Technical background
Detect the polycomponent in the biased sample in the immunoassay, adopt parallel single component analytical method usually, this method need be carried out a plurality of testing processes synchronously, the test set complexity, and reagent consumption is many, and workload is excessive.How in single flow process, to realize the synchronous detection of a plurality of indexs, become the research focus of immunoassay in recent years.
The polynary synchronous immunodetection that sees report at present mainly is divided into: spatial discrimination pattern and multi-tracer pattern.The spatial discrimination pattern need adopt comparatively complicated array apparatus, and difficulty of processing is bigger, and the technique for fixing of immunoreagent is had relatively high expectations, and requires each array point uniformity, and can not be inter-adhesive; Especially when using electrochemical mode to detect, the requirement of counter electrode array is more strict, and the distance that needs to keep enough is to avoid cross reaction; This method is also higher to the requirement of detector, needs to adopt expensive array detector, therefore, is subjected to certain restriction on use range.
In the multi-tracer pattern, when marking thing with enzyme, different markers has distinct optimum analysis condition usually, in same analysis system, unite simply and use multiple marker, can only in the optimum analysis condition of different markers, select a tendency towards compromise condition, caused the reduction of analytical effect thus, marker is many more, selects difficulty big more; And in such as optics or Electrochemical Detection mode, the resolution efficient of different markers is often limited, and the signal overlap problem is sometimes very outstanding, and therefore, the research temperature of this method greatly reduces in recent years.
Summary of the invention
The technology of preparing that the purpose of this invention is to provide a kind of magnetically encoded nano-wire, and provide a kind of brand-new marker for polynary synchronous immunodetection with this nano wire.
Magnetically encoded nano-wire of the present invention adopts electrochemical deposition technique, and in the nanometer blind hole on anodic oxidation aluminium formwork, the electrochemical deposition of magnetic/nonmagnetic substance is carried out in layering respectively.
Described magneticsubstance is a kind of in iron, the nickel; Described nonmagnetic substance is the copper metal.
The nanometer blind hole of described anodic oxidation aluminium formwork is evenly distributed, and is parallel to each other and perpendicular to template surface, and the regular hexagon that is high-sequential is arranged.Nanometer blind hole mean diameter is 50nm, and nanometer blind hole density is 10
6Individual/mm
2, the nanometer blind hole road degree of depth is 10 μ m.
Described magnetically encoded nano-wire diameter is 50nm, and the degree of depth is 750nm.
A kind of preparation method of magnetically encoded nano-wire is characterized in that adopting following steps:
A) preparation anodic oxidation aluminium formwork: aluminium foil is cut into suitable dimension, is placed in the 0.3M oxalic acid solution ice bath, voltage 40V anodic oxidation 1 hour through electrochemical etching, cleaning; Then the template after the oxidation was soaked 1 hour in phosphoric acid and chromic acid mixture, repeat the identical oxidation step in front again; Carry out the ladder step-down at last and handle, obtain porous anodic alumina template, the mol ratio of phosphoric acid and chromic acid is 3: 1 in the mixed solution;
B) preparation electrolytic solution: prepare electrolytic iron liquid respectively, nickel electrolyte, copper electrolyte is standby, wherein:
Electrolytic iron liquid is by FeSO
47H
2O 100g/L, (NH
4)
2SO
415g/L, MgSO
430g/L, xitix 1g/L, glycerol 2mg/L mixes, adjust pH to 3;
Nickel electrolyte is by NiSO
46H
2O 60g/L, boric acid 30g/L, MgSO
430g/L, Na
2SO
45g/L mixes adjust pH to 3;
Copper electrolyte is by CuSO
420g/L, MgSO
420g/L, C
6H
8O
7H
2O 15g/L mixes adjust pH to 3;
C) preparation magnetically encoded nano-wire: respectively with anodic oxidation aluminium formwork and stereotype as anode and negative electrode, adopt the 50Hz alternating-current, in corresponding electrolytic solution, carry out electrochemical deposition according to the metallic substance of making nano wire, remove the template aluminum substrate that is obtained behind the electrochemical deposition with saturated tin chloride then, template is immersed dissolving removal in the sodium hydroxide solution, with deionized water clean repeatedly, magnetic support is settled down to pH value of solution=7.0, after supersound process, obtain homodisperse relevant nanometer line solution.
Described ladder step-down was behind secondary oxidation, reduced once oxidation voltage every 10 minutes, and each step-down 2V is till voltage drops to 10V, with the blocking layer of attenuate and homogenizing anodic oxidation aluminium formwork; The metallic substance that is used to make nano wire is selected magneticsubstance iron or nickel for use, and nonmagnetic substance copper.
A kind of purposes of magnetically encoded nano-wire is characterized in that: the multi-tracer that this magnetically encoded nano-wire is used as immunodetection.
Compare with prior art, advantage of the present invention is: this magnetically encoded nano-wire thing that serves as a mark is used for polynary synchronous immunodetection, and is simple in structure, cost is lower, the serviceable indicia thing is more, the analysis condition unanimity of different coding marker.
Embodiment
Embodiment 1:
The preparation anodic oxidation aluminium formwork
Aluminium foil is cut into suitable dimension, is placed in the 0.3M oxalic acid solution ice bath, voltage 40V anodic oxidation 1 hour through electrochemical etching, cleaning; Then the template after the oxidation was soaked 1 hour in phosphoric acid and chromic acid mixing solutions (mol ratio 10: 3), repeat the identical oxidation step in front again; Carry out the ladder step-down at last and handle, can obtain porous anodic alumina template.
Preparation electrolytic solution
Electrolytic iron liquid is by FeSO
47H
2O 100g/L, (NH
4)
2SO
415g/L, MgSO
430g/L, xitix 1g/L, glycerol 2mg/L mixes, adjust pH to 3;
Copper electrolyte is by CuSO
420g/L, MgSO
420g/L, C
6H
8O
7H
2O 15g/L mixes adjust pH to 3.
The preparation Fe nanowire
Respectively with anodic oxidation aluminium formwork and stereotype as two electrodes, adopt the 50Hz alternating-current, in electrolytic iron liquid, carry out electrochemical deposition, deposition voltage 15V, depositing time 5min makes Fe nanowire.
Preparation iron/copper nano-wire
Respectively with anodic oxidation aluminium formwork and stereotype as two electrodes, adopt the 50Hz alternating-current, at first in electrolytic iron liquid, carry out electrochemical deposition, deposition voltage 15V, depositing time 4min; Then, ultrasonic washing was carried out supersound process 5 minutes in copper electrolyte, carry out electrochemical deposition in copper electrolyte, deposition voltage 15V, and depositing time 1min obtains iron/copper nano-wire.
Preparation copper/iron/copper nano-wire
Respectively with anodic oxidation aluminium formwork and stereotype as two electrodes, adopt the 50Hz alternating-current, at first in copper electrolyte, carry out electrochemical deposition, deposition voltage 15V, depositing time 0.5min; Then, ultrasonic washing, supersound process is 5 minutes in electrolytic iron liquid, carries out electrochemical deposition in electrolytic iron liquid, deposition voltage 15V, depositing time 4min; After the ultrasonic washing, supersound process is 5 minutes in copper electrolyte, carries out electrochemical deposition in copper electrolyte, deposition voltage 15V, and depositing time 0.5min obtains copper/iron/copper nano-wire.
Remove the template aluminum substrate that is obtained after the different galvanic deposit with saturated tin chloride respectively, template immersed in the 1mol/L sodium hydroxide solution remove, with deionized water clean repeatedly, magnetic support is settled down to pH value of solution=7.0, obtain homodisperse iron, iron/copper, copper/iron/copper nano-wire solution through supersound process.
Embodiment 2:
The preparation anodic oxidation aluminium formwork
Aluminium foil is cut into suitable dimension, is placed in the 0.3M oxalic acid solution ice bath, voltage 40V anodic oxidation 1 hour through electrochemical etching, cleaning; Then the template after the oxidation was soaked 1 hour in phosphoric acid and chromic acid mixing solutions (mol ratio 10: 3), repeat the identical oxidation step in front again; Carry out the ladder step-down at last and handle, can obtain required anodic oxidation aluminium formwork.
Preparation electrolytic solution
Nickel electrolyte is by NiSO
46H
2O 60g/L, boric acid 30g/L, MgSO
430g/L, Na
2SO
45g/L mixes adjust pH to 3;
Copper electrolyte is by CuSO
420g/L, MgSO
420g/L, C
6H
8O
7H
2O 15g/L mixes adjust pH to 3.
Preparation nickel nano wire
Respectively with anodic oxidation aluminium formwork and stereotype as two electrodes, adopt the 50Hz alternating-current, in nickel electrolyte, carry out electrochemical deposition, deposition voltage 15V, depositing time 5min obtains the nickel nano wire.
Preparation nickel/copper nano-wire
Respectively with anodic oxidation aluminium formwork and stereotype as two electrodes, adopt the 50Hz alternating-current, at first in nickel electrolyte, carry out electrochemical deposition, deposition voltage 15V, depositing time 4min; Then, ultrasonic washing was carried out supersound process 5 minutes in copper electrolyte, carry out electrochemical deposition in copper electrolyte, deposition voltage 15V, and depositing time 1min obtains nickel/copper nano-wire.
Preparation copper/nickel/copper nano-wire
Respectively with anodic oxidation aluminium formwork and stereotype as two electrodes, adopt the 50Hz alternating-current, at first in copper electrolyte, carry out electrochemical deposition, deposition voltage 15V, depositing time 0.5min; Then, ultrasonic washing, supersound process is 5 minutes in nickel electrolyte, carries out electrochemical deposition in nickel electrolyte, deposition voltage 15V, depositing time 4min; At last, ultrasonic washing, supersound process is 5 minutes in copper electrolyte, carries out electrochemical deposition in copper electrolyte, deposition voltage 15V, depositing time 0.5min obtains copper/nickel/copper nano-wire.
Remove the template aluminum substrate that is obtained after the different galvanic deposit with saturated tin chloride respectively, template immersed in the 1mol/L sodium hydroxide solution remove, with deionized water clean repeatedly, magnetic support is settled down to pH value of solution=7.0, obtain homodisperse nickel, nickel/copper, copper/nickel/copper nano-wire solution through supersound process.