CN102419319A - Silicon nano wire based sensor having selective fluorescence response on nitric oxide - Google Patents
Silicon nano wire based sensor having selective fluorescence response on nitric oxide Download PDFInfo
- Publication number
- CN102419319A CN102419319A CN2011102661626A CN201110266162A CN102419319A CN 102419319 A CN102419319 A CN 102419319A CN 2011102661626 A CN2011102661626 A CN 2011102661626A CN 201110266162 A CN201110266162 A CN 201110266162A CN 102419319 A CN102419319 A CN 102419319A
- Authority
- CN
- China
- Prior art keywords
- silicon
- silicon nanowires
- nanowire array
- amino
- nanowires
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention belongs to the field of fluorescence chemical sensor with nanostructure and especially relates to a silicon nano wire based chemical sensor having selective fluorescence response on nitric oxide, and a preparation method and application thereof. According to the invention, silicon nano wire and silicon nano wire array prepared from a chemical vapor deposition method and from a chemical etching method are treated with a surface treatment; then an organic micromolecule substance amino fluorescein is used to carry out covalent modification on the silicon nano wire and the silicon nano wire array to obtain the silicon nano wire based sensor having selective fluorescence response on nitric oxide. The silicon nano wire based chemical sensor having selective fluorescence response on nitric oxide can be used in detection of solution and biosystem.
Description
Technical field
The invention belongs to the fluorescence chemical sensor field of nanostructured, particularly to nitrogen monoxide have the selectivity fluorescence response based on chemical sensor of silicon nanowires and preparation method thereof, and the application of this chemical sensor.
Background technology
Finding that nitrogen monoxide is discharged by endothelium in vivo in 1987 produces, and in the immune response of human body, it is the cardiovascular system important information transmission person in the nervous system that unifies.Since then, nitric oxide production physiological action has caused many biologists and chemist's concern.
At present, it is a lot of to be applied to nitric oxide production detection means, mainly contains electrochemical process and electron paramagnetic resonance spectrum (EPR) method etc.But above method is owing to low spatial resolution, even the comparatively complicated instrument of needs, and this makes its application in cell imaging obtain restriction.Yet fluorescent technique is because high sensitivity of self and high spatial resolution make it in cell and in the imaging of extracellular nitrogen monoxide, demonstrate great advantage.
Reported the multiple reagent that can be used for the nitrogen monoxide fluorescence imaging in the document, like Yang Y J, J.Am.Chem.Soc.2010,132,13114; Lippard S J, Acc.Chem.Res.2007,40,41; Lim M H, Nature Chem.Biol.2006,2,375; Duarte AJ, Sensors, 2010,10,1661.Yet these methods in use all have some limitation, and the most widely used reagent is exactly the diamido fluorescein derivative, and it not only reacts with nitrogen monoxide, and other molecule that discharges in the cell also can be participated in reaction.And the imaging agents of most of report all need use in organic solvent, and this might cause cell generation pathology.Other limitation that these imaging agents exist also comprises and is easy to photobleaching, is difficult to synthetic and in use needs hydrolysis etc.
Therefore the synthetic suitable fluorescent reagent that can be applied to nitrogen monoxide detection in the living things system has very important significance.
Summary of the invention
One of the object of the invention provides the chemical sensor based on silicon nanowires that nitrogen monoxide is had the selectivity fluorescence response.
Two of the object of the invention provides the preparation method based on the chemical sensor of silicon nanowires who nitrogen monoxide is had the selectivity fluorescence response.
Three of the object of the invention provide to nitrogen monoxide have the selectivity fluorescence response based on the chemical sensor of silicon nanowires in the application aspect the biological detection.
The chemical sensor based on silicon nanowires that nitrogen monoxide is had a selectivity fluorescence response of the present invention is by the finishing that 3-aminopropyltriethoxywerene werene and the silicon nanowires that has carried out surface clean or silicon nanowire array reaction obtain amino silicon nanowires or silicon nanowire array to be arranged; Obtain silicon nanowires or the silicon nanowire array (amino on silicon nanowires or the silicon nanowire array and glutaraldehyde reaction) that finishing has aldehyde radical with the glutaraldehyde reaction; Be modified at the amino luciferin that is obtained by the aldehyde radical on silicon nanowires or the silicon nanowire array and amino luciferin reaction on silicon nanowires or the silicon nanowire array through reduction (with sodium borohydride as reductive agent), obtaining finishing has the chemical sensor as the silicon nanowires of going back ortho states amino luciferin that nitrogen monoxide is had the selectivity fluorescence response.
Described silicon nanowires is the silicon nanowires by chemical vapour deposition technique or the resulting different size of chemical etching method.
The preparation method based on the chemical sensor of silicon nanowires that nitrogen monoxide is had a selectivity fluorescence response of the present invention may further comprise the steps:
1) will use silicon nanowires that chemical vapour deposition technique prepares is that 70～95 ℃ volume ratio is 1: 1～8: 1 the concentrated sulphuric acid (mass concentration is 50%～98%) and H in temperature
2O
2Heated 30～90 minutes in the mixed liquor of (mass concentration is 5%～30%), be cooled to room temperature afterwards, take out also repeated water washing is immersed in H to neutral under room temperature
2O: H
2O
2(mass concentration is 5%～30%): NH
4The volume ratio of OH is in 3: 1: 1～9: 1: 1 the mixed liquor and places (be 1～2.5 hour general standing time), take out and repeated water washing to neutral, vacuum drying; Or
Is that 70～95 ℃ volume ratio is 1: 1～8: 1 the concentrated sulphuric acid (mass concentration is 50%～98%) and H with the silicon nanowire array that uses chemical etching method to prepare in temperature
2O
2Heated 30～90 minutes in the mixed liquor of (mass concentration is 5%～30%); Be cooled to room temperature afterwards; Take out and repeated water washing to neutral, (treatment conditions: the mass content of oxygen is 10～100%, and voltage is 100～500 volts to place the oxygen plasma system to carry out the surface treatment of silicon nanowire array then; Time is 10 seconds～5 minutes, and temperature is 10～35 ℃);
2) in reactor, add silicon nanowires or silicon nanowire array, 5～30mL dry toluene and 0.05～0.3mL 3-aminopropyltriethoxywerene werene (0.21～1.26mmol) of the drying that 10～60mg step 1) obtains; After being heated to 80～120 ℃ under inert gas (like the nitrogen) protection; Isothermal reaction 12～36 hours; Be cooled to room temperature; Filter to collect silicon nanowires or silicon nanowire array with microfilter, with organic solvent repeatedly ultrasonic cleaning remove unreacted 3-aminopropyltriethoxywerene werene, filter and collect finishing amino silicon nanowires or silicon nanowire array are arranged;
3) with step 2) finishing that obtains has amino silicon nanowires or silicon nanowire array to place reactor; Add mass concentration and be 10%～50% glutaraldehyde water solution and (can be 1.5～10mL); Stirring at room reaction (the general reaction time is 30～120 minutes); Then with organic solvent repeatedly ultrasonic cleaning remove unreacted glutaraldehyde, filter to collect silicon nanowires or silicon nanowire array that finishing has aldehyde radical;
4) finishing that step 3) is obtained has the silicon nanowires of aldehyde radical or silicon nanowire array to place reactor; Add concentration and be that ethanolic solution or the acetone soln (general Overall Steps 3) of the amino luciferin of 1～6mol/L obtain that to use 0.5～3mL concentration through the silicon nanowires of pentanedial decoration or silicon nanowire array be the ethanolic solution or the acetone soln of the amino luciferin of 1～6mol/L); Stirring reaction under the room temperature (the general reaction time is 0.5～3 hour); Then with organic solvent repeatedly ultrasonic cleaning remove unreacted amino luciferin; Do not have fluorescence until cleansing solution, filter and collect silicon nanowires or the silicon nanowire array of modifying through amino luciferin;
5) silicon nanowires that the amino luciferin of process that step 4) is obtained is modified; Or the silicon nanowire array modified of the amino luciferin of the process that obtains from step 4) is scraped the single silicon nanowires of getting and is placed ethanol or acetone; Ultrasonic it is disperseed, get the NaBH that dispersion liquid places 0.1～5M
4Reduce in the WS, reaction under room temperature (the general reaction time is 2～24 hours), the reactant liquor room temperature preservation, obtaining finishing has as the silicon nanowires of going back the amino luciferin of ortho states that nitrogen monoxide is had the selectivity fluorescence response; The chemical sensor based on silicon nanowires that promptly obtains nitrogen monoxide is had the selectivity fluorescence response is (like the silicon nanowires of the amino luciferin modification of the process that 1～15mg step 4) is obtained; Or scrape the single silicon nanowires of getting on the silicon nanowire array of modifying through amino luciferin and place 0.2～3mL ethanol or acetone; Ultrasonic it is disperseed, get the NaBH that 0.05～0.5mL dispersion liquid places 0.1～5M
4Reduce in the WS).
The method that chemical vapour deposition technique described in the present invention prepares silicon nanowires is: under the room temperature; Silicon monoxide, mortar is put into porcelain boat after grinding; And porcelain boat is placed on the middle part of quartz ampoule, before the heating, system is evacuated to 10 with mechanical pump and molecular pump to quartz ampoule earlier
-3Pa feeds the mixed gas of argon gas (accounting for the volume 95% of mixed gas) and hydrogen (accounting for the volume 5% of mixed gas) subsequently with the flow velocity of 15～30sccm (mL/min), when pressure stability during at 700～10000Pa, system begins to heat up; System rises to 300 ℃ with 10～20 ℃/min; Be warming up to 800 ℃ with 10～20 ℃/min again, close gas check valve and pump lock this moment, is incubated 10～30 minutes continued and is warming up to 1350 ℃; 1350 ℃ of reactions down (the general reaction time is 3～7 hours); Reaction naturally cools to room temperature after finishing, and collects the product silicon nanowires in the both sides of porcelain boat.
The method that chemical etching method described in the present invention prepares silicon nanowire array is: get (100) silicon chip of different size, carry out ultrasonic cleaning (time of general ultrasonic cleaning is 5～30 minutes) with ethanol, acetone, distilled water successively; Afterwards this silicon chip being immersed in mass concentration is that soak time is 30 seconds～15 minutes in 2～5% the HF WS; Silicon chip taken out be placed on that to contain concentration be 3～8mmol/LAgNO
3With (general soak time is 1～5 minute) in the mixed aqueous solution of 2～7mol/L HF; It is 2～7mol/L HF and 0.05～0.4mol/L H that silicon chip taking-up back immersion is contained concentration
2O
2Mixed aqueous solution in, system is 30～60 ℃ water bath heat preservation by temperature; Take out silicon chip after 5～45 minutes; Put into concentrated hydrochloric acid (mass concentration is 36%): the volume ratio of red fuming nitric acid (RFNA) (mass concentration is 36%) is 3: 1 a mixed liquor; Soak and take out silicon chip after 0.5～2.5 hour; Naturally dry with (placing surface plate to treat that it dries naturally) behind the distilled water flushing, obtain the silicon nanowire array that constitutes by silicon nanowires.
The present invention can carry out step 5) again with silicon nanowires or the silicon nanowire array that the amino luciferin of the process that step 4) obtains is modified after further handling; Described processing is that silicon nanowires or silicon nanowire array that the amino luciferin of process that step 4) obtains is modified are placed reactor; It is the ethanolic solution or the acetone soln of the sodium triacetoxy borohydride of 0.01～0.06mol/L that ethanolic solution or silicon nanowires that the amino luciferin of process that acetone soln (general Overall Steps 4) obtains is modified or the silicon nanowire array that adds concentration and be the sodium triacetoxy borohydride of 0.01～0.06mol/L uses 2～12mL concentration); In temperature is 30～70 ℃ of following reacting by heating (the general reaction time is 1～8 hour); Then with organic solvent repeatedly ultrasonic cleaning remove unreacted sodium triacetoxy borohydride; Filter and collect silicon nanowires or the silicon nanowire array that obtains through amino luciferin modification, drying at room temperature.It is that the ethanolic solution or the acetone soln of the sodium triacetoxy borohydride of 0.01～0.06mol/L handled that silicon nanowires that the amino luciferin of the process that step 4) is obtained is modified or silicon nanowire array use concentration; Can be so that the more firm surface that is modified at silicon nanowires or silicon nanowire array of amino fluorescein molecule guarantees that in follow-up course of reaction it is difficult for splitting away off from the surface of silicon nanowires or silicon nanowire array.
The diameter of the silicon nanowires that described chemical vapour deposition technique prepares is 5～20nm.
The diameter of the silicon nanowires in the silicon nanowire array that described chemical etching method prepares is 100～300nm, and length is 5～40 μ m.
The dry toluene that described dry toluene preferably newly steams.
The organic solvent of described cleaning usefulness can be the organic solvent of using always, like methyl alcohol, ethanol or acetone.
The chemical sensor based on silicon nanowires that nitrogen monoxide is had a selectivity fluorescence response of the present invention can be used for the detection to nitric oxide molecule; It is when doing to detect to nitric oxide molecule with XRF or fluorescent microscope; At the bottom of the above-mentioned active group of chemical sensor of modifying through special organic molecule as fluoroscopic examination based on silicon nanowires; Coupling XRF or fluorescent microscope; In the system that has nitrogen monoxide to exist, the above-mentioned chemical sensor based on silicon nanowires can produce fluorescence, through drawing the calibration curve of nitric oxide production concentration and characteristic fluorescence characteristic peak intensity; Intensity by the above-mentioned detected characteristic fluorescence characteristic peak of the chemical sensor based on silicon nanowires is confirmed nitric oxide production concentration in the system, thereby realizes the detection to nitric oxide molecule.That is, high selectivity and sensitivity by means of at the bottom of this active group can detection by quantitative go out the concentration of nitric oxide molecule, thereby realize constructing of sensor.
Of the present invention to nitrogen monoxide have the selectivity fluorescence response based on the chemical sensor of silicon nanowires when nitric oxide molecule is detected; The described system that has nitrogen monoxide to exist; Can be directly to join and contain in the solution that nitrogen monoxide is had a selectivity fluorescence response of the present invention based on the chemical sensor of silicon nanowires with containing nitric oxide production solution; Also can be to produce nitric oxide production material and contain the solution that nitrogen monoxide is had a selectivity fluorescence response of the present invention and mix, under 37 ℃, hatch based on the chemical sensor of silicon nanowires; Also can be to produce nitric oxide production biological sample (tissue extract or cell) suitably to hatch (suitably promptly being) with the chemical sensor that nitrogen monoxide is had a selectivity fluorescence response of the present invention, carry out the test of fluorescence intensity afterwards again to guarantee that the biological sample activity is a prerequisite based on silicon nanowires.Detect with XRF or fluorescent microscope.Used excitation source be mercury lamp (excitation wavelength is 470～495nm), xenon lamp (excitation wavelength be 470～495nm) or excitation wavelength be the laser instrument of 488nm, the emission light of this sensor is green glow.
The described system that has nitrogen monoxide to exist is animal's liver extract (can the nitrogen monoxide in the animal's liver extract be detected).
The present invention carries out surface treatment with the silicon nanowires and the silicon nanowire array that prepare based on chemical vapour deposition technique and chemical etching method; And then it is carried out chemical covalent modification with the amino luciferin of organic molecule material, obtain nitrogen monoxide is had the chemical sensor based on silicon nanowires of selectivity fluorescence response.The chemical sensor based on silicon nanowires that nitrogen monoxide is had a selectivity fluorescence response of the present invention can be used for nitric oxide production detection in solution and the living things system.
Description of drawings
Fig. 1. the HRTEM photo of the silicon nanowires that the process chemical vapour deposition technique of the embodiment of the invention 1 prepares.
Fig. 2. the SEM photo of the silicon nanowire array that the process chemical etching method of the embodiment of the invention 2 prepares.
Fig. 3. at the bottom of the active group in the embodiment of the invention 1～8 1 with active group at the bottom of the Response Mechanism of 2 couples of NO; Wherein: Fig. 3 a be at the bottom of the active group 1 with active group at the bottom of 2 chemosynthesis process synoptic diagram; Fig. 3 b be at the bottom of the active group 1 with active group at the bottom of the response synoptic diagram of 2 couples of NO.
Fig. 4. 1 as detecting substrate, the linearity curve of the concentration of system fluorescence and NO at the bottom of the active group of the embodiment of the invention 1.
Fig. 5. 1 as detecting substrate at the bottom of the active group of the embodiment of the invention 1, when the change in concentration of NO in the liver extract is detected, the linearity curve of volume drafting of system fluorescence intensity and add liver extract.
Fig. 6. 2 as detecting substrates at the bottom of the active group of the embodiment of the invention 2, the fluoroscopic image that NO in the solution is detected.A is the light field photo of 2 single nano-wires that scrape at the bottom of the active group; B is the fluorescence photo of 2 single nano-wires that scrape at the bottom of the active group; C is a 0.5M hydroxyethyl piperazine second thiosulfonic acid (deoxygenation) for 2 single nano-wires that scrape at the bottom of the active group place 1mL concentration, and adding 45 μ L, to contain concentration be 0.1M HCl and 0.1MNaNO
2Mixed solution, and observable fluorescence photo is carried out in insulation after 1 hour under fluorescent microscope in water-bath (37 ℃).
Embodiment
Embodiment 1
1) under the room temperature, silicon monoxide is put into porcelain boat after mortar grinds, and porcelain boat is placed on the middle part of quartz ampoule, before the heating, system is evacuated to 10 with mechanical pump and molecular pump earlier
-3Pa feeds the mixed gas of argon gas (accounting for the volume 95% of mixed gas) and hydrogen (accounting for the volume 5% of mixed gas) subsequently with the flow velocity of 25sccm (mL/min), when pressure stability 10
3During Pa, system begins to heat up.System rises to 300 ℃ with 10 ℃/min, is warming up to 800 ℃ with 20 ℃/min again, closes gas check valve and pump lock this moment; Keep 10 minutes continued to be warming up to 1350 ℃, 1350 ℃ were reacted 5 hours down, after reaction finishes; Naturally cool to room temperature; Product is collected in both sides at porcelain boat, and resulting silicon nanowires is that the center is that diameter is the monocrystalline silicon line of 15～20nm, and there is the amorphous silica layer (see figure 1) of one deck 1～3nm the outside.
2) silicon nanowires that step 1) is obtained is that 70 ℃ volume ratio is 5: 1 the concentrated sulphuric acid (mass concentration is 98%) and H in temperature
2O
2Heating is 30 minutes in the mixed liquor of (mass concentration is 30%), is cooled to room temperature afterwards, and take out also repeated water washing is immersed in H to neutral under room temperature
2O: H
2O
2(mass concentration is 30%): NH
4The volume ratio of OH is in 7: 1: 1 the mixed liquor and placed 2.5 hours, take out and repeated water washing to neutral, vacuum drying.
The new dry toluene and the 0.2mL 3-aminopropyltriethoxywerene werene (0.84mmol) of steaming of silicon nanowires, 10mL of the drying that 3) adding 45.5mg step 2 in round-bottomed flask) obtains; After under nitrogen protection, being heated to 90 ℃; Isothermal reaction 24 hours is cooled to room temperature, filters with microfilter and collects silicon nanowires; With ethanol repeatedly ultrasonic cleaning remove unreacted 3-aminopropyltriethoxywerene werene, filter collecting finishing has amino silicon nanowires.
4) finishing that step 3) is obtained has amino silicon nanowires to place round-bottomed flask; Adding 5mL mass concentration is 50% glutaraldehyde water solution; Stirring at room reaction 1 hour; Then with ethanol repeatedly ultrasonic cleaning remove unreacted glutaraldehyde, filter to collect the silicon nanowires that finishing has aldehyde radical.
5) finishing that step 4) is obtained has the silicon nanowires of aldehyde radical to place round-bottomed flask; Adding 2mL concentration is the ethanolic solution of the amino luciferin of 5mol/L; Stirring reaction is 2 hours under the room temperature; Then with ethanol repeatedly ultrasonic cleaning remove unreacted amino luciferin, do not have fluorescence until cleansing solution, filter to collect the silicon nanowires of modifying through amino luciferin.
6) silicon nanowires that the amino luciferin of process that step 5) is obtained is modified places round-bottomed flask; Adding 6mL concentration is the ethanolic solution of the sodium triacetoxy borohydride of 0.024mol/L; In temperature is 50 ℃ of following reacting by heating 4 hours; Then with ethanol repeatedly ultrasonic cleaning remove unreacted sodium triacetoxy borohydride, filter to collect and to obtain the silicon nanowires modified through amino luciferin, drying at room temperature.
7) the silicon nanowires 6mg that the amino luciferin of the process that step 6) is obtained is modified places 1mL ethanol, ultrasonic it is disperseed, and the dispersion liquid of getting 0.2mL places the NaBH of 3mL 0.1M
4Reduce in the WS, reaction is 3 hours under room temperature, the reactant liquor room temperature preservation, and obtaining finishing has as the silicon nanowires of going back the amino luciferin of ortho states that nitrogen monoxide is had the selectivity fluorescence response; It is 1 (see figure 3) at the bottom of the active group.
With above-mentioned obtain nitrogen monoxide is had at the bottom of the active group of selectivity fluorescence response 1 as nitric oxide molecule is carried out the fluoroscopic examination substrate, the coupling XRF carries out fluoroscopic examination to the NO that produces in the solution system, excitation source is an xenon lamp.At the bottom of containing active group, in the hydroxyethyl piperazine second thiosulfonic acid WS (deoxygenation) of 1 1mL 0.5M, add 45 μ L0.1M HCl and 100 μ L0.1M NaNO
2Solution, and insulation in water-bath (37 ℃), with the optical excitation of 450nm, every first order fluorescence that carried out at a distance from 5 minutes detects, and finds the increase along with nitric oxide concentration in the system, and the intensity at 1 fluorescent characteristics peak strengthens gradually at the bottom of the above-mentioned active group.The intensity and the nitric oxide concentration at fluorescent characteristics peak are linear, thereby have drawn the linear scaled curve (see figure 4) of fluorescent characteristics peak intensity and nitric oxide concentration.
Get fresh mouse liver, 12 grams of weighing stir in the PBS of 2mL degerming damping fluid, and in 4000rpm centrifugal 15 minutes, get supernatant liquor, in 4000rpm centrifugal 15 minutes, collect clear liquid, obtain the liver extract, freezing preservation.
With at the bottom of the above-mentioned active group that obtains 1 as the fluoroscopic examination substrate, the coupling XRF carries out fluoroscopic examination to the above-mentioned liver extract for preparing.At the bottom of containing active group among 1 the 1mL PBS (degerming); Add 10 μ L liver extracts gradually,, carry out fluoroscopic examination afterwards immediately with the optical excitation of 450nm; The result shows that the fluorescent characteristics peak intensity significantly increases (see figure 5) along with the increase of liver extract in the system.
Embodiment 2
1) gets (100) silicon chip of 2cm * 0.5cm, used ethanol, acetone, each ultrasonic cleaning of distilled water successively 10 minutes; Afterwards this silicon chip being immersed in mass concentration is in 5% the HF WS 15 minutes; This silicon chip taken out be placed on that to contain concentration be 5mmol/L AgNO
3In the mixed aqueous solution of 4.8mol/L HF; Soak after 2.5 minutes to take out and put into 10mL to contain concentration be 4.8mol/L HF and 0.2mol/LH
2O
2In the mixed aqueous solution, system is in 50 ℃ of water bath heat preservations; Take out silicon chip after 35 minutes, put into the mixed liquor that fills 4.5mL concentrated hydrochloric acid (mass concentration is 36%) and 1.5mL red fuming nitric acid (RFNA) (mass concentration is 36%); Soak and take out silicon chip after 1 hour, use distilled water flushing, wash clean is placed in the surface plate and dries naturally, obtains the silicon nanowire array that is made up of silicon nanowires, and wherein the diameter of silicon nanowires is 100～200nm, and length is 25～40 μ m (see figure 2)s.
2) silicon nanowire array that step 1) is obtained is that 90 ℃ volume ratio is 5: 1 the concentrated sulphuric acid (mass concentration is 98%) and H in temperature
2O
2Heating is 50 minutes in the mixed liquor of (mass concentration is 30%); Be cooled to room temperature afterwards; Take out and repeated water washing to neutral, (treatment conditions: the mass content of oxygen is 100%, and voltage is 500 volts to place the oxygen plasma system to carry out the surface treatment of silicon nanowire array then; Time is 5 minutes, and temperature is 35 ℃).
The new dry toluene and the 0.2mL 3-aminopropyltriethoxywerene werene (0.84mmol) of steaming of silicon nanowire array, 10mL of the drying that 3) adding 45.5mg step 2 in round-bottomed flask) obtains; After under nitrogen protection, being heated to 90 ℃; Isothermal reaction 24 hours is cooled to room temperature, filters with microfilter and collects silicon nanowire array; With ethanol repeatedly ultrasonic cleaning remove unreacted 3-aminopropyltriethoxywerene werene, filter collecting finishing has amino silicon nanowire array.
4) finishing that step 3) is obtained has amino silicon nanowire array to place round-bottomed flask; Adding 5mL mass concentration is 30% glutaraldehyde water solution; Stirring at room reaction 1 hour; Then with ethanol repeatedly ultrasonic cleaning remove unreacted glutaraldehyde, filter to collect the silicon nanowire array that finishing has aldehyde radical.
5) finishing that step 4) is obtained has the silicon nanowire array of aldehyde radical to place round-bottomed flask; Adding 2mL concentration is the ethanolic solution of the amino luciferin of 5mol/L; Stirring reaction is 2 hours under the room temperature; Then with ethanol repeatedly ultrasonic cleaning remove unreacted amino luciferin, do not have fluorescence until cleansing solution, filter to collect the silicon nanowire array of modifying through amino luciferin.
6) silicon nanowire array that the amino luciferin of process that step 5) is obtained is modified places round-bottomed flask; Adding 6mL concentration is the ethanolic solution of the sodium triacetoxy borohydride of 0.024mol/L; In temperature is 50 ℃ of following reacting by heating 4 hours; Then with ethanol repeatedly ultrasonic cleaning remove unreacted sodium triacetoxy borohydride, filter to collect and to obtain the silicon nanowire array modified through amino luciferin, drying at room temperature.
7) will under the silicon nanowire array root that the amino luciferin of the process that step 6) obtains is modified, scrape the single silicon nanowires of the 2mg that gets and place 1mL ethanol, and ultrasonic it disperseed, the dispersion liquid of getting 0.2mL places the NaBH of 3mL 3M
4Reduce in the WS, reaction is 5 hours under room temperature, the reactant liquor room temperature preservation, and obtaining finishing has as the silicon nanowires of going back the amino luciferin of ortho states that nitrogen monoxide is had the selectivity fluorescence response, promptly at the bottom of the active group 2.
With above-mentioned obtain nitrogen monoxide is had at the bottom of the active group of selectivity fluorescence response 2 as nitric oxide molecule is carried out the fluoroscopic examination substrate, the coupling fluorescent microscope carries out fluoroscopic examination to the NO that produces in the solution system, excitation source is an xenon lamp.At the bottom of containing active group, in the hydroxyethyl piperazine second thiosulfonic acid WS (deoxygenation) of 2 1mL 0.5M, add 45 μ L 0.1M HCl and 0.1mL 2.6M NaNO
2Solution, and insulation in water-bath (37 ℃) were observed under fluorescent microscope after 1 hour, with the NO solution reaction after, 2 fluorescence intensity obviously strengthens (see figure 6) at the bottom of the active group.
Embodiment 3
1) under the room temperature, silicon monoxide is put into porcelain boat after mortar grinds, and porcelain boat is placed on the middle part of quartz ampoule, before the heating, system is evacuated to 10 with mechanical pump and molecular pump earlier
-3Pa feeds the mixed gas of argon gas (accounting for the volume 95% of mixed gas) and hydrogen (accounting for the volume 5% of mixed gas) subsequently with the flow velocity of 25sccm (mL/min), when pressure stability 10
3During Pa, system begins to heat up.System rises to 300 ℃ with 20 ℃/min, is warming up to 800 ℃ with 10 ℃/min again, closes gas check valve and pump lock this moment; Keep 10 minutes continued to be warming up to 1350 ℃, 1350 ℃ were reacted 5 hours down, after reaction finishes; Naturally cool to room temperature; Product is collected in both sides at porcelain boat, and resulting silicon nanowires is that central diameter is the monocrystalline silicon line of 5～15nm, and there is the amorphous silica layer of one deck 1～3nm the outside.
2) silicon nanowires that step 1) is obtained is that 70 ℃ volume ratio is 5: 1 the concentrated sulphuric acid (mass concentration is 50%) and H in temperature
2O
2Heating is 30 minutes in the mixed liquor of (mass concentration is 5%), is cooled to room temperature afterwards, and take out also repeated water washing is immersed in H to neutral under room temperature
2O: H
2O
2(mass concentration is 5%): NH
4The volume ratio of OH is in 7: 1: 1 the mixed liquor and placed 2.5 hours, take out and repeated water washing to neutral, vacuum drying.
The new dry toluene and the 0.05mL 3-aminopropyltriethoxywerene werene (0.21mmol) of steaming of silicon nanowires, 5mL of the drying that 3) adding 10mg step 2 in round-bottomed flask) obtains; After under nitrogen protection, being heated to 80 ℃; Isothermal reaction 12 hours is cooled to room temperature, filters with microfilter and collects silicon nanowires; With ethanol repeatedly ultrasonic cleaning remove unreacted 3-aminopropyltriethoxywerene werene, filter collecting finishing has amino silicon nanowires.
4) finishing that step 3) is obtained has amino silicon nanowires to place round-bottomed flask; Adding 1.5mL mass concentration is 35% glutaraldehyde water solution; Stirring at room reaction 30 minutes; Then with ethanol repeatedly ultrasonic cleaning remove unreacted glutaraldehyde, filter to collect the silicon nanowires that finishing has aldehyde radical.
5) finishing that step 4) is obtained has the silicon nanowires of aldehyde radical to place round-bottomed flask; Adding 0.5mL concentration is the acetone soln of the amino luciferin of 1mmol/L; Stirring reaction is 30 minutes under the room temperature; Then with ethanol repeatedly ultrasonic cleaning remove unreacted amino luciferin, do not have fluorescence until cleansing solution, filter to collect the silicon nanowires of modifying through amino luciferin.
6) silicon nanowires that the amino luciferin of process that step 5) is obtained is modified places round-bottomed flask; Adding 2mL concentration is (0.02mmol) ethanolic solution of the sodium triacetoxy borohydride of 0.01mol/L; In temperature is 30 ℃ of following heating reflux reactions 1 hour; Then with ethanol repeatedly ultrasonic cleaning remove unreacted sodium triacetoxy borohydride, filter to collect and to obtain the silicon nanowires modified through amino luciferin, drying at room temperature.
7) the silicon nanowires 1mg that the amino luciferin of the process that step 6) is obtained is modified places 0.2mL ethanol, ultrasonic it is disperseed, and the dispersion liquid of getting 0.05mL places the NaBH of 0.5mL 0.1mol/L
4Reduce in the WS, reaction is 2 hours under room temperature, the reactant liquor room temperature preservation, and obtaining finishing has as the silicon nanowires of going back the amino luciferin of ortho states that nitrogen monoxide is had the selectivity fluorescence response, promptly at the bottom of the active group 1.
With above-mentioned obtain nitrogen monoxide is had at the bottom of the active group of selectivity fluorescence response 1 as nitric oxide molecule is carried out the fluoroscopic examination substrate, the coupling XRF with the optical excitation of 450nm, carries out fluoroscopic examination to the NO that produces in the solution system.
Get fresh mouse liver, 6 grams of weighing stir in the PBS of 3mL degerming damping fluid, and in 4000rpm centrifugal 15 minutes, get supernatant liquor, in 4000rpm centrifugal 15 minutes, collect clear liquid, obtain the liver extract, freezing preservation.
With at the bottom of the above-mentioned active group that obtains 1 as the fluoroscopic examination substrate, the coupling XRF carries out fluoroscopic examination to the above-mentioned liver extract for preparing.At the bottom of containing active group, among 1 the 1mL PBS (degerming), add 10 μ L liver extracts gradually, carry out fluoroscopic examination afterwards immediately, with the optical excitation of 450nm, the result shows that the fluorescent characteristics peak intensity significantly increases along with the increase of liver extract in the system.
Embodiment 4
1) gets (100) silicon chip of 2cm * 0.5cm, used ethanol, acetone, each ultrasonic cleaning of distilled water successively 10 minutes; Afterwards this silicon chip being immersed in mass concentration is in 5% the HF WS 15 minutes; This silicon chip taken out be placed on that to contain concentration be 5mmol/L AgNO
3In the mixed aqueous solution of 4.8mol/L HF; Soak after 2.5 minutes to take out and put into 10mL to contain concentration be 4.8mol/L HF and 0.2mol/LH
2O
2In the mixed aqueous solution, system is in 50 ℃ of water bath heat preservations; Take out silicon chip after 15 minutes, put into the mixed liquor that fills 4.5mL concentrated hydrochloric acid (mass concentration is 36%) and 1.5mL red fuming nitric acid (RFNA) (mass concentration is 36%); Soak and take out silicon chip after 1 hour, use distilled water flushing, wash clean is placed in the surface plate and dries naturally, obtains the silicon nanowire array that is made up of silicon nanowires, and wherein the diameter of silicon nanowires is 200～300nm, and length is 5～25 μ m.
2) silicon nanowire array that step 1) is obtained is that 95 ℃ volume ratio is 4: 1 the concentrated sulphuric acid (mass concentration is 70%) and H in temperature
2O
2Heating is 90 minutes in the mixed liquor of (mass concentration is 15%); Be cooled to room temperature afterwards; Take out and repeated water washing to neutral, (treatment conditions: the mass content of oxygen is 10%, and voltage is 100 volts to place the oxygen plasma system to carry out the surface treatment of silicon nanowire array then; Time is 10 seconds, and temperature is 10 ℃);
The new dry toluene and the 0.3mL 3-aminopropyltriethoxywerene werene (1.26mmol) of steaming of silicon nanowire array, 30mL of the drying that 3) adding 10mg step 2 in round-bottomed flask) obtains; After under nitrogen protection, being heated to 120 ℃; Isothermal reaction 12 hours is cooled to room temperature, filters with microfilter and collects silicon nanowire array; With ethanol repeatedly ultrasonic cleaning remove unreacted 3-aminopropyltriethoxywerene werene, filter collecting finishing has amino silicon nanowire array.
4) surface modification that step 3) is obtained has amino silicon nanowire array to place round-bottomed flask; Adding 5mL mass concentration is 10% glutaraldehyde water solution; Stirring at room reaction 1 hour; Then with ethanol repeatedly ultrasonic cleaning remove unreacted glutaraldehyde, filter to collect the silicon nanowire array that finishing has aldehyde radical.
5) finishing that step 4) is obtained has the silicon nanowire array of aldehyde radical to place round-bottomed flask; Adding 2mL concentration is the ethanolic solution of the amino luciferin of 5mol/L; Stirring reaction is 2 hours under the room temperature; Then with ethanol repeatedly ultrasonic cleaning remove unreacted amino luciferin, do not have fluorescence until cleansing solution, filter to collect the silicon nanowire array of modifying through amino luciferin.
6) silicon nanowire array that the amino luciferin of process that step 5) is obtained is modified places round-bottomed flask; Adding 6mL concentration is the ethanolic solution of the sodium triacetoxy borohydride of 0.024mol/L; In temperature is 50 ℃ of following reacting by heating 4 hours; Then with ethanol repeatedly ultrasonic cleaning remove unreacted sodium triacetoxy borohydride, filter to collect and to obtain the silicon nanowire array modified through amino luciferin, drying at room temperature.
7) will under the silicon nanowire array root that the amino luciferin of the process that step 6) obtains is modified, scrape the single silicon nanowires of the 10mg that gets and place 5mL ethanol, and ultrasonic it disperseed, the dispersion liquid of getting 0.5mL places the NaBH of 5mL 1.85mol/L
4Reduce in the WS, reaction is 18 hours under room temperature, the reactant liquor room temperature preservation, and obtaining finishing has as the silicon nanowires of going back the amino luciferin of ortho states that nitrogen monoxide is had the selectivity fluorescence response, promptly at the bottom of the active group 2.
With above-mentioned obtain nitrogen monoxide is had at the bottom of the active group of selectivity fluorescence response 2 as nitric oxide molecule is carried out the fluoroscopic examination substrate, the coupling fluorescent microscope carries out fluoroscopic examination to the NO that produces in the solution system.With 2 placing and produce nitric oxide production solution system at the bottom of the active group, and insulation in water-bath (37 ℃) in, under fluorescent microscope, observed afterwards in 1 hour, xenon lamp as excitation source with the NO solution reaction after, 2 fluorescence intensity obviously strengthens at the bottom of the active group.
Embodiment 5
1) be that 95 ℃ volume ratio is 8: 1 the concentrated sulphuric acid (mass concentration is 80%) and H with the silicon nanowires that obtains through chemical vapour deposition technique among the embodiment 1 in temperature
2O
2Heating is 90 minutes in the mixed liquor of (mass concentration is 20%), is cooled to room temperature afterwards, and take out also repeated water washing is immersed in H to neutral under room temperature
2O: H
2O
2(mass concentration is 20%): NH
4The volume ratio of OH is in 9: 1: 1 the mixed liquor and placed 2.5 hours, take out and repeated water washing to neutral, vacuum drying.
The new dry toluene and the 0.3mL 3-aminopropyltriethoxywerene werene (1.26mmol) of steaming of silicon nanowires, 30mL of the drying that 2) adding 60mg step 1) obtains in round-bottomed flask; After under nitrogen protection, being heated to 120 ℃; Isothermal reaction 36 hours is cooled to room temperature, filters with microfilter and collects silicon nanowires; With acetone repeatedly ultrasonic cleaning remove unreacted 3-aminopropyltriethoxywerene werene, filter collecting finishing has amino silicon nanowires.
4) finishing that step 3) is obtained has amino silicon nanowires to place round-bottomed flask; Adding 10mL mass concentration is 50% glutaraldehyde water solution; Stirring at room reaction 120 minutes; Then with ethanol repeatedly ultrasonic cleaning remove unreacted glutaraldehyde, filter to collect the silicon nanowires that finishing has aldehyde radical.
5) finishing that step 4) is obtained has the silicon nanowires of aldehyde radical to place conical flask; Adding 3mL concentration is the ethanolic solution of the amino luciferin of 6mmol/L; Stirring reaction is 3 hours under the room temperature; Then with ethanol repeatedly ultrasonic cleaning remove unreacted amino luciferin, do not have fluorescence until cleansing solution, filter to collect the silicon nanowires of modifying through amino luciferin.
6) silicon nanowires that the amino luciferin of process that step 5) is obtained is modified places round-bottomed flask; Adding 12mL concentration is (0.72mmol) ethanolic solution of the sodium triacetoxy borohydride of 0.06mol/L; In temperature is 70 ℃ of following heating reflux reactions 8 hours; Then with ethanol repeatedly ultrasonic cleaning remove unreacted sodium triacetoxy borohydride, filter to collect and to obtain the silicon nanowires modified through amino luciferin, drying at room temperature.
7) the silicon nanowires 15mg that the amino luciferin of the process that step 6) is obtained is modified places 3mL ethanol, ultrasonic it is disperseed, and the dispersion liquid of getting 0.5mL places the NaBH of 5mL 0.8mol/L
4Reduce in the WS, reaction is 24 hours under room temperature, the reactant liquor room temperature preservation, and obtaining finishing has as the silicon nanowires of going back the amino luciferin of ortho states that nitrogen monoxide is had the selectivity fluorescence response, promptly at the bottom of the active group 1.
With above-mentioned obtain nitrogen monoxide is had at the bottom of the active group of selectivity fluorescence response 1 as nitric oxide molecule is carried out the fluoroscopic examination substrate, the coupling XRF carries out fluoroscopic examination to the NO that produces in the solution system.With 1 placing and can slowly produce nitric oxide production solution system at the bottom of the active group, with the optical excitation of 450nm, every first order fluorescence that carried out at a distance from 5 minutes detects, and writes down fluorescence curve.Discovery is along with the increase of nitric oxide concentration in the system, 1 fluorescent characteristics peak at the bottom of the above-mentioned active group (intensity strengthen gradually.The intensity and the nitric oxide concentration at fluorescent characteristics peak are linear, thereby have drawn the linear scaled curve of fluorescent characteristics peak intensity and nitric oxide concentration.
Embodiment 6
1) be that 70 ℃ volume ratio is 4.5: 1 the concentrated sulphuric acid (mass concentration is 90%) and H with the silicon nanowire array that obtains through chemical etching method among the embodiment 2 in temperature
2O
2Heating is 30 minutes in the mixed liquor of (mass concentration is 25%); Be cooled to room temperature afterwards; Take out and repeated water washing to neutral, (treatment conditions: the mass content of oxygen is 50%, and voltage is 300 volts to place the oxygen plasma system to carry out the surface treatment of silicon nanowire array then; Time is 3 minutes, and temperature is 25 ℃).
The new dry toluene and the 0.05mL 3-aminopropyltriethoxywerene werene (0.21mmol) of steaming of silicon nanowire array, 5mL of the drying that 2) adding 60mg step 1) obtains in round-bottomed flask; After under nitrogen protection, being heated to 120 ℃; Isothermal reaction 36 hours is cooled to room temperature, filters with microfilter and collects silicon nanowire array; With acetone repeatedly ultrasonic cleaning remove unreacted 3-aminopropyltriethoxywerene werene, filter collecting finishing has amino silicon nanowire array.
3) with step 2) finishing that obtains has amino silicon nanowire array to place round-bottomed flask; Adding 5mL mass concentration is 45% glutaraldehyde water solution; Stirring at room reaction 1 hour; Then with ethanol repeatedly ultrasonic cleaning remove unreacted glutaraldehyde, filter to collect the silicon nanowire array that finishing has aldehyde radical.
4) finishing that step 3) is obtained has the silicon nanowire array of aldehyde radical to place round-bottomed flask; Adding 2mL concentration is the ethanolic solution of the amino luciferin of 5mol/L; Stirring reaction is 2 hours under the room temperature; Then with ethanol repeatedly ultrasonic cleaning remove unreacted amino luciferin, do not have fluorescence until cleansing solution, filter to collect the silicon nanowire array of modifying through amino luciferin.
5) will under the silicon nanowire array root that the amino luciferin of the process that step 4) obtains is modified, scrape the single silicon nanowires of the 20mg that gets and place 15mL ethanol, and ultrasonic it disperseed, the dispersion liquid of getting 1.5mL places the NaBH of 18mL 5mol/L
4Reduce in the WS, reaction is 14 hours under room temperature, the reactant liquor room temperature preservation, and obtaining finishing has as the silicon nanowires of going back the amino luciferin of ortho states that nitrogen monoxide is had the selectivity fluorescence response, promptly at the bottom of the active group 2.
With above-mentioned obtain nitrogen monoxide is had at the bottom of the active group of selectivity fluorescence response 2 as nitric oxide molecule is carried out the fluoroscopic examination substrate, the coupling fluorescent microscope carries out fluoroscopic examination to the NO that produces in the solution system.With 2 placing and to produce nitric oxide production solution system at the bottom of the active group; And insulation in water-bath (37 ℃), under fluorescent microscope, to observe after 1 hour, xenon lamp is as excitation source; The result be illustrated in the NO solution reaction after, 2 fluorescence intensity obviously strengthens at the bottom of the active group.
Embodiment 7
1) with embodiment 3 through in to cross silicon nanowires that chemical vapour deposition technique obtains be that 90 ℃ volume ratio is 5.5: 1 the concentrated sulphuric acid (mass concentration is 98%) and H in temperature
2O
2Heating is 90 minutes in the mixed liquor of (mass concentration is 30%), is cooled to room temperature afterwards, and take out also repeated water washing is immersed in H to neutral under room temperature
2O: H
2O
2(mass concentration is 30%): NH
4The volume ratio of OH is in 6.5: 1: 1 the mixed liquor and placed 3 hours, take out and repeated water washing to neutral, vacuum drying.
The new dry toluene and the 0.3mL 3-aminopropyltriethoxywerene werene (1.26mmol) of steaming of silicon nanowires, 15mL of the drying that 2) adding 60mg step 1) obtains in round-bottomed flask; After under nitrogen protection, being heated to 95 ℃; Isothermal reaction 28 hours is cooled to room temperature, filters with microfilter and collects silicon nanowires; With acetone repeatedly ultrasonic cleaning remove unreacted 3-aminopropyltriethoxywerene werene, filter collecting finishing has amino silicon nanowires.
3) with step 2) finishing that obtains has amino silicon nanowires to place round-bottomed flask; Adding 7mL mass concentration is 50% glutaraldehyde water solution; Stirring at room reaction 90 minutes; Then with acetone repeatedly ultrasonic cleaning remove unreacted glutaraldehyde, filter to collect the silicon nanowires that finishing has aldehyde radical.
4) finishing that step 3) is obtained has the silicon nanowires of aldehyde radical to place conical flask; Adding 3mL concentration is the acetone soln of the amino luciferin of 5mmol/L; Stirring reaction is 3 hours under the room temperature; Then with acetone repeatedly ultrasonic cleaning remove unreacted amino luciferin, do not have fluorescence until cleansing solution, filter to collect the silicon nanowires of modifying through amino luciferin.
5) the silicon nanowires 8mg that the amino luciferin of the process that step 4) is obtained is modified places 1.5mL ethanol, ultrasonic it is disperseed, and the dispersion liquid of getting 0.5mL places the NaBH of 6mL 2.6mol/L
4Reduce in the WS, reaction is 10 hours under room temperature, the reactant liquor room temperature preservation, and obtaining finishing has as the silicon nanowires of going back the amino luciferin of ortho states that nitrogen monoxide is had the selectivity fluorescence response, promptly at the bottom of the active group 1.
With above-mentioned obtain nitrogen monoxide is had at the bottom of the active group of selectivity fluorescence response 1 as nitric oxide molecule is carried out the fluoroscopic examination substrate, the coupling XRF carries out fluoroscopic examination to the NO that produces in the solution system.With 1 placing and slowly to produce nitric oxide production system at the bottom of the active group; With xenon lamp as excitation source; With the optical excitation of 450nm, every first order fluorescence that carried out at a distance from 6 minutes detects, and the record fluorescence curve; Discovery is along with the increase of nitric oxide concentration in the system, and the intensity at 1 fluorescent characteristics peak strengthens gradually at the bottom of the above-mentioned active group.The intensity and the nitric oxide concentration at fluorescent characteristics peak are linear, thereby have drawn the linear scaled curve of fluorescent characteristics peak intensity and nitric oxide concentration.
Embodiment 8
1) be that 90 ℃ volume ratio is 3: 1 the concentrated sulphuric acid (mass concentration is 98%) and H with the silicon nanowire array that obtains through chemical etching method among the embodiment 4 in temperature
2O
2Heating is 50 minutes in the mixed liquor of (mass concentration is 30%); Be cooled to room temperature afterwards; Take out and repeated water washing to neutral, (treatment conditions: the mass content of oxygen is 80%, and voltage is 500 volts to place the oxygen plasma system to carry out the surface treatment of silicon nanowire array then; Time is 5 minutes, and temperature is 30 ℃).
The new dry toluene and the 0.3mL 3-aminopropyltriethoxywerene werene (1.26mmol) of steaming of silicon nanowire array, 10mL of the drying that 2) adding 60mg step 1) obtains in round-bottomed flask; After under nitrogen protection, being heated to 90 ℃; Isothermal reaction 36 hours is cooled to room temperature, filters with microfilter and collects silicon nanowire array; With acetone repeatedly ultrasonic cleaning remove unreacted 3-aminopropyltriethoxywerene werene, filter collecting finishing has amino silicon nanowire array.
3) with step 2) finishing that obtains has amino silicon nanowire array to place round-bottomed flask; Adding 5mL mass concentration is 50% glutaraldehyde water solution; Stirring at room reaction 1 hour; Then with ethanol repeatedly ultrasonic cleaning remove unreacted glutaraldehyde, filter to collect the silicon nanowire array that finishing has aldehyde radical.
4) finishing that step 3) is obtained has the silicon nanowire array of aldehyde radical to place round-bottomed flask; Adding 2mL concentration is the ethanolic solution of the amino luciferin of 5mol/L; Stirring reaction is 2 hours under the room temperature; Then with ethanol repeatedly ultrasonic cleaning remove unreacted amino luciferin, do not have fluorescence until cleansing solution, filter to collect the silicon nanowire array of modifying through amino luciferin.
5) silicon nanowire array that the amino luciferin of process that step 4) is obtained is modified places round-bottomed flask; Adding 6mL concentration is the ethanolic solution of the sodium triacetoxy borohydride of 0.024mol/L; In temperature is 50 ℃ of following reacting by heating 4 hours; Then with ethanol repeatedly ultrasonic cleaning remove unreacted sodium triacetoxy borohydride, filter to collect and to obtain the silicon nanowire array modified through amino luciferin, drying at room temperature.
6) will under the silicon nanowire array root that the amino luciferin of the process that step 5) obtains is modified, scrape the single silicon nanowires of the 8mg that gets and place 1mL ethanol, and ultrasonic it disperseed, the dispersion liquid of getting 0.5mL places the NaBH of 6mL 2.6mol/L
4Reduce in the WS, reaction is 10 hours under room temperature, the reactant liquor room temperature preservation, and obtaining finishing has as the silicon nanowires of going back the amino luciferin of ortho states that nitrogen monoxide is had the selectivity fluorescence response, promptly at the bottom of the active group 2.
With above-mentioned obtain nitrogen monoxide is had at the bottom of the active group of selectivity fluorescence response 2 as nitric oxide molecule is carried out the fluoroscopic examination substrate, the coupling fluorescent microscope carries out fluoroscopic examination to the NO that produces in the solution system.With 2 placing and can produce nitric oxide production solution system at the bottom of the active group, and insulation in water-bath (37 ℃), under fluorescent microscope, observe after 1 hour, xenon lamp is as light source, with the NO solution reaction after, 2 fluorescence intensity obviously strengthens at the bottom of the active group.
Claims (9)
1. chemical sensor that nitrogen monoxide is had the selectivity fluorescence response based on silicon nanowires; It is characterized in that: described chemical sensor based on silicon nanowires is by the finishing that 3-aminopropyltriethoxywerene werene and silicon nanowires or silicon nanowire array reaction obtain amino silicon nanowires or silicon nanowire array to be arranged; Obtain silicon nanowires or the silicon nanowire array that finishing has aldehyde radical with the glutaraldehyde reaction; Be modified at the amino luciferin that is obtained by aldehyde radical and amino luciferin reaction on silicon nanowires or the silicon nanowire array through reduction, obtaining finishing has the chemical sensor as the silicon nanowires of going back the amino luciferin of ortho states that nitrogen monoxide is had the selectivity fluorescence response.
2. according to claim 1 have the chemical sensor based on silicon nanowires of selectivity fluorescence response to nitrogen monoxide, it is characterized in that: described silicon nanowires is to be the silicon nanowires of 5～20nm by the diameter that chemical vapour deposition technique prepares; Or be 100～300nm by the diameter that chemical etching method prepares, length is the silicon nanowires of 5～40 μ m.
3. one kind according to claim 1 and 2ly has the preparation method based on the chemical sensor of silicon nanowires of selectivity fluorescence response to nitrogen monoxide, it is characterized in that described method may further comprise the steps:
1) will use silicon nanowires that chemical vapour deposition technique prepares is that 70～95 ℃ volume ratio is 1: 1～8: 1 the concentrated sulphuric acid and H in temperature
2O
2Mixed liquor in heated 30～90 minutes, be cooled to room temperature afterwards, take out washing to neutral, under room temperature, be immersed in H
2O: H
2O
2: NH
4The volume ratio of OH is in 3: 1: 1～9: 1: 1 the mixed liquor and places, and takes out also washing to neutral, vacuum drying; Or
Is that 70～95 ℃ volume ratio is 1: 1～8: 1 the concentrated sulphuric acid and H with the silicon nanowire array that uses chemical etching method to prepare in temperature
2O
2Mixed liquor in heated 30～90 minutes; Be cooled to room temperature afterwards, take out and wash, place the oxygen plasma system to carry out the surface treatment of silicon nanowire array then to neutral; Treatment conditions: the mass content of oxygen is 10～100%; Voltage is 100～500 volts, and the time is 10 seconds～5 minutes, and temperature is 10～35 ℃;
2) in reactor, add silicon nanowires or silicon nanowire array, 5～30mL dry toluene and 0.05～0.3mL 3-aminopropyltriethoxywerene werene of the drying that 10～60mg step 1) obtains; After under inert gas shielding, being heated to 80～120 ℃; Isothermal reaction 12～36 hours; Be cooled to room temperature; Filter and collect silicon nanowires or silicon nanowire array, remove unreacted 3-aminopropyltriethoxywerene werene, filter and collect silicon nanowires or the silicon nanowire array that finishing has amino with the organic solvent ultrasonic cleaning;
3) with step 2) finishing that obtains has amino silicon nanowires or silicon nanowire array to place reactor; The adding mass concentration is 10%～50% glutaraldehyde water solution; The stirring at room reaction; Remove unreacted glutaraldehyde with the organic solvent ultrasonic cleaning then, filter and collect silicon nanowires or the silicon nanowire array that finishing has aldehyde radical;
4) finishing that step 3) is obtained has the silicon nanowires of aldehyde radical or silicon nanowire array to place reactor; Adding concentration is the ethanolic solution or the acetone soln of the amino luciferin of 1～6mol/L; Stirring reaction under the room temperature; Then with organic solvent repeatedly ultrasonic cleaning remove unreacted amino luciferin, do not have fluorescence until cleansing solution, filter to collect silicon nanowires or the silicon nanowire array modified through amino luciferin;
5) silicon nanowires that the amino luciferin of process that step 4) is obtained is modified; Or the silicon nanowire array modified of the amino luciferin of the process that obtains from step 4) is scraped the single silicon nanowires of getting and is placed ethanol or acetone; The ultrasonic silicon nanowires that makes disperses, and gets the NaBH that dispersion liquid places 0.1～5M
4Reduce in the WS, under room temperature, react, obtaining finishing has pair nitrogen monoxide to have the chemical sensor of the silicon nanowires of going back the amino luciferin of ortho states of selectivity fluorescence response.
4. preparation method according to claim 3; It is characterized in that: silicon nanowires that the amino luciferin of the process that step 4) is obtained is modified or silicon nanowire array are through further carry out step 5) after the processing again; Described processing is that silicon nanowires or silicon nanowire array that the amino luciferin of process that step 4) obtains is modified are placed reactor; Adding concentration is the ethanolic solution or the acetone soln of the sodium triacetoxy borohydride of 0.01～0.06mol/L; In temperature is 30～70 ℃ of following reacting by heating; Then with organic solvent repeatedly ultrasonic cleaning remove unreacted sodium triacetoxy borohydride, filter to collect and to obtain silicon nanowires or the silicon nanowire array modified through amino luciferin.
5. preparation method according to claim 3 is characterized in that: the reaction time of the described stirring at room reaction of step 3) is 0.5～2 hour;
The reaction time of stirring reaction is 0.5～3 hour under the described room temperature of step 4);
The described reaction time of under room temperature, reacting of step 5) is 2～24 hours.
6. preparation method according to claim 3 is characterized in that: the diameter of the silicon nanowires that described chemical vapour deposition technique prepares is 5～20nm;
The diameter of the silicon nanowires in the silicon nanowire array that described chemical etching method prepares is 100～300nm, and length is 5～40 μ m.
7. according to claim 3 or 4 described preparation methods, it is characterized in that: described organic solvent is methyl alcohol, ethanol or acetone.
8. one kind according to any described application based on the chemical sensor of silicon nanowires that nitrogen monoxide is had the selectivity fluorescence response of claim 1～3; It is characterized in that: the described chemical sensor based on silicon nanowires that nitrogen monoxide is had a selectivity fluorescence response is used for the detection to nitric oxide molecule; In the system that has nitrogen monoxide to exist; Described chemical sensor based on silicon nanowires can produce fluorescence; Through drawing the calibration curve of nitric oxide production concentration and characteristic fluorescence characteristic peak intensity, confirm nitric oxide production concentration in the system by the intensity of the detected characteristic fluorescence characteristic peak of described chemical sensor based on silicon nanowires.
9. application according to claim 8 is characterized in that: the described system that has nitrogen monoxide to exist is the animal's liver extract.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102661626A CN102419319A (en) | 2011-09-08 | 2011-09-08 | Silicon nano wire based sensor having selective fluorescence response on nitric oxide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011102661626A CN102419319A (en) | 2011-09-08 | 2011-09-08 | Silicon nano wire based sensor having selective fluorescence response on nitric oxide |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102419319A true CN102419319A (en) | 2012-04-18 |
Family
ID=45943832
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011102661626A Pending CN102419319A (en) | 2011-09-08 | 2011-09-08 | Silicon nano wire based sensor having selective fluorescence response on nitric oxide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102419319A (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102928391A (en) * | 2012-10-11 | 2013-02-13 | 中国科学院理化技术研究所 | Silicon nanowire ordered array-based pH fluorescence sensor and manufacturing method and application thereof |
| CN103712968A (en) * | 2014-01-06 | 2014-04-09 | 中国科学院理化技术研究所 | Silicon-nanowire-based fluorescence chemical sensor with selective fluorescence response to free copper ions and complexing copper ions |
| CN103937488A (en) * | 2014-03-25 | 2014-07-23 | 中国科学院理化技术研究所 | Silicon-nanowire-based alkaline phosphatase fluorescent chemosensor, preparation method and application |
| CN104155277A (en) * | 2014-08-27 | 2014-11-19 | 中国科学院理化技术研究所 | Preparation method of sulfur ion fluorescent chemical sensor based on silicon nanowire or silicon nanowire array |
| CN104792758A (en) * | 2015-04-22 | 2015-07-22 | 中国科学院理化技术研究所 | Preparation method for hydrogen sulfide fluorescent chemosensor based on silicon nanowire or silicon nanowire array |
| CN108913120A (en) * | 2018-06-21 | 2018-11-30 | 中国科学院理化技术研究所 | A kind of single silicon nanowire fluorescence chemical sensor and its preparation method and application for detecting hypochlorite |
| CN109897630A (en) * | 2019-04-17 | 2019-06-18 | 中国科学院理化技术研究所 | A kind of nano wire, preparation method, the Ratio-type fluorescence chemical sensor comprising the nano wire and application |
| CN113686927A (en) * | 2021-08-06 | 2021-11-23 | 上海纳感医疗科技有限公司 | Method for improving biochemical detection selectivity of silicon nanowire sensor |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101191794A (en) * | 2007-08-27 | 2008-06-04 | 中国科学院理化技术研究所 | One-dimensional nanostructured fluorescent chemical biosensor and method for making same and uses |
| WO2011085533A1 (en) * | 2010-01-15 | 2011-07-21 | 大连理工大学 | Fluorescent probe compounds, preparation method and application thereof |
-
2011
- 2011-09-08 CN CN2011102661626A patent/CN102419319A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101191794A (en) * | 2007-08-27 | 2008-06-04 | 中国科学院理化技术研究所 | One-dimensional nanostructured fluorescent chemical biosensor and method for making same and uses |
| WO2011085533A1 (en) * | 2010-01-15 | 2011-07-21 | 大连理工大学 | Fluorescent probe compounds, preparation method and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| JONG-HO KIM ET AL.: "The rational design of nitric oxide selectivity in", 《NATURE CHEMISTRY》 * |
| RONG MIAO ET AL.: "Modified silicon nanowires: a fluorescent nitric oxide biosensor with enhanced", 《JOURNAL OF MATERIALS CHEMISTRY》 * |
| 穆丽璇: "基于硅纳米线的光响应化学传感器研究", 《中国博士学位论文全文数据库 工程科技I辑》 * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102928391A (en) * | 2012-10-11 | 2013-02-13 | 中国科学院理化技术研究所 | Silicon nanowire ordered array-based pH fluorescence sensor and manufacturing method and application thereof |
| CN103712968A (en) * | 2014-01-06 | 2014-04-09 | 中国科学院理化技术研究所 | Silicon-nanowire-based fluorescence chemical sensor with selective fluorescence response to free copper ions and complexing copper ions |
| CN103712968B (en) * | 2014-01-06 | 2017-01-11 | 中国科学院理化技术研究所 | Silicon-nanowire-based fluorescence chemical sensor with selective fluorescence response to free copper ions and complexing copper ions |
| CN103937488B (en) * | 2014-03-25 | 2016-03-09 | 中国科学院理化技术研究所 | Based on the alkaline phosphatase fluorescence chemical sensor of silicon nanowires and method for making and application |
| CN103937488A (en) * | 2014-03-25 | 2014-07-23 | 中国科学院理化技术研究所 | Silicon-nanowire-based alkaline phosphatase fluorescent chemosensor, preparation method and application |
| CN104155277A (en) * | 2014-08-27 | 2014-11-19 | 中国科学院理化技术研究所 | Preparation method of sulfur ion fluorescent chemical sensor based on silicon nanowire or silicon nanowire array |
| CN104792758A (en) * | 2015-04-22 | 2015-07-22 | 中国科学院理化技术研究所 | Preparation method for hydrogen sulfide fluorescent chemosensor based on silicon nanowire or silicon nanowire array |
| CN108913120A (en) * | 2018-06-21 | 2018-11-30 | 中国科学院理化技术研究所 | A kind of single silicon nanowire fluorescence chemical sensor and its preparation method and application for detecting hypochlorite |
| 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 |
| CN109897630A (en) * | 2019-04-17 | 2019-06-18 | 中国科学院理化技术研究所 | A kind of nano wire, preparation method, the Ratio-type fluorescence chemical sensor comprising the nano wire and application |
| CN109897630B (en) * | 2019-04-17 | 2021-12-14 | 中国科学院理化技术研究所 | Nanowire, preparation method thereof, ratio type fluorescence chemical sensor containing nanowire and application |
| CN113686927A (en) * | 2021-08-06 | 2021-11-23 | 上海纳感医疗科技有限公司 | Method for improving biochemical detection selectivity of silicon nanowire sensor |
| CN113686927B (en) * | 2021-08-06 | 2024-02-09 | 上海纳感医疗科技有限公司 | Method for improving biochemical detection selectivity of silicon nanowire sensor |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102419319A (en) | Silicon nano wire based sensor having selective fluorescence response on nitric oxide | |
| CN101974326B (en) | Method for preparing novel fluorescent silica nanospheres | |
| Lan et al. | A carbon dot-based fluorescence turn-on sensor for hydrogen peroxide with a photo-induced electron transfer mechanism | |
| CN103926234B (en) | A kind of monolayer nanometer gold surface reinforced Raman active substrate and preparation method thereof | |
| CN103937488B (en) | Based on the alkaline phosphatase fluorescence chemical sensor of silicon nanowires and method for making and application | |
| CN106749359B (en) | A kind of synthesis and application for detecting hydrogen peroxide novel fluorescence probe | |
| CN110272734B (en) | Preparation method and application of high-quantum-yield carbon quantum dots for NO detection | |
| CN107556305B (en) | Fluorescent probe for detecting aluminum ions, preparation method and application | |
| US20140154184A1 (en) | Time-gated fluorescence imaging with si-containing particles | |
| CN107589162A (en) | A kind of preparation method and application based on complex of iridium Photoelectrochemistrbiosensor biosensor | |
| CN109607512A (en) | It is a kind of to pass through the pyrolysis standby carbon quantum dot of system with molecular sieve for preparing and its in the application of hydrogen peroxide context of detection | |
| Zhang et al. | Synthesis of novel fluorescent carbon quantum dots from Rosa roxburghii for rapid and highly selective detection of o-nitrophenol and cellular imaging | |
| Sawalha et al. | Optical properties and photoactivity of carbon nanodots synthesized from olive solid wastes at different carbonization temperatures | |
| CN105018070A (en) | Preparation method of mercury ion rate probe based on rhodamine derivative and quantum dot fluorescent microsphere | |
| Sun et al. | A porous material excited by near-infrared light for photo/chemodynamic and photothermal dual-mode combination therapy | |
| CN110194951A (en) | Tetraphenyl ethylene derivatives fluorescent probe and preparation method thereof | |
| Liu et al. | Surface modification of gold nanoparticles by cetirizine through surface plasmon resonance and preliminary study of the in vitro cellular cytotoxicity | |
| CN101851502A (en) | Ru(bpy)3-doped Ag@SiO2 fluorescent nano particles and preparation method thereof | |
| Zhao et al. | Ultrasensitive electrochemiluminescence biosensor for permethrin based on iron oxide nanomaterials and Au nanoparticles | |
| CN103712968B (en) | Silicon-nanowire-based fluorescence chemical sensor with selective fluorescence response to free copper ions and complexing copper ions | |
| CN113201335A (en) | Enhanced fluorescent carbon dot, preparation method and application in cadmium ion detection | |
| CN102928391B (en) | Silicon nanowire ordered array-based pH fluorescence sensor and manufacturing method and application thereof | |
| Foroughi et al. | A label-free electrochemical biosensor based on 3D cubic Eu 3+/Cu 2 O nanostructures with clover-like faces for the determination of anticancer drug cytarabine | |
| CN110144049B (en) | Copper-terephthalic acid nano-particle, preparation method and application thereof | |
| CN108913120B (en) | Single silicon nanowire fluorescence chemical sensor for detecting hypochlorite, and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20120418 |