WO2013152679A1

WO2013152679A1 - Integrated micro-optical structure array device

Info

Publication number: WO2013152679A1
Application number: PCT/CN2013/073524
Authority: WO
Inventors: 王振宇; 董凌志; 梁银针; 马贵兰; 戴良
Original assignee: 无锡国盛精密模具有限公司
Priority date: 2012-04-10
Filing date: 2013-03-30
Publication date: 2013-10-17
Also published as: CN103364344A

Abstract

An integrated micro-optical structure array device comprises a solid substrate (300) integrated with micro-optical structure arrays (310) and a frame (400). Multiple micro-optical structure arrays (310) exist, and each micro-optical structure array (310) is arranged in the form of an array on a plane. The frame (400) is combined on the solid substrate (300) and separates the micro-optical structure arrays (310). A micro-optical structure (111) is a light diverting structure or is formed by integrating a light diverting structure and a light condensing structure. The light diverting structure is a circular table structure (111c), a square column structure, a hexagonal column structure, or an octahedral column structure, and the light condensing structure is a convex lens (111e) or a Fresnel lens (111d). The size of the device is the same as that of a standard microplate. One-to-one correspondence exists between the positions of micro-optical structure arrays (310) and holes on the standard microplate. The micro-optical structure improves the signal collecting efficiency, and a standard microplate structure thereof is compatible with an existing device, and can implement automation in the whole detection process.

Description

Integrated micro-optical structure array device

Technical field

The present invention relates to the field of biotechnological detection by constructing a microanalytical system by immobilizing target molecules on the surface of a micro-optical structure, and more particularly to an integrated micro-optical structure array device.

Background technique

In the prior art, the microarray technology is a planar carrier in which a plurality of cells are arranged neatly in rows and columns, each of which regularly or specifically adsorbs a gene or a protein molecule. An analytical device called a microarray should conform to: 1 is planar, 2 regular, and 3 are three requirements on the microscopic scale. The microscopic scale of the microarray allows the reaction to occur rapidly in terms of reaction kinetics, enabling rapid detection of a large number of indicators.

Currently, microarray chips usually use standard glass slides (1 inch * 3 inches). However, microarray chips using glass slides have the following problems:

1. The operation is complicated, such as the paste of the chip fence, the cleaning of the slide, and all of them are manual operation, which is not conducive to the accuracy and stability of the reaction.

2. The test is not flexible. The entire slide must be used each time. If the number of samples is small, it will be wasted.

3. The organic molecular film coated on the surface of the glass is easy to fall off, resulting in unstable experimental results.

4. The planar structure of the glass slide results in low fluorescence collection efficiency, the glass surface is not easy to be chemically treated, and the background signal of the glass is high, which makes it unfavorable for highly sensitive detection.

Summary of the invention

SUMMARY OF THE INVENTION A primary object of the present invention is to provide a novel integrated micro-optical structure array device which can improve the signal collection efficiency of a biochip, realize automatic operation, and enhance the flexibility of detection, in view of the above-mentioned drawbacks of the prior art.

In order to achieve the above object, the present invention adopts the following technical solutions:

An integrated micro-optical structure array device, comprising: a solid substrate and a frame integrated with an array of micro-optical structures; the micro-optical structure array has a plurality of groups, and each group of micro-optical structure arrays are arranged in an array on one plane The frame is bonded to a solid substrate and separates the array of micro-optical structures.

The micro-optical structure has a signal enhancement effect on signals such as fluorescence, chemiluminescence, time-resolved fluorescence, and surface plasmon resonance (SPR).

An integrated micro-optical structure array device as described above, wherein

The solid substrate is composed of a plurality of pore structures having the same structure and size, which are arranged at equal intervals; A set of micro-optical structure arrays are integrated on the bottom surface of the pore structure, and the inner surface of the pore structure is combined with an active group capable of binding with biomolecules, and the outer surface is provided with a card structure;

The frame is provided with a plurality of cells arranged in a rectangular array on the same plane, each of the cells is provided with a positioning structure, and each of the cells is respectively accommodated with one of the hole structures, and the positioning structure and the structure are The card structure of the outer surface of the hole structure is phase-locked, the solid substrate is fixed on the frame, and the micro-optical structure array on the inner bottom surface of each hole structure is located on the same plane.

The integrated micro-optical structure array device as described above, preferably, the material of the solid substrate is one of polystyrene, a cyclic olefin polymer, and a styrene/acrylonitrile copolymer.

In the integrated micro-optical structure array device as described above, preferably, the solid substrate and the micro-optical structure are integrally formed.

An integrated micro-optical structure array device as described above, wherein

The solid substrate is a planar structure, and the sets of micro-optical structure arrays are integrated on an upper surface of the solid substrate and arranged in a rectangular array;

The frame is coupled to the upper surface of the solid substrate, and is provided with a plurality of through holes, each of the through holes is arranged in a rectangular array, and the through holes are in one-to-one correspondence with the positions of the micro-optical structure array An array of optical structures is exposed from the vias;

On the upper surface of the solid substrate, an active group capable of binding to a biomolecule is bonded to a position exposed from the through hole.

In the integrated micro-optical structure array device as described above, the micro-optical structure and the solid substrate are integrally formed.

The integrated micro-optical structure array device as described above, preferably,

The frame is a rigid material that is bonded to the solid substrate by means of gluing or mechanical fastening; or

The frame is a rubber material adhered to the upper surface of the solid substrate.

The integrated micro-optical structure array device as described above, preferably, the integrated micro-optical structure array device is the same size as a standard microplate, and the positions of the sets of micro-optical structure arrays are on a standard microplate The holes correspond one by one.

An integrated micro-optical structure array device as described above, wherein the standard microplate can be a standard

384-well plates, 96-well plates, 72-well plates, 48-well plates, 8-well plates, 12-well plates, 16-well plates, and the like.

The integrated micro-optical structure array device as described above, wherein the micro-optical structure is a light-converting structure or is a combination of a light-converting structure and a light-concentrating structure.

The integrated micro-optical structure array device as described above, preferably, the light-converting structure is a truncated cone structure, One of a cylindrical structure, a square column structure, a hexahedral column structure, and an octahedral column structure, and the upper surface of the light conversion structure is planar.

In the integrated micro-optical structure array device as described above, preferably, the outer surface of the light-converting structure is plated with a reflection film, and the reflection film is one of a metal reflection film, a dielectric reflection film, and a metal dielectric reflection film.

In the integrated micro-optical structure array device as described above, preferably, the light-concentrating structure is one of a convex lens and a Fresnel lens, and the light-concentrating structure is integrated at the bottom of the light-converting structure.

The integrated micro-optical structure array device as described above, preferably, the concentrating structure and the light-converting structure are integrally formed

The beneficial effects of the invention are:

The micro-optical structure array device of the present invention adopts a micro-optical structure to improve signal collection efficiency and thus reduce the requirements of the detection device, and the micro-optical structure can also provide signal position information, simplifying software processing time and work. The microplate structure is compatible with existing equipment, which can automate the whole inspection process and improve the accuracy and stability of the detection. The detachable structure makes the detection more flexible, and each plate can realize 1-96. Detection of one sample.

DRAWINGS

1 is a schematic view showing the overall structure of Embodiment 1 of an integrated micro-optical structure array device of the present invention. Figure 2 is a longitudinal cross-sectional view showing one of the holes in Embodiment 1 of the present invention.

Figure 3 is a top plan view of a solid substrate incorporating an array of micro-optical structures of Examples 2 and 3 of the present invention.

4 is a schematic view showing the arrangement of a micro-optical structure array according to Embodiment 2 of the present invention.

Fig. 5 is a schematic plan view showing the appearance of the perforated frame according to the second embodiment of the present invention.

Fig. 6 is a longitudinal sectional view showing a perforated frame according to a second embodiment of the present invention.

Fig. 7 is a longitudinal sectional view showing another perforated frame according to a second embodiment of the present invention.

Figure 8 is a schematic view showing the structure of a pallet of Embodiment 2 of the present invention.

Fig. 9 is a schematic plan view showing the appearance of the perforated frame of the embodiment 3 of the present invention.

Figure 10 is a longitudinal cross-sectional view showing an embodiment of a micro-optical structure employed in the present invention.

Figure 1 is a longitudinal cross-sectional view showing another embodiment of the micro-optical structure employed in the present invention.

detailed description

The present invention provides an integrated micro-optical structure array device. The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. (1) Structure of integrated micro-optical structure array device Example 1

As shown in the structural schematic view of Fig. 1 and the longitudinal cross-sectional view of Fig. 2, in the present embodiment, the integrated micro-optical structure array device is composed of a slat 100 and a plate frame 200 in which an array of micro-optical structures is integrated. The structure of the slat 100 is the same as that of the ELISA (Enzyme Linked Immunosorbent Assay) used in the prior art, and can be commonly used in 8 holes, 12 holes, 16 holes, etc., and can be disassembled into arbitrary holes. The number is placed on the board frame. The single aperture structure 110 of the slats 100 can be circular or square shaped with the same aperture spacing as a standard microplate. The inner surface 112 of each pore structure 110 is surface treated to have a reactive group capable of binding to a biomolecule, such as an aldehyde group or an epoxy group capable of binding to a protein sample, and a micro-optical structure 111 integrated on the inner bottom surface. The array, and preferably, the height ratio of the pore structure 110 to the micro-optical structure 111 is greater than 100 times. The micro-optical structure 111 has a light-converting effect to increase the signal collection rate. The outer surface of each of the hole structures 110 is further provided with a latching structure 113 for fixing to the frame 200.

The plate frame 200 can be designed with reference to the specifications of existing standard microplates, such as standard 384-well plates, standard 96-well plates, standard 48-well plates, 8-hole slats, and the like. Specifically, taking a standard 96-well plate specification as an example, the plate frame 200 can accommodate 1-96 hole structures 110 having 117 (13x9) groups arranged in a standard 96-well plate layout (12x8). The petal structure 210 of the fixed pore structure. Specifically, the frame 200 includes an outer frame, and the space enclosed by the vertical stripes is separated by 11 horizontal stripes and 7 vertical stripes to form 96 square cells distributed in a grid shape. 210 is placed at four right angles of each cell. The petal structure at the intersection of the horizontal strip and the longitudinal strip is composed of four petal-shaped elastic clips; the petal structure at the intersection of the horizontal strip or the longitudinal strip and the outer frame is composed of two petal-shaped elastic clips. Each of the elastic clips has a curved surface, and the arc protrudes toward the center of the hole in which the arc is located, so that each of the four corners of each of the holes of the frame 200 is provided with an elastic clip protruding toward the center thereof. A hole structure 113 can be fixed therein.

When the slats 100 are placed on the slab 200, four elastic clips located at the four corners of each of the cells are clamped in the latch structure 113 of each of the hole structures 110, so that the hole structure 110 is realized on the slab 200. It is fixed to constitute the integrated micro-optical structure array device in this embodiment. The interaction between the elastic clip and the latching structure 113 can also ensure that the panel 200 is used when the panel frame 200 is inverted, and the slat 100 can still be fixed on the panel 200. This kind of good fixation, The integrated micro-optical structure array device is made easy to pick and place.

In this embodiment, the material of the slat 100 integrated with the micro-optical structure array may be a material such as polystyrene, a cycloolefin polymer, a styrene/acrylonitrile copolymer, etc., which is integrally processed by an injection molding process. On the inner bottom surface of each of the hole structures 113 of the slat 100 of the present invention, an array of 8*8 micro-optical structures is formed in a circumference having a diameter of about 4 mm; the micro-optical structure array having an upper surface diameter of one hundred micrometers is evenly distributed, And at the same level, and the height error of each micro-optical structure is strictly controlled within 5%.

The mold core of the injection mold of the slat 100 of the invention adopts EDM electric discharge machining mode, and the specific process route is as follows: 1 The electrode is engraved by a five-axis machining center, and the electrode is subjected to surface polishing treatment; 2 The EDM machine tool is configured with a micro-machining power supply to ensure the core The size, shape and surface roughness requirements; 3 The core is finally combined with mechanical, physical and chemical surface treatment technology to achieve a surface roughness of Ra ≤ 0.02 m without destroying the size and shape.

When the integrated micro-optical structure array device is applied for detection, the upper surface of the micro-optical structure array is spotted, and after being fixed, sealed, cleaned, etc., the micro-array device can be directly placed in the automatic enzyme-linked immunoassay workstation. Or other general-purpose automated instruments for 96-well plates for loading, incubating, washing, etc.; after the end of the operation, place them in a specific scanner for reading. Alternatively, the integrated micro-optical structure array device can be placed directly in a fully automated workstation with loading, incubation, plate washing, and scanning functions for full automation. In addition, the microarray device can also be applied to manual operations.

Example 2

As shown in Figs. 3 and 5, in the present embodiment, the integrated micro-optical structure array device is composed of a solid substrate 300 and a perforated frame 400 having an integrated micro-optical structure array on its surface.

The solid substrate 300 may be of a size of 1 inch * 3 inches, 0.72 inches * 3 inches, 2 inches * 6 inches, etc., and may be made of glass or a polymer material. The micro-optical structure array 310 is integrated on the surface of the solid substrate, and the two are integrally formed. As shown in Figure 4, the density and location of the array of micro-optical structures 310 can be as desired. The perforated frame 400 can be of a standard size of 16 holes, 24 holes, 48 holes, 72 holes or 96 holes. The holes 410 can be circular or square, and the holes are through holes and have no bottom.

The solid substrate 300 and the perforated frame 400 can be combined in various ways, and there are three preferred combinations. The first is a combination of gluing, that is, using a medical adhesive that has no effect on the reaction. The second type is a combination of a gasket, that is, a gasket is first fixed on the surface of the solid substrate 300, and then the solid substrate 300 is bonded to the perforated frame 400 through a gasket. The advantage of this method is that the perforated frame 400 can be removed from the gasket and is a detachable structure. The third method is a combination of mechanical means, that is, bonding the solid substrate 300 to the perforated frame 400 by a jig. As shown in FIG. 6, the outer edge of the lower surface of the perforated frame 400 has a downwardly projecting step structure 420 such that when it is combined with the solid substrate, the lower edge of the stepped structure 420 protrudes from the bottom surface of the solid substrate, so when During the inspection operation, the bottom surface of the solid substrate is not in contact with the operation surface, thereby preventing the solid substrate from being scratched or contaminated to affect the detection result. As shown in Fig. 7, the outer edge of the upper surface of the perforated frame 400 may also have a stepped structure 430 that projects outwardly in a horizontal plane such that the perforated frame 400 can be placed on the pallet 500 as shown in FIG. Preferably, the size of the plate frame 500 can be the same as that of the standard 96-hole plate. If the perforated frame 400 is 24 holes, the plate holder 500 can be placed with four perforated frames 400 that have been combined with the solid substrate 300. Advantages of the step structure 430 It is easy to operate and easy to automate.

The surface treatment of the solid substrate 300 may be performed before or after it is combined with the perforated frame 400 by surface treatment such that the surface of the solid substrate 300 has active groups that can bind to biomolecules. If the surface treatment is performed before the solid substrate 300 is bonded to the perforated frame 400, the solid substrate 300 is bonded to the perforated frame 400 after the surface treatment is spotted, and then the micro-optical structure array 310 integrated on the surface of the solid substrate 300 is completed. The upper surface is made of bioactive molecules. If the surface treatment is performed after the solid substrate 300 is bonded to the perforated frame 400, the bioactive molecules are spotted on the upper surface of the micro-optical structure array 310 integrated on the solid substrate surface 300 after the surface treatment is completed. After the spotting is completed, the integrated micro-optical structure array device of the embodiment can be directly placed in an instrument with an automatic function of loading, incubating, washing, scanning, etc., after being fixed, closed, cleaned, etc., Can be operated manually. Example 3

As shown in FIG. 3 and FIG. 9, in the present embodiment, the integrated micro-optical structure array device is composed of a solid substrate 300 having a micro-optical structure array integrated on its surface and a chip fence 600.

The solid substrate 300 may be of a size of 1 inch * 3 inches, 0.72 inches * 3 inches, 2 inches * 6 inches, etc., and may be made of glass or a polymer material. The micro-optical structure array 310 is integrated on the surface of the solid substrate, and the two are integrally formed. The chip fence 600 is a rubber material, which is soft in texture and easy to adhere to the surface of the solid substrate 300. Its function is mainly to isolate the sample and avoid cross-contamination, and is suitable for occasions where the sample to be tested is small.

The solid substrate 300 of the integrated micro-optical structure array 310 is first surface-treated to have active groups on the surface, and then spotted on the surface of the micro-optical structure. After the spotting is completed, after fixing, sealing, cleaning, etc., the chip fence 600 is pasted on the surface of the substrate, and then the integrated micro-optical structure array device of the embodiment can be applied to the sample loading, incubation, washing, and Manual operations such as scanning. The size and density of the holes 610 of the chip fence 600 can be flexibly selected as needed.

(ii) Structure of micro-optical structures

In the above integrated micro-optical structure array device, the micro-optical structure 111 may be a light-converting structure, and specifically, may be designed as a truncated cone structure, a cylindrical structure, a square pillar structure, a hexahedral column structure or an octahedral column structure. Through the light-converting structure, the direction of the divergent signal can be changed so that it can be detected by a signal detector placed under the chip body, thereby increasing the signal collection rate. In addition, the micro-optical structure 11 1 also It can be composed of a light-converting structure and a concentrating structure, that is, a concentrating structure, such as a convex lens structure and a Fresnel lens, is integrated at the bottom of the light-converting structure. Since the light-converting structure mainly relies on reflection to steer the signal, the reflective film can be plated on the reflective surface (ie, the outer surface) of the micro-optical structure light-conducting structure to increase signal reflectivity and reduce signal transmission loss, such as metal reflective film, dielectric reflection. Film, metal dielectric reflective film, etc.

When the micro-optical structure 111 is a light-converting structure, an array composed of the structure is distributed on the upper surface of the solid substrate; when the micro-optical structure 111 is composed of a light-converting structure and a condensing structure, the structure is from a solid substrate The upper surface extends longitudinally through the entire solid substrate from top to bottom, wherein the light-converting structure is distributed on the upper surface of the substrate, and the light-concentrating structure is located below the substrate.

Example 4

As shown in the longitudinal sectional view of Fig. 10, in the present embodiment, the micro-optical structure 111 is composed of a light-converting structure and a condensing structure. The light-converting structure is a truncated cone structure l l lc, and the concentrating structure is a Fresnel lens l l ld, which is integrated under the light-converting structure. The entire micro-optical structure 111 extends longitudinally through the entire solid substrate from the top of the solid substrate surface, wherein the truncated cone structure 111c is distributed over the surface of the solid substrate. The angle between the bottom surface and the side surface of the truncated cone structure 111c is designed to be a fixed value θ, which is between 45 and 70 degrees, depending on the materials used. The frustum interface I of the truncated cone structure 111c (i.e., the outer surface of the light-converting structure) is a surface of the plated reflective film, and the light within a certain angular range generated by the fluorescent molecular layer 111b distributed on the upper surface 111a of the frustum is totally reflected at the interface I. The reflected light is reflected at the concentrating interface II, and then the light is refracted at the concentrating interface III, and finally the light is reflected at the concentrating interface IV to be received by the detector directly under the chip body.

Example 5

As shown in the longitudinal sectional view of Fig. 11, in the present embodiment, the micro-optical structure 111 is composed of a light-converting structure and a condensing structure. The light-converting structure is a circular table structure l l lc, and the specific structural design thereof is the same as that of the fourth embodiment. The concentrating structure is a convex lens l l le , which is integrated under the light-converting structure. The entire micro-optical structure 111 extends through the entire solid substrate from the top to the bottom in a longitudinal direction from the surface of the solid substrate, wherein the truncated cone structure 111c is distributed on the surface of the solid substrate. The light within a certain angular range generated by the fluorescent molecular layer 111b distributed on the upper surface 111a of the circular table is totally reflected at the interface I of the truncated cone, and the reflected light is refracted at the collecting interface V to be received by the detector directly below the chip body. .

It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit the scope of the present invention. Those skilled in the art, under the guidance of the spirit of the technical solutions of the present invention, Modifications or equivalents are intended to be included within the scope of the appended claims.

Claims

An integrated micro-optical structure array device, characterized in that it comprises a solid substrate and a frame integrated with an array of micro-optical structures; the micro-optical structure array has a plurality of groups, and each group of micro-optical structure arrays is on a plane Arranged in an array; the frame is bonded to a solid substrate and the sets of micro-optical structures are separated.

2. The integrated micro-optical structure array device according to claim 1, wherein the solid substrate is composed of a plurality of pore structures having the same structure and the same size; the bottom surface of each of the pore structures Each of them is integrated with a set of micro-optical structure arrays, and the inner surface of the pore structure is combined with an active group capable of binding with biomolecules, and the outer surface is provided with a card structure;

The frame is provided with a plurality of cells arranged in a rectangular array on the same plane, and each of the cells is provided with a positioning structure, and each of the holes is respectively accommodated with one of the hole structures, and the positioning structure and the positioning structure are The latch structure of the outer surface of the hole structure is phase-locked, the solid substrate is fixed on the frame, and the micro-optical structure array on the inner bottom surface of each hole structure is located on the same plane. Book

The integrated micro-optical structure array device according to claim 2, wherein the material of the solid substrate is one of polystyrene, a cycloolefin polymer, and a styrene/acrylonitrile copolymer.

The integrated micro-optical structure array device according to claim 1 , wherein the solid substrate is a planar structure, and the sets of micro-optical structure arrays are integrated on an upper surface of the solid substrate and arranged in a rectangular array;

5. The integrated micro-optical structure array device according to claim 1, wherein the frame is a rigid material combined with the solid substrate and by gluing or mechanical fixing; or

The integrated micro-optical structure array device according to any one of claims 1 to 5, wherein the integrated micro-optical structure array device has the same size as a standard microplate, and the groups are The position of the micro-optical structure array corresponds one-to-one with the holes on the standard microplate.

7. The integrated micro-optical structure array device according to claim 1, wherein the micro-optical structure is a light-converting structure or is formed by a combination of a light-converting structure and a light-concentrating structure.

The integrated micro-optical structure array device according to claim 7, wherein the light-converting structure is one of a truncated cone structure, a cylindrical structure, a square pillar structure, a six-sided column structure, and an octagonal column structure. And the upper surface of the light-converting structure is flat.

9. The integrated micro-optical structure array device according to claim 8, wherein an outer surface of the light-converting structure is plated with a reflective film, and the reflective film is a metal reflective film, a dielectric reflective film, and a metal dielectric reflective film. One of them.

10. The integrated micro-optical structure array device according to claim 7, wherein the concentrating structure is one of a convex lens and a Fresnel lens, and the concentrating structure is integrated at a bottom of the light-converting structure.