CN1453365A

CN1453365A - Biochip array

Info

Publication number: CN1453365A
Application number: CN 02118330
Authority: CN
Inventors: 苏友欣; 鲁希连; 龙训民; 董忠和; 刘大章; 李世光; 林世明; 黄荣山; 林启万; 许世明; 李世元
Original assignee: GONGZHUN PRECISION INDUSTRY Co Ltd
Current assignee: GONGZHUN PRECISION INDUSTRY Co Ltd
Priority date: 2002-04-24
Filing date: 2002-04-24
Publication date: 2003-11-05
Anticipated expiration: 2022-04-24
Also published as: CN1184332C

Abstract

The present invention relates to one kind of biochip array. The biochip array includes one transparent substrate, several notched reaction zones on the substrate, each of which has several fixed attaching units in certain height with the first attaching plane on the top and the second attaching plane on the side. Therefore, the present invention can bear sample solution and maintain ultimate tag effectively, has increased action area, and is suitable for transmissive or reflective imaging detector.

Description

Array biochip

Technical field

The present invention relates to a kind of array biochip, particularly a kind of array biochip that is used for thymus nucleic acid (DNA), protein or biomass cells.

Background technology

Biochip mainly utilizes the specificity (specificity) between biomolecules, biomolecules (is mainly included nucleic acid, protein, carbohydrate, cell or tissue etc.) be fixed on particular substrate (substrate) and go up formation bioprobe (biological probe), afterwards, the biological specimen solution that desire is detected is (as blood, urine, body fluid or saliva etc.) on chip, mix, because its inside may be contained corresponding bioprobe and can carry out the target molecule (target) of molecular interaction with it, therefore, by having specificity and the bioprobe and the target molecule of ligand carry out association reaction (binding reaction) or ferment catalyzed reaction (catalytic reaction) each other, and cause that the sign signal (comprises optics, piezoelectricity, electrochemistry, the energy signal) produces, utilize proofing unit that (bioprobe-target molecule) mixture is carried out optics or the quantitative and image document discriminatory analysis of intensity of radioactivity detection again, to obtain the analytical information of detection of biological sample.

At present the bioprobe of biochip is fixed (immobilization) normal substrate that adopts and can be divided into two kinds, and a. adopts opaque substrate, as polymeric film (as nylon membrane) or be coated with the slide (glass slide) of metallic film; B. adopt transparency carrier, as the barren slide; But the two still has following problem in use:

1.a the metallic film that bioaffinity is strong in the method (as golden film) is though can be by connecting thing (linker) or forming covalent bonds with bioprobe itself, and light tight substrate such as nylon membrane also can utilize chemistry or physics (as ultraviolet commissure UVcross-linking) mode to carry out covalent bonds, but, these substrates are because of can't printing opacity, penetration (transmittant) scanner or the microscope that make colorimetric (colorimetric) or use fluorescent measure (fluorometric) usefulness can't be suitable for, only can use reflective (reflective) proofing unit, the suitability that lacks proofing unit, and limited the use of the fluorescent determination techniques of higher sensitivity, cause the inconvenience in the image analysis contrast;

2.b method adopts transparency carrier, though have the penetration of being applicable to and optical detection apparatuss such as reflective scanning mode instrument or microscope carry out the correlated advantage of image analysis, but, the fixing operation of its bioprobe is except that need carry out the pre-process of slide, often because of using physical property absorption to break away from the error that causes in qualitative, the quantitative analysis because of cleaning easily;

3. no matter the biochip made from a or b method, the small molecules terminal marking thing that in the process that detects, all is difficult for the underlined information of effective leave strip, to be applied in the ELISA detection technique on the biochip, through colorimetric or the fluorescent terminal marking thing (chromophore or fluorescent products) after the ferment catalysis, cleaning biochip surface and detaching when biochip detects, run off easily because of substrate is difficult to comprise carrying, cause the error of follow-up judged result.

Summary of the invention

Main purpose of the present invention is to solve the above problems, and avoids the existence of defective, the invention provides that a kind of bioprobe tack is stable, the proofing unit suitability strong and can effectively carry and keep the array biochip of terminal marking thing.

For achieving the above object, the present invention is a kind of array biochip, it includes a transparency carrier, on this substrate, be concaved with at least more than one reaction zone array, and a plurality of adhesion unit spaced apart are arranged at each reaction zone internal fixing, and this adhesion unit has a predetermined height and has the biomolecules affinity, and at its end face and side formation confession bioprobe fixed first attachment surface and second attachment surface, like this, reaction zone carrying sample solution also keeps the terminal marking thing, and increase the active area of bioprobe and sample solution with second attachment surface, and the transparency carrier by two adhesion unit and adhesion unit spaced apart be applicable to penetration or the reflective proofing unit that colorimetric or fluorescent detect simultaneously, constitutes an array formula biochip like this.

Relevant detailed description of the present invention and technology contents, existing as follows with regard to accompanying drawings:

Description of drawings

Fig. 1 is a floor map of the present invention;

Fig. 2-1 is a synoptic diagram of the present invention;

Fig. 2-2 is local enlarged diagrams of Fig. 2 of the present invention-1;

Fig. 3 is that the present invention uses synoptic diagram;

Fig. 4-1 is the another embodiment of the present invention synoptic diagram;

Fig. 4-2 is diagrammatic cross-sections of Fig. 4 of the present invention-1;

Fig. 4-3 is local enlarged diagrams of Fig. 4 of the present invention-2;

Fig. 4-4 is catalyzed reaction synoptic diagram of Fig. 4 of the present invention-3;

Fig. 5 makes performance synoptic diagram of the present invention.

Embodiment

See also Fig. 1,2-1,2-2, be plane of the present invention, the plane amplification of reaction zone 20 and the local enlarged diagram of Fig. 2-1, as shown in the figure: the present invention is a kind of array biochip, mainly utilize the specificity between biomolecules, predetermined biomolecules is fixed on becomes bioprobe 50 on the substrate 10, so that corresponding target molecule 61 can form mark with bioprobe 50 interactions in the sample solution 60 to be detected, carry out discriminatory analysis so that offer scanning device, it includes a transparency carrier 10 with at least more than one reaction zone 20, in this reaction zone 20, be laid with a plurality of adhesion unit 30 at interval, and this adhesion unit 30 has a predetermined height, and at its end face and side formation confession bioprobe 50 fixed, first attachment surface 31 and second attachment surface 32, therefore, the present invention can increase the active area of bioprobe 50 and sample solution 60 with second attachment surface 32, and effectively keep terminal marking thing 72 after ferment 70 effects with the gap of adhesion unit 30, and offer penetration or reflective proofing unit discriminatory analysis simultaneously by transparency carrier 10, constitute an array formula biochip like this.

Aforesaid substrate 10 is a glass, and the reaction zone 20 on it can be configured on the substrate 10 with the array way manufacturing, and its reaction zone 20 can be made becomes circle, rectangle or other geometrical shape, in addition, above-mentioned adhesion unit 30 is to be made with the high material of biomolecules affinity, and the material that its biomolecules affinity is high can be a heavy metal, as gold or silver-colored, but with Jin Weijia, and the square section of its adhesion unit 30 can be circle, rectangle or other geometrical shape.

During manufacturing, can utilize the interface layer 40 that the configuration one earlier of micro electronmechanical photolithography technology can both engage with adhesion unit 30 and substrate 10, and on interface layer 40, make adhesion unit 30, and this adhesion unit 30 repeats to be distributed on the reaction zone 20 with a predetermined spacing, according to present manufacturing technology, this spacing can be 1,5,10,25,50,100 μ m, but its spacing is good with little person, in order to increase the total area of second attachment surface, the predetermined height of this adhesion unit 30 is then at least greater than 50nm, make adhesion unit 30 lateral second attachment surfaces 32 have more abundant biomolecules bond area, can be in conjunction with more target molecule 61, it highly can be 100,300,500,1000nm, but so that the spacing near adhesion unit 30 is good more more.

During use, see also Fig. 3, for the present invention uses synoptic diagram, inject on the reaction zone 20 with sample solution 60, the target molecule 61 that is linked with marker 62 (as fluorescent substance FITC) in the sample solution 60 promptly begins and first of adhesion unit 30, two attachment surfaces 31, bioprobe 50 on 32 interacts, and the marker 62 that utilizes target molecule 61 to be connect produces label information, can utilize the label information of first attachment surface 31 to carry out discriminatory analysis like this, and can utilize the label information of second attachment surface 32 to carry out discriminatory analysis for the penetration proofing unit for reflective proofing unit.

See also Fig. 4-1,4-2,4-3,4-4, be another embodiment of the present invention, the diagrammatic cross-section of Fig. 4-1, the part amplification of Fig. 4-2 and the catalyzed reaction synoptic diagram of Fig. 4-3, as shown in the figure: this reaction zone 20 is arranged with on substrate 10 ' with a drop section 11, should avoid destroying bioprobe 50 when not acting on sample solution 60 detaching, in addition, as when using the ELISA detection technique, be subjected to ferment 70 catalytic marks to be subjected to matter 71, the terminal marking thing 72 that produces with colorimetric or fluorescent effect, can be carried and be retained in the drop section 11, make that terminal marking thing 72 is unlikely to run off because of the difficult carrying of base material when biochip detects detaching, influence the result of subsequent detection.

At last, seeing also Fig. 5, is performance synoptic diagram of the present invention, as shown in the figure, reaction zone 20 on the substrate 10,10 ' can arrange with 2 * 2,3 * 3,4 * 4 or 5 * 5 array way, and the spacing of this reaction zone 20 can cooperate existing automatization titration apparatus and designs.

Claims

1. array biochip, utilize interbiotic specificity, predetermined biomolecules is fixed on becomes bioprobe (50) on the substrate (10), so that corresponding target molecule (61) can form mark with bioprobe (50) interaction in the sample solution to be detected (60), carry out discriminatory analysis so that offer proofing unit, it is characterized in that:

Described biochip includes a transparency carrier (10), which is provided with at least more than one reaction zone (20);

A plurality of interior and adhesion unit (30) spaced apart of above-mentioned reaction zone (20) that are fixed on, described adhesion unit (30) has a predetermined height, and forms for bioprobe (50) fixed first attachment surface (31) and second attachment surface (32) at its end face and side;

Like this, second attachment surface (32) increases the active area of bioprobe (50) and sample solution (60), and with the marker (62) behind the effective stick effect in gap of adhesion unit (30), and offer penetration or reflective proofing unit discriminatory analysis simultaneously by transparency carrier (10), constitute an array formula biochip like this.

2. array biochip according to claim 1 is characterized in that, described adhesion unit (30) is to be distributed on the reaction zone (20) with a predetermined spacing, and described spacing can be 1,5,10,25,50,100 μ m, with the little person of spacing for well.

3. array biochip according to claim 1 is characterized in that, the predetermined height of described adhesion unit (30) is at least greater than 50nm, can be 100,300,500,1000nm, and is good with the spacing near adhesion unit (30).

4. array biochip according to claim 1 is characterized in that, its square section of described adhesion unit (30) can be circle, rectangle or other geometrical shape.

5. array biochip according to claim 1 is characterized in that, described reaction zone (20) can be configured on the substrate (10) with matrix-style.

6. array biochip according to claim 1 is characterized in that, described reaction zone (20) can be circle, rectangle or other geometrical shape.

7. array biochip according to claim 1 is characterized in that, described adhesion unit (30) is to make with the high material of biomolecules affinity.

8. array biochip according to claim 1 is characterized in that, described adhesion unit (30) is to utilize micro electronmechanical photolithography fabrication techniques on substrate (10).

9. array biochip according to claim 1 is characterized in that, described substrate (10) is the glass of light-permeable.

10. array biochip according to claim 1 is characterized in that, described reaction zone (20) is arranged with on substrate (10 ') with a drop section (11).