NO concentration in organism, there is open duct on the sensor fibre, and NO fluorescence probes are fixed in the surface in open duct by the way of chemical bonding, the NO fluorescence probes are the quantum dot being bonded on open duct, and quantum dot is the semiconductor-quantum-point containing Cd elements.Functionalization (hydroxylating and silanization) is carried out to the opening channel surfaces of optical fiber, makes its surface that there is the group that can be chemically bonded with NO fluorescence probes, NO fluorescence probes is directly fixed on open channel surfaces.The problems such as fluorescence probe that generally use is generated by means of physical methods such as electrostatic interaction, Van der Waals forces is easily leaked, detached can effectively be solved.The NO fluorescence probes that the present invention selects are the homemade high performance semiconductor-quantum-point of one kind (CdTe quantum, CdS quantum dot, CdTe/CdS nuclear shell structure quantum points), substantially increase stability and the sensitivity of NO sensors.
Description
Technical field
The invention belongs to biological detection and technical field of clinical medical detection, are related to a kind of dense for detecting NO in organism
The micro optical fiber biosensor of degree, the more particularly to a kind of preparation of NO fluorescence probes.
Background technology
Nitric oxide (NO) is used as a kind of unique vaso-active substance, participates in and a variety of physiology of mediating central nervous system
Process and pathologic process are a kind of important messenger molecule and effector molecule in organism.NO levels can be used as research god
Through the important of a variety of disease pathologies such as different physiological roles and cardiovascular and cerebrovascular, diabetes, tumour, obesity such as, immune, digestion
One of index.If the direct measurement to NO contents in human body can be realized, so that it may regulate and control several important diseases physiology, disease to disclose NO
The mechanism of reason process, and internal NO levels can be adjusted by NO donors medicine or enzyme inhibitor, this is to the mankind's a variety of diseases early stage
Diagnosing and treating has particularly important meaning.
NO is free radical micro-molecular gas, simple in structure and extremely unstable, have it is fat-soluble, can be freely through cell
Film, experiment find to have the Concentration Testing of NO in human body greatly difficult.This is because under normal circumstances, NO amounts are seldom
, average each cell only discharges 1~200attomol (1attomol=10-18Mol), and metabolism it is too fast (half-life period 5~
60s), it is detected again after obtaining sample it is difficult to reflect the expression of NO at that time.Currently, detection NO concentration divides two methods：(1) between
Detection method is connect, its metabolite nitrous acid or nitrous acid ester are detected using NO synthesis enzyme inductions.Due to the induction table of synzyme
The concentration for needing the regular hour up to the generation with metabolite, therefore detecting synzyme and metabolite cannot reflect completely to be worked as
When NO concentration.(2) direct Detection Method, including：Electron paramagnetic resonance spectrum (EPR) method (EPR), chemoluminescence method, mass spectrography, fluorescence
Method, electrochemical sensor etc..The universal sensitivity of first three detection method is relatively low, trivial operations, sample treatment require high, time-consuming length
And it is of high cost；Although latter two method can monitor NO concentration in real time, preparation process requirement is very high, signal detection system is multiple
It is miscellaneous, stability is poor etc., practical application is greatly limited.
Fiber-optic biosensor technology is a kind of high sensitivity risen in recent ten years, can carry out analyzing in real time at a distance
Trace analysis.Optical fiber biosensor is the biological information that will selectively analyze object, is continuously changed into analytical instrument
The analysis measuring device of the optical signalling easily measured can carry out real-time, on-line checking, it has also become various to single (more) kind objects
The important detection means of biomass, molecule, ion.It has the characteristics that accuracy of detection is high, response is fast, strong antijamming capability.Through
After development for many years, identify that sensitive layer (i.e. sensing layer), the optical fiber NO sensors of development obtain by modifying NO in optical fiber end
Substantive progress.
Currently, there are many optical fiber NO sensor of document report and are related to multi-disciplinary intersection, include mainly：Chemistry
Light emitting-type, absorption-type and fluorescent type etc..Compared to other types of optical fiber NO sensors, fluorescent type optical fiber NO transducer sensitivities are more
High, response faster, also is more conducive to be commercialized, and is one of the effective means realized NO high sensitivity and detected real-time.However, existing
There is fluorescent type optical fiber NO sensors commonly existing problems with：1, NO is horizontal in Relative biological body, and detection sensitivity is low.Usually
It is to fix NO in optical fiber end to identify sensitive layer, since fiber end face effective area is small, the load capacity of fluorescence probe is limited；And
Fluorescence signal is to excite generation by the evanscent field of fiber end face, and the energy of evanscent field is smaller under normal circumstances causes its excitation
Fluorescence signal it is also fainter, the sensitivity of sensor receives larger limitation.2, lack the knowledge highly selective to analyte
Other ability.Application NO fluorescence probes the most universal are organic dyestuff at present, and in most cases their excitation spectrum is all
Relatively narrow, wider and intensity is low again for fluorescent characteristics spectrum；The defect or photochemical stability of most serious be poor, photobleaching and photodissociation,
Cause optical fiber NO sensor stabilities poor.3, object and sensing probe are detected due to being in open environment, is spread in molecule
Under function influence, fluorescence probe easily leaks, and causes sensor stabilization performance poor.
With the development of optical fiber transmission theory and preparation process, microstructured optical fibers (Microstructure optical
Fiber, MOF) as the research hotspot of sensory field of optic fibre.By the increase with the used duct number of microstructured optical fibers,
Difficulty for fixing biological sensitive materials on its inner wall be consequently increased and the selection of sensitive material fixing means also more
Add and is restricted.Recent years, researcher simplified the multi-pore channel of traditional microstructured optical fibers, ran through whole volume
Micropore provide good reacting environment for biomass detection, to prepare of new generation high sensitivity, high stability, quick response
Optical fiber biosensor provide possibility.Compared to traditional optical fiber biosensor, based on the life for simplifying microstructured optical fibers
Object sensor has the advantages that extremely prominent：(1) sensitivity is significantly improved.Simplifying microstructured optical fibers has sizable internal ratio table
Area can be used for the load of sensitive material, and sensitive material has sufficiently large contact area with determinand, can effectively carry
The sensitivity of high optical fiber sensing probe；(2) stability is significantly improved.Determinand is carried out inside the microchannel of MOF sensing probes
Detection, the collection capacity of sample can be greatly reduced, it is exposed in outer easy damaged, easily let out to avoid traditional fiber sensing probe sensitive layer
The problems such as leakage；(3) distribution of flexible design MOF internal gutters, diameter, duty ratio are to optimize sensing capabilities.
Invention content
The present invention is prepared for high performance quantum for the problems such as existing fiber NO transducer sensitivities are low, stability is poor
Point is used as NO fluorescence probes；Select the method fixed amount for simplifying microstructured optical fibers and opening channel surfaces using chemical bonding at it
It is sub-, develop novel NO fibre optical sensors.
Technical solution of the present invention includes the following contents：
A kind of sensor fibre for detecting NO concentration in organism is provided, there is open duct on the sensor fibre, and
NO fluorescence probes are fixed in the surface in open duct by the way of chemical bonding, which is to be bonded on open duct
Quantum dot, quantum dot be the semiconductor-quantum-point containing Cd elements.
Above-mentioned technical proposal is connect, fluorescence probe is CdTe quantum, CdS quantum dot or CdTe/CdS core-shell quanta dots.
Above-mentioned technical proposal is connect, there are two symmetrically arranged open ducts for tool on the sensor fibre.
Above-mentioned technical proposal is connect, there are one open ducts for tool on the sensor fibre.
The present invention also provides a kind of optical fiber biosensors for detecting NO concentration in organism, which is characterized in that
Including light source, sensor fibre and information collection and data processing system, the sensor fibre is the sense light of above-mentioned technical proposal
It is fine.
This law additionally provides a kind of preparation method for detecting the sensor fibre of NO concentration in organism, and feature exists
In including the following steps：
1) quantum dot for being bonded on open duct is prepared, quantum dot is the semiconductor-quantum-point containing Cd elements；
2) two kinds of microstructured optical fibers with open duct are selected, hydroxylating and silanization are carried out to open bore road surface；
3) under the action of crosslinking agent, the quantum dot of preparation is directly anchored to open duct table by way of chemical bond
Face.
Above-mentioned technical proposal is connect, the quantum dot is CdTe quantum, CdS quantum dot or CdTe/CdS nucleocapsid quantum
Point.
Above-mentioned technical proposal is connect, CdTe quantum preparation method is as follows：
1) it is passed through high-purity N2Half an hour fully excludes the air in condensation reflux unit；
2) by CdCl2Solution is added in three-neck flask, adds 180~220mg citrate dihydrate trisodiums under magnetic stirring,
It is sequentially added into 6~8mL Na2TeO3Solution, the water of 90~100mL, 95~105mg L-cysteines and 125~135mg
NaBH4, obtain mixed solution；
3)N2Under protection, mixture solution temperature is raised to 70~90 DEG C, in spiral reflux condense 40~
50min；
4) after condensing reflux, cooled to room temperature, obtains CdTe quantum under nitrogen protection；
Above-mentioned technical proposal is connect, the preparation method of CdS quantum dot is as follows：
1) it is passed through high-purity N2Half an hour fully excludes the air in condensation reflux unit；
2) by CdCl2Solution is added in three-neck flask, adds 180~220mg citrate dihydrate trisodiums under magnetic stirring；
It is sequentially added into 1~2mmol thio ureas, the water of 90~100mL, 95~105mg L-cysteines and 125~135mg again
NaBH4, obtain mixed solution；
3)N2Under protection, mixture solution temperature is raised to 70~90 DEG C, in spiral reflux condense 40~
50min；
4) after condensing reflux, cooled to room temperature, obtains CdS quantum dot under nitrogen protection.
Above-mentioned technical proposal is connect, CdTe/CdS core-shell quanta dots preparation methods are as follows：
1) it is passed through high-purity N2Half an hour fully excludes the air in condensation reflux unit；
2) by CdCl2Solution is added in three-neck flask, adds 180~220mg citrate dihydrate trisodiums under magnetic stirring；
It is sequentially added into 6~8mLNa again2TeO3Solution, the water of 90~100mL, 95~105mg L-cysteines and 125~135mg
NaBH4, obtain mixed solution；
3)N2Under protection, mixture solution temperature is raised to 70~90 DEG C, in spiral reflux condense 30~
40min, by a small amount of deionized water dissolving 1~2mmol thio ureas, the 20~30min that is injected into reaction unit that the reaction was continued；
4) after condensing reflux, cooled to room temperature, obtains CdTe/CdS nuclear shell structure quantum points under nitrogen protection.
Beneficial effects of the present invention：
(1) semiconductor-quantum-point that the present invention prepares a kind of high performance water-soluble is improved as NO fluorescence probes to NO's
Sensitive property, to increase the sensitivity detected to NO.
(2) present invention selects the simplification microstructured optical fibers for concentrating different structure, to the opening channel surfaces of microstructured optical fibers
Functionalization (hydroxylating and silanization) is carried out, makes its surface that there is the quantum dot that can contain Cd with NO fluorescence probes chemistry occurs
NO fluorescence probes are directly fixed on open channel surfaces by the group of bonding, it is possible to prevente effectively from generally use is by means of electrostatic
Fluorescence probe the problems such as easily leaking, detaching that the physical methods such as effect, Van der Waals force generate, improves the stability and again of sensor
Renaturation.
(3) the NO fibre optical sensors developed of the present invention, using simplify the big air hole paths of opening of microstructured optical fibers as
Micro reaction pool makes the fluorescence probe for being fixed on its surface quickly be contacted with NO and reacts, and improves the response time；Significantly increase simultaneously
Add sensitive layer effective area, improves the sensitivity of sensor.
Description of the drawings
Present invention will be further explained below with reference to the attached drawings and examples, in attached drawing：
Fig. 1 a, Fig. 1 b are the sectional views of two kinds of simplified microstructured optical fibers.
Fig. 2 is that quantum dot is fixed on optical fiber opening channel surfaces by way of chemical bond.
Fig. 3 is integrally fixed at the CdTe/CdS core-shell quanta dots detection NO schematic diagrames that microstructured optical fibers open hole surface.
Fig. 4 is based on the NO sensor experiment installation drawings for simplifying microstructured optical fibers.
Specific implementation mode
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to the accompanying drawings and embodiments, right
The present invention is further elaborated.It should be appreciated that described herein, specific examples are only used to explain the present invention, not
For limiting the present invention.
The present invention is used to detect the sensor fibre of NO concentration in organism, as shown in figure 3, having open bore on sensor fibre
Road, and NO fluorescence probes are fixed in the surface in open duct by the way of chemical bonding, which is to be bonded in opening
Quantum dot on duct, quantum dot are the semiconductor-quantum-point containing Cd elements.Cd elements are very sensitive for NO, containing this
Quantum dot and the NO contact of element can react quickly.Therefore the semiconductor-quantum-point for containing Cd elements is visited as NO fluorescence
Needle can be such that the fluorescence probe for being fixed on open channel surfaces is quickly contacted with NO and react, and improve the response time；Simultaneously significantly
Increase sensitive layer effective area, improves the sensitivity of sensor.
In the embodiment of the present invention, fluorescence probe is CdTe quantum, CdS quantum dot or CdTe/CdS core-shell quanta dots
(quantum dot of i.e. two kinds Material claddings, existing CdTe, also there is CdS).
As shown in Figure 1a, there are one open ducts for tool on the sensor fibre that one embodiment of the invention is selected.As shown in Figure 1 b,
There are two symmetrically arranged open ducts for tool on the sensor fibre that another embodiment of the present invention is selected.
As shown in figure 4, optical fiber biosensor of the present invention for detecting NO concentration in organism, including light source are (optional
With laser), sensor fibre and information collection and data processing system (selecting spectrometer), the sensor fibre be above-mentioned implementation
The sensor fibre of example.Laser is sent out from light source through filter, and a part of light enters micro-structure sense light by micro objective
Fine (MOF), a part of light enter spectrometer.The sensor fibre is placed in environment to be measured, the NO concentration in environment can influence solid
It is scheduled on the quantum dot that sensor fibre opens duct, fluorescence signal generates variation, this signal intensity can be shown on spectrometer
Out, to achieve the purpose that detect the NO concentration in organism.
The present invention is used to detect the preparation method of the sensor fibre of NO concentration in organism, as shown in Fig. 2, including following step
Suddenly：
1) quantum dot for being bonded on open duct is prepared, quantum dot is the semiconductor-quantum-point containing Cd elements；
2) two kinds of microstructured optical fibers with open duct are selected, hydroxylating and silanization are carried out to open bore road surface；
3) under the action of crosslinking agent, the quantum dot of preparation is directly anchored to open duct table by way of chemical bond
Face.
Wherein, CdTe quantum, CdS quantum dot or CdTe/CdS core-shell quanta dots can be selected in quantum dot.
Embodiment 1
The preparation process of each quantum dot introduced below：
(1) preparation of CdTe quantum.
It is passed through high-purity N2Half an hour fully excludes the air in condensation reflux unit.By CdCl2Solution is added to three necks
In flask, add 200mg citrate dihydrate trisodiums under magnetic stirring；It is sequentially added into 8mLNa again2TeO3Solution, 92mL water,
100mg L-cysteines and 130mg NaBH4。N2Said mixture solution temperature is being raised to 80 DEG C under protection, in spiral
50min is condensed in reflux.After condensing reflux, cooled to room temperature, obtains CdTe quantum under nitrogen protection.
(2) NO fluorescence probes open the fixation of channel surfaces in microstructured optical fibers.
The semiconductor-quantum-point prepared in step (1) is fixed on to the opening duct table of microstructured optical fibers shown in Fig. 1 (a)
Face.Functionalization, i.e. hydroxylating and silanization are carried out to the opening channel surfaces of microstructured optical fibers first；Then in the work of crosslinking agent
Under, the semiconductor-quantum-point of preparation is directly anchored to open channel surfaces, specific steps such as Fig. 2 by way of chemical bond
It is shown.
(3) detection of the hypersensitive NO sensors to NO based on microstructured optical fibers.
Based on the Experimental equipment of microstructured optical fibers hypersensitive NO sensors, as shown in Figure 4.473nm is excited into optical coupling
Fluorescence is sent out to fiber-optic sensor probe vitalizing semiconductor quantum dot sensitive layer to transmit along optical fiber, when semiconductor-quantum-point and NO are sent out
Its fluorescence signal (intensity or phase shift) changes after raw reaction.Using spectrometer real-time tracking and record sensor probe
The highly sensitive real-time monitoring to NO concentration is realized in the variation of fluorescence signal.
Embodiment 2
Except microstructured optical fibers shown in Fig. 1 (a) of step (2) in embodiment 1 are changed to shown in Fig. 1 (b) there are two tools
The microstructured optical fibers in open atmosphere hole, other implementations are the same as embodiment 1.
The microstructured optical fibers of Fig. 1 (b) have 2 open big air hole paths as micro reaction pool, can make to be fixed on it
The fluorescence probe on surface, which is quickly contacted with NO, to react, and improves the response time；Sensitive layer effective area is dramatically increased simultaneously, is carried
The sensitivity of high sensor.
Embodiment 3
Except the CdTe quantum of step (1) in embodiment 1 is changed to CdS quantum dot, other implementations are the same as embodiment 1.
The preparation method of CdS quantum dot：It is passed through high-purity N2Half an hour fully excludes the air in condensation reflux unit.
By CdCl2Solution is added in three-neck flask, adds 200mg citrate dihydrate trisodiums under magnetic stirring；It is sequentially added into again
0.0974g thio ureas, the water of 92mL, 100mg L-cysteines and 130mg NaBH4。N2By above-mentioned mixing under protection
Object solution temperature is raised to 80 DEG C, and 50min is condensed in spiral reflux.It is naturally cold under nitrogen protection after condensing reflux
But to room temperature, CdS quantum dot is obtained.
Embodiment 4
Except the CdTe quantum of step (1) in embodiment 1 is changed to CdTe/CdS nuclear shell structure quantum points, other embodiment party
Method is the same as embodiment 1.
The preparation of CdTe/CdS nuclear shell structure quantum points：It is passed through high-purity N2Half an hour fully excludes condensation reflux unit
In air.By CdCl2Solution is added in three-neck flask, adds 200mg citrate dihydrate trisodiums under magnetic stirring；It presses again suitable
8mLNa is added in sequence2TeO3Solution, the water of 92mL, 100mg L-cysteines and 130mg NaBH4。N2Will be above-mentioned under protection
Mixture solution temperature is raised to 80 DEG C, and 30min is condensed in spiral reflux.By a small amount of deionized water dissolving 0.0974g
Thio urea (about 1.28mmol), the 20min that is injected into reaction unit that the reaction was continued.After condensing reflux, under nitrogen protection
Cooled to room temperature obtains CdTe/CdS nuclear shell structure quantum points.
It should be understood that for those of ordinary skills, it can be modified or changed according to the above description,
And all these modifications and variations should all belong to the protection domain of appended claims of the present invention.
Claims (10)
1. a kind of sensor fibre for detecting NO concentration in organism, which is characterized in that have open bore on the sensor fibre
Road, and NO fluorescence probes are fixed in the surface in open duct by the way of chemical bonding, which is to be bonded in opening
Quantum dot on duct, quantum dot are the semiconductor-quantum-point containing Cd elements.
2. the sensor fibre according to claim 1 for detecting NO concentration in organism, which is characterized in that fluorescence probe
For.
3. the sensor fibre according to claim 1 for detecting NO concentration in organism, which is characterized in that the sense light
There are two symmetrically arranged open ducts for tool on fibre.
4. the sensor fibre according to claim 1 for detecting NO concentration in organism, which is characterized in that the sense light
There are one open ducts for tool on fibre.
5. a kind of optical fiber biosensor for detecting NO concentration in organism, which is characterized in that including light source, sensor fibre
With information collection and data processing system, the sensor fibre is the sensor fibre described in any one of claim 1-4.
6. a kind of preparation method for detecting the sensor fibre of NO concentration in organism, which is characterized in that include the following steps：
1）The quantum dot for being bonded on open duct is prepared, quantum dot is the semiconductor-quantum-point containing Cd elements；
2）Two kinds of microstructured optical fibers with open duct are selected, hydroxylating and silanization are carried out to open bore road surface；
3）Under the action of crosslinking agent, the quantum dot of preparation is directly anchored to open channel surfaces by way of chemical bond.
7. preparation method according to claim 6, which is characterized in that the quantum dot is CdTe quantum, CdS quantum dot
Or CdTe/CdS core-shell quanta dots.
8. preparation method according to claim 7, which is characterized in that CdTe quantum preparation method is as follows：
1）It is passed through high-purity N2Half an hour fully excludes the air in condensation reflux unit；
2）By CdCl2Solution is added in three-neck flask, adds 180 ~ 220 mg citrate dihydrate trisodiums under magnetic stirring, by suitable
6 ~ 8mL Na are added in sequence2TeO3Solution, the water of 90 ~ 100 mL, 95 ~ 105mg L-cysteines and 125 ~ 135mg NaBH4,
Obtain mixed solution；
3）N2Under protection, mixture solution temperature is raised to 70 ~ 90 DEG C, 40 ~ 50min is condensed in spiral reflux；
4）After condensing reflux, cooled to room temperature, obtains CdTe quantum under nitrogen protection.
9. preparation method according to claim 7, which is characterized in that the preparation method of CdS quantum dot is as follows：
1）It is passed through high-purity N2Half an hour fully excludes the air in condensation reflux unit；
2）By CdCl2Solution is added in three-neck flask, adds 180 ~ 220 mg citrate dihydrate trisodiums under magnetic stirring；It presses again
1 ~ 2 mmol thio ureas, the water of 90 ~ 100mL, 95 ~ 105mg L-cysteines and 125 ~ 135mg is added in sequence
NaBH4, obtain mixed solution；
3）N2Under protection, mixture solution temperature is raised to 70 ~ 90 DEG C, 40 ~ 50min is condensed in spiral reflux；
4）After condensing reflux, cooled to room temperature, obtains CdS quantum dot under nitrogen protection.
10. preparation method according to claim 7, which is characterized in that CdTe/CdS core-shell quanta dots preparation methods are as follows：
1）It is passed through 2 half an hour of high-purity N, fully excludes the air in condensation reflux unit；
2）By CdCl2Solution is added in three-neck flask, adds 180 ~ 220 mg citrate dihydrate trisodiums under magnetic stirring；It presses again
6 ~ 8mLNa is added in sequence2TeO3Solution, the water of 90 ~ 100 mL, 95 ~ 105mg L-cysteines and 125 ~ 135mg
NaBH4, obtain mixed solution；
3）N2Under protection, mixture solution temperature is raised to 70 ~ 90 DEG C, 30 ~ 40 min are condensed in spiral reflux, will be lacked
1 ~ 2 mmol thio ureas of deionized water dissolving of amount, 20 ~ 30 min that are injected into reaction unit that the reaction was continued；
4）After condensing reflux, cooled to room temperature, obtains CdTe/CdS nuclear shell structure quantum points under nitrogen protection.
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|---|---|---|---|---|
| US20030066998A1 (en) * | 2001-08-02 | 2003-04-10 | Lee Howard Wing Hoon | Quantum dots of Group IV semiconductor materials |
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