CN104089999A - Carbon quantum dot-nanowire array-based cardiomyocyte signal molecule sensor and preparation method thereof - Google Patents
Carbon quantum dot-nanowire array-based cardiomyocyte signal molecule sensor and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of photoelectrochemical sensors, and particularly relates to a carbon quantum dot-nanowire array-based cardiomyocyte signal molecule sensor based on sunlight drive and a preparation method thereof. The preparation method comprises the following steps: under a hydrothermal system, growing a titanium dioxide nanowire array on a conductive glass substrate; obtaining silane-functionalized nitrogen-doped carbon dots (N-CDots) at one step in situ through using a microwave method; then placing titanium dioxide nanowires processed by concentrated sulfuric acid into an N-CDot ethanol solution, adding stronger ammonia water, standing and reacting, and defining as N-CDot-TiO2. The photoelectric current of the N-CDot-TiO2 is improved by almost two times compared with that of pure TiO2, and the prepared photoelectric current on-off sensor can be used for dynamically detecting the content of H2S star molecules of cardiomyocytes in real time in situ. The carbon quantum dot-nanowire array-based cardiomyocyte signal molecule sensor is smart in design and wide in the source of raw materials, the preparation method is simple, environment-friendly, low in price, fast to response, wide in linear range, high in selectivity, and beneficial to popularization and application.
Description
Technical field
The invention belongs to Optical Electro-Chemistry sensor technical field, the carbon quantum dot-nano-wire array that is specifically related to a kind of sunshine driving is applied to signaling molecule sensor of cardiac muscle cell and preparation method thereof.
Background technology
In recent years; becoming increasingly conspicuous of current the faced energy crisis of the mankind and problem of environmental pollution; the demand for development of society is developed efficiently; free of contamination clean energy resource; the energy that supports human survival and development is just from oil, and coal etc. are that the main pollution consumption-orientation energy turns to the clean regenerative energy.PhotoelectrochemicalTechnique Technique is usingd its room temperature deep reaction and can directly utilize sun power to drive the special performances such as reaction as light source, and becomes a kind of desirable environmental pollution treatment technology and clear energy sources production technology.Wherein, semiconductor nano material TiO especially
2nano material is owing to having unique photoelectrochemical behaviour, excellent thermal stability, biologically inert, nontoxic and simple for production etc., make no matter it is aspect solar energy photoelectric conversion, or aspect the degradation treatment of refuse, having good actual application prospect.Yet, nano-TiO
2also there are problems in itself, such as: forbidden band is wide, and electronics one hole of generation is short and very easily compound to the life-span, and spectral response range is narrower, and these have all suppressed nano-TiO to a great extent
2the active and application of Optical Electro-Chemistry.For current nano-TiO
2these defects that exist, can adopt sensitization, semiconductor is compound, nano-electrode builds in order and the approach such as novel electrolytes design to nano-TiO
2optical Electro-Chemistry system is carried out modification and optimization.TiO
2photoelectric property and chemical modification become a general study hotspot.
In view of TiO
2existing spectral response range is narrow, the compound serious shortcoming of photo-generated carrier, to TiO
2modify the focus that becomes current research with modification.Modification and modification mainly concentrate on two aspects, and the first is carried out the control of pattern to it, and second it be that it is carried out to chemical modification.TiO
2that material has is cheap, stable in properties, be easy to the advantages such as preparation, and it is carried out to further chemical modification, can improve the defect that its spectral response range is narrow, charge carrier recombination rate is low, and it is applied among Optical Electro-Chemistry reaction system better.Based on TiO
2optical Electro-Chemistry to detect principle be to utilize test substance to be fixed on TiO
2electrode surface, and the analysis of quantitative and qualitative analysis is carried out in the variation of system Optical Electro-Chemistry signal during by monitoring test substance generation particular organisms chemical reaction (as photocurrent etc.) to determinand.As Willner etc. utilizes the variation of CdS Optical Electro-Chemistry signal, realized the detection of DNA, and Chen Da etc. utilize the reaction of horseradish peroxidase (HRP) catalyzing hydrogen peroxide for accompanying TiO
2nanometer pipe array electrode has been realized the detection to hydrogen peroxide under visible ray in the variation of 400 nm place photocurrents.
Optical Electro-Chemistry grows up in the 1980s as a kind of sensitive detection means, and is further being promoted in recent years.The whole system that Optical Electro-Chemistry detects adopts light to excite, detection be electric signal, both can phase mutual interference, therefore the analytical approach based on Optical Electro-Chemistry has very high sensitivity; Compare with electrochemical detection method, Optical Electro-Chemistry analytical approach can detect by the pattern of array owing to adopting light to excite therefore; In addition, Optical Electro-Chemistry adopts electron detection device, and non-light detects, so instrument volume is little, and cost is low, and testing cost is other detection modes and Yan Yao little relatively.The ultimate principle that Optical Electro-Chemistry detects is exactly that based semiconductor material or Optical Electro-Chemistry signal response molecule are under the illumination of suitable wavelength, absorb after photon energy the electronic transition of semiconductor valence band to conduction band or signal response molecule from ground state transition to excited state, produce photogenerated charge, and be further transferred to electrode surface formation photocurrent, by detecting the variation (size, direction) of photo-signal, obtain corresponding detected molecular information.The people such as Weber use ruthenium dipyridine compound as Optical Electro-Chemistry signaling molecule, to being present in the Ru (bpy) in serum
3 2+carried out Optical Electro-Chemistry detection, the in the situation that of uric acid and oxalic acid existence, the detectability that the method detects ruthenium compound can reach 10
-10mol/mL; Lacourse etc. utilize photoelectrochemical method to detect the small-molecule substances such as acetone, phenol, and its detectability can reach 20-30ppm in 200uL system.Weetall laboratory utilizes PhotoelectrochemicalMethod Method to detect DNA first, has started and has utilized PhotoelectrochemicalTechnique Technique to carry out the macromolecular epoch of detection of biological.Afterwards, many researchers are attempted directly utilizing the photoelectrochemical behaviour of semiconductor material to detect biomolecule.For example, calendar year 2001, wiliner etc. utilize the Optical Electro-Chemistry response success of CdS quantum dot to detect DNA biomacromolecule; The grand seminar of the Guo Liang of Ecotope Centre, Chinese Academy of Sciences utilizes Optical Electro-Chemistry detection technique to interact and carried out quantitative detection biotin one microbiotic (biotin-avidin), and they adopt ruthenium dipyridine sensitization SnO
2semiconductor nano electrode is as Optical Electro-Chemistry response interface, then microbiotic is adsorbed onto electrode surface by the effect of Electrostatic Absorption, along with the biotin adding and electrode surface microbiotic interact, also can there is corresponding variation in photo-signal, obtain thus biological respinse and successfully utilize TiO
2the photoelectrochemical behaviour of nano-electrode, do not introducing under the prerequisite of any other Optical Electro-Chemistry signaling molecule, directly the heterozygosis process of DNA has been carried out to Optical Electro-Chemistry detection, thereby provide a kind of unmarked molecule, highly sensitive, easy and novel detection method cheaply for the detection of DNA heterozygosis process.
As a kind of novel fluorescent carbon nano material, there is good fluorescence stability, the performance of a series of excellences such as unglazed scintillation, good water-soluble and biocompatibility and hypotoxicity and excitation wavelength and emission wavelength can regulate and control, making fluorescent carbon point replace the status of quantum dot in biomedicine becomes possibility.In numerous methods of preparing carbon point, adopt the expensive carbon source materials such as carbon nano-tube or Graphene, use the expensive equipment such as laser pulse instrument, or the mode modified carbon point such as doped metal ion and surface passivation PEG, the carbon point compared with high-fluorescence quantum yield can be obtained.Yet because above-mentioned these preparation method's processing steps are loaded down with trivial details, the more high factor of raw materials cost, what cause fluorescent carbon point is syntheticly difficult to large-scale production.In addition utilize glucose, polyglycol etc. as carbon source, under strong acid condition, microwave or ultrasound wave are assisted while decomposing, and subsidiary reaction more complicated causes detachment process loaded down with trivial details time-consuming, and fluorescent carbon point output is not high, so and the application of the method is restricted.Utilize respectively glycerine and tributyl phosphate to prepare the carbon point that fluorescent emission wavelength excites dependence, for structure and the luminescence mechanism of research carbon point provides good, a new material simultaneously.A kind of method of easy microwave-heating is successfully used in the preparation of carbon point.The principle that microwave assisting method is prepared carbon point is generally under the high-energy microwave action that organic molecule provides in moment carbonization occurs, crosslinked nucleation, thus form the carbon point with fluorescence property.The inventive method is simple and efficient to handle, and raw material is easy to get, and environmental friendliness adopts simple microwave reactor can successfully realize the preparation of carbon point.
Sulfuretted hydrogen is considered to a kind of toxic gas with rotten egg smell always, in recent years, has caused the exploration interest of vast researcher as the third gaseous signal molecule after NO, CO.In ox, mouse and people's brain, there is the Endogenous Hydrogen Sulfide of high concentration, up to 10 ~ 600 mM.In vivo, the various physiology courses of sulfuretted hydrogen wide participation, have different physiological roles, comprise vasodilation, anti-oxidant, anti-apoptosis and antiinflammation etc.H in body
2the change of S concentration is relevant with various diseases, as sick in Alzheimer Alzheimer (being senile dementia), Down's syndrome, diabetes, cirrhosis, chronic obstructive pulmonary disease etc., therefore the biological study for sulfuretted hydrogen has become current hot fields.For the relation between Study on Endogenous sulfuretted hydrogen and relevant disease more in depth, to the action site of body and molecule mechanism thereof, realization seems very important to accurate, quick, sensitive, the in situ detection of sulfuretted hydrogen in body, thereby analytical work person is developed to new biological tissue sulfurated hydrogen detection strategy and technology has proposed new challenge.Traditional detection H
2s method mainly comprises colourimetry, electrochemical process, vapor-phase chromatography, metal inducement sulphur sedimentation etc., yet these methods need to be carried out complexity, lengthy and tedious pre-service to sample, make cannot realize H in body
2the Real-Time Monitoring of S level and space monitoring.And sulfuretted hydrogen kalabolism is quick in vivo, cause the fluctuation of its level very large, make that its accurate detection is had to great difficulty.The endogenous H of current bibliographical information
2s concentration range is 10 to 100 μ M.By fluorescent small molecule probe, carrying out fluoroscopic examination and fluorescence imaging is at present for the online the most attractive molecular imaging technology detecting of biomolecule, mainly gives the credit to its high sensitivity, good selectivity, not damaged and is applicable to active somatic cell, tissue and toy.Therefore, develop new fluorescence probe for H
2the detection of S and imaging, can provide very important means to its research for us.Up to now, design detects H
2the fluorescence probe of S is mainly with its reductibility, nucleophilicity and the strong complexing power to copper ion.
H
2s and copper ion complexing, Nagano seminar has reported the copper ion Complex probe HSip-1 of the large ring of a kind of nitrogen-containing hetero, and the copper ion of sulfuretted hydrogen in probe pulled out copper ion after being combined from probe, discharge fluorophor, the fluorescence of probe is recovered.This class probe can be realized the high selectivity to sulfuretted hydrogen, high responsive detection, and can be used for detecting in active somatic cell.Yu adopts dinaphthol as fluorescence signal group, in connections can with the part of copper ion complexing, developed a kind of multi-functional fluorescence probe, detection when can realize sulphion in aqueous solution and copper ion.These probes are to H above
2improvement and perfection has been carried out in one or more aspects such as S selectivity, response time, detectability, serum detection, cell and imaging of tissue, has obtained good progress.Yet great majority still adopt common fluorescent dye as signal reporter group, excite with emission wavelength in ultraviolet-visible district, for cell and living imaging, there is many drawbacks, as tissue damaged is severe, penetration capacity is not strong etc.
The present invention has designed the carbon quantum dot-nano-wire array of novel sunshine driving for cardiac muscle cell's signaling molecule sensor, for Real-time and Dynamic Detection H according to above research background
2s.
Summary of the invention
The object of this invention is to provide cardiac muscle cell's signaling molecule sensor of a kind of carbon quantum dot-nano-wire array driving based on sunshine and its preparation method and application.
Cardiac muscle cell's signaling molecule transducer production method of a kind of carbon quantum dot-nano-wire array driving based on sunshine provided by the invention, concrete steps are:
(1) water is mixed to 0.1 – with concentrated acid after 0.5 hour, add wherein titanium source, Xi Yuan, under stirring, react 5 – 10 min, then transfer in reactor, the substrate of suitable size (glass sheet) face down is put into still, under 140 180 ℃ of – conditions, react 4 – 20 hours; The product that above-mentioned steps is made is rinsed well, is dried, roasting 1 – 2 hours under 550 ℃ of temperature conditions of 450 – in air atmosphere, then roasting 0.5 – 2 hours under 550 ℃ of temperature conditions of 350 – in inert gas and reducibility gas mixed atmosphere, obtains and mixes titanium dioxide nanowire array;
(2) titanium dioxide nanowire array is placed in to concentrated acid soaked overnight, makes hydroxyl on its surface band;
(3) the nitrogenous carbon point of microwave method one-step synthesis, 0.25 g citric acid, be carbon source, the melamine of 0-20 mg is nitrogenous source, utilizes 5 ml organosilanes for coupling agent and functionalized reagent, and single stage method original position obtains the carbon point (being designated as N-CDots) of the nitrogen doping of silane-functionalized;
(4) then by the titanium dioxide nano thread of processing through the concentrated sulphuric acid, be put in the ethanolic solution of N-CDot, add strong aqua, standing 24-48 hour makes it reaction, is defined as N-CDot-TiO
2; Substrate N
2dry up, stand-by;
N-CDot-TiO
2than pure TiO
2photocurrent has improved and has approached 2 times;
(5) by cancellation metal ions M (as Cu
2+) cancellation fluorescence, and then N-CDot-TiO
2the photocurrent of+M has been got back to again and pure TiO
2similar size, has at this moment added again S
2-and H
2s competes out by M again, and then fluorescence recovers again, photocurrent get back to again before N-CDot-TiO
2size, like this preparation a sensor that photocurrent is opened and closed, can be for real-time in-situ detection of dynamic cardiac muscle cell's H
2the content size of S star molecule.
In the present invention, described carbon source is one or more in citric acid, polyglycol or polystyrene sphere.
In the present invention, described nitrogenous source is one or more in melamine, urea or ethylenediamine.
In the present invention, described silane is N-β-(aminoethyl)-γ-aminopropyltrimethoxysilane (AEAPMS) or 3-aminopropyl triethoxysilane (APTES).
Described cancellation metal ions M can be Cu
2+, Zn
2+or Fe
3+.
In the present invention, in cardiac muscle cell's signaling molecule sensor of the carbon quantum dot-nano-wire array driving based on sunshine, lowest detection is limited to 10 nM, highly sensitive, and selectivity is good.
Novel nano linear array of the present invention also can be applicable to Optical Electro-Chemistry sensor: take carbon quantum dot-nano-wire array as working electrode, Ag/AgCl electrode is contrast electrode, platinum filament is to electrode, pH7.4 PBS solution is reaction electrolytic solution, irradiating light intensity is a sunshine, and irradiating sectional area is can be according to 0.02-0.06 cm
2size regulates.
Carbon quantum dot-nano-wire array of the present invention is that first hydro-thermal method is prepared titanium oxide nano wire, by the synthetic nitrating carbon point of a step microwave method, finally by the silane coupled step original position nitrating carbon point of growing on titanium oxide nano wire.By copper ion, carry out cancellation fluorescence, again by sulphion or H
2s comes competitively copper ion to be cemented out, thereby fluorescence recovers again, has formed the sensor of photocurrent switch.It is simple that carbon quantum dot-TiOx nano linear array has manufacture craft, cost is low, good stability, high sensitivity and low detectability environment amenable feature, being applicable to large area produces, for significant on Optical Electro-Chemistry ability sensor research, in photoelectricity bioanalysis field, also there is good application prospect.
Accompanying drawing explanation
Fig. 1: the carbon point SEM photo that a microwave method is synthetic; B is for amplifying crystal lattice pattern; C is the fluorescence intensity peak of the carbon point of different itrogen content of getter with nitrogen dopeds; D is that side SEM figure and the e of carbon quantum dot-nano-wire array is SEM front view (FV); F is the TEM figure of carbon quantum dot-nano-wire array.
Fig. 2: a is the photoelectricity flow graph of pure zirconia titanium, carbon point sensitization titanium dioxide, nitrogen-doped carbon point sensitization titanium dioxide, copper ion cancellation nitrogen-doped carbon point sensitization titanium dioxide etc.; B nitrogen-doped carbon point sensitization titanium dioxide is along with add the variation diagram of the photocurrent of copper ion and sulphion along with the time in solution.
Fig. 3 is the signaling molecule sensor diagram that the carbon quantum dot-nano-wire array of sunshine driving is applied to cardiac muscle cell.
Embodiment
Below, by following examples, the invention will be further described, and it will contribute to understand the present invention, but not limit content of the present invention.
Under hydrothermal system, take tetra-n-butyl titanate as titanium source, water is solvent, and butter of tin is Xi Yuan, and concentrated hydrochloric acid is acid source, containing the titanium dioxide nanowire array of growing in the substrate of fluorine tin oxide (FTO) electro-conductive glass.Again by the nitrogenous carbon point of microwave method one-step synthesis, citric acid is carbon source, the melamine of different proportion is nitrogenous source, utilizing organosilane N-β-(aminoethyl)-γ-aminopropyltrimethoxysilane (AEAPMS) is coupling agent and functionalized reagent, and single stage method original position has obtained the carbon point (N-CDots) of the nitrogen doping of silane-functionalized.Then by the titanium dioxide nano thread of processing through the concentrated sulphuric acid, be put in the ethanolic solution of N-CDot, add strong aqua, within standing 24 hours, make it reaction, be defined as N-CDot-TiO
2.N-CDot-TiO
2than pure TiO
2photocurrent has improved and has approached 2 times, passes through Cu
2+can cancellation fluorescence, and then N-CDot-TiO
2+ Cu
2+photocurrent got back to again and pure TiO
2similar size, has at this moment added again S
2-and H
2s is again by Cu
2+out, and then fluorescence recovers again in competition, photocurrent get back to again before N-CDot-TiO
2size, prepare like this sensor that a photocurrent is opened and closed, can be for real-time in-situ detection of dynamic cardiac muscle cell's H
2the content size of S star molecule.The present invention designs ingenious, starting material wide material sources, and preparation method is simple, environmental protection, cheap, responds quick, the wide range of linearity, high selectivity, is conducive to promote.
As shown in Figure 3, take carbon quantum dot-nano-wire array as working electrode, Ag/AgCl electrode is contrast electrode, and platinum filament is to electrode, and pH 7.4 PBS solution are reaction electrolytic solution, and irradiating light intensity is a sunshine, and irradiated area is 1 cm
2.Under 0 V condition, survey its photoelectricity flow valuve along with the time changes (as Fig. 2 b).
Claims (6)
1. a preparation method for cardiac muscle cell's signaling molecule sensor of the carbon quantum dot-nano-wire array driving based on sunshine, is characterized in that concrete steps are:
(1) water is mixed to 0.1 – with concentrated acid after 0.5 hour, add wherein titanium source, Xi Yuan, reacts 5 – 10 min under stirring, then transfer in reactor, and substrate front surface is put into still down, under 140 180 ℃ of – conditions, reacts 4 – 20 hours; The product that above-mentioned steps is made is rinsed well, is dried, roasting 1 – 2 hours under 550 ℃ of temperature conditions of 450 – in air atmosphere, then roasting 0.5 – 2 hours under 550 ℃ of temperature conditions of 350 – in inert gas and reducibility gas mixed atmosphere, obtains and mixes titanium dioxide nanowire array;
(2) titanium dioxide nanowire array is placed in to concentrated acid soaked overnight, makes hydroxyl on its surface band;
(3) the nitrogenous carbon point of microwave method one-step synthesis, 0.25 g citric acid, be carbon source, the melamine of 0-20 mg is nitrogenous source, utilizes 5 ml organosilanes for coupling agent and functionalized reagent, single stage method original position obtains the carbon point of the nitrogen doping of silane-functionalized, is designated as N-CDots;
(4) then by the titanium dioxide nano thread of processing through the concentrated sulphuric acid, be put in the ethanolic solution of N-CDot, add strong aqua, standing 24-48 hour makes it reaction, is defined as N-CDot-TiO
2; Glass sheet N
2dry up, stand-by;
(5) by cancellation metal ions M cancellation fluorescence, and then N-CDot-TiO
2the photocurrent of+M is got back to again and pure TiO
2similar size, at this moment adds S
2-and H
2s, competes out by M again, and then fluorescence recovers again, photocurrent get back to again before N-CDot-TiO
2size, be prepared into like this H for real-time in-situ detection of dynamic cardiac muscle cell
2the sensor that the photocurrent of the content size of S star molecule is opened and closed.
2. preparation method as claimed in claim 1, is characterized in that described carbon source is one or more in citric acid, polyglycol or polystyrene sphere.
3. preparation method as claimed in claim 1, is characterized in that described nitrogenous source is one or more in melamine, urea or ethylenediamine.
4. preparation method as claimed in claim 1, is characterized in that described silane is N-β-(aminoethyl)-γ-aminopropyltrimethoxysilane or 3-aminopropyl triethoxysilane.
5. the preparation method as described in one of claim 1-4, is characterized in that described cancellation metal ions M is Cu
2+, Zn
2+or Fe
3+.
6. cardiac muscle cell's signaling molecule sensor of the carbon quantum dot-nano-wire array driving based on sunshine being prepared by the described preparation method of one of claim 1-5, lowest detection is limited to 10 nM.
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