CN218167070U - Microarray chip - Google Patents
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- CN218167070U CN218167070U CN202220223733.1U CN202220223733U CN218167070U CN 218167070 U CN218167070 U CN 218167070U CN 202220223733 U CN202220223733 U CN 202220223733U CN 218167070 U CN218167070 U CN 218167070U
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Abstract
The utility model relates to a microarray chip, its preparation method and application, it includes base plate and at least one detection array, and detection array is including electrode and detection site, and the material of detection site is porotic graphite alkene and/or porotic graphene oxide, and the base plate is all located to electrode and detection site, and the electrode is connected with the detection site electricity, and detection array is the orderly arrangement. The utility model discloses a microarray chip is owing to be provided with the electrode of connecting the detection site on the base plate, through appling or partially positive voltage or partially negative voltage to the electrode to the promotion target object that is detected gathers on the graphite alkene of the detection site that also has the probe to the electrode direction, increases the sensitivity and the lower limit that detect, and preparation method is simple and easy, is particularly useful for the detection of the microorganism that the content is lower in the sample.
Description
Technical Field
The utility model relates to a biochemistry detects the apparatus field, especially relates to a microarray chip for biochemistry detects.
Background
Since graphene, a novel material, was first discovered in 2004 and prepared in the laboratory, it has unique physical or chemical properties, and is widely used in various fields, and plays an increasingly important role in many fields. The focus of development of graphene materials in various fields is how to fully utilize unique attributes of graphene, integrate the graphene into the field, solve some problems in the field, or optimize the prior art method in the field, and improve the working efficiency. In the field of biochemistry, the intervention of graphene is mainly applied to a certain extent as a biosensor at present, for example, various biomolecules are connected to the surface of graphene and are fluorescently labeled, and the interaction between the graphene and a target object is utilized to quench fluorescence, so as to achieve the purpose of detection.
Microarray chips or biochips have a small volume, high detection sensitivity, and relatively low requirements for sample size and the like, and are therefore rapidly developed. Fig. 11 to 13 show a conventional microarray, taking the detection of pathogenic microorganisms as an example, in which a substrate A1 of a microarray chip is provided with detection sites a11, an antibody T is immobilized on the detection sites a11, and when a target R in a sample approaches or contacts the antibody T on the detection sites a11, the antibody T interacts with the target R to capture the target R, and then the detection result is shown by an electrochemical signal or a fluorescent signal.
As shown in fig. 11 to 13, the conventional microarray is a flat surface, the antibody T is laid on each detection site a11, and when the microarray chip performs detection, if the sample itself contains a small amount of target R or the concentration of the target R is too low, the detection result may be false negative, and the like.
SUMMERY OF THE UTILITY MODEL
The utility model aims to improve current microarray chip, increase microarray chip and treat the capture ability of the target object that detects, promote microarray chip's detectability to the solution is in the detectability that increases and improve microarray chip, can be so that its manufacturing cost's increase control to a certain extent, establishes the basis for large-scale production, also uses microarray chip to detect the condition that the germ content of pathogenic microorganism is less than normal standard simultaneously.
In order to solve the technical problem mentioned above, increase microarray chip and examine time measuring to the sample, improve the rate of accuracy that detects, the utility model discloses an one of the technical scheme do, provide a microarray chip, it is including:
a substrate;
at least one detection array disposed on the substrate and including:
an electrode is arranged on the base plate and is provided with a plurality of electrodes,
at least one detection site, wherein the material of the detection site is porous graphene and/or porous graphene oxide,
a recognition substance, wherein the recognition substance is arranged at the detection site and can specifically interact with a target object in the sample;
the detection site is electrically connected with the electrode;
the detection arrays are arranged in order.
Preferably, the microarray chip further comprises at least one verification array disposed on the substrate, wherein the verification array comprises at least one of the detection sites and the identifier.
Preferably, the microarray chip as described above, wherein the target is a microorganism, and the bacteria content of the microorganism in the sample is: 10 2 cells/ml to 10 6 cells/ml。
Preferably, the microarray chip as described above, wherein the detection arrays are arranged in a staggered manner.
Preferably, the voltage of at least one of the detection arrays is a positive voltage or a negative voltage, wherein the positive voltage is 0.2v to 0.8v, and the negative voltage is-0.2 v to-0.8 v.
The utility model provides a technical scheme, a preparation method as aforementioned microarray chip, it includes following step:
s1, photoetching an electrode wiring pattern on a substrate;
s2, evaporating and plating a film on the substrate obtained in the step S1, and removing the photoresist to prepare electrode wiring after film forming;
s3, spraying a graphene oxide solution on the specific site of the electrode wiring obtained in the step S2;
s4, freeze-drying the substrate obtained in the step S3;
and S5, annealing the substrate obtained in the step S4 under the conditions that the environmental pressure is less than one atmosphere and the freeze-drying temperature is-50 ℃ to-120 ℃, heating to 400 ℃ to 600 ℃ in an environment containing argon and hydrogen to reduce partial graphene oxide, and obtaining the porous graphene or the composition of the porous graphene and the porous graphene oxide.
Preferably, in the preparation method, in the step S2, a material of the electrode wire is selected from the group consisting of: titanium, chromium, aluminum or nickel and any combination thereof; the thickness of the film is 50 to 200 nm.
Preferably, in the step S3, the graphene oxide solution is prepared by a Hummers method, and the amount of the graphene oxide solution sprayed on the specific site is 0.5 μ l to 1 μ l.
The third technical solution provided by the utility model provides a detection method of microarray chip, which is suitable for the range of the bacteria content of the microorganism in the sample to be 10 2 cells/ml to 10 6 The microbiological detection is carried out on the samples of cells/ml, which comprises the following steps: the detection site of the microarray chip is positively charged at a voltage of 0.2 to 0.8 volts.
Preferably, the detection method comprises detecting with the microarray chip.
The utility model discloses an useful part lies in:
1. because the whole individual of target object all generally appears to carry certain positive charge or negative charge, consequently, the utility model discloses a detection array of microarray chip is provided with the electrode, makes each detection site can be electrified with the help of electrode connection to according to the nature of the electric charge that the target object carried, select to exert positive voltage or negative voltage to the electrode, and adjust voltage to suitable size simultaneously, make the electric charge that each detection site carried can be opposite with the electric charge that the target object carried, consequently can increase the attraction and the gathering ability of detection array to the target object. For example, pathogenic bacteria or single-stranded DNA are usually negatively charged, and a positive voltage can be applied to the electrodes to attract more negatively charged targets to the detection array, so as to bind the probes (probes) or antibodies (antibodies) pre-arranged on the detection sites, thereby improving the detection sensitivity.
2. The utility model discloses a microarray chip preparation method is simple, through photoetching, coating by vaporization film forming, spraying oxidation graphite alkene and freeze-drying step, realizes increasing the electrode to microarray chip to the arrangement of detection array is convenient for adjust, and the yield is high and the price is controllable, easily realizes commercial mass production.
3. The utility model discloses a detection method can be obvious promote the detectivity and the detection lower limit of current microarray chip, especially obviously lower to the content of target object in some samples to under the easy condition that produces false negative of conventional microarray detection, can be with the help of the production appeal of applied voltage to the target object, consequently, the low target object of the commonly contained bacterial volume of specially adapted detection, especially pathogenic microorganism.
4. The utility model discloses a detection method range of application is extensive, can combine together with current ripe microarray detection chip and use, improves current microarray chip's detectability through increasing the electrode to the array.
Drawings
Fig. 1 is a perspective view of a first embodiment of the present invention.
Fig. 2 is a side view of a first embodiment of the present invention.
Fig. 3 is a top view of the first embodiment of the present invention.
Fig. 4 is an enlarged detail view of the area a in fig. 3.
Fig. 5 is an enlarged detail view of the region B in fig. 4.
Fig. 6 is a schematic diagram of a second embodiment of the present invention.
Fig. 7 to 9 are schematic arrangement diagrams of the detection array according to the third embodiment of the present invention.
Fig. 10 is a schematic view of a first preparation method of the present invention.
FIG. 11 is a schematic top view of a conventional microarray.
Fig. 12 is an enlarged detail view of a' portion in fig. 11.
Fig. 13 is an enlarged detail view of the region B' in fig. 12.
Detailed Description
The following description will be made in conjunction with the accompanying drawings and the preferred embodiments of the present invention to further illustrate the technical means adopted to achieve the objects of the present invention.
The utility model relates to a microarray chip, it is including base plate and detection array, and the base plate preferred adopts glass or quartz material, and detection array sets up in the base plate, and it is including electrode and a plurality of detection site, and the material of detection site is porotic graphite alkene or porotic graphene oxide, perhaps is the combination of its both different proportions, and the electrode is connected with a plurality of detection site electricity, is connected with the discernment thing on the detection site, and detection array is the orderly arrangement. In the present invention, unless otherwise specified, the identifier refers to a substance capable of specifically interacting with a target in a sample, and mainly includes, but is not limited to, antibodies, nucleic acid probes, and various RNAs, DNAs, and the like. The means of interaction includes physical or chemical bonding, physical or chemical reaction, and the like. The target is organic or inorganic substances to be detected in the sample, such as antigens, DNA, ion groups or various types of microorganisms, bacteria, fungi or viruses.
The utility model discloses a microarray chip, the aforesaid discernment thing that its testing site is connected generally all passes through fluorescence labeling to the antibody is the example particularly, through fluorescence labeling on the antibody, after detecting the target object and taking place the interaction, arouse fluorescence signal's change to further turn into the testing result and demonstrate out, no longer give unnecessary details.
The specific arrangement mode of the detection array is not limited, and may be equidistant row-column arrangement, or other arrangement modes, and specifically, the detection array may be appropriately selected according to the type and property of the target object to be detected, for example, the detection array may also be cross-arranged, so as to facilitate detection. In the following preferred examples, the description will be given taking an equidistant row-column arrangement as an example.
The number of the detection arrays may be plural or one, and is generally set to two to six, and the number is not limited herein. The electrodes of the detection array are arranged on the substrate and electrically connected with each detection site of the detection array. In addition, the microarray chip of the present invention may further comprise a verification array, i.e., a verification array in which the detection sites are not connected to electrodes, so that the detection array and the verification array exist on one substrate at the same time, and are used under special circumstances.
Example one
Referring to fig. 1 and 2, a first preferred embodiment of the present invention includes a glass substrate 1, detection arrays G1 and G2 are disposed on the glass substrate 1, the detection arrays G1 and G2 include electrodes 2 and detection sites 3, the detection sites 3 are formed by porous graphene oxide, and the electrodes 2 are electrically connected to the detection sites 3. Referring also to FIG. 3, in the preferred embodiment, the detection sites 3 and the electrodes 2 are electrically connected in such a manner that each electrode 2 is laid on the glass substrate 1 in a fork shape, and the detection sites 3 are respectively disposed on the electrodes 2.
In the first embodiment, two detection arrays are provided in total, the power ends 21 of the electrodes 2 of the two detection arrays are far away from each other and located at two ends of the glass substrate 1, and the power ends 21 located at two ends of the glass substrate 1 are beneficial to laying of a circuit, so that the interference on a sample is reduced to the greatest extent. Detection sites 3 are respectively arranged on the fork-shaped wishbone of each electrode 2 at equal intervals. The uniform arrangement of the detection sites 3 is beneficial to increasing the probability that each detection site 3 contacts the target object R in the sample, and the following situations are prevented: in the region where the amount of the target substance R is large in the sample, detection omission or detection result distortion occurs due to the deficiency or absence of the detection site 3. Those skilled in the art can set more detection arrays G1 or G2 according to actual needs, and can further group and partition the detection arrays G1 and G2, and the distribution pattern of the electrodes 2 is not limited to the fork-shaped structure shown in fig. 3. The material of the detection site 3 may be porous graphene or porous graphene oxide, or porous reduced graphene oxide, or a combination of porous graphene and porous graphene oxide.
As shown in fig. 1 and 4, when the material of the detection site 3 in the first embodiment is preferably porous graphene or porous graphene oxide, or a combination of the two at different ratios, the porous graphene or porous graphene oxide can increase the specific surface area of the detection site 3, so that when a recognition substance is connected to the detection site 3, for example, in the first embodiment, the recognition substance is preferably a fluorescent-labeled antibody T, the spatial distribution level and number of the antibody T can be significantly increased (fig. 4 and 5).
In contrast, in the conventional microarray shown in FIGS. 11 to 13, the antibodies T on the glass substrate A1 are located on the same plane as each of the detection sites A11, the number of the antibodies T is much lower, and the spatial distribution is almost none. The quantity and spatial distribution of the antibodies T can directly influence the contact and interaction probability of the antibodies T and the target object R, and further influence the detection efficiency. Therefore, because the utility model discloses a detection site 3 is porotic graphite alkene or porotic graphite oxide, or is porotic graphite alkene and porotic graphite oxide's composition, and it can further improve the detectivity of microarray, improves the rate of accuracy that detects.
In practical operation of the microarray chip, a certain voltage may be applied to the electrodes 2 of the detection arrays G1 and G2, for example, when the target R is a pathogenic bacterium or a single-stranded RNA, cDNA or ssDNA, since the target R generally has a negative charge as a whole, a bias voltage may be applied to the electrodes 2, preferably, in the first embodiment, the bias voltage is 0.2 to 0.8 volts, and the voltage in this range can achieve the best detection result through experimental detection. After the voltage is applied, the positively charged electrode 2 positively charges the detection site 3 to which it is connected, because the different charges attract each other, and thus more target object R such as bacteria can be attracted and accumulated to the vicinity of the detection site 3, enabling the antibody T connected to the detection site 3 to approach the target object, increasing the ability of the antibody T to capture the target object R. Since the antibody T is fluorescently labeled, the detection result can be further shown by the change of the fluorescence signal (not shown in the figure). Of course, the detection site 3 can be fixed to other macromolecules besides the binding antibody T, such as DNA fragments, enzymes or protein fragments, etc., and will not be described herein.
When the microarray chip of the first embodiment is applied to the detection of the target object R with positive charge, for example, some positively charged ion groups, some metal ions, etc., a negative voltage can be applied to the electrodes 2 of the detection arrays G1 and G2, which can increase the attraction of the detection sites 3 to the positive ion groups, as described above, thereby improving the detection efficiency, and generally, the magnitude of the negative voltage is from-0.2 to-0.8 volts, which is the best detection effect, and is too large or too small, which is not good for the attraction of the target object R.
Further, in the same microarray chip, different detection arrays may be respectively set to be charged with positive and negative electricity, thereby satisfying some needs to simultaneously detect the target R appearing to be charged with different charges, respectively. With continued reference to FIG. 1, the two sense arrays G1 and G2 of embodiment one are set with different voltages. When the target object R to be detected includes both pathogenic bacteria and specific micro metal ions or a certain kind of positively charged ionic groups, the electrodes 2 of the detection array G1 may be set to be applied with a positive bias voltage, and correspondingly, the electrodes 2 of the detection array G2 may be applied with a negative bias voltage. Thus, the detection array G1 is beneficial to attracting and gathering pathogenic bacteria, and the detection array G2 is beneficial to attracting and gathering positively charged groups, so that the detection capability of different targets R can be simultaneously increased. In the first embodiment, only two detection arrays are provided, and when more detection arrays are provided, the other detection arrays can be alternately arranged in a cross manner, which is beneficial to improving the detection capability of the diversified targets R at the same time.
Example two
As shown in fig. 6, the second embodiment is similar to the first embodiment, except that three detection arrays are provided, which are detection arrays G3, G4 and G5, respectively, and a new verification array G6 is provided, and the detection sites 3 of the verification array G6 are not provided with electrodes 2, nor are they electrically connected to any of the electrodes 2, so that the verification array G6 is not electrically connected. The microarray chip of example two can be used for the following purposes, in addition to having a detection function similar to that of example one.
The microarray chip of the second embodiment can be used to detect or verify whether the target R carries positive charges, negative charges or exhibits neutrality as a whole. Thus, the electrodes 2 of the detection arrays G3 and G4 of the second example were applied with a positive voltage and the electrodes 2 of the detection array G5 were applied with a negative voltage, while the detection site 3 of the verification array G6 was rendered neutral by not connecting either electrode as a control. When a sample containing only the target substance R to be measured is detected, if the positive signals of G3 and G4 in the detection result are much stronger than those of G5 which is used as the control G6 and is more negative than the negative voltage, it indicates that the target substance R carries a negative charge as a whole. If the detection result is opposite to this, the target R carries a positive charge as a whole. If the detection results of G3, G4, G5 and G6 are not very different, it indicates that the target R is a neutral group to a large extent, or is not electrolyzed in the liquid, and the like.
EXAMPLE III
The detection array of the third embodiment is different from the arrangement mode of the respective arrangement of the partitions of each detection array of the first embodiment and the second embodiment, but preferably adopts three staggered arrangement modes, and those skilled in the art can also adopt other staggered arrangement modes according to needs.
Referring to fig. 7 to 9, the detection arrays according to the third embodiment of the present invention are arranged in a staggered manner. FIG. 7 shows a fully staggered arrangement of rows and columns, i.e., each detection array is interdigitated with rows of another detection array and the columns are staggered. Specifically, in the staggered arrangement of fig. 7, the detection array G7 and the detection array G8 are inserted into each other in a staggered manner, and G7 and G8 are each provided with three rows of detection sites 3. Taking fig. 7 as an example, a first row of detection sites of G7 is a first row, a second row of detection sites of G8 is a second row, and then a second row of detection sites of G7 is sequentially arranged in turn. Furthermore, each detection site of each row of G7 and each detection site of each row of G8 are not on the same line, but are staggered with each other. The advantage of this arrangement is that the most dense number of detection sites can be obtained on the chip.
Fig. 8 shows an alternate staggered arrangement of rows, that is, the rows of each detection array and the rows of the other detection array are alternately staggered, and the columns are in the same column as a whole, which is very similar to the staggered arrangement of the detection arrays shown in fig. 7, except that the detection sites of each row of the detection array G9 and the detection sites of each row of the detection array G10 are in the same straight line or in the same column as seen in a plan view. When the detection sites are orderly arranged, which is beneficial to the laser scanning chip, the laser element actuates the smoothness of the platform movement to further improve the detection speed, thereby achieving the advantage of the design.
Fig. 9 shows an alternate staggered arrangement of the detection arrays, which is different from the arrangement shown in fig. 7 and 8. The detection arrays are alternately staggered to form a staggered arrangement of the whole detection array, in short, after the detection array G11 is arranged, the detection array G12 is arranged below, then the detection array G13 is arranged, namely, the first two rows are the detection array G11, the middle two rows are the detection array G12, and the last two rows are the detection array G13. The staggered arrangement of the detection arrays of fig. 7-9 is merely an example, and the actual situation may not be as shown with only three rows or two rows. The arrangement design is favorable for arbitrarily selecting whether the detection array is applied with voltage or not, and the array without voltage can be used as a control group for detection, so the method has the advantage of being more flexible in implementation.
The arrangement modes of the different detection arrays can be applied to different scenes. For example, when the same kind of target object R is precisely detected, the arrangement shown in fig. 7 may be preferably adopted, and this staggered arrangement enables the most dense and uniform arrangement of the detection sites 3, and the target object can be detected to the greatest extent.
The utility model discloses another provide a technical solution for preparation microarray chip.
The general steps of the preparation method are as follows:
s1, manufacturing an electrode wiring pattern on a substrate by using a photoetching process (lithography).
S2, manufacturing electrode wiring by using an evaporation film forming technology (evaporator), wherein the electrode material is selected from one of the group consisting of the following metals: titanium (Ti), chromium (Cr), aluminum (Al), nickel (Ni), or any combination thereof. The total thickness of the formed film is 50-200nm, and the photoresist is removed after the film is formed.
S3, spraying a graphene oxide solution on the specific positions of the electrode wiring prepared in the step S2, wherein the graphene oxide solution is manufactured by a Hummers method, and the spraying amount of each position is 0.5-1 mu l.
S4, carrying out freeze-drying (freeze-drying) on the materials, quickly removing moisture in the graphene oxide solution to obtain porous graphene oxide, wherein the environmental pressure is less than one atmosphere, and the freeze-drying temperature is-50 DEG O C to-120 O And C.
S5, annealing the substrate in a gas containing argon (Ar) and hydrogen (H) 2 ) Is heated to 400 DEG under the environment O C-600 O And C, reducing partial graphene oxide to obtain porous graphene/graphene oxide.
S6, modifying the antibody of the pathogenic bacteria (such as escherichia coli) which is subjected to fluorescent labeling on each detection site.
Preparation example 1
Preparation example one was used to prepare the microarray chip shown in example one. Referring to fig. 10, it should be noted that fig. 10 is only an exemplary illustration and is not an actual product, for example, fig. 10 shows 2 detection arrays, each detection array includes three rows of detection sites, each row of detection sites has only two detection sites, but the actual product is far more than this number. The specific steps of preparation example one are as follows:
s1, carrying out electrode wiring on a glass substrate 1 by using a photolithography process (lithography) to manufacture an electrode wiring pattern, wherein two detection arrays are arranged, the electrode 2 of each detection array is provided with about 6-8 rows, the distance between each row and the previous row and the next row is 2-3mm, and the length of each row is about 3.5cm.
And S2, further manufacturing the evaporation film of the electrode wiring according to the pattern of the electrode wiring manufactured in the step S1 by using an evaporation film forming technology (evaporator). The electrode material is selected from titanium (20 nm) as a bottom adhesion layer, then nickel (100 nm) is used as a conductive layer for forming alloy with graphene, the total thickness of the formed film is about 120nm, and photoresist is removed after the film is formed.
S3, preparing 100ml of graphene oxide solution through Hummers method, and spraying the graphene oxide solution on the electrode wiring at intervals of 2-3mm to form detection sites 3, wherein the diameter of the detection sites 3 is about 1mm. Each test site 3 was sprayed with about 0.5. Mu.l.
S4, carrying out freeze-drying on the whole glass substrate 1 obtained in the step S3, quickly removing moisture in the graphene oxide solution to obtain porous graphene oxide, wherein the environmental pressure is 0.05mbar (40 mtorr), and the freeze-drying temperature is about-50 DEG C O C。
S5, annealing the glass substrate 1 obtained in the step S4, and carrying out annealing on the glass substrate containing argon (Ar) and hydrogen (H) 2 ) Is heated to 500 deg.C O And C, reducing partial graphene oxide to obtain porous graphene/graphene oxide.
S6, modifying the antibody T of pathogenic bacteria (such as escherichia coli) which is subjected to fluorescent labeling on each detection site 3, thereby finally completing the preparation of the porous oxidation microarray chip of the utility model.
Example of detection
As described above, the microarray chip of the present invention can significantly improve the detection capability due to the addition of the electrodes, and particularly, for the detection of pathogenic microorganisms, most of the pathogenic microorganisms are carried with negative electricity as a whole, and when a positive voltage is applied, the microarray chip can exert its effect. For example, in general clinical tests for the detection of microorganisms, the minimum requirement for the bacterial content in a sample is 10 6 cells/ml, samples with a bacterial content below this level, usually fail to detect a result or give false negative results even if the target pathogen to be detected is still present. By the technology of the utility model, the bacteria content in the sample is only over 10 2 The cells/ml can detect the signal, thus greatly increasing the limit of detection (LOD), which is very beneficial to the treatment and interpretation of infection test.
Detection example 1
The microarray chip of preparation example I is used for detection, and for detection of Escherichia coli, the detection site of the microarray chip is modified with an Escherichia coli antibody with a fluorescent marker (capable of detecting Escherichia coli with a length of about 5-20 μm), and the method mainly comprises the following steps:
1. adding the colony to be detected into a 5ml test tube (if the volume of the colony to be detected reaches 5ml, the step can be omitted);
2. placing into a centrifuge, and rotating at 1800 (rpm) speed for 5 minutes;
3. the upper suspension is poured off, 0.5ml of precipitate is taken and averagely dropped on the detection site of the chip;
4. standing for 5 minutes;
5. applying positive voltage to the electrodes of the chip, wherein the voltage is 0.2-0.8 volt and lasts for 1 minute in total;
6. releasing the voltage, and washing the surface of the chip by using a buffer solution;
7. draining off water;
8. the chip is placed into a laser Scanner (Scanner) for scanning, and the detection result is calculated according to the intensity of the fluorescence signal.
The utility model discloses a microarray chip is owing to adopted the new material of porosity graphite alkene and increased the structure of electrode 2, and the quantity and the spatial distribution level of the probe on the test site 3 have obtained obvious promotion. Further, when the counter electrode 2 is set to a voltage opposite to that of the target R according to actual needs, it is possible to attract and aggregate more of the target R to the detection site 3 so as to interact with the probe such as the antibody T on the detection site 3, thereby improving the sensitivity of detection and the lower limit of detection. Further, as a result of experiments and practical work, it was concluded that the detection sensitivity was higher as a whole when the bias voltage applied to the electrode 2 was 0.2V to 0.8V or the bias voltage was-0.2V to-0.8V, but it was needless to say that fine adjustment was required within the above-mentioned voltage range for the target R, which is specific pathogenic bacteria, nucleic acids, antigens, DNA. The utility model discloses a preparation method of microarray chip selects to arrange electrode 2 on glass substrate 1 with the mode of coating by vaporization film-forming earlier, then the preparation of spraying graphite alkene is detection site 3 on electrode 2's wiring, later freeze-drying is poromeric graphene oxide again, and its step is simple and easy, can realize electrode 2 and detection site 3's combination, is suitable for the batch production of scale. The detection method of the microarray chip of the present invention is characterized in that the detection site of the microarray is provided with a voltage, and the voltage is opposite to the voltage of the individual microorganism, and the voltage is specially suitable for the detection method of the microarray chip in which the content of the microorganism in the sample is less than 10 6 Under the condition of cells/ml, the limit of detection (LOD) is obviously improved, and the detection of pathogenic bacteria is further expanded.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and although the present invention has been disclosed with reference to the above preferred embodiment, but not to limit the present invention, any person skilled in the art can make modifications or changes to equivalent embodiments without departing from the scope of the present invention, and any simple modification, equivalent change and modification made to the above embodiments by the technical spirit of the present invention still fall within the scope of the present invention.
Claims (5)
1. A microarray chip, comprising:
a substrate;
at least one detection array disposed on the substrate and including:
an electrode is arranged on the base plate and is provided with a plurality of electrodes,
at least one detection site, wherein the material of the detection site is porous graphene or porous graphene oxide,
a recognition substance, wherein the recognition substance is arranged at the detection site and can specifically interact with a target object in the sample;
the detection site is electrically connected with the electrode;
the detection arrays are arranged in order.
2. The microarray chip of claim 1 further comprising at least one validation array disposed on said substrate, said validation array comprising at least one of said test site and said identifier.
3. The microarray chip of claim 1, wherein said detection array is arranged in a staggered manner.
4. The microarray chip of claim 1 wherein at least one of said detection arrays has a positive voltage of 0.2 volts to 0.8 volts or a negative voltage of-0.2 volts to-0.8 volts.
5. The microarray chip of claim 4, wherein the staggered arrangement is a full staggered arrangement of rows and columns, a staggered arrangement of rows and columns, and/or a staggered arrangement of detection arrays.
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