CN101726528A - Test chip, detection apparatus, and method for detecting analyte - Google Patents
Test chip, detection apparatus, and method for detecting analyte Download PDFInfo
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- CN101726528A CN101726528A CN200910205180A CN200910205180A CN101726528A CN 101726528 A CN101726528 A CN 101726528A CN 200910205180 A CN200910205180 A CN 200910205180A CN 200910205180 A CN200910205180 A CN 200910205180A CN 101726528 A CN101726528 A CN 101726528A
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Abstract
A test chip for detecting an analyte modified with a modulator releasing electrons upon photoexcitation, comprising: a semiconductor electrode part including a metal layer formed on a semiconductor layer; a probe immobilized on the metal layer, the probe trapping the analyte; and a counter electrode part including a conductive layer. A detection apparatus and a method for detecting an analyte are also disclosed.
Description
Technical field
The electric current that the present invention relates to use the light stimulus of the modification material by modifying analyte to take place detects the detection method of inspection chip, pick-up unit and the analyte of analyte.
Background technology
In the clinical examination of disease and diagnosis, come the gene of the disease origin that comprised in the detection of biological body sample and protein etc. with gene detection method, immunologic detection method.Particularly can enumerate immune colloidal gold chromatography method (Immunochromato), latex agglutination (Latex Agglutination), ferment immunization, chemiluminescence immunoassay method and gene amplification PCR method etc.
But these detection methods all have room for improvement from arbitrary viewpoint of simplification, rapidity and cost.
Thereby the electric current that has proposed to produce by the light stimulus of sensitizing coloring matter in the world discloses No. 2007/037341 pamphlet is used in the method that analyte detects.In this method, at first on electrode, form semiconductor layer, and on this semiconductor layer, fix the probe that can combine with analyte.Then, after utilizing probe substance to capture the analyte of using the sensitizing coloring matter modified, the sensitizing coloring matter irradiation of modifying analyte is made the light of sensitizing coloring matter excitation.Its result, just from modifying the sensitizing coloring matter generation electronics of analyte, the electronics that passes through to be taken place is admitted by semiconductor layer, and produces electric current.The electric current that is produced is detected.Here use bridging agents such as silane coupling agent, stationary probe on semiconductor layer.But, thereby since the lower detection efficiency that makes electric current of its electric conductivity of silane coupling agent reduce, so lower and so on the problem points of the detection sensitivity of analyte is just arranged.
Summary of the invention
That is, the invention provides following technical scheme:
(1) a kind of inspection chip, the analyte after being used for detecting modification material that utilization produces electronics by light stimulus and modifying comprises:
The semi-conducting electrode portion that possesses the metal level that is formed on the semiconductor layer;
Catch the probe on the above-mentioned metal level of being fixed on of analyte; And
Possess conductive layer to utmost point portion.
(2) in the inspection chip that technical scheme (1) is put down in writing, above-mentioned metal level is made up of the metal that dissolves by electrolytic solution.
(3) in the inspection chip that technical scheme (2) is put down in writing, above-mentioned electrolytic solution comprises iodine or iodide.
(4) in technical scheme (1)～(3) in any inspection chip of being put down in writing, above-mentioned metal level is made up of the metal that carries out chemisorption with above-mentioned probe.
(5) in the inspection chip that technical scheme (4) is put down in writing, the metal that carries out chemisorption with above-mentioned probe is a gold.
(6) in the inspection chip that technical scheme (4) is put down in writing, above-mentioned probe has mercapto as to carry out combine of chemisorption basic with above-mentioned metal level.
(7) a kind of pick-up unit is used for detecting and uses the analyte of modification material through modifying that produces electronics by light stimulus, comprising:
Received portion can receive the inspection chip checking the mode of chip and constitute, wherein this inspection chip comprises: the semi-conducting electrode portion that possesses the metal level that is formed on the semiconductor layer; Catch the probe on the above-mentioned metal level of being fixed on of analyte; With possess conductive layer to utmost point portion;
To modifying the light source that material carries out light stimulus, this trim analyte that above-mentioned inspection chip received in the above-mentioned inspection chip that is inserted in the portion of verifying is modified; And
Amperometric determination portion measures the electric current that flows out from the modification material of analyte being modified by the light stimulus of above-mentioned light source.
(8) in the pick-up unit that technical scheme (7) is put down in writing, above-mentioned light source sends the light of the wavelength of the modification material that excitation modifying analyte.
(9) a kind of detection method is used for detecting and uses the analyte of modification material through modifying that produces electronics by light stimulus, may further comprise the steps:
Comprising the semi-conducting electrode portion that possesses the metal level that is formed on the semiconductor layer; Catch the probe on the above-mentioned metal level of being fixed on of analyte; With possess conductive layer to using the sample comprise analyte on the inspection chip of utmost point portion, catch analyte with the above-mentioned probe that is fixed on the above-mentioned metal level thus;
To modify material and import analyte;
Irradiation is to modifying the light that material encourages; And
The electric current that detection is produced from the modification material through blasting.
(10) in the detection method that technical scheme (9) is put down in writing, also comprise: in above-mentioned semi-conducting electrode portion with above-mentionedly add the electrolyte medium be used to flow through electric current between to utmost point portion.
Description of drawings
Fig. 1 is the stereographic map as the pick-up unit 1 of an embodiment of the invention.
Fig. 2 is the block diagram of pick-up unit 1.
Fig. 3 is the stereographic map of the inspection chip 4 of use on pick-up unit 1.
Fig. 4 is the stereographic map that expression has the top plate of the semi-conducting electrode portion 15 that checks chip 4.
Fig. 5 is that expression has the stereographic map to the bottom plate of utmost point portion 16 of checking chip 4.
Fig. 6 is the stereographic map of the inspection chip 4 when upper substrate 13 is unloaded.
Fig. 7 is the sectional view that the structure of chip 4 is checked in expression.
Fig. 8 is that expression is checked the semi-conducting electrode portion 15 of chip 4 and to the synoptic diagram of the structure of utmost point portion 18.
Fig. 9 is the process flow diagram that a corpse or other object for laboratory examination and chemical testing is injected into the method for checking chip 4 that expression is undertaken by the user.
Figure 10 is the process flow diagram of the detection course of action of expression pick-up unit 1.
When Figure 11 is hybridization reaction and the synoptic diagram of the semi-conducting electrode portion 15 of electrolytic solution when adding.
Figure 12 is the chart of expression by the photoelectricity flow valuve that mensuration obtained of embodiment 1 and comparative example 1.
Figure 13 is the chart of photoelectricity flow valuve detected in embodiment 2 and comparative example 2.
Figure 14 is the chart that is illustrated among the data that obtained in embodiment 2 and the comparative example 2, derives from the current value of modifying material.
Figure 15 is the chart of photoelectricity flow valuve detected in embodiment 3, comparative example 3 and comparative example 4.
Figure 16 is the chart of photoelectricity flow valuve detected in embodiment 4, comparative example 5 and comparative example 6.
Figure 17 is the chart of the S/N ratio under each thickness detected in embodiment 5.
Embodiment
Below, record and narrate the present invention based on embodiment shown in the drawings.In addition, this disclosed embodiment should be considered in all respects as example rather than restrictive.Scope of the present invention is not the explanation of above-mentioned embodiment but represented and then also comprise and the meaning of the scope equalization of claim and all changes in its scope by the scope of claim.
(structure of pick-up unit)
Fig. 1 is the stereographic map of the related pick-up unit of expression an embodiment of the invention.This pick-up unit is to detect the device that the nucleic acid take from biological cell or synthesized with manual type, protein, peptide etc. have the analyte of specific bond.This pick-up unit 1 for example can detect the mRNA of human papilloma virus as the reason virus of cervix cancer (below be referred to as HPV) from corpse or other object for laboratory examination and chemical testing sample.
The pick-up unit 1 of present embodiment possesses chip that insert to check chip 4 and is received portion 3 and show the display 2 of testing result.In addition, check that chip 4 also possesses sample inlet 11.
This checks that chip 4 is chips that disposable HPV detects usefulness, and the chip that is inserted into pick-up unit 1 is received portion 3.Check that chip 4 has following function, promptly, catch the mRNA of the HPV that uses the modification material modified that produces electronics by means of light stimulus by injecting corpse or other object for laboratory examination and chemical testing sample from sample inlet 11.
Fig. 2 is the block diagram of the structure of expression pick-up unit 1.Pick-up unit 1 possesses light source 5, galvanometer 6, power supply 32, A/D transformation component 7, control part 8 and display 2.
(checking the structure of chip 4)
Use Fig. 3～Fig. 8 to describe with regard to the structure of employed inspection chip 4 on the pick-up unit 1.
Fig. 3 is a stereographic map of checking chip 4.Check that chip 4 possesses: infrabasal plate 16, be arranged at infrabasal plate 16 tops upper substrate 13, be sandwiched in the silicon rubber 12 in infrabasal plate 16 and the upper substrate 13.On upper substrate 13, also be provided with in addition and lead to inner sample inlet 11.
Fig. 4 be make Fig. 3 inspection chip 4 in the horizontal direction to right rotation 90 degree, and the stereographic map of the upper substrate 13 under the state of Rotate 180 degree in vertical direction.On the surface of this upper substrate 13, be formed with: semi-conducting electrode portion 15 and the contact conductor 14 that is connected to semi-conducting electrode portion 15.Upper substrate 13 is formed by silicon dioxide (SiO2), and contact conductor 14 is two-layer and form by tin indium oxide (ITO) and antimony dopant tin oxide (ATO).Use Fig. 8 to narrate in the back about semi-conducting electrode portion 15.
Fig. 5 is inspection chip 4 stereographic map of the infrabasal plate 16 under the state of right rotation 90 degree in the horizontal direction that makes Fig. 3.On the surface of infrabasal plate 16, be formed with respectively: to utmost point portion 18, be connected to contact conductor 17, reference electrode 31, be connected to the contact conductor 30 of reference electrode 31 utmost point portion 18.
Fig. 6 is the stereographic map of the inspection chip 4 of the upper substrate 13 with the inspection chip 4 of Fig. 3 when unloading up.Silicon rubber 12 is configured on the infrabasal plate 16 in the mode of surrounding utmost point portion 18 and reference electrode portion 31 as shown in Figure 6.Be connected to the contact conductor 17 of utmost point portion 18 and the contact conductor 30 that is connected to reference electrode portion 31 are extended to outside the frame in the frame of silicon rubber 12.This extends to the outer contact conductor 17 of frame and contact conductor 30 couples together with power supply 32.
The sample inlet 11 that is arranged at upper substrate 13 is the holes that connect upper substrate 13.Corpse or other object for laboratory examination and chemical testing sample and electrolytic solution described later are injected in the frame of silicon rubber 12 from this sample inlet 11.
Fig. 7 is the sectional view of A-A cross section structure of the inspection chip 4 of presentation graphs 3.As shown in Figure 7, check that the upper substrate 13 and the infrabasal plate 16 that are comprised on the chip 4 dispose across silicon rubber 12.Between upper substrate 13 and infrabasal plate 16, be formed with space 25.Across this space 25, be formed on the upper substrate 13 semi-conducting electrode portion 15 be formed on infrabasal plate 16 on mutually opposed to utmost point portion 18 and reference electrode portion 31 (not shown).On this space 25, inject corpse or other object for laboratory examination and chemical testing sample and electrolytic solution described later via sample inlet 11.
As shown in Figure 7, the contact conductor 14 that is connected to semi-conducting electrode portion 15 extends to outside the space 25 along upper substrate 13, is connected to contact conductor in the utmost point portion 18 17 and the contact conductor 30 (not shown) that is connected to reference electrode portion 31 are extended to outside the space 25 along infrabasal plate 16.This contact conductor 14 is connected to galvanometer 6, and contact conductor 17 and contact conductor 30 are connected to power supply 32.
In addition, in the present embodiment, surface at upper substrate 13 forms semi-conducting electrode portion 15, form utmost point portion 18 and reference electrode portion 31 on the surface of infrabasal plate 16, but semi-conducting electrode portion 15, to the configuration relation of utmost point portion 18, reference electrode portion 31 so long as each electrode and other electrodes are configured in the frame of silicon rubber 12 does not non-contiguously then limit especially.For example, can also be on same substrate configuring semiconductor electrode part 15, to utmost point portion 18 and reference electrode portion 31.
Here, further be described in detail with regard to semi-conducting electrode portion 15 shown in Figure 4.Fig. 8 is an expression semi-conducting electrode portion 15 and to the synoptic diagram of the structure of utmost point portion 18.
Fixing the probe 23 that seizure uses the mRNA24 of the HPV after the modification material 22 that produces electronics by means of light stimulus is modified to use on the metal level 19 that in semi-conducting electrode portion 15, is comprised.This modification material 22 is ruthenium fluor-complexs, modifies material 22 and combines and modify mRNA by carrying out peptide with mRNA.
The contact conductor 14 that is connected in the semi-conducting electrode portion 15 is connected to galvanometer 6, and the contact conductor 30 that is connected in the contact conductor 17 that connected in the utmost point portion 18 and the reference electrode portion 31 is connected to power supply 32.Galvanometer 6 couples together with power supply 32, is determined at semi-conducting electrode portion 15 and the electric current to flowing through between the utmost point portion 18 with this galvanometer 6.
The conductive layer 21 that is comprised in the semi-conducting electrode portion 15 is by this two-layer composition of layer of the layer of the tin indium oxide that forms by sputter (ITO) and the antimony dopant tin oxide (ATO) that forms by sputter on this ITO layer.Semiconductor layer 20 is made up of the layer of the titanium dioxide that forms by sputter (TiO2).The layer of the gold (Au) that metal level 19 is formed by general evaporation is formed.Utmost point portion 18 is made up of the layer of the platinum that forms by sputter.
(having adopted the detection method of HPV pick-up unit)
Just adopted the detection method of pick-up unit 1 to describe with reference to Fig. 9～Figure 11 with above-mentioned formation.Fig. 9 is the process flow diagram that a corpse or other object for laboratory examination and chemical testing is injected the method for checking chip 4 that the expression user carries out.Figure 10 is the process flow diagram of the detection course of action of expression pick-up unit 1.Figure 11 is that hybridization is when (hybridization) and the synoptic diagram of the semi-conducting electrode portion 15 of electrolytic solution when adding.
According to the process flow diagram of Fig. 9, corpse or other object for laboratory examination and chemical testing sample is injected from the sample inlet 11 of checking chip 4 step S1 user.This corpse or other object for laboratory examination and chemical testing sample is to homogenize and extract to handle and through refining mRNA from the uterine cervix cell.By this step S1 as shown in Figure 11, the probe 23 on the metal level 19 is caught the mRNA24 of the HPV in the corpse or other object for laboratory examination and chemical testing sample by hybridization.
At step S2, the user will check that the solution in the chip 4 discharge from sample inlet 11, and with the hybridization cleansing solution to washing in the inspection chip 4.
At step S3, the user injects the modification material 22 that comprises the base assortment that can combine with the mRNA24 of HPV from sample inlet 11.22 couples of mRNA24 that captured with probe 23 of the modification material that is injected modify.
At step S4, the user will check that the solution in the chip 4 discharge from sample inlet 11, and with washing with damping fluid to washing in the inspection chip 4.
At step S5, the user injects electrolytic solution from sample inlet 11.This electrolytic solution comprises iodine with as electrolyte, comprises the tetrapropyl ammonium iodide with as supporting electrolyte, and comprises acetonitrile and ethylene carbonate are mixed into 6: 4 organic solvent by volume with as solvent.Then be included in iodine in the electrolytic solution with regard to dissolution of metals layer 19 if add electrolytic solution.
Use Figure 11 to describe with regard to the dissolving of this metal level 19.Figure 11 is the synoptic diagram of when hybridization and the electrolytic solution semi-conducting electrode portion 15 when adding.
The gold atom of mercapto that probe 23 has by probe 23 (SH yl) and metal level 19 carries out covalent bond, and is fixed on the metal level 19.Covalent bond is firm combination.Therefore, can prevent that probe 23 is peeled off from metal level 19 when step S1 makes the operation of washing of the operation of its hybridization and step S2.
If the iodine that interpolation electrolytic solution then is included in the electrolytic solution just makes metal level 19 dissolvings of being made up of gold (Au), probe 23 is configured on the semiconductor layer 20.Thus, the electronics that is produced from the modification material 22 that is encouraged by means of the rayed of light source 5 just is fed into semiconductor layer 20 expeditiously.
Figure 10 is the process flow diagram of the detection course of action of expression pick-up unit 1.Carried out the user after the flow process of Fig. 9, the user will check that chip 4 is inserted into the chip insertion mouth 3 of pick-up unit shown in Figure 11, and beginning is measured in indication on display 2.
At step S6, the contact conductor 14,17,31 that is inserted into the inspection chip 4 of pick-up unit 1 is connected to galvanometer 6 and power supply 32.Then, power supply 32 applies the 0V current potential with reference electrode portion 31 as benchmark in semi-conducting electrode portion 15.
At step S7, light source 5 shines the modification material 22 that the mRNA24 to HPV modifies with laser light, so that modify material 22 excitations.Electronics takes place in the modification material 22 that is energized, and the electronics that is taken place is transported to semiconductor layer 20.Its result just flows through electric current in semi-conducting electrode portion 15 and between to utmost point portion 18.
At step S8, measure with galvanometer 6 and to result from that the electronics of step S5 moves and in semi-conducting electrode portion 15 and the electric current that flows through between to utmost point portion 18.Because the current value that galvanometer 6 is measured has correlativity with the number of modifying material 22, so can carry out the quantitative measurement of HPV based on the current value of being measured.
At step S9, at first, the current value that passes through digital conversions by A/D transformation component 7 is imported into control part 8.Then, control part 8 is based on the inspection amount line of the relation of expression current value that is made in advance and HPV amount, according to the HPV amount of estimating through the current value of digital conversion in the corpse or other object for laboratory examination and chemical testing sample.Then, control part 8 is created the testing result picture that is used for showing the HPV amount of being estimated on display 2.
At step S10, the testing result picture of being created by control part 8 is sent on the display 2 and is presented at display 2.
In addition, though analyte is the mRNA24 of HPV in the present embodiment, can also adopt nucleic acid, protein or the peptide etc. taked from biological cell or synthesized with manual type as analyte.At this moment, probe 23 for example can adopt nucleic acid, protein or peptide etc. so long as can catch the material of analyte and get final product.
And, though used the ruthenium fluor-complex in the present embodiment as modifying material 22, so long as then do not limit especially by means of the material of light source 5 excitations and generation electronics.For example can enumerate metal fluor-complex, organic pigment and quantum dot etc.Particularly can list the metal phthalein cyanine, the ruthenium fluor-complex, the osmium fluor-complex, the iron fluor-complex, the zinc fluor-complex, 9-benzene oxa anthracenes pigment, cyanine class pigment, metal cyanine class pigment, the oxa anthracenes pigment, the triphenylmethane pigment, acridine pigment oxazine class pigment, the Coumarins pigment, merocyanine class pigment, if red cyanines class pigment, polymethine class pigment, porphyrin class pigment, phthalocyanine class pigment, rhodamine class pigment, the oxa anthracenes pigment, chlorophyll class pigment, eosin class pigment, mercurochrome class pigment, indigo class pigment or cadmium selenide etc.
In addition, in the present embodiment, light source 5 so long as send does not then limit the light source of the light of the wavelength that the material of modifying analyte encourages especially.For example can enumerate laser, light emitting diode, inorganic electroluminescent element, organic electroluminescent device, white light source, possess the white light source of optical filter etc.
Represented after probe 23 captures the mRNA24 of HPV the example of modifying with the mRNA24 that modifies 22 couples of HPV of material in the present embodiment.Also can be to modify the mRNA24 of HPV, and the mRNA24 that catches HPV by probe 23 carry out the detection of the mRNA24 of HPV with modifying material 22.In addition, when analyte and probe are nucleic acid, can also enumerate to make and modify material and be combined in method between the double-strandednucleic acid that the probe that captures analyte and analyte form based on interpolation (Intercalation).
In addition, though used gold as metal level 19 in the present embodiment, metal level 19 is so long as can get final product with the metal that probe 23 combines.Preferably can with probe 23 covalently bound metals.And then the metal that preferably can combine with the mercapto of probe 23.But for example illustration gold, platinum, silver, palladium, nickel, mercury, rhodium, ruthenium, copper or their alloy etc.Though used evaporation as making metal level 19 be formed on method on the semiconductor layer 20 in addition in the present embodiment, also can use sputter, mint-mark, screen printing, electroplating processes or sol-gal process to wait and form.
Though in the present embodiment, semiconductor layer 20 uses titanium dioxide (TiO2), and semiconductor layer 20 so long as can obtain can be accepted to be got final product by the material of the energy level of the electronics that excitation produced of modifying material 22.For example, can enumerate semiconductors such as silicon, germanium, titanium dioxide (TiO2), indium oxide (In2O3), tin oxide (SnO2), zinc paste (ZnO), cadmium selenide (CdSe), cadmium sulfide (CdS), gallium nitride (GaN), titanium nitride compound semiconductors such as (TiN) or organism semiconductor etc.
Though in the present embodiment, conductive layer 21 has used with tin indium oxide (ITO) and the formed conductive layer of antimony dopant tin oxide (ATO), so long as conductive material does not then limit especially.For example can enumerate metals such as platinum, gold, silver, copper, electric conductivity pottery or metal oxide etc.In addition, self also bring into play under the function situation as conductive material at semiconductor layer 20, conductive layer 21 can omit.
In addition, though in the present embodiment, used usefulness platinum formed to utmost point portion 18, so long as conductive material does not then limit especially to utmost point portion.For example can enumerate metals such as gold, silver, copper, electric conductivity pottery or metal oxide etc.
Though in above-mentioned present embodiment, used iodine as making metal level 19 dissolved substances and electrolyte, making metal level 19 dissolved substances and electrolyte also can be different materials.
In addition, though probe 23 directly is combined on the metal level 19 in the present embodiment, also can between probe 23 and metal level 19, there be bridging agents such as dithioglycol.
In addition, pick-up unit 1 in the present embodiment and inspection chip 4 also can make a distinction stationary probe 23 one by one by a plurality of zones separated from one another on metal level 19, and each zone are utilized individually the rayed of light source 5.Thus, just can measure a plurality of samples with a semi-conducting electrode portion 15.By by the fixing multiple probe in each zone, check that with regard to enough one of energy chip 4 carries out the evaluation of many corpse or other object for laboratory examination and chemical testing, multinomial range estimation is fixed.
And though in the present embodiment, power supply 32 has applied the 0V current potential with reference electrode portion 31 as benchmark in semi-conducting electrode portion 15, and this reference electrode portion 31 can omit.In the case, power supply 32 is just to apply the 0V current potential to utmost point portion 18 as benchmark in semi-conducting electrode portion 15.
[embodiment]
Embodiment 1 (research that whether forms at the metal level of semi-conducting electrode)
(making of semi-conducting electrode portion)
Semi-conducting electrode portion forms conductive layer, semiconductor layer and metal level and makes on substrate.Conductive layer is to have formed the tin indium oxide (ITO) of 100nm thickness and the layer of antimony dopant tin oxide (ATO) by sputter on the substrate that is made of silicon dioxide (SiO2).Semiconductor layer is the layer that has formed the titanium dioxide (TiO2) of 10nm thickness on conductive layer by sputter.Metal level is the layer that has formed the gold thin film of 10nm thickness on semiconductor layer by evaporation.Semiconductor layer comprises the semiconductor of titanium or chromium by employing, and the bounding force of metal level and semiconductor layer is improved.Being connected with the semi-conducting electrode that is used for being connected with galvanometer in this semi-conducting electrode portion goes between.
(to the making of utmost point portion)
To utmost point portion is that the platinum film that has formed 200nm thickness by sputter on the substrate that is made of silicon dioxide (SiO2) is made.Be connected be used for being connected the utmost point is gone between at this on to utmost point portion with galvanometer.
(fixing of probe)
At first, semi-conducting electrode portion is immersed in makes the aqueous solution (concentration and probe concentration 10 μ M) 18 hours that probe disperses.This probe is the DNA (24base) with mercapto.After having carried out dipping,, and make its dry 30 minutes with ultrapure water washing semi-conductor electrode part.Its result, the mercapto that has by probe combines with the gold atom of metal level, and probe just is fixed on the metal level.
(making of analyte)
Analyte be make modify material be incorporated into comprise with probe mutually complementary base assortment DNA and make.It is Pulsar650 (バ イ オ サ one チ テ Network ノ ロ ジ one ズ ジ ヤ パ Application company) that the modification material has used sensitizing coloring matter.This sensitizing coloring matter is the ruthenium fluor-complex, and the DNA peptide is incorporated on this ruthenium fluor-complex.
(hybridization of analyte and probe)
At first, will be configured as the next door with silicon rubber (thickness 0.2mm) around the semi-conducting electrode portion.Injecting 10 μ L hybridization solution with the formed space of this silicon rubber.The solution that will utilize the DNA (concentration 10 μ M) that modifies after the material modification and hybridization buffer (Affymetrix company) to mix has been used in this hybridization with solution.
Then, the glass sheet that closes the lid on silicon rubber becomes the moist state of solution and hybridizes.At this moment, the probe in the semi-conducting electrode portion is caught with the analyte of modifying the material modified.Hybridization be with 45 ℃, leave standstill and it reacted in 1 hour.Behind the hybridization reaction, come the washing semi-conductor electrode part with washing with buffering agent (WashbufferA, Affymetrix company) and ultrapure water, and make its drying with fan blower.
(modulation of electrolytic solution)
Electrolytic solution be with the 4-propyl ammonium chloride of 0.6M (NPr4Cl) as supporting electrolyte salt, as electrolyte it is dissolved in the iodine of 0.06M and acetonitrile (AN) and ethylene carbonate (EC) are mixed into volume ratio in 6: 4 the solvent and modulate.
(photocurrent mensuration)
The substrate that will have the semi-conducting electrode portion that makes analyte and probe hybridization is configured to sidewall with silicon rubber (thickness 0.2mm) on every side.Injecting 10 μ L electrolytic solution with the formed space of this silicon rubber.Then, seal with the substrate that has utmost point portion from substrate top with the semi-conducting electrode portion that has been filled electrolytic solution.Thus, just become semi-conducting electrode portion and utmost point portion is contacted with the state of electrolytic solution.
Then, be connected to galvanometer with the semi-conducting electrode lead-in wire with to utmost point lead-in wire.Then from semi-conducting electrode portion direction towards to utmost point portion from light source irradiation light.Light source has used the LASER Light Source of wavelength 473nm, intensity 13mW.Its result modifies material and is energized, and the electronics that is taken place from the modification material through blasting is transported to semiconductor layer, flows through electric current in semi-conducting electrode portion and between to utmost point portion.This current value is measured.
Comparative example 1
Comparative example 1 is measured with method similarly to Example 1 except the operation of formation metal level on the semiconductor layer.
(result)
Figure 12 is the chart of expression by the current value that mensuration obtained of embodiment 1 and comparative example 1.In the method for embodiment 1, obtained the current value of 229nA.Then obtained the current value of 80nA in the method for comparative example 1 with respect to this.
As can be known by form metal level on semiconductor layer, obtain about 3 times current value according to this fact, the detection sensitivity of electric current has improved.
Embodiment 2 (based on modifying the detection of material) through the amperometric determination of the analyte of modification
(making of semi-conducting electrode portion)
Make of method similarly to Example 1.
(to the making of utmost point portion)
Make of method similarly to Example 1.
(fixing of probe)
Carry out with method similarly to Example 1.
(modulation of analyte)
Preparation make modify material be incorporated into comprise with probe mutually the analyte (analyte A) and making of the DNA of complementary base assortment modify material be incorporated into do not comprise with probe mutually the analyte (analyte B) of the DNA of complementary base assortment as analyte.
The modification material has used the Pulsar650 (バ イ オ サ one チ テ Network ノ ロ ジ one ズ ジ ヤ パ Application company) as sensitizing coloring matter.This sensitizing coloring matter is the ruthenium fluor-complex, combines with DNA by the peptide combination.
(utilizing the seizure of the analyte of probe)
Carry out hybridization reaction with the probe on analyte A or analyte B and the metal level.At first, will be configured to the next door with silicon rubber (thickness 0.2mm) around the semi-conducting electrode portion.Injecting 10 μ L hybridization solution with the formed space of this silicon rubber.The solution that will utilize the DNA (concentration 10 μ M) that modifies after the material modification and hybridization buffer (Affymetrix company) to mix has been used in this hybridization with solution.
Then, the glass sheet that closes the lid on silicon rubber becomes the moist state of solution and hybridizes.Hybridization be with 45 ℃, leave standstill and it reacted in 1 hour.Behind the hybridization reaction, after washing with buffering agent (WashbufferA, Affymetrix company) and ultrapure water with washing, make its drying with fan blower.
(modulation of electrolytic solution)
Carry out with method similarly to Example 1.
(amperometric determination)
The substrate that will have the semi-conducting electrode portion that makes analyte A or analyte B and probe hybridization is configured to sidewall with silicon rubber (thickness 0.2mm) on every side.Injecting 10 μ L electrolytic solution with the formed space of this silicon rubber, and sealing with the substrate that has utmost point portion from substrate top with the semi-conducting electrode portion of having filled electrolytic solution.Thus, just become semi-conducting electrode portion and utmost point portion is contacted with the state of electrolytic solution.
Then, be connected to galvanometer with the semi-conducting electrode lead-in wire with to utmost point lead-in wire.From semi-conducting electrode portion direction towards to utmost point portion from light source irradiation light.This light source has used the LASER Light Source of wavelength 473nm, intensity 13mW.Thus, the modification material of modifying analyte is energized, and the electronics that is taken place from the modification material through blasting is transported to semiconductor layer, flows through electric current in semi-conducting electrode portion and between to utmost point portion.This current value is measured.
In addition, in order to measure the current value that derives from electrode, do not make the probe hybridization on analyte and the metal level operation measured electric current.The electric current that derives from electrode is meant that electrode self is energized the electric current that is taken place by rayed.
Comparative example 2
Except the operation of formation metal level on the semiconductor layer, carry out the detection of analyte similarly to Example 2.
(result)
Figure 13 is the chart at the detected current value of embodiment 2 and comparative example 2.
In embodiment 2, have with the DNA (analyte A) of the complementary mutually base assortment of probe in use and to have carried out under the situation of hybridization, detected current value is 36.3nA.
In addition, use do not have with probe mutually the DNA (analyte B) of complementary base assortment carried out under the situation of hybridization, current value is 24.7nA.This current value is equal with the current value 24.9nA that derives from electrode.Can confirm according to this fact, make current value detected under the situation of its hybridization with analyte A is not by the current value that non-specific adsorption produced of analyte to semi-conducting electrode portion, but by identifying the current value that specific detection produced of assortment.
Figure 14 is the chart that is expressed among the data that obtained in embodiment 2 and the comparative example 2, derives from the current value of modifying material.Derive from the current value of modifying material and be meant the value that derives from the current value of electrode from deduction the current value that mensuration obtained of analyte.
Derive from the current value of modifying material as can be known, used situation (embodiment a 2) side of the semi-conducting electrode portion that comprises metal level bigger approximately 4.5 times than the situation of having used the semi-conducting electrode portion (comparative example 2) that does not comprise metal level.
Embodiment 3 (having used the effect of the semi-conducting electrode of the modification material that carries out long wavelength's excitation)
(making of semi-conducting electrode portion)
Semi-conducting electrode portion forms conductive layer, semiconductor layer and metal level and makes on substrate.Conductive layer is the layer that has formed the tin indium oxide (ITO) of 100nm thickness on the substrate that is made of silicon dioxide (SiO2) by sputter.Semiconductor layer is the layer that has formed the indium oxide (In2O3) of 10nm thickness on conductive layer by sputter.Metal level is the layer that has formed the gold thin film of 10nm thickness on semiconductor layer by evaporation.Being connected with the semi-conducting electrode that is used for being connected with galvanometer in this semi-conducting electrode portion goes between.
Then, the semi-conducting electrode portion of made is carried out sintering (150 ℃) in oxygen atmosphere, so that the bounding force of metal level and semiconductor layer improves.
(to the making of utmost point portion)
To utmost point portion is that the platinum film that has formed 200nm thickness by sputter on the substrate of being made up of silicon dioxide (SiO2) is made.Be connected be used for being connected the utmost point is gone between at this on to utmost point portion with galvanometer.
(fixing of probe substance)
At first, semi-conducting electrode portion is immersed in makes the aqueous solution (concentration and probe concentration 10 μ M) 18 hours that probe disperses.This probe is the DNA (24base) with mercapto.After having carried out dipping,, and make its dry 30 minutes with ultrapure water washing semi-conductor electrode part.Its result, the mercapto that has by probe combines with the gold atom of metal level, and probe just is fixed on the metal level.
(making of analyte)
Make to make as analyte and modify material and be incorporated into and comprise and the probe analyte of the DNA of complementary base assortment mutually.The modification material has used the AlexaFluor750 (イ Application PVC ト ロ ジ エ Application company) as organic pigment.The DNA peptide is incorporated on this organic pigment.
(hybridization of analyte and probe)
At first, will be configured as the next door with silicon rubber (thickness 0.2mm) around the semi-conducting electrode portion.Injecting 10 μ L hybridization solution with the formed space of this silicon rubber.The solution that will utilize the DNA (concentration 10 μ M) that modifies after the material modification and hybridization buffer (Affymetrix company) to mix has been used in this hybridization with solution.
Then, the glass sheet that closes the lid on silicon rubber becomes the moist state of solution and hybridizes.At this moment, the probe in the semi-conducting electrode portion is caught with the analyte of modifying after material is modified.Hybridization be with 45 ℃, leave standstill and it reacted in 1 hour.Behind the hybridization reaction, come the washing semi-conductor electrode part with washing with buffering agent (WashbufferA, Affymetrix company) and ultrapure water, and make its drying with fan blower.
(modulation of electrolytic solution)
Electrolytic solution be with the 4-propyl ammonium chloride of 0.6M (NPr4Cl) as supporting electrolyte salt, as electrolyte it is dissolved in the iodine of 0.06M and acetonitrile (AN) and ethylene carbonate (EC) are mixed into volume ratio in 6: 4 the solvent and modulate.
(photocurrent mensuration)
The substrate that will have the semi-conducting electrode portion that makes analyte and probe hybridization is configured in the mode of surrounding with silicon rubber (thickness 0.2mm) on every side.Injecting 10 μ L electrolytic solution with the formed space of this silicon rubber.Then, seal with the substrate that has utmost point portion from substrate top with the semi-conducting electrode portion that has been filled electrolytic solution.Thus, just become semi-conducting electrode portion, utmost point portion and reference electrode portion be contacted with the state of electrolytic solution.
Then, the current potential with reference electrode applies the 0V current potential as benchmark on semi-conducting electrode.Then from semi-conducting electrode portion direction towards to utmost point portion from light source irradiation light.On this light source, used the LASER Light Source (Cube785, コ ヒ レ Application ト company) of wavelength 785nm, intensity 13mW.Thus, modify material and be energized, the electronics that is taken place from the modification material through blasting is transported to semiconductor layer, flows through electric current in semi-conducting electrode portion and between to utmost point portion.This current value is measured.
In addition, in order to measure the current value that derives from electrode, do not make the probe hybridization on analyte and the metal level operation measured electric current.The electric current that derives from electrode is meant that electrode self encourages the electric current that is taken place by rayed.
Comparative example 3
Comparative example 3 is measured by operation similarly to Example 3 except the operation of formation metal level on the semiconductor layer.
Comparative example 4
Comparative example 4 uses silane coupling agent (aminopropyl triethoxysilane: APTES) dna probe is fixed.
(making of semi-conducting electrode portion)
Semi-conducting electrode portion forms the thin layer of conductive layer, semiconductor layer and silane coupling agent and makes on substrate.Conductive layer is the layer that has formed the tin indium oxide (ITO) of 100nm thickness on the substrate of being made up of silicon dioxide (SiO2) by sputter.Semiconductor layer is the layer that has formed the indium oxide (In2O3) of 10nm thickness on conductive layer by sputter.The thin layer of silane coupling agent be on the semiconductor layer by with silane coupling agent (aminopropyl triethoxysilane: APTES) be dissolved in the solution of toluene, be immersed in the electrode that has formed conductive layer and semiconductor layer on the substrate and the layer that forms with 1% concentration.After electrode having been carried out heating, in toluene, carry out three ultrasonics washings (5 minutes) repeatedly then, the silane coupling agent that is not combined in the semi-conducting electrode surface is as yet removed by washing with dewatered ethanol with 110 ℃.Like this, semi-conducting electrode portion is made.Being connected with the semi-conducting electrode that is used for being connected with galvanometer in this semi-conducting electrode portion goes between.
(to the making of utmost point portion)
Make of method similarly to Example 3.
(fixing of probe substance)
Make the probe stationary that constitutes by DNA (24base) on semiconductor layer.At first, with the reagent of aqueous solution (concentration and probe concentration 100 μ M) that probe is disperseed and the crosslinked usefulness of UV (Microarraycrosslinking reaget D, Amersham) solution that has carried out mixing with 1: 9 the mixing ratio 6 μ L that on semi-conducting electrode, drip.Afterwards, with UV crosslinking chemical (FS-1500, Off Na コ シ) irradiation UV light (160mJ), wash and make its dry 10 minutes with ultrapure water.Its result, UV is crosslinked just to become the bridging agent of DNA and silane coupling agent with reagent, and probe is fixed on the semiconductor layer.
(modulation of analyte)
Modulate with method similarly to Example 3.
(hybridization of analyte and probe)
Carry out with method similarly to Example 3.
(modulation of electrolytic solution)
Modulate with method similarly to Example 3.
(photocurrent mensuration)
Carry out with method similarly to Example 3.
(result)
Figure 15 is the chart that is illustrated in photoelectricity flow valuve detected in embodiment 3, comparative example 3 and the comparative example 4.
In embodiment 3, have with the DNA of the complementary mutually base assortment of probe in use and to have carried out under the situation of hybridization reaction, detected photoelectricity flow valuve is 158nA.Under the situation of only having fixed dna probe, detected photoelectricity flow valuve is 0.082nA in addition.S/N=158/0.082=1930 in view of the above.
In comparative example 3, have with the DNA of the complementary mutually base assortment of probe in use and to have carried out under the situation of hybridization reaction, detected photoelectricity flow valuve is 0.24nA.Under the situation of only having fixed dna probe, detected photoelectricity flow valuve is 0.028nA in addition.S/N=0.24/0.028=8.6 in view of the above.Improve 660 times as if comparing as can be known with previous embodiment 3 by the photoelectricity flow valuve of using metal level to derive from the modification material, S/N is than 220 times of raisings.
In addition, in comparative example 4, have with the DNA of the complementary mutually base assortment of probe in use and to have carried out under the situation of hybridization reaction, detected photoelectricity flow valuve is 19nA.Under the situation of only having fixed dna probe, detected photoelectricity flow valuve is 0.021nA in addition.S/N=19/0.021=900 in view of the above.If compare with the foregoing description 3,8 times of photoelectricity flow valuve raisings, 2 times of the S/N raisings of modifying material by using metal level to derive from.In comparative example 4, find to derive from the raising of the photoelectricity flow valuve of modifying material and the raising of S/N similarly.
According to above explanation, if make metal level be formed on semi-conducting electrode portion, the detection sensitivity of electric current will improve.As the essential factor that the detection sensitivity of current value improves, can think to make (1) DNA fixed amount increase the raising etc. that (2) electric conductivity improves the caused light-to-current inversion efficient of plasma excitation in (3) metal level because of forming metal level.
Embodiment 4 (having used the research of the non-special absorption of a base many types of (SNP))
(making of semi-conducting electrode portion)
Semi-conducting electrode portion forms conductive layer, semiconductor layer and metal level and makes on substrate.Conductive layer is the layer that has formed the tin indium oxide (ITO) of 100nm thickness on the substrate that is made of silicon dioxide (SiO2) by sputter.Semiconductor layer is the layer that has formed the indium oxide (In2O3) of 10nm thickness on conductive layer by sputter.Metal level is the layer that has formed the gold thin film of 2nm thickness on semiconductor layer by evaporation.Being connected with the semi-conducting electrode that is used for being connected with galvanometer in this semi-conducting electrode portion goes between.
(to the making of utmost point portion)
To utmost point portion is that the platinum film that has formed 200nm thickness by sputter on the substrate of being made up of silicon dioxide (SiO2) is made.Be connected be used for being connected the utmost point is gone between at this on to utmost point portion with galvanometer.
(fixing of probe substance)
At first, semi-conducting electrode portion is immersed in makes the aqueous solution (concentration and probe concentration 10 μ M) 16 hours that probe disperses.This probe is the DNA (24base) with mercapto.After having carried out dipping,, and make its dry 10 minutes with ultrapure water washing semi-conductor electrode part.Its result, the mercapto that has by probe combines with the gold atom of metal level, and probe just is fixed on the metal level.
(making of analyte)
Making makes and modifies the analyte that material is incorporated into the DNA that comprises the base assortment not complementary with probe (only 1 base is not complementary) as analyte.The modification material has used the Alexa Fluor750 (イ Application PVC ト ロ ジ エ Application company) as organic pigment.The DNA peptide is incorporated on this organic pigment.
(hybridization of analyte and probe)
At first, will be configured as the next door with silicon rubber (thickness 0.2mm) around the semi-conducting electrode portion.Injecting 10 μ L hybridization solution with the formed space of this silicon rubber.The solution that will utilize the DNA (concentration 10 μ M) that modifies after the material modification and hybridization buffer (Affymetrix company) to mix has been used in this hybridization with solution.
Then, the glass sheet that closes the lid on silicon rubber becomes the moist state of solution and hybridizes.At this moment, the probe in the semi-conducting electrode portion is caught with the analyte of modifying after material is modified.Hybridization be with 45 ℃, leave standstill and it reacted in 1 hour.Behind the hybridization reaction, after washing with buffering agent (Wash bufferA, Affymetrix company) and ultrapure water with washing, make its drying with fan blower.
(modulation of electrolytic solution)
Electrolytic solution be with the 4-propyl ammonium chloride of 0.6M (NPr4Cl) as supporting electrolyte salt, as electrolyte it is dissolved in the iodine of 0.06M and acetonitrile (AN) and ethylene carbonate (EC) are mixed into volume ratio in 6: 4 the solvent and modulate.
(photocurrent mensuration)
The substrate that will have the semi-conducting electrode portion that makes analyte and probe hybridization is configured in the mode of surrounding with silicon rubber (thickness 0.2mm) on every side.Injecting 10 μ L electrolytic solution with the formed space of this silicon rubber.Then, seal with the substrate that has utmost point portion from substrate top with the semi-conducting electrode portion that has been filled electrolytic solution.Thus, just become semi-conducting electrode portion, utmost point portion and reference electrode portion be contacted with the state of electrolytic solution.
Then, the current potential with reference electrode applies the 0V current potential as benchmark on semi-conducting electrode.Then from semi-conducting electrode portion direction towards to utmost point portion from light source irradiation light.On this light source, used the LASER Light Source (Cube785, コ ヒ レ Application ト company) of wavelength 785nm, intensity 13mW.Thus, modify material and be energized, the electronics that is taken place from the modification material through blasting is transported to semiconductor layer, flows through electric current in semi-conducting electrode portion and between to utmost point portion.This current value is measured.
In addition, in order to measure the current value that derives from electrode, do not make the probe hybridization on analyte and the metal level operation measured electric current.The electric current that derives from electrode is meant that electrode self encourages the electric current that is taken place by rayed.
Comparative example 5
Comparative example 5 except used as analyte with the complementary mutually base assortment of probe, measure by operation similarly to Example 4.
Comparative example 6
Comparative example 6 is measured by operation similarly to Example 4 except not making analyte hybridization.
(result)
Figure 16 is the chart of photoelectricity flow valuve detected in embodiment 4, comparative example 5 and comparative example 6.
In embodiment 4, use the DNA have with the not complementary base assortment of probe to carry out under the situation of hybridization reaction, detected photoelectricity flow valuve is 1.7nA.In comparative example 5, use have with probe mutually the DNA of complementary base assortment carried out under the situation of hybridization reaction, detected photoelectricity flow valuve is 195nA.Only fixed in comparative example 6 under the situation of dna probe, detected photoelectricity flow valuve is 0.067nA.Can confirm that in view of the above the non-DNA that is adsorbed on specifically on the gold thin film is less, can assortment detect analyte specifically.
Embodiment 5 (the thickness dependence of gold thin film)
(making of semi-conducting electrode portion)
Semi-conducting electrode portion forms conductive layer, semiconductor layer and metal level and makes on substrate.Conductive layer is the layer that has formed the tin indium oxide (ITO) of 100nm thickness on the substrate that is made of silicon dioxide (SiO2) by sputter.Semiconductor layer is the layer that has formed the indium oxide (In2O3) of 10nm thickness on conductive layer by sputter.Metal level is the layer that has formed the gold thin film of 0.2nm thickness on semiconductor layer by evaporation.Being connected with the semi-conducting electrode that is used for being connected with galvanometer in this semi-conducting electrode portion goes between.
(to the making of utmost point portion)
To utmost point portion is that the platinum film that has formed 200nm thickness by sputter on the substrate that is made of silicon dioxide (SiO2) is made.Be connected be used for being connected the utmost point is gone between at this on to utmost point portion with galvanometer.
(fixing of probe substance)
At first, semi-conducting electrode portion is immersed in makes the aqueous solution (concentration and probe concentration 10 μ M) 16 hours that probe disperses.This probe is the DNA (24base) with mercapto.After having carried out dipping,, and make its dry 10 minutes with ultrapure water washing semi-conductor electrode part.Its result, the mercapto that has by probe combines with the gold atom of metal level, and probe just is fixed on the metal level.
(making of analyte)
Make to make as analyte and modify material and be incorporated into and comprise and the probe analyte of the DNA of complementary base assortment mutually.The modification material has used the AlexaFluor750 (イ Application PVC ト ロ ジ エ Application company) as organic pigment.The DNA peptide is incorporated on this organic pigment.
(hybridization of analyte and probe)
At first, will be configured as the next door with silicon rubber (thickness 0.2mm) around the semi-conducting electrode portion.Injecting 10 μ L hybridization solution with the formed space of this silicon rubber.The solution that will utilize the DNA (concentration 1 μ M) that modifies after the material modification and hybridization buffer (Affymetrix company) to mix has been used in this hybridization with solution.
Then, the glass sheet that closes the lid on silicon rubber becomes the moist state of solution and hybridizes.At this moment, the probe in the semi-conducting electrode portion is caught with the analyte of modifying after material is modified.Hybridization be with 45 ℃, leave standstill and it reacted in 1 hour.Behind the hybridization reaction, after washing with buffering agent (WashbufferA, Affymetrix company) and ultrapure water with washing, make its drying with fan blower.
(modulation of electrolytic solution)
Electrolytic solution be with the 4-propyl ammonium chloride of 0.6M (NPr4Cl) as supporting electrolyte salt, as electrolyte it is dissolved in the iodine of 0.06M and acetonitrile (AN) and ethylene carbonate (EC) are mixed into volume ratio in 6: 4 the solvent and modulate.
(photocurrent mensuration)
The substrate that will have the semi-conducting electrode portion that makes analyte and probe hybridization is configured in the mode of surrounding with silicon rubber (thickness 0.2mm) on every side.Injecting 10 μ L electrolytic solution with the formed space of this silicon rubber.Then, seal with the substrate that has utmost point portion from substrate top with the semi-conducting electrode portion that has been filled electrolytic solution.Thus, just become semi-conducting electrode portion, utmost point portion and reference electrode portion be contacted with the state of electrolytic solution.
Then, the current potential with reference electrode applies the 0V current potential as benchmark on semi-conducting electrode.Then from semi-conducting electrode portion direction towards to utmost point portion from light source irradiation light.On this light source, used the LASER Light Source (Cube785, コ ヒ レ Application ト company) of wavelength 785nm, intensity 13mW.Thus, modify material and be energized, the electronics that is taken place from the modification material through blasting is transported to semiconductor layer, flows through electric current in semi-conducting electrode portion and between to utmost point portion.This current value is measured.
And then in the making of semi-conducting electrode portion, the current value when changing over 1nm, 2nm and 5nm about the thickness of gold thin film is all similarly measured with above-mentioned.
In addition, in order to measure the current value that derives from electrode, do not make the probe hybridization on analyte and the metal level operation measured electric current.The electric current that derives from electrode is meant that electrode self encourages the electric current that is taken place by rayed.
(result)
Figure 17 is the chart of the S/N ratio under each thickness detected in embodiment 5.When gold thin film is 1nm, obtain best S/N ratio as can be known.
In addition, during more than or equal to 5nm, gold thin film will be peeled off from semiconductor layer by washing procedure at the thickness of gold thin film.Therefore, during more than or equal to 5nm, just need to use the semiconductor layer (embodiment 1 and 2) that comprises titanium or chromium, perhaps use the semiconductor layer (embodiment 3) that has carried out sintering processes, so that the raising of the bounding force of gold thin film and semiconductor layer at the thickness of gold thin film.
Claims (10)
1. check chip for one kind, be used for detecting utilization and produce the analyte of the modification material modified of electronics, it is characterized in that comprising by light stimulus:
The semi-conducting electrode portion that possesses the metal level that is formed on the semiconductor layer;
Catch the probe on the above-mentioned metal level of being fixed on of analyte; And
Possess conductive layer to utmost point portion.
2. the inspection chip of putting down in writing according to claim 1 is characterized in that:
Above-mentioned metal level is made up of the metal that dissolves by electrolytic solution.
3. the inspection chip of putting down in writing according to claim 2 is characterized in that:
Above-mentioned electrolytic solution comprises iodine or iodide.
4. according to any inspection chip of being put down in writing in the claim 1～3, it is characterized in that:
Above-mentioned metal level is made up of the metal that carries out chemisorption with above-mentioned probe.
5. the inspection chip of putting down in writing according to claim 4 is characterized in that:
The metal that carries out chemisorption with above-mentioned probe is a gold.
6. the inspection chip of putting down in writing according to claim 4 is characterized in that:
Above-mentioned probe has mercapto as to carry out combine of chemisorption basic with above-mentioned metal level.
7. pick-up unit is used for detecting utilization and produces the analyte of the modification material modified of electronics by light stimulus, it is characterized in that comprising:
Received portion can receive the inspection chip checking the mode of chip and constitute, wherein this inspection chip comprises: the semi-conducting electrode portion that possesses the metal level that is formed on the semiconductor layer; Catch the probe on the above-mentioned metal level of being fixed on of analyte; With possess conductive layer to utmost point portion;
To modifying the light source that material carries out light stimulus, this trim analyte that above-mentioned inspection chip received in the above-mentioned inspection chip that is inserted in the portion of verifying is modified; And
Amperometric determination portion measures the electric current that flows out from the modification material of analyte being modified by the light stimulus of above-mentioned light source.
8. the pick-up unit of putting down in writing according to claim 7 is characterized in that:
Above-mentioned light source sends the light of the wavelength that the modification material of modifying analyte is encouraged.
9. detection method is used for detecting utilization and produces the analyte of the modification material modified of electronics by light stimulus, it is characterized in that may further comprise the steps:
Comprising the semi-conducting electrode portion that possesses the metal level that is formed on the semiconductor layer; Catch the probe on the above-mentioned metal level of being fixed on of analyte; With possess conductive layer to using the sample comprise analyte on the inspection chip of utmost point portion, catch analyte with the above-mentioned probe that is fixed on the above-mentioned metal level thus;
To modify material and import to analyte;
Irradiation is to modifying the light that material encourages; And
The modification material electric current that produced of detection after the excitation.
10. the detection method of putting down in writing according to claim 9 is characterized in that also comprising:
Be used to electrolyte medium that electric current is flow through in above-mentioned semi-conducting electrode portion and above-mentioned the interpolation between to utmost point portion.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
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| JP2008-281584 | 2008-10-31 | ||
| JP2008281584 | 2008-10-31 |
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| CN101726528A true CN101726528A (en) | 2010-06-09 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102539506A (en) * | 2010-11-30 | 2012-07-04 | 希森美康株式会社 | Method of electrochemically detecting a sample substance |
| CN102590304A (en) * | 2010-12-28 | 2012-07-18 | 希森美康株式会社 | Photocurrent detection electrode, manufacturing method, and working electrode substrate |
| CN105829876A (en) * | 2013-12-16 | 2016-08-03 | 美国圣戈班性能塑料公司 | Electrode and method for making an electrode |
| US10794851B2 (en) | 2016-11-30 | 2020-10-06 | Saint-Gobain Performance Plastics Corporation | Electrode and method for making an electrode |
-
2009
- 2009-10-16 CN CN200910205180A patent/CN101726528A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102539506A (en) * | 2010-11-30 | 2012-07-04 | 希森美康株式会社 | Method of electrochemically detecting a sample substance |
| CN102539506B (en) * | 2010-11-30 | 2014-11-05 | 希森美康株式会社 | Method of electrochemically detecting a sample substance |
| CN102590304A (en) * | 2010-12-28 | 2012-07-18 | 希森美康株式会社 | Photocurrent detection electrode, manufacturing method, and working electrode substrate |
| CN105829876A (en) * | 2013-12-16 | 2016-08-03 | 美国圣戈班性能塑料公司 | Electrode and method for making an electrode |
| US10794851B2 (en) | 2016-11-30 | 2020-10-06 | Saint-Gobain Performance Plastics Corporation | Electrode and method for making an electrode |
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