CN217781104U

CN217781104U - Double-hole sample-adding microfluidic biochip card for NanoSPR detection

Info

Publication number: CN217781104U
Application number: CN202220716281.0U
Authority: CN
Inventors: 黄丽萍; 王易; 刘钢; 曾少奇
Original assignee: Liangzhun Shanghai Medical Devices Co ltd
Current assignee: Liangzhun Shanghai Medical Devices Co ltd
Priority date: 2022-03-30
Filing date: 2022-03-30
Publication date: 2022-11-11
Anticipated expiration: 2032-03-30

Abstract

The utility model discloses a double-hole sample-adding micro-fluidic biological chip card for NanoSPR detection, belonging to the technical field of nano plasma resonance; the device specifically comprises a bottom plate, wherein a flow channel is arranged on the bottom plate; one end of the flow channel is provided with a waste liquid pool, and the other end of the flow channel is respectively connected with a first sample adding pool and a second sample adding pool with different volumes through connecting channels; the depths of the second sample adding pool and the first sample adding pool are deeper than the flow channel; a chip groove is arranged in the middle of the flow channel, and a biological chip is placed in the chip groove; the bottom plate is also provided with a through hole, and the position of the through hole corresponds to the chip slot; the surface sealing laminating of runner and waste liquid pond has hydrophilic membrane, the waste liquid pond is equipped with water absorbing material. The utility model provides a micro-fluidic biochip card which can be specially applied to the micro-fluidic biochip detection technology that reagents with different volumes are added in sequence; simple structure, the application of sample is convenient, need not additionally use syringe, flow pump application of sample.

Description

Double-hole sample-adding microfluidic biochip card for NanoSPR detection

Technical Field

The utility model belongs to the technical field of nanometer plasma resonance, especially a diplopore application of sample micro-fluidic biological chip card for NanoSPR detects.

Background

Microfluidic Surface Plasmon Resonance (SPR) is commonly used in a variety of biochemical detection methods. The SPR technique does not require exogenous labeling of the sample and can also be used to measure biochemical reaction kinetics, thus obtaining affinity and thermodynamic parameters. The working detection principle of most of the currently used SPR systems based on microfluidic flow cells is as follows: the sample is transferred to the flow cell by an injection or pulling method by using an automatic sampler, reacts with a Microfluidic Chip (Microfluidic Chip) in the flow cell, and then is placed into an enzyme-labeling instrument and other instruments for detection.

For example, chinese granted patent CN213875425U provides a biosensor cartridge and a biosensor device, in which one or more flow cells, a sensor chip and a sampler are integrated together to form an independent biosensor cartridge which can be conveniently detached and replaced, and the biosensor cartridge can be replaced after a test, so that the problems of complex structure and the like of the existing biosensor analysis instrument based on surface plasmon resonance are solved, the working efficiency is improved, and the use cost is reduced. However, the analysis cartridge and the related devices also need to be matched with other devices and instruments such as a syringe, a flow pump and the like to realize the flow of the sample in the flow channel and the flow cell, so that the analysis cartridge and the related devices are inconvenient to carry and use; and the combined contact area of the sample liquid and the sensor chip is limited, which is not favorable for obtaining the detection accuracy and the sensitivity is limited.

Disclosure of Invention

In order to solve the problem of the prior art, the utility model provides a diplopore application of sample micro-fluidic biological chip card for NanoSPR detects realizes through following technical scheme.

The double-hole sample-adding microfluidic biochip card for detecting NanoSPR comprises a bottom plate, wherein a flow channel is arranged on the bottom plate; one end of the flow channel is provided with a waste liquid pool, and the other end of the flow channel is respectively connected with a first sample adding pool and a second sample adding pool with different volumes through connecting channels; the depths of the second sample adding pool and the first sample adding pool are deeper than the flow channel; a chip groove is arranged in the middle of the flow channel, and a biological chip is placed in the chip groove; the bottom plate is also provided with a through hole, and the position of the through hole corresponds to the chip slot; the surface sealing laminating of runner and waste liquid pond has hydrophilic membrane, the waste liquid pond is equipped with water absorbing material.

The utility model provides a two-hole application of sample micro-fluidic biological chip card is to the chip card of the brand-new structure of nanometer surface plasmon resonance technical invention, and this chip card can be used to test specific NanoSPR detection method application that intermolecular interaction affinity etc. need successively additionally add the plus reagent of different quantity. The most important innovation point of the chip card is that the sample adding pools are arranged into two, namely a first sample adding pool and a second sample adding pool, and = the depths of the first sample adding pool and the second sample adding pool are deeper than the flow channel, and the volumes of the first sample adding pool and the second sample adding pool are larger or smaller.

Taking the test of the intermolecular affinity as an example, a sample application reservoir with a large volume is provided for first adding a ligand (capable of specifically binding to a target in a sample, generally 200. Mu.l) to bind to a biochip, and a sample application reservoir with a small volume is provided for later adding a sample to be tested (generally 30. Mu.l). The specific using method comprises the following steps:

(1) A ligand reagent is dripped into a first sample adding pool (with larger volume), ligand reagent liquid flows into a flow channel and flows into a chip groove along the flow channel through the guiding action of a hydrophilic membrane and the capillary action of the flow channel, the ligand reagent liquid contacts with the biochip and infiltrates the surface of the biochip to start reaction, and the ligand is combined to the surface of the biochip;

(2) The reacted ligand reagent waste liquid continuously flows towards the waste liquid pool along with the flow channel and is absorbed by the water absorption material in the waste liquid pool;

(3) Then adding a sample to be detected into the second sample adding pool (with a smaller volume), wherein the liquid of the sample to be detected also flows into the flow channel and flows into the chip groove along the flow channel under the guiding action of the hydrophilic membrane and the capillary action of the flow channel; if the target object to be detected exists in the sample to be detected, the target object to be detected and the ligand are combined and fixed on the biochip; the waste liquid of the sample to be detected after reaction continuously flows towards the waste liquid pool along with the flow channel and is absorbed by the water absorption material in the waste liquid pool;

(4) After the reaction is finished, putting the mixture into an enzyme-labeling instrument and other instruments for qualitative and quantitative detection.

The liquid in the microfluidic biochip card can flow mainly by means of the hydrophilic action of the hydrophilic membrane and the active driving action of the capillary action of the flow channel. The hydrophilic membrane can be made of common materials, for example, finished products (products of the company such as Desha/TESA) can be directly purchased, and the hydrophilic membrane is generally adhered to the bottom plate by using waterproof double-sided adhesive (such as 3M waterproof double-sided adhesive) so as to seal and isolate the flow channel and the chip groove from the outside; certainly, the hydrophilic film can be pasted by coating waterproof glue with low fluidity, but the surface of the hydrophilic film is smooth after pasting. The water-absorbing material can be common water-absorbing paper or sponge and other materials. In order to slow down the flow velocity of liquid, the depth of the first sample adding pool and the second sample adding pool is set to be larger than the depth of the flow channel, so that the first sample adding pool and the second sample adding pool can overflow the sample adding pool and slowly flow into the flow channel only after a certain amount of liquid is added into the first sample adding pool and the second sample adding pool.

After the first sample adding pool and the second sample adding pool are respectively and independently arranged, the respective connecting channels have various design forms communicated with the flow channels, so that the first sample adding pool and the second sample adding pool can be communicated with the flow channels from different positions and can be communicated with the flow channels at the same position. When two application of sample ponds communicate with the runner in same position, the utility model discloses an applicant confirms through the product to assembly production, if add reagent and infiltration runner back in (the great) first application of sample pond of volume, add the reagent again in second application of sample pond and will get into the runner because hydrophilic membrane's from driving force and hydrophilicity, can not pour back into first application of sample pond.

The above-mentioned test of intermolecular affinity is only one example of the use of the biochip card of the present invention, and the chip card may be used for qualitative and quantitative tests in which different reagents are added successively, for example, ELISA method, colloidal gold method, etc.

Preferably, the first sample adding pool and the second sample adding pool have the same projection shape, and the depth of the first sample adding pool is larger than that of the second sample adding pool. This is one of the design modes of the first sample adding pool and the second sample adding pool.

More preferably, the bottom surface of the first sample addition cell and/or the second sample addition cell is a slope inclined toward the flow channel. As an arrangement mode of the first/second sample adding pool, the inclined plane can be used for meeting the aim of reducing the flow speed of liquid.

Preferably, the first sample adding pool and/or the second sample adding pool comprise an overflow area and a sample adding area, the overflow area is communicated with the connecting channel, the depth of the overflow area is the same as that of the flow channel (i.e. the connecting channel), and the depth of the sample adding area is greater than that of the flow channel. As a second setting mode of the first/second sample adding pool, a mode of setting an overflow area in a step shape is adopted, liquid is added into the sample adding area, and the liquid enters the overflow area after overflowing and then flows into the flow channel.

Preferably, the flow passages are distributed in a U shape. The flow channel is arranged in a U shape, so that the length of the flow channel is prolonged, the flowing time and the flowing speed of the liquid in the flow channel are increased, and the purpose of accommodating more liquid is realized.

Preferably, the flow channel comprises a main flow channel and 2 secondary flow channels, wherein the 2 secondary flow channels are arranged on two sides of the main flow channel, and the chip groove is positioned on the main flow channel; the first sample adding pool and the second sample adding pool are positioned at one end of the main flow channel, the 2 secondary flow channels are respectively connected to the other end of the main flow channel, and the tail ends of the secondary flow channels are provided with waste liquid pools. The main flow channel and the 2 secondary flow channels form an M-shaped structure, the main flow channel is arranged in the middle, and the 2 secondary flow channels are arranged on two sides of the main flow channel. After the liquid flows into the main flow channel and reacts with the biochip, the waste liquid is divided into two paths along 2 secondary flow channels and flows into corresponding waste liquid pools.

Preferably, the connecting channel is smoothly connected with the first sample adding pool, the second sample adding pool and the flow channel. The smooth connection enables the liquid to flow more smoothly in the connecting channel and the flow passage.

Preferably, the waste liquid pool is provided with a vent hole. The purpose of the vent hole is to facilitate the air in the flow channel and the chip groove to be discharged, and ensure the liquid to flow normally in the flow channel.

Preferably, the inlet and the outlet of the chip groove connected with the flow channel are both in a horn structure. The chip groove and the runner are connected by an inlet and an outlet. The inflow port is arranged in a horn shape, so that the inflowing liquid can be spread out to infiltrate all areas of the biochip, and the liquid is prevented from flowing out of the chip groove only in the middle area of the biochip to affect the detection effect. The outflow port is arranged to be horn-shaped, so that dead angles can be reduced, air bubbles in the flow channel and the chip groove can be completely extruded, and the influence of the air bubbles on the refraction or reflection effect of light emitted by the probe is avoided.

Preferably, the bottom plate is of a sandwich structure, a water absorbing material is arranged in the sandwich structure, and the waste liquid pool is communicated with the water absorbing material. The water absorbing material (such as water absorbing paper) is arranged in the interlayer of the bottom plate, and the waste liquid pool is communicated with the water absorbing material, so that more waste liquid can be absorbed.

Compared with the prior art, the beneficial effects of the utility model are that: the utility model provides a microfluidic biological chip card, which has simple structure and convenient sample adding, and does not need to additionally use an injector or a flow pump for sample adding; the microfluidic biochip detection technology can be specially applied to the microfluidic biochip detection technology which needs to add reagents with different volumes in sequence, for example, the microfluidic biochip detection technology is used for researching the interaction process among various biomolecules (such as polypeptides, proteins, oligonucleotides, viruses, bacteria and small molecular compounds); specific antibody detection or quality control, disease mechanism and drug screening; the method belongs to the technical field of relevant pharmacokinetics real-time monitoring, ligand fishing, immunoregulation, structure-function relation, epitope identification and the like.

Drawings

Fig. 1 is a top view of the double-well sample-adding microfluidic biochip card for NanoSPR detection provided by the present invention;

FIG. 2 is a top view of a first, preferably double-well sample-loading microfluidic biochip card;

FIG. 3 isbase:Sub>A cross-sectional view taken along line A-A of FIG. 2;

FIG. 4 is a cross-sectional view of a second preferred dual-well sample-loading microfluidic biochip card;

FIG. 5 is a cross-sectional view of a third preferred dual-well sample-loading microfluidic biochip card;

FIG. 6 is a top view of a fourth preferred dual-well sample loading microfluidic biochip card;

FIG. 7 is a top view of a fifth preferred dual-well sample-loading microfluidic biochip card;

FIG. 8 is a partial top view of a chip slot of a sixth, preferably double well loading microfluidic biochip card;

FIG. 9 is a cross-sectional view of a seventh preferred dual-well sample-loading microfluidic biochip card.

In the figure: 1. a base plate; 2. a flow channel; 201. a connecting channel; 202. a main flow passage; 203. a secondary flow passage; 3. a waste liquid tank; 4. a first sample addition pool; 5. a second sample addition pool; 6. a chip slot; 7. a biochip; 8. a through hole; 9. a hydrophilic membrane; 10. an overflow area; 11. a sample adding area; 12. absorbent paper.

Detailed Description

The technical solutions of the embodiments in this patent will be described clearly and completely with reference to the accompanying drawings, and it is obvious that the described embodiments are only some embodiments, not all embodiments, of this patent. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the patent without making creative efforts, shall fall within the protection scope of the patent.

In the description of this patent, it is noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", "top", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, and are only for convenience in describing the patent and for simplicity in description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore, should not be construed as limiting the patent. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.

In the description of this patent, it is noted that, unless expressly stated or limited otherwise, the terms "mounted," "connected," and "in communication with" are to be construed broadly and include, for example, fixed connections, removable connections, or integral connections; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. It is to be noted that all the figures are exemplary representations. The meaning of the above terms in this patent may be specifically understood by those of ordinary skill in the art.

The patent is described in further detail below with reference to specific embodiments and with reference to the attached drawings.

Example 1

As shown in fig. 1, the double-well sample-adding microfluidic biochip card for NanoSPR detection provided in this embodiment includes a bottom plate 1, where a flow channel 2 is arranged on the bottom plate 1; one end of the flow channel 2 is provided with a waste liquid pool 3, and the other end is respectively connected with a first sample adding pool 4 with a larger volume and a second sample adding pool 5 with a smaller volume through a connecting channel 201; the depths of the second sample adding pool 5 and the first sample adding pool 4 are deeper than the flow channel 2; a chip groove 6 is arranged in the middle of the flow channel 2, and a biochip 7 is placed in the chip groove 6; the bottom plate 1 is also provided with a through hole 8, and the position of the through hole 8 corresponds to the chip slot 6; the surface sealing laminating of runner 2 and waste liquid pond 3 has hydrophilic membrane 9, waste liquid pond 3 is equipped with water absorbing material.

As shown in fig. 2 and 3, the first sample adding pool 4 and the second sample adding pool 5 have the same projection shape, and the depth of the first sample adding pool 4 is greater than that of the second sample adding pool 5.

As shown in FIG. 4, the bottom surfaces of the first sample addition cell 4 and the second sample addition cell 5 are inclined surfaces inclined toward the flow channel 2.

As shown in fig. 5, the first sample addition reservoir 4 and the second sample addition reservoir 5 include an overflow area 10 and a sample addition area 11, the overflow area 10 is communicated with the connecting channel 201, the depth of the overflow area 10 is the same as that of the flow channel 2, and the depth of the sample addition area 11 is greater than that of the flow channel 2.

As shown in fig. 6, the flow channels 2 are distributed in a U shape.

As shown in fig. 7, the flow channel 2 includes a

main flow channel

202 and 2

sub-flow channels

203,2 of the sub-flow channels 203 are provided on both sides of the main flow channel 202, and the chip tray 6 is located on the main flow channel 202; the first sample adding pool 4 and the second sample adding pool 5 are positioned at one end of the

main flow channel

202, 2 of the auxiliary flow channels 203 are respectively connected at the other end of the main flow channel 202, and the tail ends of the auxiliary flow channels 203 are provided with waste liquid pools 3.

As shown in fig. 8, the inlet and the outlet of the chip groove 6 connected to the flow channel 2 are both in a horn shape.

As shown in fig. 9, the bottom plate 1 is a sandwich structure, and the sandwich structure is provided with water absorbent paper 12 therein, and the waste liquid tank 3 is communicated with the water absorbent paper 12.

The above embodiments describe the implementation of the present invention in detail, however, the present invention is not limited to the specific details of the above embodiments. Within the scope of the claims and the technical idea of the present invention, various simple modifications and changes can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

Claims

1. The double-hole sample-adding microfluidic biochip card for detecting NanoSPR is characterized by comprising a bottom plate, wherein a flow channel is arranged on the bottom plate; one end of the flow channel is provided with a waste liquid pool, and the other end of the flow channel is respectively connected with a first sample adding pool and a second sample adding pool with different volumes through connecting channels; the depths of the second sample adding pool and the first sample adding pool are deeper than the flow channel; a chip groove is formed in the middle of the flow channel, and a biological chip is placed in the chip groove; the bottom plate is also provided with a through hole, and the position of the through hole corresponds to the chip slot; the surface sealing laminating of runner and waste liquid pond has hydrophilic membrane, the waste liquid pond is equipped with water-absorbing material.

2. The double-hole sample-adding microfluidic biochip card for nanoSPR detection according to claim 1, wherein the first sample-adding pool and the second sample-adding pool have the same projection shape, and the depth of the first sample-adding pool is greater than that of the second sample-adding pool.

3. The double-well sample-loading microfluidic biochip card for NanoSPR detection according to claim 2, wherein the bottom surface of the first sample loading well and/or the second sample loading well is a slope inclined toward the flow channel.

4. The double-well sample-adding microfluidic biochip card for nanoSPR detection according to claim 1, wherein the first sample-adding reservoir and/or the second sample-adding reservoir comprises an overflow area and a sample-adding area, the overflow area is communicated with the connecting channel, the depth of the overflow area is the same as that of the flow channel, and the depth of the sample-adding area is greater than that of the flow channel.

5. The double-well sample-loading microfluidic biochip card for NanoSPR detection according to any one of claims 1 to 4, wherein the flow channels are distributed in a U-shape.

6. The double-hole sample-adding microfluidic biochip card for nanoSPR detection according to claim 1, wherein the flow channel comprises a main flow channel and 2 secondary flow channels, wherein the 2 secondary flow channels are arranged on two sides of the main flow channel, and the chip slot is positioned on the main flow channel; the first sample adding pool and the second sample adding pool are positioned at one end of the main flow channel, the 2 secondary flow channels are respectively connected to the other end of the main flow channel, and the tail ends of the secondary flow channels are provided with waste liquid pools.

7. The double-well sample-loading microfluidic biochip card for NanoSPR detection according to claim 1, wherein the connecting channel is smoothly connected to the first sample-loading reservoir, the second sample-loading reservoir and the flow channel.

8. The double-hole sample-adding microfluidic biochip card for nanoSPR detection according to claim 1, wherein the waste liquid pool is provided with a vent hole.

9. The double-hole sample-adding microfluidic biochip card for NanoSPR detection according to claim 1, wherein the inlet and outlet of the chip slot connected to the flow channel are horn-shaped.

10. The micro fluidic chip card with double-hole sample feeding for NanoSPR detection according to claim 1, wherein the bottom plate is of a sandwich structure, a water absorbing material is arranged in the sandwich structure, and the waste liquid pool is communicated with the water absorbing material.