CN103712968B - Silicon nanowire-based fluorescence chemical sensor with selective fluorescence response to free-state copper ions and complex-state copper ions - Google Patents
Silicon nanowire-based fluorescence chemical sensor with selective fluorescence response to free-state copper ions and complex-state copper ions Download PDFInfo
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- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention relates to a silicon nanowire-based fluorescence chemical sensor with selective fluorescence response to free copper ions and complex copper ions, and a preparation method and application thereof. The invention activates the surface of the silicon nanowire or the silicon nanowire ordered array, and then sequentially reacts with 3-2- (2-aminoethylamino) ethylaminopropyl trimethoxy silane and dansyl chloride to realize the surface functional modification of the silicon nanowire or the silicon nanowire ordered array, thereby obtaining the fluorescent chemical sensor. The fluorescence chemical sensor can be used for detecting the free copper ions and the complex copper ions in a common solution system and a biological system containing the free copper ions and the complex copper ions, and can also be used as a substrate for cell growth to construct an activity detection chip carrying cells and observe the release of the free copper ions and the complex copper ions in the apoptosis process in real time and in situ.
Description
Technical field
The invention belongs to the fluorescence chemical sensor field of nanostructured, particularly to free state copper ion
With copper in complex state have selectivity fluorescence response fluorescence chemical sensor based on silicon nanowires and
Preparation method, and the application of this fluorescence chemical sensor.
Background technology
As one of metal ion that content in the middle of human body is most, copper ion is in regulation person's physiological processes
Play very important effect.In organism, most of copper ions are all to exist with complex state form:
A part of copper ion and some specific protein bound, a part of and numerous little molecular nutrition material knots
Close.These copper ion complex serve vital effect during living things catalysis.Although it is biological
In body, the total concentration of copper ion is substantially up to 100 micromoles, but the copper ion of free state is the most considerably less,
Less than one in the most each cell.The most in succession develop numerous fluorescence for detecting copper ion
Sensor, but these sensors are mostly aimed at the detection of free state copper ion, to complex state copper from
The detection method of son is the deficientest.Therefore, in order to preferably study copper ion work in living things system
With, it is far from being enough for only leaning on high-sensitive free state copper ion fluorescent optical sensor, it is often more important that development
The excellent fluorescent optical sensor of copper in complex state detection can be applied to.Copper in complex state can be applied to
One of effective ways of detection, it is simply that design one and free state copper ion is had sufficiently strong complexing power
Fluorescent optical sensor.When this fluorescence chemical sensor is present in the system containing copper in complex state,
It can take by force from copper complex easily copper ion and and these copper ion generation complexings, enter one
Step promotes the fluorescence signal of this fluorescence chemical sensor self to change, thus believes according to sensor fluorescence
Number change realize the detection of copper in complex state.This fluorescent optical sensor is to the detection of copper ion also simultaneously
Should have the advantages such as high selectivity, high sensitivity, it is possible to be prevented effectively from other metal ion and living things system
In active biomolecule interference that free state copper ion and copper in complex state are detected.
Recent studies indicate that, by having organic molecule or the biology point of high Selective recognition ability
Son is fixed to the surface of monodimension nanometer material, can improve greatly some fluorescent optical sensors selectivity and
Sensitivity.In numerous monodimension nanometer materials, silicon nanowires is owing to having avirulence, bio-compatibility
The advantage such as good and the most integrated, has been widely used in constructing of fluorescent optical sensor, and these
Sensor has all shown good detection performance.The oldered array of silicon nanowires is possible not only to spontaneous simultaneously
Permeates cell membranes, but also the adhesion between cell and substrate can be strengthened and play the work of fixing cell
With, therefore it is successfully applied in the fields such as gene transfer, supporting structure and anti-biotic material.Based on
Above consideration, the present invention is first by small molecule organic compound 3-2-(2-aminoethylamino) ethylamino
Propyl trimethoxy silicane modifies silicon nanowires or the surface of silicon nanowires oldered array, passes through the most again
Surface is modified with 3-2-(2-aminoethylamino) silicon nanowires of diethylaminobutyyl trimethoxy silane or
The amino on silicon nanowires oldered array surface and dansyl Cl reaction, obtained through surface modify to trip
Amorph copper ion and copper in complex state have the fluorescence chemical based on silicon nanowires of selectivity fluorescence response
Sensor, this fluorescence chemical sensor can not only be applied to free state copper in regular solution and living things system
Ion and the detection of copper in complex state, and load can be constructed further as the substrate of cell growth
There is an Activity determination chip of cell, and utilize this chip in real time, in situ detection apoptosis process discharges
Free state copper ion and copper in complex state.
Summary of the invention
An object of the present invention is to provide that free state copper ion and copper in complex state are had selectivity is glimmering
The fluorescence chemical sensor based on silicon nanowires of photoresponse.
The two of the purpose of the present invention are to provide that free state copper ion and copper in complex state are had selectivity is glimmering
The preparation method of the fluorescence chemical sensor based on silicon nanowires of photoresponse.
The three of the purpose of the present invention are to provide that free state copper ion and copper in complex state are had selectivity is glimmering
The application in terms of biological detection of the fluorescence chemical sensor based on silicon nanowires of photoresponse.
The present invention free state copper ion and copper in complex state are had selectivity fluorescence response based on silicon
The fluorescence chemical sensor of nano wire, is by silicon nanowires (available chemical vapour deposition technique prepares)
Or after silicon nanowires oldered array (available chemical etching method prepares) carries out surface active, more successively
With 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy silane and dansyl Cl react,
Realize the surface-functionalized modification of silicon nanowires or silicon nanowires oldered array, thus obtain free state copper
Ion and copper in complex state have fluorescence chemical based on the silicon nanowires sensing of selectivity fluorescence response
Device.The present invention free state copper ion and copper in complex state are had selectivity fluorescence response based on silicon
It is the most molten that the fluorescence chemical sensor of nano wire can be used for containing free state copper ion and copper in complex state
Free state copper ion in liquid system and living things system and the detection of copper in complex state, it is also possible to will be to trip
Amorph copper ion and copper in complex state have the fluorescence chemical based on silicon nanowires of selectivity fluorescence response
The substrate that sensor grows as cell, constructs the Activity determination chip being loaded with cell, and utilize its in real time,
Free state and the release of copper in complex state in home position observation apoptosis process.
The present invention free state copper ion and copper in complex state are had selectivity fluorescence response based on silicon
The fluorescence chemical sensor of nano wire, be by the silicon nanowires of surface active or silicon nanowires oldered array and
3-2-(2-aminoethylamino) reaction of diethylaminobutyyl trimethoxy silane, surface must be arrived and be modified with 3-2-
The silicon nanowires of (2-aminoethylamino) diethylaminobutyyl trimethoxy silane or the orderly battle array of silicon nanowires
Row, and utilize 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy silane modify silicon nanometer
The amino on line or silicon nanowires oldered array surface and the acyl chlorides key in dansyl Cl molecule occur to replace instead
Should, the surface obtained is modified with 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy silane
Silicon nanowires or silicon nanowires oldered array react with dansyl Cl, make 3-2-(2-aminoethylamino)
The product of diethylaminobutyyl trimethoxy silane and dansyl Cl is modified in silicon nanowires or silicon nanometer
Obtain behind the surface of linear array.
Silicon nanowires in the silicon nanowires of described surface active is to be obtained not by chemical vapour deposition technique
With the silicon nanowires of size, the most a diameter of 5～the silicon nanowires of 15nm.
Silicon nanowires oldered array in the silicon nanowires oldered array of described surface active is to be carved by chemistry
The various sizes of silicon nanowires oldered array that erosion method obtains, the preferably silicon in silicon nanowires oldered array are received
A diameter of the 100 of rice noodle～250nm, a length of 5～30 μm.
The present invention free state copper ion and copper in complex state are had selectivity fluorescence response based on silicon
The preparation method of the fluorescence chemical sensor of nano wire comprises the following steps:
1) silicon nanowires (available chemical vapour deposition technique prepares) being placed in temperature is 75～90 DEG C
Volume ratio is concentrated sulphuric acid (mass concentration is 98%) and the H of 1:2～9:12O2Mixed liquor in heat 30～180
Minute, it is cooled to room temperature, takes out silicon nanowires and be washed to neutrality；Under room temperature, silicon nanowires is placed in body
Long-pending than the H for 2:1:1～8:1:12O:H2O2:NH4In the mixed liquor of OH, (general silicon nanowires is at mixed liquor
In time be 0.5～3 hour), then take out silicon nanowires and be washed to neutrality, vacuum drying, obtain
The silicon nanowires that surface is activated；
Or silicon nanowires oldered array (available chemical etching method prepare) is placed in temperature is 75～95 DEG C
The concentrated sulphuric acid that volume ratio is 1:1～9:1 (mass concentration is 98%) and H2O2Mixed liquor in heat
30～180 minutes, it is cooled to room temperature, takes out silicon nanowires oldered array and be washed to neutrality；Then by silicon
Nanometer line ordered array is placed in oxygen plasma system and carries out processing (treatment conditions: the quality of oxygen contains
Amount is 5～100%, and voltage is 100～700 volts, and the time is 5 seconds～8 minutes, and temperature is 15～45 DEG C),
Obtain the silicon nanowires oldered array that surface is activated；
2) the silicon nanometer that surface that the 5～100mg step 1) that are dried obtain is activated is added in the reactor
Line or the dry toluene of silicon nanowires oldered array, 2.5～50mL and the 3-2-(2-ammonia of 0.05～0.6mL
Base ethylamino) diethylaminobutyyl trimethoxy silane, under inert gas shielding, it is heated to 75～125 DEG C
After, isothermal reaction 10～40 hours, it is cooled to room temperature, collects silicon nanowires or silicon nanowires oldered array,
Unreacted 3-2-(2-aminoethylamino is removed with organic solvent ultrasonic cleaning) diethylaminobutyyl front three
TMOS, must arrive surface and be modified with 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy silicon
The silicon nanowires of alkane or silicon nanowires oldered array；
3) by 5～100mg steps 2) surface that obtains is modified with 3-2-(2-aminoethylamino) ethyl
The silicon nanowires of TSL 8330 or silicon nanowires oldered array are placed in and are mounted with 5～15mL
Dichloromethane reactor in, be subsequently added the triethylamine of 0.05～1.0mL, be stirred at room temperature and (typically stir
The time mixed is 5～80 minutes), two solutions of chlorine 1～10mL(being added dropwise over dansyl Cl afterwards are the reddest
The concentration of sulfonic acid chloride is 0.05～8mmol/L), (time of general stirring reaction is in stirring reaction under room temperature
0.5～6 hour), by 3-2-(2-aminoethylamino) modification of diethylaminobutyyl trimethoxy silane
The amino on silicon nanowires or silicon nanowires oldered array surface takes with the acyl chlorides key in dansyl Cl molecule
Generation reaction, must arrive surface and be modified with 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy silane
Silicon nanowires or silicon nanowires oldered array with the product of dansyl Cl；Then anti-with organic solvent
Multiple ultrasonic cleaning removes unreacted dansyl Cl, obtains free state copper ion and copper in complex state tool
The fluorescence chemical sensor based on silicon nanowires of selective fluorescence response.
Silicon nanowires in step 1) is the various sizes of silicon nanowires obtained by chemical vapour deposition technique,
The most a diameter of 5～the silicon nanowires of 15nm.
Silicon nanowires oldered array in step 1) is the various sizes of silicon nanometer obtained by chemical etching method
A diameter of 100～250nm of silicon nanowires in line oldered array, preferably silicon nanowires oldered array, long
Degree is 5～30 μm.
The newest dry toluene steamed of described dry toluene.
The organic solvent of described cleaning can be conventional organic solvent, such as methanol, ethanol or acetone.
Heretofore described is prepared the method for silicon nanowires by chemical vapour deposition technique: under room temperature,
Silicon monoxide is put in porcelain boat after mortar grinder, and porcelain boat is placed on the middle part of quartz ampoule, push true
In empty tube furnace；System is first evacuated to 10 with mechanical pump and molecular pump-3Pa, subsequently with 10～40sccm
(mL/min) flow velocity is passed through argon (accounting for the volume 95% of mixed gas) and (accounts for mixed gas with hydrogen
Volume 5%) mixed gas, when pressure stability is in 750～10000Pa, system starts to warm up；
System rises to 300 DEG C with 10～25 DEG C/min, then is warming up to 800 DEG C with 10～25 DEG C/min, now closes
Gas check valve and pump lock, be continuously heating to 1350 DEG C after being incubated 10～40 minutes, reacts 3～10 at 1350 DEG C
After hour, naturally cool to room temperature, collect product silicon nanowires in the both sides of porcelain boat.
Heretofore described is prepared the method for silicon nanowires oldered array by chemical etching method: take not
With (100) silicon chip of size, carry out ultrasonic cleaning 5～35 minutes with ethanol, acetone, distilled water successively；
Afterwards this silicon chip is placed in mass concentration be 1～6% HF aqueous solution in 25 seconds～20 minutes；Take out silicon chip
It is placed on containing the AgNO that concentration is 3～10mmol/L3Mixed aqueous solution with the HF of 2～8mol/L
In 1～10 minute；Immerse after taking out silicon chip containing the HF and 0.05～0.8mol/L that concentration is 1～10mol/L
H2O2Mixed aqueous solution in, the water bath heat preservations 5 of 25～65 DEG C～55 minutes；The silicon chip of taking-up is put
Enter concentrated hydrochloric acid (mass concentration is 36%): the volume ratio of concentrated nitric acid (mass concentration is 36%) is 3:1's
In mixed liquor, soak 0.5～3 hour, take out silicon chip and naturally dry with after distilled water flushing, obtaining by silicon
The silicon nanowire array that nano wire is constituted.
The present invention free state copper ion and copper in complex state are had selectivity fluorescence response based on silicon
It is the most molten that the fluorescence chemical sensor of nano wire can be used for containing free state copper ion and copper in complex state
Free state copper ion in liquid system or living things system (liver extracting solution or cell culture fluid) and complex state
The detection of copper ion；Can also ring free state copper ion and copper in complex state being had selectivity fluorescence
The substrate that the fluorescence chemical sensor based on silicon nanowires answered grows as cell, described Fluoresceinated
Learn the cell obtaining growth on the surface of sensor, construct the Activity determination chip being loaded with cell, and utilize
This chip in real time, the free state copper ion of release and copper in complex state in situ detection apoptosis process.
Described is used for the regular solution system containing free state copper ion and copper in complex state or organism
Free state copper ion in system's (liver extracting solution or cell culture fluid) or the detection of copper in complex state,
It is described fluorescence chemical sensor to be placed in regular solution system to be detected or living things system (liver carries
Take liquid or cell culture fluid) in as the Activity determination substrate of fluoroscopic examination, by fluorescence spectrophotometer or swash
The detection of light Laser Scanning Confocal Microscope is at regular solution system to be detected or living things system (liver extracting solution or thin
Born of the same parents' culture fluid) described in the fluorescent characteristics peak intensity of fluorescence chemical sensor, by comparing to be detected
Regular solution system or living things system (liver extracting solution or cell culture fluid) described in fluorescence chemical
The fluorescent characteristics peak intensity of sensor and this fluorescence chemical sensor to free state copper ion and complex state copper from
Fluorescent characteristics peak intensity in the calibration curve of son detection, detection by quantitative goes out regular solution system to be detected
Or the free state copper ion concentration in living things system (liver extracting solution or cell culture fluid) and complex state copper
Ion concentration.That is, in the system with the presence of free state copper ion or copper in complex state, described base
Fluorescence in the chemical sensor of silicon nanowires can be quenched, by drawing free state copper ion or complex state
The concentration of copper ion and the calibration curve of fluorescent characteristics peak intensity, by as this of Activity determination substrate
The high selectivity of bright fluorescence chemical sensor and sensitivity, detected by described fluorescence chemical sensor
The intensity at fluorescent characteristics peak determine free state copper ion and the concentration of copper in complex state in system, permissible
Detection by quantitative goes out regular solution system to be detected or living things system (liver extracting solution or cell culture fluid)
In free state copper ion concentration or copper in complex state concentration, the concentration range of detection is 0nm～5 μMs.
Result shows that the present invention's has selectivity fluorescence response to free state copper ion and copper in complex state
The fluorescence intensity of fluorescence chemical sensor based on silicon nanowires along with regular solution system or living things system
In free state copper ion and the rising of copper in complex state concentration and reduce.Free state is obtained by drafting
The fluorescence chemical sensor based on silicon nanowires of copper ion and the concentration of copper in complex state and the present invention
The relation of fluorescent characteristics peak intensity, obtains this fluorescence chemical sensor to free state copper ion and complex state copper
The calibration curve of ion detection.
Excitation source used in described fluorescence spectrophotometer is xenon lamp, and excitation wavelength is 350～500nm.
The excitation wavelength of the described laser instrument in laser confocal microscope is 405nm.
Described regular solution system can be phosphate buffer or HEPES buffer etc..
Described living things system can be animal livers extracting solution or cell culture fluid etc..
The free state copper ion of release and complex state copper in described real-time, in situ detection apoptosis process
Ion, be the Activity determination chip upside down that described constructing is loaded with cell in copolymerization Jiao's culture dish, so
Rear addition can make cell digest, the biological reagent of apoptosis or chemical reagent, makes apoptosis, carefully
Born of the same parents can gradually discharge copper ion (including free state and complex state) during apoptosis, thus causes institute
The green fluorescence that the fluorescence chemical sensor stated is launched is quenched, thus can by observe biological reagent or
The green fluorescence that fluorescence chemical sensor described in chemical reagent is launched is quenched, and in real time, examines in situ
Survey free state copper ion and the copper in complex state of release in apoptosis process.
Described biological reagent such as trypsin.
The organic solvents such as described chemical reagent such as ethanol, methanol, acetone or toluene.
Described copper in complex state can be the complex such as (EDTA-Cu) containing copper ion, it is also possible to
It it is the bioactive molecule (such as superoxide dismutase) containing copper ion.
The present invention free state copper ion and copper in complex state are had selectivity fluorescence response based on silicon
The fluorescence chemical sensor of nano wire, no matter in free state copper ion or the detection of copper in complex state
All show good selectivity and high susceptiveness, it is possible to achieve copper ion (free state in system to be measured
And complex state) detection by quantitative, in further postgraduate's objects system copper ion physiological action provide
Good convenience.The selectivity fluorescence that has free state copper ion and copper in complex state of the present invention rings
The fluorescence chemical sensor based on silicon nanowires answered, due to structure includes copper ion is had strong
The group of complexing power, therefore, it is possible to effectively have an effect with copper ion, thus avoids other metal ion
Interference free state copper ion and copper in complex state detected with bioactive molecule.
Accompanying drawing explanation
Fig. 1. the HRTEM photo of the silicon nanowires that in the embodiment of the present invention 1, chemical vapour deposition technique obtains.
Fig. 2. the SEM photograph of the silicon nanowire array that chemical etching method obtains in the embodiment of the present invention 2.
Fig. 3. the silicon nanowires of the embodiment of the present invention 1～4 or the modification schematic diagram of silicon nanowire array.
Fig. 4. the fluorescence chemical sensor based on silicon nanowires of the embodiment of the present invention 3 is in the trip of variable concentrations
Fluorescent characteristics peak curve in amorph copper ion solution.
Fig. 5. in the embodiment of the present invention 3 with the fluorescent characteristics of fluorescence chemical sensor based on silicon nanowires
Peak is characterized peak, makees standard curve with free state copper ion concentration, obtains based on silicon nanowires Fluoresceinated
Learn the fluorescent characteristics peak intensity linearity curve with the change of free state copper ion concentration of sensor.
Fig. 6. the fluorescence chemical sensor based on silicon nanowires of the embodiment of the present invention 1 is at the network of variable concentrations
Close the fluorescent characteristics peak curve in state copper ion (superoxide dismutase SOD) solution.
Fig. 7. the fluorescent characteristics peak of the fluorescence chemical sensor based on silicon nanowires in the embodiment of the present invention 1
It is characterized peak, makees standard curve with copper in complex state concentration (superoxide dismutase SOD), obtain
The fluorescent characteristics peak intensity of fluorescence chemical sensor based on silicon nanowires is (super with copper in complex state concentration
Superoxide dismutase SOD) the linearity curve of change.
Fig. 8. the fluorescent characteristics peak of the fluorescence chemical sensor based on silicon nanowires in the embodiment of the present invention 3
It is characterized peak, makees standard curve with the volume of liver extracting solution, obtain fluorescence chemical based on silicon nanowires
The fluorescent characteristics peak intensity of sensor is with the linearity curve of the change of the volume of liver extracting solution.
The release of copper ion in real-time in-situ monitoring apoptosis process in Fig. 9 a. embodiment of the present invention 4
Fluorescent confocal images；Wherein in figure a), b) and c) be cell after adding trypsin 0 minute, 10
Minute and fluorescence photo after 20 minutes；D), e) and f) being activity chip is adding after trypsin 0 point
Confocal microscope photo behind clock, 10 minutes and 20 minutes.
Fig. 9 b. is blue-fluorescence (cell fluorescence) intensity versus time curve in Fig. 9 a.
Fig. 9 c. is Fig. 9 a Green fluorescence (activity chip fluorescence) intensity versus time curve.
Detailed description of the invention
Embodiment 1
1) preparation of silicon nanowires:
Under room temperature, silicon monoxide is put in porcelain boat after mortar grinder, and porcelain boat is placed on quartz ampoule
Middle part, pushes in vacuum tube furnace；System is first evacuated to 10 with mechanical pump and molecular pump-3Pa, subsequently with 10
Sccm(mL/min) flow velocity is passed through argon (accounting for the volume 95% of mixed gas) and (accounts for hydrogen and mix
The volume 5% of gas) mixed gas, when pressure stability is at 10000Pa, system starts to warm up；System
System rises to 300 DEG C with 25 DEG C/min, then is warming up to 800 DEG C with 25 DEG C/min, now close gas check valve and
Pump lock, is continuously heating to 1350 DEG C after being incubated 10 minutes, after reacting 10 hours at 1350 DEG C, from
So be cooled to room temperature, the both sides of porcelain boat collect product silicon nanowires, a diameter of the 5 of silicon nanowires～15nm,
The HRTEM photo of silicon nanowires is as shown in Figure 1；
2) surface active of silicon nanowires:
The silicon nanowires that step 1) chemical vapour deposition technique prepares is placed in the body that temperature is 75 DEG C
Amass than concentrated sulphuric acid (mass concentration is 98%) and the H being 9:12O2Mixed liquor in heat 180 minutes, cold
But to room temperature, take out silicon nanowires and be washed to neutrality；It is 8:1:1 that silicon nanowires is placed under room temperature volume ratio
H2O:H2O2:NH4In the mixed liquor of OH 3 hours, then take out silicon nanowires and be washed to neutrality,
Vacuum drying, obtains the silicon nanowires that surface is activated；
3) modification (modification schematic diagram is as shown in Figure 3) of silicon nanowires:
Add the step 2 that is dried of 5mg in the reactor) the activated silicon nanowires in the surface that obtains, 2.5
The dry toluene of mL and the 3-2-(2-aminoethylamino of 0.05mL) diethylaminobutyyl trimethoxy
Silane, after being heated to 125 DEG C under argon shield, isothermal reaction 10 hours, it is cooled to room temperature, collects
Silicon nanowires, removes unreacted 3-2-(2-aminoethylamino with methanol ultrasonic cleaning) ethylamino third
Base trimethoxy silane, must arrive surface and be modified with 3-2-(2-aminoethylamino) diethylaminobutyyl front three
The silicon nanowires of TMOS；
Surface obtained above for 5mg is modified with 3-2-(2-aminoethylamino) diethylaminobutyyl three
The silicon nanowires of methoxy silane is placed in the reactor of the dichloromethane being mounted with 5mL, is subsequently added
The triethylamine of 0.05mL, is stirred at room temperature 5 minutes, is added dropwise over two solutions of chlorine 1mL of dansyl Cl afterwards
(concentration of dansyl Cl is 0.05mmol/L), stirring reaction 0.5 hour under room temperature.Pass through 3-2-(2-
Aminoethylamino) amino of surface of silicon nanowires modified of diethylaminobutyyl trimethoxy silane is with red
Acyl chlorides key generation substitution reaction in sulfonic acid chloride molecule, thus surface must be arrived and be modified with 3-2-(2-amino-ethyl
Amino) silicon nanowires of product of diethylaminobutyyl trimethoxy silane and dansyl Cl；Use first
Alcohol ultrasonic cleaning repeatedly removes unreacted dansyl Cl, obtain to free state copper ion and complex state copper from
Son has the fluorescence chemical sensor based on silicon nanowires of selectivity fluorescence response.
By the base that free state copper ion and copper in complex state are had selectivity fluorescence response obtained above
In the fluorescence chemical sensor of silicon nanowires as the substrate of fluoroscopic examination, it is combined fluorescence spectrophotometer, to tool
The solution system having different free state copper ion concentration and copper in complex state concentration carries out fluoroscopic examination, swashs
Luminous source is xenon lamp.Described fluorescence chemical sensor is dispersed in the HEPES buffering that pH value is 7.4
In liquid, in this system, the most gradually add copper in complex state (superoxide dismutase), and to above-mentioned
There is the solution system of variable concentrations copper in complex state and carry out fluoroscopic examination, find along with complex state copper from
The increase of sub-concentration, the intensity at the fluorescent characteristics peak of above-mentioned solution system gradually weakens, as shown in Figure 6,
Wherein vertical coordinate is the fluorescent characteristics peak intensity of sensor, and abscissa is the wavelength launching light.Fluorescent characteristics
The intensity at peak and the concentration of copper in complex state are linear, thus depict the intensity at fluorescent characteristics peak
With the linear scaled curve of copper in complex state concentration, as it is shown in fig. 7, wherein vertical coordinate is described glimmering
The fluorescent characteristics peak intensity of Optochemical sensor, abscissa is the concentration of copper in complex state.According to described
The intensity at fluorescent characteristics peak that detects in scattered solution system of fluorescence chemical sensor determine tested
The concentration of the copper in complex state in survey solution system, thus realize copper in complex state in solution system
The detection of concentration.
Take fresh mouse liver, weigh 6.5 grams, stir evenly in the PBS that 2mL is degerming, in
4000rpm is centrifuged 15 minutes, takes the supernatant, is centrifuged 15 minutes in 4000rpm, collects clear liquid,
Obtain liver extracting solution, freezen protective.
By the base that free state copper ion and copper in complex state are had selectivity fluorescence response obtained above
In the fluorescence chemical sensor of silicon nanowires as the substrate of fluoroscopic examination, it is combined fluorescence spectrophotometer, to dynamic
Copper ion in thing liver extracting solution carries out fluoroscopic examination, and excitation source is xenon lamp.By described Fluoresceinated
Sensor distribution, in the HEPES buffer that pH value is 7.4, the most gradually adds in this system
Animal livers extracting solution, and the above-mentioned solution system with different liver extract concentration is carried out fluorescence inspection
Surveying, find along with the increase of liver extracting solution, the intensity at the fluorescent characteristics peak in above-mentioned solution system is gradually
Weaken.
Embodiment 2
1) preparation of silicon nanowires oldered array:
Take (100) silicon chip of 0.5cm × 3cm, carry out ultrasonic cleaning with ethanol, acetone, distilled water successively
5～35 minutes；Afterwards this silicon chip is placed in the HF aqueous solution that mass concentration is 1% 20 minutes；Take out
It is the AgNO of 3mmol/L that silicon chip is placed on containing concentration3With in the mixed aqueous solution of the HF of 2mol/L
10 minutes；The H containing HF and 0.05mol/L that concentration is 1mol/L is immersed after taking out silicon chip2O2's
In mixed aqueous solution, at the water bath heat preservation 55 minutes of 65 DEG C；The silicon chip of taking-up is put into concentrated hydrochloric acid (quality
Concentration is 36%): in the mixed liquor that volume ratio is 3:1 of concentrated nitric acid (mass concentration is 36%), soak
3 hours, take out silicon chip and naturally dry with after distilled water flushing, obtaining the silicon nanometer being made up of silicon nanowires
Line oldered array, the SEM photograph of silicon nanowires oldered array is as in figure 2 it is shown, wherein: the figure on the left side is
The top view of silicon nanowire array, the figure on the right is the side view of silicon nanowire array；Silicon nanowires is orderly
A diameter of 100 of silicon nanowires in array～250nm, a length of 5～30 μm；
2) surface active of silicon nanowires oldered array:
The silicon nanowires oldered array prepared by step 1) chemical etching method is placed in temperature
The volume ratio of 75～95 DEG C is concentrated sulphuric acid (mass concentration is 98%) and the H of 1:12O2Mixed liquor in heat 30
Minute, it is cooled to room temperature, takes out silicon nanowires oldered array and be washed to neutrality；Then by silicon nanowires
Oldered array be placed in oxygen plasma system and carry out processing (treatment conditions: the mass content of oxygen is 5%,
Voltage is 100 volts, and the time is 8 minutes, and temperature is 15 DEG C), the silicon nanowires obtaining surface activated has
Sequence array；
3) modification (modification schematic diagram is as shown in Figure 3) of silicon nanowire array:
Adding the step 2 that is dried of 55mg in the reactor) the activated silicon nanowires in the surface that obtains is orderly
The 3-2-(2-aminoethylamino of array, the dry toluene of 2.5mL and 0.05mL) ethylamino third
Base trimethoxy silane, after being heated to 75 DEG C under argon shield, isothermal reaction 10 hours, it is cooled to room
Temperature, collects silicon nanowires oldered array, cleans with EtOH Sonicate and removes unreacted 3-2-(2-amino-ethyl
Amino) diethylaminobutyyl trimethoxy silane, surface must be arrived and be modified with 3-2-(2-aminoethylamino)
The silicon nanowires oldered array of diethylaminobutyyl trimethoxy silane.
Surface obtained above is modified with 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy
The silicon nanowires oldered array of silane is placed in the reactor of the dichloromethane being mounted with 5mL, is subsequently added
The triethylamine of 0.05mL, is stirred at room temperature 80 minutes, is added dropwise over two solutions of chlorine 1mL of dansyl Cl afterwards
(concentration of dansyl Cl is 0.05mmol/L), stirring reaction 0.5 hour under room temperature；Pass through 3-2-(2-
Aminoethylamino) the silicon nanowires oldered array surface modified of diethylaminobutyyl trimethoxy silane
Amino and the acyl chlorides key generation substitution reaction in dansyl Cl molecule, thus surface must be arrived and be modified with 3-2-(2-
Aminoethylamino) the silicon nanometer of product of diethylaminobutyyl trimethoxy silane and dansyl Cl
Line oldered array；Remove unreacted dansyl Cl with ethanol ultrasonic cleaning repeatedly, obtain free state copper
Ion and copper in complex state have the fluorescence chemical based on silicon nanowire array of selectivity fluorescence response and pass
Sensor.
By the base that free state copper ion and copper in complex state are had selectivity fluorescence response obtained above
Fluorescence chemical sensor in silicon nanowire array is cut into the square and dense by quality of 0.5 cm x 0.5 centimetre
Degree is the alcohol disinfecting of 75%, takes therein a piece of, is placed in a hole of 24 porocyte culture plates,
And 1 milliliter of cell concentration of addition is 10 in this hole6The A549 cell suspending liquid of individual cells/ml, will
Tissue Culture Plate is placed in (CO in cell culture incubator2: 5%；Temperature: 37 DEG C) carry out cell cultivation, 15
After hour, taking out growth has the active chip of A549 cell, with degerming PBS (pH7.4)
Wash three times.For the ease of the observation of cell, growth have the active chip of A549 cell be transferred to afterwards
In the hole that another one is clean, and in this hole, add 1 milliliter of DMEM cell culture fluid, then to this hole
The DAPI aqueous solution of middle addition 1.5 microlitre 2 mcg/ml, (CO in cell culture incubator2: 5%；Temperature
Degree: 37 DEG C) in hatch 15 minutes after, wash five times with degerming PBS (pH7.4),
Obtain growing the active chip having the A549 cell through DAPI dyeing.
The active chip that growth obtained above has A549 cell is inverted in that to fill 1mL tryptic
In copolymerization Jiao's culture dish, then it is combined laser confocal microscope, to copper ion in A549 apoptosis process
Release process carry out in real time, detection in situ.With the laser excitation of 405nm, detect immediately along with time
Between change, A549 cell and the change of substrate fluorescence, find to increase over time, A549 cell
Gradually apoptosis, the fluorescence intensity of activity chip gradually weakens.
Embodiment 3
1) preparation of silicon nanowires:
Under room temperature, silicon monoxide is put in porcelain boat after mortar grinder, and porcelain boat is placed on quartz ampoule
Middle part, pushes in vacuum tube furnace；System is first evacuated to 10 with mechanical pump and molecular pump-3Pa, subsequently with 40
Sccm(mL/min) flow velocity is passed through argon (accounting for the volume 95% of mixed gas) and (accounts for hydrogen and mix
The volume 5% of gas) mixed gas, when pressure stability is at 750Pa, system starts to warm up；System
Rise to 300 DEG C with 10 DEG C/min, then be warming up to 800 DEG C with 10 DEG C/min, now close gas check valve and pump
Lock, is continuously heating to 1350 DEG C after being incubated 40 minutes, when reacting 3 at 1350 DEG C after, natural cooling
To room temperature, collect product silicon nanowires, a diameter of the 5 of silicon nanowires～15nm in the both sides of porcelain boat；
2) surface of silicon nanowires activation:
The silicon nanowires prepared with chemical vapour deposition technique is placed in the volume ratio that temperature is 90 DEG C is
The concentrated sulphuric acid (mass concentration is 98%) of 9:1 and H2O2Mixed liquor in heat 30 minutes, be cooled to room temperature,
Take out silicon nanowires and be washed to neutrality；It is 2:1:1's that silicon nanowires is placed under room temperature volume ratio
H2O:H2O2:NH4In the mixed liquor of OH 0.5 hour, then take out silicon nanowires and be washed to neutrality, very
Empty dry, obtain the silicon nanowires that surface is activated；
3) modification (modification schematic diagram is as shown in Figure 3) of silicon nanowires:
Add the step 2 that is dried of 100mg in the reactor) the activated silicon nanowires in the surface that obtains,
The dry toluene of 50mL and the 3-2-(2-aminoethylamino of 6mL) diethylaminobutyyl trimethoxy
Silane, after being heated to 75 DEG C under argon shield, isothermal reaction 40 hours, it is cooled to room temperature, collects silicon
Nano wire, removes unreacted 3-2-(2-aminoethylamino with acetone ultrasonic cleaning) diethylaminobutyyl
Trimethoxy silane, must arrive surface and be modified with 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy
The silicon nanowires of base silane；
Surface obtained above for 100mg is modified with 3-2-(2-aminoethylamino) diethylaminobutyyl
The silicon nanowires of trimethoxy silane is placed in the reactor of the dichloromethane being mounted with 15mL, is subsequently added
The triethylamine of 1mL, is stirred at room temperature 80 minutes, is added dropwise over two solutions of chlorine 10mL of dansyl Cl afterwards
(concentration of dansyl Cl is 8mmol/L), stirring reaction 6 hours under room temperature.By 3-2-(2-amino
Ethylamino) amino of surface of silicon nanowires modified of diethylaminobutyyl trimethoxy silane and red sulphonyl
Acyl chlorides key generation substitution reaction in chlorine molecule, thus surface must be arrived and be modified with 3-2-(2-aminoethylamino)
The silicon nanowires of the product of diethylaminobutyyl trimethoxy silane and dansyl Cl；With acetone repeatedly
Ultrasonic cleaning removes unreacted dansyl Cl, obtains having free state copper ion and copper in complex state
The fluorescence chemical sensor based on silicon nanowires of selectivity fluorescence response.
By the base that free state copper ion and copper in complex state are had selectivity fluorescence response obtained above
In the fluorescence chemical sensor of silicon nanowires as the substrate of fluoroscopic examination, it is combined fluorescence spectrophotometer, to tool
The solution system having different free state copper ion concentration and copper in complex state concentration carries out fluoroscopic examination, swashs
Luminous source is xenon lamp.Described fluorescence chemical sensor is dispersed in the HEPES buffering that pH value is 7.4
In liquid, in this system, the most gradually add free state copper ion, and to above-mentioned, there is different free state copper
The solution system of ion concentration carries out fluoroscopic examination, finds along with the increase of free state copper ion concentration, on
The intensity at the fluorescent characteristics peak stating solution system gradually weakens, and as shown in Figure 4, wherein vertical coordinate is sensing
The fluorescent characteristics peak intensity of device, abscissa is the wavelength launching light.The intensity at fluorescent characteristics peak and free state
The concentration of copper ion is linear, thus depicts the intensity at fluorescent characteristics peak and free state copper ion is dense
The linear scaled curve of degree, as it is shown in figure 5, wherein vertical coordinate is the glimmering of described fluorescence chemical sensor
Light characteristic peak intensity, abscissa is the concentration of free state copper ion.According to described fluorescence chemical sensor
The intensity at the fluorescent characteristics peak detected in scattered solution system determines the trip in detected solution system
The concentration of amorph copper ion, thus realize the detection of free state copper ion concentration in solution system.
Take fresh mouse liver, weigh 10 grams, stir evenly in the PBS that 2mL is degerming, in
4000rpm is centrifuged 15 minutes, takes the supernatant, is centrifuged 15 minutes in 4000rpm, collects clear liquid,
Obtain liver extracting solution, freezen protective.
By the base that free state copper ion and copper in complex state are had selectivity fluorescence response obtained above
In the fluorescence chemical sensor of silicon nanowires as the substrate of fluoroscopic examination, it is combined fluorescence spectrophotometer, to dynamic
Copper ion in thing liver extracting solution carries out fluoroscopic examination, and excitation source is xenon lamp.By described Fluoresceinated
Sensor distribution, in the HEPES buffer that pH value is 7.4, the most gradually adds in this system
Animal livers extracting solution, and the above-mentioned solution system with different liver extract concentration is carried out fluorescence inspection
Surveying, find along with the increase of liver extracting solution, the intensity at the fluorescent characteristics peak of above-mentioned solution system gradually subtracts
Weak, as shown in Figure 8, wherein vertical coordinate is the fluorescent characteristics peak intensity of described fluorescence chemical sensor,
Abscissa is the wavelength launching light.
Embodiment 4
1) preparation of silicon nanowires oldered array:
Take (100) silicon chip of 1cm × 4cm, carry out ultrasonic cleaning with ethanol, acetone, distilled water successively
35 minutes；Afterwards this silicon chip is placed in the HF aqueous solution that mass concentration is 6% 25 seconds；Take out silicon chip
Being placed on containing concentration is the AgNO of 10mmol/L3With in the mixed aqueous solution of the HF of 8mol/L 1
Minute；The H containing HF and 0.8mol/L that concentration is 10mol/L is immersed after taking out silicon chip2O2Mixed
In Heshui solution, at the water bath heat preservation 55 minutes of 25 DEG C；The silicon chip of taking-up is put into concentrated hydrochloric acid, and (quality is dense
Degree is 36%): in the mixed liquor that volume ratio is 3:1 of concentrated nitric acid (mass concentration is 36%), soak
0.5 hour, taking out silicon chip and naturally dry with after distilled water flushing, the silicon obtaining being made up of silicon nanowires is received
Rice noodle oldered array；A diameter of 100 of silicon nanowires in silicon nanowires oldered array～250nm, length
It is 5～30 μm；
2) surface active of silicon nanowires oldered array:
It is 95 DEG C that the silicon nanowires oldered array prepared by step 1) chemical etching method is placed in temperature
The concentrated sulphuric acid that volume ratio is 9:1 (mass concentration is 98%) and H2O2Mixed liquor in heat 180 minutes,
It is cooled to room temperature, takes out silicon nanowires oldered array and be washed to neutrality.Then by silicon nanowires oldered array
It is placed in oxygen plasma system and carries out processing (treatment conditions: the mass content of oxygen is 100%, voltage
Being 700 volts, the time is 5 seconds, and temperature is 45 DEG C), obtain the silicon nanowires oldered array that surface is activated；
3) modification (modification schematic diagram is as shown in Figure 3) of silicon nanowires oldered array:
Adding the step 2 that is dried of 45mg in the reactor) the activated silicon nanowires in the surface that obtains is orderly
The 3-2-(2-aminoethylamino of array, the dry toluene of 50mL and 0.6mL) diethylaminobutyyl
Trimethoxy silane, after being heated to 125 DEG C under inert gas shielding, isothermal reaction 40 hours, cooling
To room temperature, collect silicon nanowires oldered array, clean with EtOH Sonicate and remove unreacted 3-2-(2-amino
Ethylamino) diethylaminobutyyl trimethoxy silane, must arrive surface and be modified with 3-2-(2-amino-ethyl ammonia
Base) the silicon nanowires oldered array of diethylaminobutyyl trimethoxy silane.
Surface obtained above is modified with 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy
The silicon nanowires oldered array of silane is placed in the reactor of the dichloromethane being mounted with 15mL, is subsequently added
The triethylamine of 1mL, is stirred at room temperature 5 minutes, is added dropwise over two solutions of chlorine 10mL of dansyl Cl afterwards
(concentration of dansyl Cl is 0.8mmol/L), stirring reaction 5 hours under room temperature.By 3-2-(2-ammonia
Base ethylamino) ammonia on silicon nanowires oldered array surface modified of diethylaminobutyyl trimethoxy silane
Base and the acyl chlorides key generation substitution reaction in dansyl Cl molecule, thus surface must be arrived and be modified with 3-2-(2-
Aminoethylamino) the silicon nanometer of product of diethylaminobutyyl trimethoxy silane and dansyl Cl
Line oldered array；Remove unreacted dansyl Cl with ethanol ultrasonic cleaning repeatedly, obtain free state copper
Ion and copper in complex state have the fluorescence chemical based on silicon nanowire array of selectivity fluorescence response and pass
Sensor.
By the base that free state copper ion and copper in complex state are had selectivity fluorescence response obtained above
Fluorescence chemical sensor in silicon nanowire array is cut into the square and dense by quality of 0.5 cm x 0.5 centimetre
Degree is the alcohol disinfecting of 75%, takes therein a piece of, is placed in a hole of 24 porocyte culture plates,
And 1 milliliter of cell concentration of addition is 10 in this hole6The A549 cell suspending liquid of individual cells/ml, will
Tissue Culture Plate is placed in (CO in cell culture incubator2: 5%；Temperature: 37 DEG C) carry out cell cultivation, 15
After hour, taking out growth has the active chip of A549 cell, with degerming PBS (pH7.4)
Wash three times.For the ease of the observation of cell, growth have the active chip of A549 cell be transferred to afterwards
In the hole that another one is clean, and in this hole, add 1 milliliter of DMEM cell culture fluid, then to this hole
The DAPI aqueous solution of middle addition 1.5 microlitre 2 mcg/ml, (CO in cell culture incubator2: 5%；Temperature
Degree: 37 DEG C) in hatch 15 minutes after, wash five times with degerming PBS (pH7.4),
Obtain growing the active chip having the A549 cell through DAPI dyeing.
The active chip that growth obtained above has A549 cell is inverted in that to fill 1mL tryptic
In copolymerization Jiao's culture dish, then it is combined laser confocal microscope, to copper ion in A549 apoptosis process
Release process carry out in real time, detection in situ.With the laser excitation of 405nm, detect immediately along with time
Between change, A549 cell and the change of substrate fluorescence.Discovery increases over time, A549 cell
Gradually apoptosis, the fluorescence intensity of activity chip gradually weakens, as shown in Fig. 9 a, Fig. 9 b and Fig. 9 c, its
In: Fig. 9 a is the fluorescence photo that growth has the active chip of A549 cell, and wherein a-c is A549 cell
The fluorescence photo of apoptotic process, d-e is that the fluorescence that the fluorescence intensity of described fluorescence chemical sensor weakens shines
Sheet；Fig. 9 b is in A549 apoptosis process, and the fluorescence intensity of A549 cell is the most bent
Line；Fig. 9 c is during A549 apoptosis, the fluorescence intensity of described fluorescence chemical sensor
Versus time curve.
Embodiment 5
1) preparation of silicon nanowires:
Under room temperature, silicon monoxide is put in porcelain boat after mortar grinder, and porcelain boat is placed on quartz ampoule
Middle part, pushes in vacuum tube furnace；System is first evacuated to 10 with mechanical pump and molecular pump-3Pa, subsequently with 20
Sccm(mL/min) flow velocity is passed through argon (accounting for the volume 95% of mixed gas) and (accounts for hydrogen and mix
The volume 5% of gas) mixed gas, when pressure stability is at 8000Pa, system starts to warm up；System
System rises to 300 DEG C with 20 DEG C/min, then is warming up to 800 DEG C with 20 DEG C/min, now close gas check valve and
Pump lock, is continuously heating to 1350 DEG C after being incubated 30 minutes, after reacting 6 hours at 1350 DEG C, natural
It is cooled to room temperature, collects product silicon nanowires, a diameter of the 5 of silicon nanowires～15nm in the both sides of porcelain boat；
2) surface active of silicon nanowires:
The silicon nanowires that step 1) chemical vapour deposition technique prepares is placed in the body that temperature is 80 DEG C
Amass than concentrated sulphuric acid (mass concentration is 98%) and the H being 5:12O2Mixed liquor in heat 100 minutes, cold
But to room temperature, take out silicon nanowires and be washed to neutrality；It is 5:1:1 that silicon nanowires is placed under room temperature volume ratio
H2O:H2O2:NH4In the mixed liquor of OH 2 hours, then take out silicon nanowires and be washed to neutrality,
Vacuum drying, obtains the silicon nanowires that surface is activated；
3) modification of silicon nanowires:
Add the step 2 that is dried of 50mg in the reactor) the activated silicon nanowires in the surface that obtains, 8mL
Dry toluene and the 3-2-(2-aminoethylamino of 0.5mL) diethylaminobutyyl trimethoxy silane,
After being heated to 100 DEG C under inert gas shielding, isothermal reaction 15 hours, it is cooled to room temperature, collects silicon
Nano wire, cleans with EtOH Sonicate and removes unreacted 3-2-(2-aminoethylamino) diethylaminobutyyl
Trimethoxy silane, must arrive surface and be modified with 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy
The silicon nanowires of base silane；
Surface obtained above for 65mg is modified with 3-2-(2-aminoethylamino) diethylaminobutyyl three
The silicon nanowires of methoxy silane is placed in the reactor of the dichloromethane being mounted with 10mL, is subsequently added
The triethylamine of 0.2mL, is stirred at room temperature 15 minutes, is added dropwise over two solutions of chlorine 5mL of dansyl Cl afterwards
(concentration of dansyl Cl is 0.4mmol/L), stirring reaction 2.5 hours under room temperature.Pass through 3-2-(2-
Aminoethylamino) amino of surface of silicon nanowires modified of diethylaminobutyyl trimethoxy silane is with red
Acyl chlorides key generation substitution reaction in sulfonic acid chloride molecule, thus surface must be arrived and be modified with 3-2-(2-amino-ethyl
Amino) silicon nanowires of product of diethylaminobutyyl trimethoxy silane and dansyl Cl；Use second
Alcohol ultrasonic cleaning repeatedly removes unreacted dansyl Cl, obtain to free state copper ion and complex state copper from
Son has the fluorescence chemical sensor based on silicon nanowires of selectivity fluorescence response.
By the base that free state copper ion and copper in complex state are had selectivity fluorescence response obtained above
In the fluorescence chemical sensor of silicon nanowires as the substrate of fluoroscopic examination, it is combined fluorescence spectrophotometer, to tool
The solution system having different free state copper ion concentration and copper in complex state concentration carries out fluoroscopic examination, swashs
Luminous source is xenon lamp.Described fluorescence chemical sensor is dispersed in the HEPES buffering that pH value is 7.4
In liquid, in this system, the most gradually add copper in complex state (superoxide dismutase), and to above-mentioned
There is the solution system of variable concentrations copper in complex state and carry out fluoroscopic examination, find along with complex state copper from
The increase of sub-concentration, the intensity at the fluorescent characteristics peak of above-mentioned solution system gradually weakens.Fluorescent characteristics peak
The concentration of intensity and copper in complex state is linear, thus depicts intensity and the network at fluorescent characteristics peak
Close the linear scaled curve of state copper ion concentration.According to described fluorescence chemical sensor in solution system
The intensity at the fluorescent characteristics peak detected determines the concentration of the copper in complex state in detected solution system,
Thus realize the detection of complex state copper ion concentration in solution system.
Take fresh mouse liver, weigh 7.5 grams, stir evenly in the PBS that 2.5mL is degerming,
It is centrifuged 15 minutes in 4000rpm, takes the supernatant, be centrifuged 15 minutes in 4000rpm, collect clear liquid,
Obtain liver extracting solution, freezen protective.
By the base that free state copper ion and copper in complex state are had selectivity fluorescence response obtained above
In the fluorescence chemical sensor of silicon nanowires as the substrate of fluoroscopic examination, it is combined fluorescence spectrophotometer, to dynamic
Copper ion in thing liver extracting solution carries out fluoroscopic examination, and excitation source is xenon lamp.By described Fluoresceinated
Sensor distribution, in the HEPES buffer that pH value is 7.4, the most gradually adds in this system
Animal livers extracting solution, and the above-mentioned solution system with different liver extract concentration is carried out fluorescence inspection
Surveying, find along with the increase of liver extracting solution, the intensity at the fluorescent characteristics peak of above-mentioned solution system gradually subtracts
Weak.
Embodiment 6
1) preparation of silicon nanowires oldered array:
Take (100) silicon chip of 2cm × 3cm, carry out ultrasonic cleaning with ethanol, acetone, distilled water successively
25 minutes；Afterwards this silicon chip is placed in the HF aqueous solution that mass concentration is 4% 10 minutes；Take out silicon
It is the AgNO of 5mmol/L that sheet is placed on containing concentration3With in the mixed aqueous solution of the HF of 4mol/L 6
Minute；The H containing HF and 0.1mol/L that concentration is 6mol/L is immersed after taking out silicon chip2O2Mixing
In aqueous solution, at the water bath heat preservation 35 minutes of 45 DEG C；The silicon chip of taking-up is put into concentrated hydrochloric acid (mass concentration
It is 36%): in the mixed liquor that volume ratio is 3:1 of concentrated nitric acid (mass concentration is 36%), soak 1.5
Hour, take out silicon chip and naturally dry with after distilled water flushing, obtaining the silicon nanometer being made up of silicon nanowires
Line oldered array；A diameter of 100 of silicon nanowires in silicon nanowires oldered array～250nm, a length of
5～30 μm；
2) surface active of silicon nanowire array:
It is 65 DEG C that the silicon nanowires oldered array prepared by step 1) chemical etching method is placed in temperature
The concentrated sulphuric acid that volume ratio is 4:1 (mass concentration is 98%) and H2O2Mixed liquor in heat 90 minutes,
It is cooled to room temperature, takes out silicon nanowires oldered array and be washed to neutrality；Then by orderly for silicon nanowires battle array
Row are placed in oxygen plasma system and carry out processing (treatment conditions: the mass content of oxygen is 50%, electricity
Pressure is 400 volts, and the time is 4 minutes, and temperature is 25 DEG C), the silicon nanowires obtaining surface activated is orderly
Array；
3) modification of silicon nanowires oldered array:
Adding the step 2 that is dried of 35mg in the reactor) the activated silicon nanowires in the surface that obtains is orderly
The 3-2-(2-aminoethylamino of array, the dry toluene of 20mL and 0.3mL) diethylaminobutyyl
Trimethoxy silane, after being heated to 110 DEG C under argon shield, isothermal reaction 30 hours, it is cooled to room
Temperature, collects silicon nanowires oldered array, cleans with EtOH Sonicate and removes unreacted 3-2-(2-amino-ethyl
Amino) diethylaminobutyyl trimethoxy silane, surface must be arrived and be modified with 3-2-(2-aminoethylamino)
The silicon nanowires oldered array of diethylaminobutyyl trimethoxy silane；
Surface obtained above is modified with 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy
The silicon nanowires of silane or silicon nanowires oldered array are placed in the reactor of the dichloromethane being mounted with 12mL
In, it is subsequently added the triethylamine of 0.6mL, is stirred at room temperature 45 minutes, is added dropwise over dansyl Cl afterwards
The concentration of two solutions of chlorine 4mL(dansyl Cls is 0.6mmol/L), stirring reaction 4 hours under room temperature.Logical
Cross 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy silane modify the orderly battle array of silicon nanowires
The amino in list face and the acyl chlorides key generation substitution reaction in dansyl Cl molecule, thus obtain surface and modify
Have 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy silane and the product of dansyl Cl
Silicon nanowires oldered array；Removing unreacted dansyl Cl with ethanol ultrasonic cleaning repeatedly, it is right to obtain
Free state copper ion and copper in complex state have the based on silicon nanowire array glimmering of selectivity fluorescence response
Optochemical sensor.
By the base that free state copper ion and copper in complex state are had selectivity fluorescence response obtained above
Fluorescence chemical sensor in silicon nanowire array is cut into the square and dense by quality of 0.5 cm x 0.5 centimetre
Degree is the alcohol disinfecting of 75%, takes therein a piece of, is placed in a hole of 24 porocyte culture plates,
And 1 milliliter of cell concentration of addition is 10 in this hole6The A549 cell suspending liquid of individual cells/ml, will
Tissue Culture Plate is placed in (CO in cell culture incubator2: 5%；Temperature: 37 DEG C) carry out cell cultivation, 15
After hour, taking out growth has the active chip of A549 cell, with degerming PBS (pH7.4)
Wash three times.For the ease of the observation of cell, growth have the active chip of A549 cell be transferred to afterwards
In the hole that another one is clean, and in this hole, add 1 milliliter of DMEM cell culture fluid, then to this hole
The DAPI aqueous solution of middle addition 1.5 microlitre 2 mcg/ml, (CO in cell culture incubator2: 5%；Temperature
Degree: 37 DEG C) in hatch 15 minutes after, wash five times with degerming PBS (pH7.4),
Obtain growing the active chip having the A549 cell through DAPI dyeing.
The active chip that growth obtained above has A549 cell is inverted in that to fill 1mL tryptic
In copolymerization Jiao's culture dish, then it is combined laser confocal microscope, to copper ion in A549 apoptosis process
Release process carry out in real time, detection in situ.With the laser excitation of 405nm, detect immediately along with time
Between change, A549 cell and the change of substrate fluorescence.Discovery increases over time, A549 cell
Gradually apoptosis, the fluorescence intensity of activity chip gradually weakens.
Claims (9)
1. one kind has receiving based on silicon of selectivity fluorescence response to free state copper ion and copper in complex state
The fluorescence chemical sensor of rice noodle, is characterized in that: described fluorescence chemical sensor is by surface active
Silicon nanowires or silicon nanowires oldered array and 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy
Base silane reacts, and the surface obtained is modified with 3-2-(2-aminoethylamino) diethylaminobutyyl three
Silicon nanowires or the silicon nanowires oldered array of methoxy silane react with dansyl Cl, make 3-2-(2-amino
Ethylamino) product of diethylaminobutyyl trimethoxy silane and dansyl Cl modifies in silicon nanometer
Obtain behind the surface of line or silicon nanowire array；
Described has receiving based on silicon of selectivity fluorescence response to free state copper ion and copper in complex state
The fluorescence chemical sensor of rice noodle prepares by comprising the following steps:
1) silicon nanowires is placed in the concentrated sulphuric acid that volume ratio is 1:2～9:1 and the H that temperature is 75～90 DEG C2O2
Mixed liquor in heat 30～180 minutes, be cooled to room temperature, take out silicon nanowires and be also washed to neutrality；Room
Under temperature, silicon nanowires is placed in the H that volume ratio is 2:1:1～8:1:12O:H2O2:NH4In the mixed liquor of OH,
Then take out silicon nanowires and be washed to neutrality, vacuum drying, obtain the silicon nanowires that surface is activated；
Or silicon nanowires oldered array is placed in the concentrated sulphuric acid that volume ratio is 1:1～9:1 that temperature is 75～95 DEG C
With H2O2Mixed liquor in heat 30～180 minutes, be cooled to room temperature, take out silicon nanowires oldered array
And it is washed to neutrality；Then silicon nanowires oldered array is placed in oxygen plasma system and processes,
Obtain the silicon nanowires oldered array that surface is activated；
2) the 5～100mg steps 1 that are dried are added in the reactor) the activated silicon nanometer in the surface that obtains
Line or the dry toluene of silicon nanowires oldered array, 2.5～50mL and 3-2-(the 2-ammonia of 0.05～0.6mL
Base ethylamino) diethylaminobutyyl trimethoxy silane, under inert gas shielding, it is heated to 75～125 DEG C
After, isothermal reaction 10～40 hours, it is cooled to room temperature, collects silicon nanowires or silicon nanowires oldered array,
Unreacted 3-2-(2-aminoethylamino) diethylaminobutyyl front three is removed with organic solvent ultrasonic cleaning
TMOS, must arrive surface and be modified with 3-2-(2-aminoethylamino) diethylaminobutyyl trimethoxy silicon
The silicon nanowires of alkane or silicon nanowires oldered array；
3) by 5～100mg steps 2) surface that obtains is modified with 3-2-(2-aminoethylamino) ethyl
The silicon nanowires of TSL 8330 or silicon nanowires oldered array are placed in and are mounted with 5～15mL
Dichloromethane reactor in, be subsequently added the triethylamine of 0.05～1.0mL, be stirred at room temperature, afterwards by
Being added dropwise to two solutions of chlorine 1～10mL of dansyl Cl, under room temperature, stirring reaction, must arrive surface and be modified with 3-2-
The silicon of the product of (2-aminoethylamino) diethylaminobutyyl trimethoxy silane and dansyl Cl is received
Rice noodle or silicon nanowire array；Then unreacted dansyl Cl is removed with organic solvent ultrasonic cleaning,
To free state copper ion and copper in complex state being had the based on silicon nanowires glimmering of selectivity fluorescence response
Optochemical sensor.
It is the most according to claim 1 that free state copper ion and copper in complex state are had selectivity is glimmering
The fluorescence chemical sensor based on silicon nanowires of photoresponse, is characterized in that: the silicon of described surface active
Silicon nanowires in nano wire is a diameter of 5～the silicon nanowires of 15nm obtained by chemical vapour deposition technique；
Silicon nanowires oldered array in the silicon nanowires oldered array of described surface active is to be carved by chemistry
The silicon nanowires oldered array that erosion method obtains, the silicon nanowires in silicon nanowires oldered array a diameter of
100～250nm, a length of 5～30 μm.
It is the most according to claim 1 that free state copper ion and copper in complex state are had selectivity is glimmering
The fluorescence chemical sensor based on silicon nanowires of photoresponse, is characterized in that: described puts silicon nanowires
In the H that volume ratio is 2:1:1～8:1:12O:H2O2:NH4Time in the mixed liquor of OH is 0.5～3 hour；
Described being placed in oxygen plasma system by silicon nanowires oldered array processes, treatment conditions:
The mass content of oxygen is 5～100%, and voltage is 100～700 volts, and the time is 5 seconds～8 minutes, temperature
It it is 15～45 DEG C.
It is the most according to claim 1 that free state copper ion and copper in complex state are had selectivity is glimmering
The fluorescence chemical sensor based on silicon nanowires of photoresponse, is characterized in that: the two of described dansyl Cl
In solutions of chlorine, the concentration of dansyl Cl is 0.05～8mmol/L.
It is the most according to claim 1 that free state copper ion and copper in complex state are had selectivity is glimmering
The fluorescence chemical sensor based on silicon nanowires of photoresponse, is characterized in that: described organic solvent is first
Alcohol, ethanol or acetone.
6. free state copper ion and copper in complex state are had described in a Claims 1 to 5 any one
The application of the fluorescence chemical sensor based on silicon nanowires of selective fluorescence response, is characterized in that: institute
The fluorescence chemical sensor stated is for the regular solution system containing free state copper ion and copper in complex state
Or the free state copper ion in living things system and the detection of copper in complex state；
Or the substrate that described fluorescence chemical sensor grown as cell, pass at described fluorescence chemical
Obtain the cell of growth on the surface of sensor, construct the Activity determination chip being loaded with cell, and utilize this core
Sheet in real time, the free state copper ion of release and copper in complex state in situ detection apoptosis process；
Described regular solution system is phosphate buffer or HEPES buffer；
Described living things system is animal livers extracting solution or cell culture fluid.
Application the most according to claim 6, is characterized in that: described free state based on silicon nanowires
Copper ion and the application of copper in complex state fluorescence chemical sensor, is characterized in that: described be used for containing
Free state copper ion in the regular solution system of free state copper ion and copper in complex state or living things system
Or the detection of copper in complex state, it is that described fluorescence chemical sensor is placed in regular solution to be detected
As the Activity determination substrate of fluoroscopic examination in system or living things system, it is total to fluorescence spectrophotometer or laser
Focusing microscope detection fluorescence chemical sensing described in regular solution system to be detected or living things system
The fluorescent characteristics peak intensity of device, by comparing described in regular solution system to be detected or living things system
The fluorescent characteristics peak intensity of fluorescence chemical sensor and this fluorescence chemical sensor to free state copper ion and
Fluorescent characteristics peak intensity in the calibration curve of copper in complex state detection, detection by quantitative goes out to be detected general
Free state copper ion concentration in logical solution system or living things system and copper in complex state concentration；
The free state copper ion of release and complex state copper in described real-time, in situ detection apoptosis process
Ion, be the Activity determination chip upside down that described constructing is loaded with cell in copolymerization Jiao's culture dish, so
Rear addition can make cell digest, the biological reagent of apoptosis or chemical reagent, makes apoptosis, logical
Cross that to observe the green fluorescence that the fluorescence chemical sensor described in biological reagent or chemical reagent launched sudden
Go out, in real time, the free state copper ion of release and copper in complex state in situ detection apoptosis process.
Application the most according to claim 7, is characterized in that: described biological reagent is trypsin；
Described chemical reagent is ethanol, methanol, acetone or toluene.
9. according to the application described in claim 6 or 7, it is characterized in that: described copper in complex state is to contain
There are the complex of copper ion or the bioactive molecule containing copper ion.
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| CN108913120B (en) * | 2018-06-21 | 2021-04-02 | 中国科学院理化技术研究所 | Single silicon nanowire fluorescence chemical sensor for detecting hypochlorite, and preparation method and application thereof |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102030309A (en) * | 2010-11-10 | 2011-04-27 | 中国科学院理化技术研究所 | Mn27Si47-Si heterostructure nanowire arrays or Mn27Si47Method for preparing nanowire array |
| CN102419319A (en) * | 2011-09-08 | 2012-04-18 | 中国科学院理化技术研究所 | Silicon nanowire-based sensor with selective fluorescent response to nitric oxide |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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| CN102419319A (en) * | 2011-09-08 | 2012-04-18 | 中国科学院理化技术研究所 | Silicon nanowire-based sensor with selective fluorescent response to nitric oxide |
Non-Patent Citations (3)
| Title |
|---|
| Fluorescence quenching amplification in silica nanosensors for metal ions;Marco Montalti et al.;《Journal of Materials Chemistry》;20050503;第15卷;2810-2814 * |
| Silicon Nanowires-Based Fluorescence Sensor for Cu(II);Lixuan Mu et al.;《Nano Letters》;20081231;第8卷(第1期);104-109 * |
| 基于硅纳米线的光响应化学传感器研究;穆丽璇;《中国博士学位论文全文数据库 工程科技I辑》;20081015(第10期);3，6，29-32，62-70 * |
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