CN112808338A

CN112808338A - Micro-fluidic chip

Info

Publication number: CN112808338A
Application number: CN202110178071.0A
Authority: CN
Inventors: 黄茗; 张赛; 王刚; 钱纯亘; 胡鹍辉
Original assignee: Shenzhen Yhlo Biotech Co Ltd
Current assignee: Shenzhen Yhlo Biotech Co Ltd
Priority date: 2021-02-09
Filing date: 2021-02-09
Publication date: 2021-05-18

Abstract

The invention relates to a microfluidic chip with a plurality of distribution reaction structures. Each distribution reaction structure comprises a sample adding hole in the area A, a distribution cavity in the area A and a reaction unit which are sequentially communicated. The reaction unit comprises a first micro-channel of the first region and a reaction cavity. The reaction cavity is communicated with the first micro-channel of the first area and the first distribution cavity of the first area. When the microfluidic chip is used for detection, a sample liquid is added into the sample adding hole in the area A, enters the distribution cavity in the area A, enters the plurality of reaction cavities in the corresponding distribution reaction structure from the distribution cavity in the area A through the first micro-channel in the area A, and is mixed and reacted with a storage object in the reaction cavities. The distribution reaction structures can detect a plurality of samples, the number of the reaction units is multiple in each distribution reaction structure, and the reaction units can detect a plurality of items, so that the multi-item detection can be simultaneously carried out on the samples on the same chip, and the multi-sample and multi-item joint detection is realized.

Description

Micro-fluidic chip

Technical Field

The invention relates to the technical field of microfluidics, in particular to a microfluidic chip.

Background

Chemiluminescence immunoassay refers to a technology which generates photons by chemical reaction without excitation of external light source, heat or electric field, and can combine high-sensitivity chemiluminescence detection technology with high-specificity antigen-antibody immunoreaction to detect the content of antigen or antibody in a sample.

Point-of-care testing (POCT), which refers to clinical testing and bedside testing performed near a patient, is usually performed on-site at the time of sampling, and thus a complex processing procedure of a specimen during laboratory testing is omitted, and a testing result is obtained quickly.

The microfluidic chip is a hotspot field developed by the current micro total analysis systems (micro total analysis systems), and can be applied to the instant detection of chemiluminescence immunoassay. The microfluidic chip integrates a series of experiments related in the fields of biology, chemistry, medicine and the like, including sample pretreatment, sample reaction, result reading and the like, on a chip with a micro-nano size structure, and has the characteristics of less sample requirement, less reagent consumption, rapid reaction, accurate result and the like. The centrifugal microfluidic chip is one of microfluidic chips, and refers to a type of chip that uses centrifugal force as power to drive a sample or a reagent to move in a chip microchannel so as to perform detection. Centrifugal microfluidic chips often achieve different centrifugal forces by adjusting the centrifugal speed to control the movement of liquid on the chip. The centrifugal microfluidic chip has the advantages of highly symmetrical structure, large measurement sample, small requirement on external power drive, small supporting facilities, high automation degree, accurate measurement structure, good repeatability and the like.

The traditional centrifugal microfluidic chip is generally used for multi-item joint inspection of a single sample or multi-sample joint inspection of a single item, and can not meet the detection requirements of multiple samples and multiple items.

Disclosure of Invention

Based on this, it is necessary to provide a microfluidic chip capable of satisfying the detection requirements of multiple samples and multiple items.

The scheme of the invention is as follows:

a microfluidic chip having a center of rotation, the microfluidic chip having a plurality of distribution reaction structures distributed and spaced around the center of rotation; the distribution reaction structure comprises a sample adding hole in the area A, a distribution cavity in the area A and a reaction unit; the first-zone sample adding hole is communicated with the first-zone distribution cavity, the reaction unit comprises a first micro-channel of the first zone and a reaction cavity, and the reaction cavity is communicated with the first-zone distribution cavity through the first micro-channel of the first zone; first district distribution chamber encircles the rotation center extends, it is a plurality of among the distribution reaction structure reaction unit, it is a plurality of reaction unit follows the extending direction who first district distribution chamber distributes, first district distribution chamber compare in reaction chamber is closer to the rotation center.

In one embodiment, the distribution reaction structure further includes a separation chamber and a first-zone second microchannel, the separation chamber being communicated with the first-zone distribution chamber through the first-zone second microchannel, and the separation chamber being closer to the rotation center than the first-zone distribution chamber.

In one embodiment, the first-region second microchannel is a U-shaped microchannel, and a U-shaped port of the first-region second microchannel faces away from the rotation center.

In one embodiment, the distribution reaction structure further includes a first-region fourth microchannel and a waste liquid storage chamber, the reaction chamber is communicated with the waste liquid storage chamber through the first-region fourth microchannel, and the waste liquid storage chamber is communicated with the reaction chamber and is further away from the rotation center than the reaction chamber.

In one embodiment, the reaction chamber stores magnetic beads labeled with immune components, and the immune components are antigens or antibodies.

In one embodiment, the reaction unit further includes a first reagent inlet and an eighth microchannel, the first reagent inlet is communicated with the reaction chamber through the eighth microchannel, and the first reagent inlet is closer to the rotation center than the reaction chamber.

In one embodiment, the microfluidic chip comprises a common reagent layer and a reaction layer which are stacked, and each distribution reaction structure is arranged in the reaction layer;

the common reagent layer is provided with a zone B sample adding hole, a common reagent storage cavity, a zone B solvent adding hole, a zone B distribution cavity and a shunting unit; the solvent adding hole in the area B is communicated with the public reagent storage cavity, the public reagent storage cavity is communicated with the area B distribution cavity, and the area B distribution cavity is arranged around the rotating center; the flow distribution unit comprises a first micro flow channel of the second zone and a first connecting port of the second zone, the first connecting port of the second zone is communicated with the distribution cavity of the second zone through the first micro flow channel of the second zone, the flow distribution unit is provided with a plurality of flow distribution units, and the flow distribution units are distributed along the extending direction of the distribution cavity of the second zone; the distances between the common reagent storage cavity, the second area distribution cavity and the flow distribution unit and the rotation center are sequentially increased;

the flow distribution units are in one-to-one correspondence with the reaction units in the reaction layer, the first connecting port of the area B is communicated with the first reagent of the area A through a fifth interlayer channel, and the sample adding hole of the area B is communicated with the sample adding hole of the area A through a sixth interlayer channel.

In one embodiment, the reaction unit further includes a first reagent inlet and a first ninth microchannel, the first reagent inlet is communicated with the reaction chamber through the first ninth microchannel, and the first reagent inlet is closer to the rotation center than the reaction chamber.

In one embodiment, the common reagent storage chamber stores a first reagent.

In one embodiment, the common reagent storage chamber comprises a plurality of sub-storage chambers distributed and spaced around the rotation center, and each sub-storage chamber is communicated with the second zone distribution chamber.

In one embodiment, each of the plurality of sub-storage cavities is communicated with the second zone distribution cavity through a reagent output channel, the reagent output channel comprises a first centrifugal force flow channel, the first centrifugal force flow channel extends away from the rotation center after being led out from the common reagent storage cavity, and the first centrifugal force flow channel is provided with a first discharge micro-flow valve.

In one embodiment, the reagent output channel corresponding to at least one of the sub-storage cavities further includes a delay unit, the delay unit includes a first turning flow channel, a capillary flow channel, a second turning flow channel, and a second centrifugal flow channel, the first centrifugal flow channel extends from the common reagent storage cavity away from the rotation center and is connected to one end of the first turning flow channel, the other end of the first turning flow channel is connected to one end of the capillary flow channel, the capillary flow channel extends from the first turning flow channel and is close to the rotation center, the other end of the capillary flow channel is connected to one end of the second turning flow channel, the other end of the second turning flow channel is connected to the second centrifugal flow channel, and the second centrifugal flow channel extends from the second turning flow channel and is away from the rotation center, the second centrifugal force flow channel is provided with a second discharging micro-flow valve.

In one embodiment, the reagent output channel corresponding to at least one of the sub-storage cavities has a plurality of delay units, and the delay units are sequentially communicated.

In one embodiment, the number of the delay units in the reagent output channel corresponding to each of the sub-storage cavities is different from the number of the delay units in the reagent output channels corresponding to the other sub-storage cavities.

In one embodiment, the common reagent storage cavity includes a first sub storage cavity, a second sub storage cavity and a third sub storage cavity, the number of the time delay units in the reagent output channels corresponding to the first sub storage cavity, the second sub storage cavity and the third sub storage cavity increases in sequence, the first sub storage cavity stores a cleaning agent, the second sub storage cavity stores a pre-activator, and the third sub storage cavity stores an activator.

In one embodiment, the microfluidic chip further comprises a sample solvent addition layer, and the sample solvent addition layer, the common reagent layer and the reaction layer are sequentially stacked;

the sample solvent adding layer is provided with a third-zone sample adding hole, a third-zone solvent adding hole, a third-zone first micro-channel, a third-zone distribution cavity, a third-zone second micro-channel, a third-zone first connecting port and a storage unit; the third-zone distribution cavity is arranged around the rotation center, the third-zone solvent adding hole is communicated with the third-zone distribution cavity through the third-zone first micro-channel, and the third-zone solvent adding hole is communicated with the third-zone first connecting port through the third-zone second micro-channel; the storage unit comprises a third micro-channel of a third zone, a material storage cavity of the third zone, a fourth micro-channel of the third zone and a second connecting port of the third zone, the material storage cavity of the third zone is communicated with the distribution cavity of the third zone through the third micro-channel, and the material storage cavity of the third zone is communicated with the second connecting port of the third zone through the fourth micro-channel of the third zone; the storage units are distributed along the extension direction of the third zone distribution cavity, the distances between the third zone solvent adding hole, the third zone distribution cavity, the third zone material storage cavity and the third zone second connecting port and the rotation center are sequentially increased, and the third zone first connecting port is far away from the rotation center compared with the third zone solvent adding hole;

in the common reagent layer, the flow dividing unit further comprises a second connection port of the second zone, a second micro-channel of the second zone and a third connection port of the second zone; the second connection port of the second zone is communicated with the third connection port of the second zone through the second micro-channel of the second zone, and the second connection port of the second zone is far away from the rotation center compared with the third connection port of the second zone;

the plurality of storage units in the sample solvent adding layer correspond to the plurality of flow dividing units in the common reagent layer one by one, and the third-zone sample adding hole is communicated with the sample adding hole through a first interlayer channel; the first connecting port of the third zone is communicated with the solvent adding hole of the second zone through a second interlayer channel; the second connecting port of the third zone is communicated with the second connecting port of the second zone through a third interlayer channel; and the third connecting port of the second zone is communicated with the second reagent inlet of the first zone through a fourth interlayer channel.

In one embodiment, the second microchannel in the second zone b is a U-shaped microchannel, and a U-shaped port of the second microchannel in the second zone b faces away from the rotation center.

In one embodiment, the third distribution cavity comprises a plurality of sub-distribution cavities distributed and spaced around the rotation center, and each sub-distribution cavity is communicated with a plurality of storage units.

In one embodiment, the storage unit further comprises a reagent addition groove, a feed permeation hole, a feed microchannel and a first feed microfluidic valve channel, wherein the reagent addition groove is opened on one side surface of the microfluidic chip, the reagent addition groove is communicated with the feed microchannel through the feed permeation hole, and the feed microchannel is communicated with the third-zone material storage cavity through the first feed microfluidic valve channel; the microfluidic chip is provided with a rotation center, and the third-zone material storage cavity is far away from the rotation center compared with the reagent sample adding port.

In one embodiment, the third zone material storage cavity stores a second reagent.

In one embodiment, the second reagent is a chemical labeled antibody.

Compared with the prior art, the micro-fluidic chip has the following beneficial effects:

the microfluidic chip is provided with a plurality of distribution reaction structures which are distributed around a rotation center and are arranged at intervals. Each distribution reaction structure comprises a sample adding hole in the area A, a distribution cavity in the area A and a reaction unit which are sequentially communicated. The reaction unit comprises a third micro-channel of the A area and a reaction cavity. The reaction cavity is communicated with the first-zone distribution cavity through a third micro-channel of the first zone. When the microfluidic chip is used for detection, a sample liquid is added into the sample adding hole in the area A, enters the distribution cavity in the area A, enters the plurality of reaction cavities in the corresponding distribution reaction structure from the distribution cavity in the area A through the third microchannel in the area A, and is mixed and reacted with a storage substance in the reaction cavities. The distribution reaction structures can detect a plurality of samples, the number of the reaction units is multiple in each distribution reaction structure, and the reaction units can detect a plurality of items, so that the multi-item detection can be simultaneously carried out on the samples on the same chip, and the multi-sample and multi-item joint detection is realized.

Drawings

FIG. 1 is a schematic structural diagram of a microfluidic chip according to an embodiment of the present invention;

FIG. 2 is a schematic structural diagram of a reaction layer in the microfluidic chip shown in FIG. 1;

FIG. 3 is a schematic structural diagram of a distribution reaction structure of a reaction layer in the microfluidic chip shown in FIG. 1;

FIG. 4 is a schematic diagram showing the positional relationship among a sample solvent addition layer, a common reagent layer and a reaction layer in the microfluidic chip shown in FIG. 1;

FIG. 5 is a schematic diagram of a common reagent layer in the microfluidic chip shown in FIG. 1;

FIG. 6 is an enlarged view of a portion of FIG. 5;

FIG. 7 is a diagram showing the connection of a common reagent storage chamber to a reagent output channel without a delay unit;

FIG. 8 is a diagram showing the connection of a common reagent storage chamber to a reagent output channel having a delay unit;

FIG. 9 is a schematic diagram of a delay unit;

FIG. 10 is a diagram showing the connection of a common reagent storage chamber to a reagent output channel having two delay units;

fig. 11 is a schematic structural view of a sample solvent addition layer in the microfluidic chip shown in fig. 1;

FIG. 12 is a schematic structural diagram of a storage unit;

FIG. 13 is a schematic diagram of a partial structure of a storage unit;

fig. 14 is a cross-sectional view of a sample solvent addition layer at a partial location shown in fig. 13.

Description of reference numerals:

10. a microfluidic chip; 11. a center of rotation; 100. a sample solvent addition layer; 101. c, adding a solvent into a hole; 103. third zone sample addition wells; 104. a third zone first connecting port; 105. a third zone is a first micro-flow valve; 106. a second microfluidic valve in zone C; 107. a third zone distribution chamber; 108. third microflow valve in third area; 109. a third-zone material storage cavity; 110. a third microfluidic valve; 111. a second connection port of the third zone; 112. a third vent hole; 113. a reagent addition tank; 114. a feed infiltration opening; 115. feeding a micro flow channel; 116. a first feed micro-flow valve; 1161. a first section; 1162. a second section; 1163. a third section; 117. a second feed micro-flow valve; 121. a third zone is a first micro-channel; 122. a second micro-channel in the third zone; 123. third micro-channel of third zone; 124. the third zone is a fourth micro-channel; 125. the third zone is a fifth micro-channel; 130. a storage unit;

200. a common reagent layer; 201. a zone B solvent addition port; 202. a common reagent storage chamber; 2021. a first sub-storage chamber; 2022. a second sub-storage chamber; 2023. a third sub-storage chamber; 203. a connecting port; 204. a zone b sample addition well; 207. a zone B distribution cavity; 208. a first microfluidic valve in zone B; 209. a first connection port of the second zone; 210. a second connection port of the second zone; 211. a second microchannel in zone B; 212. a third connector of the second zone; 230. a shunting unit; 221. a first microchannel in zone B; 260. a reagent output channel; 261. a first centrifugal force flow channel; 262. a first discharge microflow valve; 263. a delay unit; 2631. a first steering runner; 2632. a capillary flow channel; 2633. a second diverting flow path; 2634. a second centrifugal force flow channel; 2635. a second discharge microflow valve;

300. a reaction layer; 330. distributing the reaction structure; 301. a zone a sample addition well; 302. a separation chamber; 3021. a first side surface; 3022. a second side surface; 3023. a third side; 304. a first zone distribution chamber; 332. a reaction unit; 305. a first microfluidic valve in region a; 306. a reaction chamber; 307. a first reagent inlet; 308. a first reagent inlet in zone a; 309. a second microfluidic valve in region a; 310. a waste liquid storage chamber; 312. an anti-backflow valve; 321. a first microchannel in the first zone; 322. a second microchannel in the first zone; 3221. a first shunt passage; 3222. a second branch flow channel; 3223. a third shunting passage; 323. a third microchannel in the first zone; 324. a fourth microchannel in the first zone; 325. a fifth microchannel in the first zone; 326. a sixth micro-channel in the first area; 327. a seventh microchannel in the first zone; 328. the eighth micro-channel of the first area; 329. the ninth micro-channel of the first area; 401. a first vent hole; 402. a second vent.

Detailed Description

To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.

It will be understood that when an element is referred to as being "in communication with" another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "left," "right," and the like as used herein are for illustrative purposes only and do not represent the only embodiments.

In the description of the present invention, it is to be understood that the terms "a zone", "b zone", "c zone", "first", "second", "third", etc. are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number or order of technical features indicated.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

As shown in fig. 1, the present invention provides a microfluidic chip 10. The microfluidic chip 10 has a center of rotation 11 with a specially designed cavity structure 12. When the microfluidic chip 10 is subjected to rotational centrifugal operation, the microfluidic chip 10 rotates around the rotation center 11. The fluid can flow in the pore structure 12 under the action of centrifugal force or capillary force, and the like, so that the purpose of test detection is realized.

Referring to fig. 2 to 3, a microfluidic chip 10 according to an embodiment of the present invention includes a reaction layer 300.

The reaction layer 300 has a plurality of distribution reaction structures 330. The plurality of distribution reaction structures 330 are distributed and spaced around the rotation center 11.

The distribution reaction structure 330 includes a first-zone sample addition hole 301, a separation chamber 302, a first-zone second microchannel 322, a first-zone distribution chamber 304, and a reaction unit 332. The first zone sample addition hole 301 communicates with the separation chamber 302. More specifically, the a-zone sample addition hole 301 communicates with the separation chamber 302 through the a-zone third micro flow channel 323. The separation chamber 302 communicates with the zone a distribution chamber 304 via a zone a second microchannel 322. The reaction unit 332 includes a first microchannel 321 in the first region and a reaction chamber 306. The reaction chamber 306 is communicated with the first micro flow channel 321 of the first zone and the first distribution chamber 304 of the first zone.

In each distribution reaction structure 330, the first zone distribution chamber 304 extends around the center of rotation 11. For example, the first zone distribution chamber 304 may be an arc chamber centered on the center of rotation 11. In each distribution reaction structure 330, there is a plurality of reaction units 332, and the plurality of reaction units 332 are distributed along the extending direction of the first distribution chamber 304. The separation chamber 302, the first zone distribution chamber 304 and the reaction chamber 306 are sequentially increased in distance from the rotation center 11.

The microfluidic chip 10 has a plurality of distribution reaction structures 330, and the distribution reaction structures 330 are disposed around the rotation center 11 and spaced apart from each other. Each distribution reaction structure 330 includes a first-zone sample addition hole 301, a first-zone distribution chamber 304, and a reaction unit 332, which are connected in this order. The reaction unit 332 includes a first microchannel 321 in the first region and a reaction chamber 306. The reaction chamber 306 is communicated with the first micro flow channel 321 of the first zone and the first distribution chamber 304 of the first zone. When the microfluidic chip 10 is used for detection, a blood sample can be added into the sample adding hole 301 in the first region, enter the first region distribution cavity 304, then enter the plurality of reaction cavities 306 in the corresponding distribution reaction structure 330 from the first region distribution cavity 304 through the first micro-channel 321 in the first region, and mix and react with the stored substances in the reaction cavities 306. The plurality of distribution reaction structures 330 can detect a plurality of samples, in each distribution reaction structure 330, the number of the reaction units 332 is multiple, and the plurality of reaction units 332 can detect a plurality of items, so that the plurality of samples can be simultaneously detected on the same chip in a plurality of items, and the multi-sample and multi-item joint detection is realized.

The number of distribution reaction structures 330 is at least two, and may be, for example, 3 to 10. In the particular example illustrated, the microfluidic chip 10 has 3 dispensing reaction structures 330.

In each distribution reaction structure 330, the number of the reaction units 332 is at least two, and may be, for example, 3 to 15. It is understood that the number of reaction units 332 may be the same or different in different dispensing reaction structures 330. In the particular example illustrated, there are 5 reaction units 332 in each distribution reaction structure 330.

In one example, the distribution reaction structure 330 further includes a separation chamber 302 and a first-zone second microchannel 322, the separation chamber 302 communicating with the first-zone distribution chamber 304 through the first-zone second microchannel 322, the separation chamber 302 being closer to the rotation center 11 than the first-zone distribution chamber 304. The whole blood sample may be separated from plasma as it passes through the separation chamber 302 and into the first zone distribution chamber 304.

In one example, the dispensing reaction structure 330 further includes a fourth microchannel 324 in the first region and a waste reservoir 310. The reaction chamber 306 is in communication with the waste reservoir 310 via the fourth microchannel 324 in the first zone. The waste liquid storage chamber 310 communicates further away from the rotation center 11 than the reaction chamber 306.

In one example, a second microfluidic valve 309 is disposed on the first-zone fourth microchannel 324. Second microfluidic valve 309 in the first region may be a trap, capillary valve, or the like.

In one example, the microfluidic chip 10 further includes a first vent 401. One end of the first vent 401 is communicated with the waste liquid storage cavity 310, and the other end is opened on one side surface of the microfluidic chip 10.

In one example, a first microfluidic channel 321 is provided with a first microfluidic valve 305. The first microfluidic valve 305 in the first region may be a trap, a capillary valve, or the like.

In one example, the first-zone second microchannel 322 is a U-shaped microchannel, and the first-zone second microchannel 322 is open away from the rotation center 11. More specifically, the second micro flow channel 322 in the first region includes a first sub flow channel 3221, a second sub flow channel 3222, and a third sub flow channel 3223, which are connected in sequence. The first branch flow path 3221 extends from the separation chamber 302 to near the rotation center 11 and is communicated with one end of the second branch flow path 3222, the other end of the second branch flow path 3222 is communicated with the third branch flow path 3223, and the third branch flow path 3223 extends from the second branch flow path 3222 to far away from the rotation center 11 and is communicated with the first distribution chamber 304. More specifically, the first branch flow path 3221 is connected to a side surface of the separation chamber 302, which is a surface connecting a side surface of the separation chamber 302 close to the rotation center 11 and a side surface far from the rotation center 11. For example, in the illustrated specific example, the separation chamber 302 is a fan-shaped annular chamber extending along the rotation circumference of the microfluidic chip 10, and the first shunt path 3221 is connected to one side plane of the separation chamber 302, which is a plane connecting the arc surface of the separation chamber 302 close to the rotation center 11 and the arc surface far away from the rotation center 11.

The separation chamber 302 preferably has a wide width in the radial direction of the microfluidic chip 10, which is advantageous for improving the separation degree of the sample.

In one example, in each distribution reaction structure 330, the separation chamber 302 communicates with the waste liquid storage chamber 310 through the fifth microchannel 325 in the first region. Excess fluid in the separation chamber 302 may be drained through the fifth microchannel 325 in the first zone to the waste reservoir 310. Further, the fifth microchannel 325 in the first region is a U-shaped microchannel, and a U-shaped opening of the fifth microchannel 325 in the first region faces away from the rotation center 11. Preferably, the fifth micro flow channel 325 in the first region is led out from the side of the separation chamber 302 close to the rotation center 11, which facilitates the discharge of the separated waste into the waste liquid storage chamber 310. Further preferably, the fifth micro flow channel 325 in the first region is led out from one end of the side surface of the separation chamber 302 close to the rotation center 11, which facilitates the discharge of the separated waste into the waste liquid storage chamber 310. In the particular example illustrated, the connection of the first-zone second fluidic channel 322 to the first-zone fifth fluidic channel 325 is at opposite ends of the separation chamber 302.

The first distribution chamber 304 preferably has a narrower width in the radial direction of the microfluidic chip 10 than the separation chamber 302, which is advantageous for filling the separation chamber 302 with the separated plasma sample and improving the uniformity of the amount of the sample delivered from the first distribution chamber 304 to the plurality of reaction chambers 306.

In one example, the a-zone distribution chamber 304 extends circumferentially along the rotation of the microfluidic chip 10. Preferably, the width of the nail zone distribution chamber 304 in the radial direction is uniform in the direction of extension thereof.

In one example, in each distribution reaction structure 330, the first-zone distribution chamber 304 communicates with the waste fluid storage chamber 310 via the first-zone sixth microchannel 326. Excess fluid in the first distribution chamber 304 may be drained to the waste reservoir chamber 310 through the first sixth microchannel 326. Further, the sixth micro flow channel 326 in the first region is a U-shaped micro flow channel, and a U-shaped opening of the sixth micro flow channel 326 in the first region faces away from the rotation center 11. Preferably, the sixth microchannel 326 of the zone A exits from the side of the distribution chamber 304 of the zone A near the center of rotation 11. Further preferably, the sixth micro flow channel 326 in the first region is led out from one end of the side surface of the separation chamber 302 close to the rotation center 11, which is beneficial for discharging the excessive liquid into the waste liquid storage chamber 310.

In the illustrated embodiment, the first-zone fifth microchannel 325 and the first-zone sixth microchannel 326 are connected to one end of a first-zone seventh microchannel 327, and the other end of the first-zone seventh microchannel 327 is connected to the waste liquid storage chamber 310. The seventh micro flow channel 327 of the first zone is provided with a backflow prevention valve 312. The backflow prevention valve 312 can prevent the waste fluid in the waste fluid storage chamber 310 from flowing backward.

Preferably, in the distribution reaction structure 330, a plurality of reaction units 332 are uniformly distributed along the extending direction of the first distribution chamber 304.

In one example, the reaction unit 332 further includes a first reagent inlet 308 and an eighth microchannel 328. The first reagent inlet 308 of the first zone is communicated with the reaction chamber 306 through an eighth micro-channel 328 of the first zone. The first reagent inlet 308 is closer to the center of rotation 11 than the reaction chamber 306.

In the above example, the desired reagent may be added to reaction chamber 306 through first reagent inlet 308 in the first zone.

Referring further to fig. 4 and 5, in one example, the microfluidic chip 10 includes a common reagent layer 200 and a reaction layer 300 stacked on each other. Each of the distribution reaction structures 330 is disposed in the reaction layer 300. The common reagent layer 200 may be used to store reagents required for multiple assays in the reaction layer 300.

The common reagent layer 200 has a zone b sample addition hole 204, a common reagent storage chamber 202, a zone b solvent addition hole 201, a zone b distribution chamber 207, and a flow dividing unit 230. The zone b solvent addition port 201 communicates with the common reagent storage chamber 202. The common reagent storage chamber 202 communicates with the zone b dispensing chamber 207. Zone b distribution chamber 207 is disposed about center of rotation 11. The flow dividing unit 230 includes a first microchannel 221 in zone b and a first connection port 209 in zone b. The first connection port 209 of the second zone is communicated with the distribution chamber 207 of the second zone through a first micro flow channel 221 of the second zone. The number of the shunt unit 230 is plural. The plurality of flow dividing units 230 are distributed along the extending direction of the zone b distribution cavity 207. The distances from the common reagent storage chamber 202, the second zone distribution chamber 207 and the flow distribution unit 230 to the rotation center 11 are increased in sequence.

The plurality of flow dividing units 230 correspond to the plurality of reaction units 332 in the reaction layer 300 one to one. The first second connector 209 communicates with the first reagent inlet 308 via a fifth interlayer channel (not shown). The zone b sample addition hole 204 communicates with the zone a sample addition hole 301 through a sixth interlayer passage (not shown in the figure).

In the above example, the sample liquid is added from the zone b sample addition hole 204, enters the zone a sample addition hole 301 in the reaction layer 300 via the sixth interlayer channel, and enters the separation chamber 302. The common reagent storage chamber 202 may store a desired reagent. During centrifugation, the reagent stored in the common reagent storage chamber 202 enters the second zone distribution chamber 207, then reaches the second zone first connection port 209 through the second zone first microchannel 221, then reaches the first reagent inlet 308 in the reaction layer 300 through the fifth inter-channel, and enters the reaction chamber 306 through the first zone eighth microchannel 328.

In one example, a first microfluidic valve 208 is disposed on the first microchannel 221.

As shown in fig. 2, in one example, the reaction unit 332 further includes a first-region second reagent inlet 307 and a first-region ninth micro flow channel 329. The second reagent inlet 307 of the first zone is communicated with the reaction chamber 306 through a ninth micro-channel 329 of the first zone. The second reagent inlet 307 in the first region is closer to the rotation center 11 than the reaction chamber 306.

In the above example, the desired reagent may be added to reaction chamber 306 through a second reagent inlet 307 in the first zone.

In one example, the b-zone distribution cavity 207 is a circular ring with the microfluidic chip 10 as a center.

In one example, the common reagent storage chamber 202 stores a first reagent.

Alternatively, the first reagent may be a single reagent, or may include multiple reagents. The first reagent stored in the common reagent storage chamber 202 may be a fluid or may be lyophilized.

The lyophilization may be performed after the first reagent in the fluid form is stored in the common reagent storage chamber 202.

Wherein, the freeze-drying treatment refers to a drying method of freezing the wet material below a freezing point (eutectic point), then sublimating and drying under proper vacuum conditions to remove ice crystals, and then carrying out resolution drying after sublimation is finished to remove part of bound water. The freeze-dried product has the following advantages: the product is dried under the condition of low temperature and vacuum, so the decomposition rate is very low and the purity is high; the volume of the original solution is basically kept when the solution is frozen, and the solution is loose and porous, has beautiful appearance and uniform color; is easy to dissolve in water, and immediately recovers the original drug property; the pollution chance is less, foreign matters are less, the dissolving performance of the medicine can be improved, and the clarity of the preparation can be improved; the water content of the freeze-dried product is lower than 8%, and the freeze-dried product can be stored for a long time and is convenient to transport.

As shown in FIG. 6, in one example, the common reagent storage chamber 202 includes a plurality of sub-storage chambers. The plurality of sub-storage cavities are distributed around the rotation center 11 and are arranged at intervals, and each sub-storage cavity is communicated with the second zone distribution cavity 207. By providing a plurality of sub-reservoirs, a plurality of reagents can be delivered to the reaction chamber 306.

Referring to fig. 6 to 10, in one example, each of the plurality of sub-storage chambers is connected to the reagent output channel 260 and the reagent distribution chamber 207 through a first centrifugal force channel 261, and the first centrifugal force channel 261 extends from the common reagent storage chamber 202 and away from the rotation center 11. The first centrifugal-force flow channel 261 has a first outlet microfluidic valve 262 to lock the reagent in the sub-reservoirs in the absence of centrifugal force.

As shown in fig. 8 and 9, in one example, the reagent output channel 260 corresponding to at least one of the plurality of sub-storage cavities further includes a time delay unit 263. The delay unit 263 includes a first turn flow path 2631, a capillary force flow path 2632, a second turn flow path 2633, and a second centrifugal force flow path 2634. The first centrifugal force flow path 261 extends from the common reagent storage chamber 202 away from the rotation center 11 and communicates with one end of the first diverting flow path 2631. The other end of the first diverting flow path 2631 communicates with one end of the capillary flow path 2632. The capillary flow path 2632 extends from the first diverting flow path 2631 to be close to the rotation center 11. The other end of the capillary flow passage 2632 communicates with one end of the second diverting flow passage 2633. The other end of the second turn flow passage 2633 communicates with the second centrifugal force flow passage 2634. The second centrifugal force flow path 2634 extends away from the rotation center 11 after being drawn from the second turn flow path 2633. The second centrifugal flow channel 2634 has a second outlet microfluidic valve 2635.

The delay unit 263 of the microfluidic chip 10 of the above example can delay the output time of the reagent from the corresponding sub-storage chamber to the other liquid path outside the reagent output channel 260. In the test, by increasing the centrifugal speed, the reagent in the sub-storage chamber breaks through the first centrifugal force flow path 261 on the first centrifugal force flow path 261 and enters the first diverting flow path 2631 of the delay unit 263, and at this time, the reagent is temporarily retained in the first diverting flow path 2631 because the centrifugal force is greater than the capillary force. The centrifugal speed is reduced, and at this time, the centrifugal force is smaller than the capillary force, and the reagent enters the capillary force flow channel 2632 from the first diverting flow channel 2631 and then enters the second diverting flow channel 2633 to reach the front of the second discharging microfluidic valve 2635 on the second centrifugal force flow channel 2634. The centrifugal speed is increased, and the reagent breaks through the second outlet microfluidic valve 2635 and is output from the second centrifugal flow channel 2634. Thus, after several rounds of centrifugal speed transition, reagent can be output through the reagent output channel 260. The microfluidic chip 10 is suitable for reactions requiring control of reagent output time and sequence, and particularly for reactions requiring sequential application of multiple reagents, can control the application time of the multiple reagents, and perform multi-step reactions. Therefore, the microfluidic chip 10 of the above example is capable of performing a relatively complicated reaction, and avoids operational complexity at the time of the experiment.

As shown in fig. 10, in one example, the reagent output channel 260 corresponding to at least one of the plurality of sub-storage cavities has a plurality of delay units 263, and the plurality of delay units 263 are sequentially communicated.

Further, in one example, the number of the delay units 263 in the reagent output channel 260 corresponding to each sub-storage chamber is different from the number of the delay units 263 in the reagent output channels 260 corresponding to the other sub-storage chambers.

In the illustrated embodiment, the common reagent storage chamber 202 includes a first sub-storage chamber 2021, a second sub-storage chamber 2022, and a third sub-storage chamber 2023. The number of the delay units 263 in the reagent output channel 260 corresponding to the first sub-storage chamber 2021, the second sub-storage chamber 2022 and the third sub-storage chamber 2023 increases in sequence. More specifically, the number of the delay units 263 in the reagent output channel 260 corresponding to the first sub-storage chamber 2021 is zero, the number of the delay units 263 in the reagent output channel 260 corresponding to the second sub-storage chamber 2022 is one, and the number of the delay units 263 in the reagent output channel 260 corresponding to the third sub-storage chamber 2023 is two. The first sub-storage chamber 2021 stores cleaning agent, the second sub-storage chamber 2022 stores pre-activator, and the third sub-storage chamber 2023 stores activator. In this manner, the order in which the cleaning agent, pre-activator, and activator are delivered into reaction chamber 306 can be precisely controlled.

The extending directions of the first diverting flow path 2631 and the second diverting flow path 2633 are the same as or close to the rotating circumference of the microfluidic chip 10. The first and second diverting

flow paths

2631 and 2633 may be, but not limited to, arc-shaped, linear, etc.

In one example, the first steering channel 2631 is an arc centered on the center of rotation 11.

In one example, the second diverting flow path 2633 is an arc centered on the center of rotation 11.

The common reagent layer 200 in the microfluidic chip 10 of the above example is used for storing a common reagent for multi-sample and multi-item detection, and can perform chemical reactions in multiple steps, thereby greatly improving the complexity of the chemical reactions in the microfluidic chip 10, expanding the category of the chemical reactions in the microfluidic chip 10, and improving the use value of the microfluidic chip 10 in the POCT field.

The traditional chip often needs an external reagent, so that a matched instrument is large in size, the detection requirements of multiple samples and multiple items are difficult to meet, and the result of orderly releasing the reagent integrated on the chip cannot be realized. In addition, in the microfluidic chip 10 of the above example, during an experiment, the operation of adding a common reagent is not required, and the reagent can be automatically and orderly released on the chip, so that supporting instruments can be reduced, and the detection of multiple samples and multiple items can be realized.

It is understood that the common reagent layer 200 may be omitted if the addition of the common reagent (first reagent) is not required.

Referring further to fig. 11, in one example, the microfluidic chip 10 further includes a sample solvent addition layer 100. The sample solvent addition layer 100, the common reagent layer 200, and the reaction layer 300 are stacked in this order. The sample solvent addition layer 100 may be used to add a desired material, such as a chemical-labeled antibody, to the reaction layer 300.

The sample solvent addition layer 100 has a third zone sample addition hole 103, a third zone solvent addition hole 101, a third zone first microchannel 121, a third zone distribution chamber 107, a third zone second microchannel 122, a third zone first connection port 104, and a storage unit 130. The third zone distribution chamber 107 is arranged around the centre of rotation 11. The third zone solvent adding hole 101 is communicated with the third zone distribution chamber 107 through a third zone first micro flow channel 121. The third zone solvent adding hole 101 is communicated with the third zone first connecting port 104 through the third zone second microchannel 122.

The storage unit 130 includes a third microchannel 123, a material storage chamber 109, a fourth microchannel 124, and a second connection port 111. The third material storage cavity 109 is communicated with the third distribution cavity 107 through a third micro-channel. The third-zone material storage cavity 109 is communicated with the third-zone second connecting port 111 through a third-zone fourth micro-channel 124. The storage units 130 are plural, and the plural storage units 130 are distributed along the extending direction of the third partition chamber 107.

The distances between the third-zone solvent adding hole 101, the third-zone distribution cavity 107, the third-zone material storage cavity 109 and the third-zone second connecting port 111 and the rotation center 11 are sequentially increased. The third-zone first connection port 104 is further away from the rotation center 11 than the third-zone solvent addition hole 101.

In the common reagent layer 200, the flow distribution unit 230 further includes a second connection port 210 in the second zone, a second microchannel 211 in the second zone, and a third connection port 212 in the second zone. The second connection port 210 of the second zone is communicated with the third connection port 212 of the second zone through a second micro channel 211 of the second zone. The second connection port 210 of zone b is further away from the center of rotation 11 than the third connection port 212 of zone b.

The plurality of storage units 130 in the sample solvent adding layer 100 correspond to the plurality of flow dividing units 230 in the common reagent layer 200 one to one. The third zone sample addition hole 103 communicates with the sample addition hole 201 through a first interlayer channel (not shown in the figure). The third-zone first connection port 104 communicates with the second-zone solvent addition hole 201 through a second interlayer passage (not shown). The second connection port 111 of the third zone communicates with the second connection port 210 of the second zone via a third interlayer channel (not shown). The second zone third port 212 communicates with the first zone second reagent inlet 307 via a fourth inter-layer channel (not shown).

In the above example, the sample liquid is added from the third zone sample addition hole 103, reaches the sample addition hole 201 in the common reagent layer 200 via the first interlayer channel, enters the first zone sample addition hole 301 in the reaction layer 300 via the sixth interlayer channel, and enters the separation chamber 302. The solvent is added from the third zone solvent adding hole 101, one path of the solvent enters the third zone distribution cavity 107 through the third zone first micro-channel 121, then enters the third zone material storage cavity 109 through the third micro-channel, then reaches the third zone second connecting port 111 through the third zone fourth micro-channel 124, then reaches the second connecting port 210 of the second zone in the common reagent layer 200 through the third interlayer channel, then reaches the third connecting port 212 of the second zone through the second zone second micro-channel 211, then reaches the first zone second reagent inlet 307 in the reaction layer 300 through the fourth interlayer channel, and enters the reaction cavity 306 through the first zone ninth micro-channel 329 to provide the solvent for the material stored in the reaction cavity 306; the other path of solvent reaches the first third-zone connection port 104 through the second third-zone micro flow channel 122, and then enters the common reagent storage cavity 202 through the second interlayer channel and the second-zone solvent adding hole 201, so as to provide the solvent for the material stored in the common reagent storage cavity 202.

In one example, the microfluidic chip 10 further has a second vent 402. One end of the second vent 402 is connected to the common reagent storage chamber 202, and the other end is open to a side surface of the microfluidic chip 10. More specifically, the common reagent storage chamber 202 has a connection port 203, and the second vent hole 402 communicates with the common reagent storage chamber 202 through the connection port 203. The second vent 402 may be used to add reagent to the common reagent storage chamber 202, and the second vent 402 may also function as a vent during testing.

In one example, the second microchannel 211 is a U-shaped microchannel. The U-shaped opening of the second microchannel 211 in zone B faces away from the center of rotation 11.

In one example, the third-zone first microfluidic channel 121 is provided with a third-zone first microfluidic valve 105.

In one example, the third microfluidic channel 122 is provided with a third microfluidic valve 106.

In one example, a third microfluidic valve 108 is disposed on the third microchannel 123.

In one example, the third microfluidic channel 124 is provided with a third microfluidic valve 110.

In one example, the storage unit 130 further includes a third microfluidic channel 125 and a third vent 112. One end of the third zone fifth microchannel 125 is communicated with the third microchannel 123 of the third zone, and the other end is communicated with the third vent 112. The third vent hole 112 is opened on one side surface of the microfluidic chip 10.

As shown in fig. 11, in one example, third distribution chamber 107 includes a plurality of sub-distribution chambers distributed and spaced around center of rotation 11. Each sub-distribution chamber is connected to a plurality of storage units 130. In the illustrated embodiment, the third distribution chamber 107 is divided into 3 sub-distribution chambers, each of which is connected to 5 storage units 130.

As shown in fig. 12 to 14, in one example, the storage unit 130 further includes a reagent addition tank 113, a feed permeation hole 114, a feed microchannel 115, and a first feed microfluidic valve 116. The reagent addition groove 113 is opened on one side surface of the microfluidic chip 10. The reagent addition tank 113 communicates with the feed microchannel 115 through the feed permeation hole 114. The feeding micro-channel 115 is connected to the third-zone material storage chamber 109 through a first feeding micro-flow valve 116. The third zone material storage chamber 109 is further away from the center of rotation 11 than the reagent loading port.

The microfluidic chip 10 of the above example has a reagent storage function, and is capable of pre-storing a reagent before a detection test. When the reagent is pre-stored, the reagent to be pre-stored is added into the reagent adding groove 113, the reagent enters the feeding micro-channel 115 through the feeding permeation hole 114, and the reagent breaks through the first feeding micro-flow valve 116 and enters the third-zone material storage cavity 109 through rotation and centrifugation. The first feed microfluidic valve 116 prevents backflow of reagent from the third zone material storage chamber 109, thereby allowing storage of reagent. The micro-fluidic chip 10 encapsulates reagents required for detection into the micro-fluidic chip 10 through structural design, and it can be understood that the third-zone material storage cavity 109 can be communicated with a detection liquid path, and when detection is required, the reagents are driven to enter the detection liquid path by increasing centrifugal force. The microfluidic chip 10 realizes the integrated microfluidic item detection of the reagent chip, can save the operation of adding corresponding reagents in the test link, is convenient to use and saves time.

In one example, the end of feed penetration hole 114 communicating with reagent addition groove 113 is open to the bottom of the groove of reagent addition groove 113, facilitating better entry of reagent into feed penetration hole 114.

In one example, the feed permeation holes 114 extend perpendicular to the disk surface of the microfluidic chip 10, which facilitates better reagent entry into the feed microchannels 115.

In one example, the first microfluidic feed valve 116 is open at the side wall of the third material storage chamber 109 at the end communicating with the third material storage chamber 109. Thus, by rotating the centrifuge, the reagent can be better introduced into the zone C material storage chamber 109.

In one example, first feed microfluidic valve 116 is a hydrophobic valve or a capillary valve.

As shown in fig. 13, in one example, the reagent storage structure further comprises a second feed microfluidic valve 117, and the feed-permeation pore 114 communicates with the feed microchannel 115 through the second feed microfluidic valve 117.

In the above example, the second feeding microfluidic valve 117 is disposed between the feeding permeation hole 114 and the feeding microchannel 115, so that when the reagent is added dropwise, the reagent is first retained in the reagent adding groove 113, and when the centrifugal device is rotated, the reagent enters the feeding microchannel 115 at the same time, thereby reducing the difference; on the other hand, backflow can be prevented during subsequent drying, which corresponds to one more safety.

As shown in fig. 13, in one example, the first feed microfluidic valve 116 includes a first section 1161, a second section 1162, and a third section 1163 in serial communication. One end of the first section 1161 is in communication with the feed microchannel 115 and one end of the third section 1163 is in communication with the third zone material storage chamber 109. The first and

third segments

1161, 1163 extend away from the center of rotation 11, and the third segment 1163 is further away from the center of rotation 11 than the first segment 1161. The first section 1161 and the third section 1163 are respectively disposed at an angle with the second section 1162. For example, the first block 1161 and the third block 1163 are perpendicular to the second block 1162. Therefore, on one hand, the extending direction of the feeding micro-channel 115 deviates from the third zone material storage cavity 109 and is staggered with other liquid paths communicated with the third zone material storage cavity 109, and on the other hand, the reagent can be better prevented from flowing back from the third zone material storage cavity 109.

As shown in fig. 14, in one example, the sample solvent addition layer 100 includes a base plate 151 and a cover plate 152, a groove structure is provided on the cover plate 152, and the base plate 151 and the cover plate 152 are butted such that the groove structure forms a cavity structure.

In one example, the third zone material storage chamber 109 stores a second reagent. The second reagent may be, but is not limited to, a chemical-labeled antibody, such as an acridinium-labeled antibody. The second reagent may be a fluid or may be lyophilized. The freeze-drying process may be performed after the second reagent in the form of fluid is stored in the third material storage chamber 109.

It is understood that the sample solvent addition layer 100 may be omitted if no second reagent is added.

In one example, reaction chamber 306 stores a third reagent.

In one example, the reaction chamber 306 stores magnetic beads labeled with an immune component, which is an antigen or an antibody, and may be lyophilized. For example, the reaction chamber 306 stores freeze-dried CTNI/NT-proBNP/D-dimer/MYO/CKMB coated magnetic beads. The magnetic beads may be magnetically immobilized so that they remain in the reaction chamber 306 and are prevented from being thrown into the waste reservoir 310. In one example, the reaction layer 300 includes a bottom plate and a well plate, the well plate is connected to the common reagent layer 200, and the bottom plate and the well plate are butted to form a cavity structure in the reaction layer 300. During manufacture, the magnetic beads marked with the immune components can be fixed on the bottom plate in a dispensing manner, and then the rest parts including the groove plate are butted with the groove plate.

The third zone material storage cavity 109 stores corresponding chemical substance labeled antibody, and can be subjected to freeze-drying treatment. For example, a lyophilized acridine labeled CTNI/NT-pro BNP/D-dimer/MYO/CKMB antibody. The chemical substance labeled antibody enters the reaction cavity 306 and then reacts with the antigen on the magnetic bead, and after incubation, the structure of the antibody-antigen-labeled antibody is formed.

The wash agent may comprise a surfactant that washes away unbound antibody and drains into the waste reservoir 310. The pre-excitation liquid can be NaOH, the excitation liquid can be hydrogen peroxide, and the acridinium ester marked on the labeled antibody can emit light under the systems of the pre-excitation liquid and the excitation liquid so as to detect a light-emitting signal.

In one example, the sample solvent addition layer 100, the common reagent layer 200, and the reaction layer 300 are integrally connected. In another example, the sample solvent addition layer 100, the common reagent layer 200, and the reaction layer 300 may be separately manufactured and then connected to each other.

The present invention will be further described below by taking as an example a method of detecting by using the microfluidic chip 10 of the specific example shown in the drawing.

The microfluidic chip 10 according to a specific example of the present invention includes a sample solvent addition layer 100, a common reagent layer 200, and a reaction layer 300, which are sequentially stacked.

The microfluidic chip 10 includes a sample solvent addition layer 100, a common reagent layer 200, and a reaction layer 300, which are stacked.

Sample solvent addition layer 100:

the sample solvent addition layer 100 has a third zone sample addition hole 103, a third zone solvent addition hole 101, a third zone first microchannel 121, a third zone distribution chamber 107, a third zone second microchannel 122, a third zone first connection port 104, and a storage unit 130. The third zone distribution chamber 107 is arranged around the centre of rotation 11. The third zone solvent adding hole 101 is communicated with the third zone distribution chamber 107 through a third zone first micro flow channel 121. The third-zone first micro-flow channel 121 is provided with a third-zone first micro-flow valve 105. The third zone solvent adding hole 101 is communicated with the third zone first connecting port 104 through the third zone second microchannel 122. The third-zone second micro flow channel 122 is provided with a third-zone second micro flow valve 106.

The storage unit 130 includes a third microchannel 123, a material storage chamber 109, a fourth microchannel 124, a second connection port 111, a fifth microchannel 125 and a third vent 112. The third material storage cavity 109 is communicated with the third distribution cavity 107 through a third micro-channel. The third microflow channel 123 of the third zone is provided with a third microflow valve 108 of the third zone. The third-zone material storage cavity 109 is communicated with the third-zone second connecting port 111 through a third-zone fourth micro-channel 124. The third zone fourth microfluidic channel 124 is provided with a third zone fourth microfluidic valve 110.

One end of the third zone fifth microchannel 125 is communicated with the third microchannel 123 of the third zone, and the other end is communicated with the third vent 112. The third vent hole 112 is opened on one side surface of the microfluidic chip 10.

The storage units 130 are plural, and the plural storage units 130 are distributed along the extending direction of the third partition chamber 107. Third zone distribution chamber 107 comprises a plurality of sub-distribution chambers distributed and spaced around center of rotation 11. Each sub-distribution chamber is connected to a plurality of storage units 130.

The storage unit 130 further includes a reagent addition tank 113, a feed permeation hole 114, a feed microchannel 115, a first feed microfluidic valve 116, and a second feed microfluidic valve 117. The reagent addition groove 113 is opened on one side surface of the microfluidic chip 10. The reagent addition tank 113 communicates with the feed microchannel 115 through the feed permeation hole 114. The feeding micro-channel 115 is connected to the third-zone material storage chamber 109 through a first feeding micro-flow valve 116. The third zone material storage chamber 109 is further away from the center of rotation 11 than the reagent loading port. The feed-permeation pore 114 communicates with the feed microchannel 115 through the second feed microfluidic valve 117.

One end of the feed penetration hole 114 communicating with the reagent addition tank 113 is opened to the bottom of the reagent addition tank 113. The feed permeation holes 114 extend perpendicular to the disk surface of the microfluidic chip 10. The first feeding micro-flow valve 116 is connected to the third material storage chamber 109 at one end and opens at the side wall of the third material storage chamber 109.

The first feed microfluidic valve 116 includes a first section 1161, a second section 1162, and a third section 1163 in serial communication. One end of the first section 1161 is in communication with the feed microchannel 115 and one end of the third section 1163 is in communication with the third zone material storage chamber 109. The first and

third segments

1161, 1163 extend away from the center of rotation 11, and the third segment 1163 is further away from the center of rotation 11 than the first segment 1161. The first and

third segments

1161 and 1163 are perpendicular to the second segment 1162.

The third-zone material storage cavity 109 is stored with acridine labeled CTNI/NT-pro BNP/D-dimer/MYO/CKMB antibody freeze-drying.

Common reagent layer 200:

the common reagent layer 200 has a zone b sample addition hole 204, a common reagent storage chamber 202, a zone b solvent addition hole 201, a second micro flow channel, a zone b distribution chamber 207, and a flow distribution unit 230. The zone b solvent addition port 201 communicates with the common reagent storage chamber 202. The common reagent storage chamber 202 communicates with the zone b distribution chamber 207 through the second microchannel. Zone b distribution chamber 207 is disposed about center of rotation 11.

The flow dividing unit 230 includes a first microchannel 221 in zone b, a first connection port 209 in zone b, a second connection port 210 in zone b, a second microchannel 211 in zone b, and a third connection port 212 in zone b. The first connection port 209 of the second zone is communicated with the distribution chamber 207 of the second zone through a first micro flow channel 221 of the second zone. The first micro flow channel 221 of the second area is provided with a first micro flow valve 208 of the second area. The second connection port 210 of the second zone is communicated with the third connection port 212 of the second zone through a second micro channel 211 of the second zone. The second microchannel 211 in zone b is a U-shaped microchannel. The U-shaped opening of the second microchannel 211 in zone B faces away from the center of rotation 11. The second connection port 210 of zone b is further away from the center of rotation 11 than the third connection port 212 of zone b.

The shunt unit 230 is plural. The plurality of flow dividing units 230 are distributed along the extending direction of the zone b distribution cavity 207. The distances from the common reagent storage chamber 202, the second zone distribution chamber 207 and the flow distribution unit 230 to the rotation center 11 are increased in sequence.

The common reagent storage chamber 202 includes a first sub-storage chamber 2021, a second sub-storage chamber 2022, and a third sub-storage chamber 2023. The plurality of sub-storage cavities are distributed around the rotation center 11 and are arranged at intervals, and each sub-storage cavity is communicated with the second zone distribution cavity 207.

Each of the plurality of sub-reservoirs is in communication with zone B distribution chamber 207 via a reagent outlet channel 260. The reagent output channel 260 includes a first centrifugal force flow path 261, and the first centrifugal force flow path 261 extends away from the rotation center 11 after being drawn from the common reagent storage chamber 202. The first centrifugal-force flow path 261 has a first discharge micro-fluidic valve 262.

Reagent output channel 260 also includes a delay element 263. The delay unit 263 includes a first turn flow path 2631, a capillary force flow path 2632, a second turn flow path 2633, and a second centrifugal force flow path 2634. The first centrifugal force flow path 261 extends from the common reagent storage chamber 202 away from the rotation center 11 and communicates with one end of the first diverting flow path 2631. The other end of the first diverting flow path 2631 communicates with one end of the capillary flow path 2632. The capillary flow path 2632 extends from the first diverting flow path 2631 to be close to the rotation center 11. The other end of the capillary flow passage 2632 communicates with one end of the second diverting flow passage 2633. The other end of the second turn flow passage 2633 communicates with the second centrifugal force flow passage 2634. The second centrifugal force flow path 2634 extends away from the rotation center 11 after being drawn from the second turn flow path 2633. The second centrifugal flow channel 2634 has a second outlet microfluidic valve 2635.

The number of the delay units 263 in the reagent output channel 260 corresponding to the first sub-storage chamber 2021 is zero, the number of the delay units 263 in the reagent output channel 260 corresponding to the second sub-storage chamber 2022 is one, and the number of the delay units 263 in the reagent output channel 260 corresponding to the third sub-storage chamber 2023 is two.

The first sub-storage chamber 2021 stores a detergent for freeze-drying, the second sub-storage chamber 2022 stores a pre-activator for freeze-drying, and the third sub-storage chamber 2023 stores an activator for freeze-drying.

Reaction layer 300:

The distribution reaction structure 330 includes a first-zone sample addition hole 301, a separation chamber 302, a first-zone second microchannel 322, a first-zone distribution chamber 304, a reaction unit 332, a first-zone fourth microchannel 324, and a waste liquid storage chamber 310. The first-zone sample addition hole 301 communicates with the separation chamber 302 through the first-zone third microchannel 323. The separation chamber 302 communicates with the zone a distribution chamber 304 via a zone a second microchannel 322.

The reaction unit 332 comprises a first micro-channel 321 in the first region, a reaction chamber 306, an eighth micro-channel 328 in the first region, a first reagent inlet 308 in the first region, a ninth micro-channel 329 in the first region, and a second reagent inlet 307 in the first region. The reaction chamber 306 is communicated with the first micro flow channel 321 of the first zone and the first distribution chamber 304 of the first zone. The reaction chamber 306 is in communication with the waste reservoir 310 via the fourth microchannel 324 in the first zone. The first reagent inlet 308 of the first zone is communicated with the reaction chamber 306 through an eighth micro-channel 328 of the first zone. The reaction cavity 306 is stored with CTNI/NT-proBNP/D-dimer/MYO/CKMB coated magnetic beads. The first reagent inlet 308 is closer to the center of rotation 11 than the reaction chamber 306. The second reagent inlet 307 of the first zone is communicated with the reaction chamber 306 through a ninth micro-channel 329 of the first zone. The second reagent inlet 307 in the first region is closer to the rotation center 11 than the reaction chamber 306.

In each distribution reaction structure 330, the separation chamber 302 is a sector-ring-shaped chamber extending along the rotation circumference of the microfluidic chip 10. In each distribution reaction structure 330, there are a plurality of reaction units 332, and the plurality of reaction units 332 are uniformly distributed along the extending direction of the first distribution chamber 304. The separation chamber 302, the first zone distribution chamber 304, the reaction chamber 306 and the waste liquid storage chamber 310 are sequentially spaced at increasing distances from the rotation center 11.

The second micro flow channel 322 in the first area includes a first sub flow channel 3221, a second sub flow channel 3222, and a third sub flow channel 3223, which are sequentially connected. The first branch flow path 3221 extends from the separation chamber 302 to near the rotation center 11 and is communicated with one end of the second branch flow path 3222, the other end of the second branch flow path 3222 is communicated with the third branch flow path 3223, and the third branch flow path 3223 extends from the second branch flow path 3222 to far away from the rotation center 11 and is communicated with the first distribution chamber 304. The first branch flow path 3221 is connected to the first side 30213021 of the separation chamber 302, and the first side 30213021 is a plane connecting the second side 30223022 of the separation chamber 302 close to the rotation center 11 and the third side 30233023 far from the rotation center 11.

In each of the distribution reaction structures 330, the separation chamber 302 communicates with the waste liquid storage chamber 310 through the fifth microchannel 325 in the first region. The fifth microchannel 325 in the first region is a U-shaped microchannel, and a U-shaped opening of the fifth microchannel 325 in the first region faces away from the rotation center 11. The first-region fifth microchannel 325 is led out from one end of the separation chamber 302 close to the second side 30223022 of the rotation center 11, and the connection position of the first-region second microchannel 322 and the first-region fifth microchannel 325 on the separation chamber 302 is located at the opposite ends of the separation chamber 302.

The first distribution chamber 304 extends circumferentially along the rotation of the microfluidic chip 10. The width of the first zone distribution chamber 304 in the radial direction is uniform in the direction of extension thereof. The first zone distribution chamber 304 has a narrower width in the radial direction of the microfluidic chip 10 than the separation chamber 302.

In each distribution reaction structure 330, the first-zone distribution chamber 304 communicates with the waste liquid storage chamber 310 through the first-zone sixth micro flow channel 326. The first-zone sixth microchannel 326 is led out from one end of the side surface of the separation chamber 302 near the rotation center 11.

The first-zone fifth micro-channel 325 and the first-zone sixth micro-channel 326 are converged and communicated with one end of a first-zone seventh micro-channel 327, and the other end of the first-zone seventh micro-channel 327 is communicated with the waste liquid storage cavity 310. The seventh micro flow channel 327 of the first zone is provided with a backflow prevention valve 312.

The first-zone fourth micro-channel 324 is provided with a first-zone second micro-flow valve 309.

The first microfluidic channel 321 of the first region is provided with a first microfluidic valve 305 of the first region.

The plurality of storage units 130 in the sample solvent adding layer 100 correspond to the plurality of flow dividing units 230 in the common reagent layer 200 one to one. The third zone sample addition hole 103 communicates with the sample addition hole 201 through the first interlayer channel. The third-zone first connection port 104 is communicated with the second-zone solvent addition hole 201 through a second interlayer channel. The second connection port 111 of the third zone is communicated with the second connection port 210 of the second zone through a third interlayer channel. The second zone third port 212 communicates with the first zone second reagent inlet 307 via a fourth inter-layer channel.

The plurality of flow dividing units 230 in the common reagent layer 200 correspond to the plurality of reaction units 332 in the reaction layer 300 one to one. Zone b first connection port 209 communicates with zone a first reagent inlet 308 via a fifth inter-layer channel. The zone b sample addition hole 204 communicates with the zone a sample addition hole 301 through a sixth interlayer channel.

The microfluidic chip 10 further comprises a second vent 402 and a first vent 401. One end of the second vent 402 is connected to the common reagent storage chamber 202, and the other end is open to a side surface of the microfluidic chip 10. One end of the first vent 401 is communicated with the waste liquid storage cavity 310, and the other end is opened on one side surface of the microfluidic chip 10.

The method for detecting by using the microfluidic chip 10 of the above specific example comprises the following steps:

(1) to the third zone sample addition well 103 of the sample solvent addition layer (1) was added the whole blood sample to be tested. The microfluidic chip 10 is added to a matched centrifugal device. Pure water was dropped into the third-zone solvent addition hole 101. The rotation is started, and the sample reaches the separation cavity 302 of the reaction layer 300 through the first interlayer channel, the sixth interlayer channel and the third micro-channel 323 of the A area for separation. The pure water reaches the third-zone second micro-flow valve 106 between the third-zone solvent adding hole 101 and the third-zone first connecting port 104 and the third-zone first micro-flow valve 105 between the third-zone solvent adding hole 101 and the third-zone distribution cavity 107.

(2) The centrifugal speed is increased, and the pure water breaks through the first micro-flow valve 105 in the third area and the second micro-flow valve 106 in the third area. The centrifugation speed is reduced and the plasma separated from the whole blood sample enters the zone a distribution chamber 304 through the zone a second microchannel 322 by capillary force. Pure water enters the third zone distribution cavity 107 of the sample solvent adding layer (1) and the public reagent storage cavity 202 in the public reagent layer 200, and the cleaning solution freeze-drying, the pre-excitation liquid freeze-drying and the excitation liquid freeze-drying stored in the three public reagent storage cavities 202 are dissolved in water.

3, the centrifugal speed is increased, and the pure water breaks through the third microfluidic valve 108 in the third zone behind the third zone distribution chamber 107 in the sample solvent addition layer 100 and enters the chemical substance labeled antibody unit 109. The chemical substance labeled antibody in the chemical substance labeled antibody unit 109 is lyophilized to be water-soluble. The plasma breaks through the first microfluidic valve 305 in the reaction layer 300 in the first area after the first area distribution chamber 304 and enters the reaction chamber 306.

(4) And (4) variable speed centrifugation, and mixing actions are simultaneously carried out in the chemical substance labeled antibody unit 109, the common reagent unit 202 and the reaction cavity 306.

(5) The centrifugal speed is increased, and the chemical substance labeled antibody breaks through the third zone fourth micro-flow valve 110 behind the chemical substance labeled antibody unit 109 and enters the second micro-flow channel 211 of the second zone through the second connecting port 111 of the third zone and the second connecting port 210 of the second zone in the common reagent layer 200. The centrifugal speed is reduced, and the chemical substance labeled antibody reaches the reaction cavity 306 in the reaction layer 300 through the second micro flow channel 211 of the second zone and the third connecting port 212 of the second zone.

(6) And (4) variable speed centrifugation, wherein the chemical substance labeled antibody in the reaction cavity 306 is uniformly mixed with the blood plasma and the magnetic beads.

(7) The centrifugal speed is increased, the mixture in the reaction chamber 306 passes through the second micro-flow valve 309 in the first region behind the reaction chamber 306 to the waste liquid storage chamber 310, which indicates that the reaction chamber 306 is filled with the reagent and the cleaning liquid in the common reagent layer 200 reaches the distribution chamber 207 in the second region. The centrifugal speed is reduced and the cleaning solution fills the zone b distribution chamber 207.

(8) The centrifugation speed is increased, the cleaning solution reaches the reaction chamber 306 through the first connection port 209 of the second zone, the fifth interlayer channel and the first reagent inlet 308 of the first zone, and the pre-excitation solution reaches the distribution chamber 207 of the second zone. The centrifugation speed is reduced and the pre-excitation liquid fills the zone b distribution chamber 207.

(9) Increasing the centrifugal speed, the cleaning solution reaches the waste solution storage chamber 310, the pre-excitation solution reaches the reaction chamber 306, and the excitation solution reaches the second zone distribution chamber 207. The centrifugation speed is reduced and the excitation liquid fills the zone b distribution chamber 207.

(10) Increasing the centrifugal speed, leading the exciting liquid to reach the reaction cavity 306, carrying out variable speed centrifugation, and mixing uniformly.

(11) Chemiluminescence captures optical signals, and numerical values are read to finish detection.

The rotation speed required in the above steps can be 100-10000r/min, such as 500r/min, 1000r/min, 2000r/min, etc. The rotation time can be 1s-10min, such as 10s, 30s, 1min, 5min, etc.

The microfluidic chip 10 of this specific example integrates detection of multiple items of multiple samples, and during detection, only a tester needs to drop a blood sample to a sample hole, and a machine drops a redissolution solvent uniformly after the tester is installed, so that multiple processes of reagent redissolution, uniform mixing, centrifugation, constant volume, reaction, elution and luminescence are automatically performed in an instrument.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. A microfluidic chip, wherein the microfluidic chip has a rotation center, and the microfluidic chip has a plurality of distribution reaction structures distributed around the rotation center and spaced apart from each other; the distribution reaction structure comprises a sample adding hole in the area A, a distribution cavity in the area A and a reaction unit; the first-zone sample adding hole is communicated with the first-zone distribution cavity, the reaction unit comprises a first micro-channel of the first zone and a reaction cavity, and the reaction cavity is communicated with the first-zone distribution cavity through the first micro-channel of the first zone; first district distribution chamber encircles the rotation center extends, it is a plurality of among the distribution reaction structure reaction unit, it is a plurality of reaction unit follows the extending direction who first district distribution chamber distributes, first district distribution chamber compare in reaction chamber is closer to the rotation center.

2. The microfluidic chip according to claim 1, wherein the distribution reaction structure further comprises a separation chamber and a first-region second microchannel, the separation chamber communicating with the first-region distribution chamber through the first-region second microchannel, the separation chamber being closer to the rotation center than the first-region distribution chamber.

3. The microfluidic chip according to claim 1 or 2, wherein the reaction unit further comprises a first reagent inlet of the first region and an eighth microchannel of the first region, the first reagent inlet of the first region communicating with the reaction chamber through the eighth microchannel of the first region, the first reagent inlet of the first region being closer to the center of rotation than the reaction chamber.

4. The microfluidic chip according to claim 3, wherein the microfluidic chip comprises a common reagent layer and a reaction layer stacked, and each of the distribution reaction structures is disposed in the reaction layer;

the flow distribution units are in one-to-one correspondence with the reaction units in the reaction layer, the first connecting port of the area B is communicated with the first reagent inlet of the area A through a fifth interlayer channel, and the sample adding hole of the area B is communicated with the sample adding hole of the area A through a sixth interlayer channel.

5. The microfluidic chip according to claim 4, wherein the reaction unit further comprises a first-zone second reagent inlet and a first-zone ninth microchannel, the first-zone second reagent inlet communicating with the reaction chamber through the first-zone ninth microchannel, the first-zone second reagent inlet being closer to the rotation center than the reaction chamber.

6. The microfluidic chip according to claim 4 or 5, wherein the common reagent storage chamber comprises a plurality of sub-storage chambers distributed around the rotation center and spaced apart from each other, and each of the sub-storage chambers is connected to the second-zone distribution chamber.

7. The microfluidic chip of claim 6, wherein each of the plurality of sub-reservoirs is in communication with the second zone distribution chamber via a reagent output channel, the reagent output channel comprising a first centrifugal force channel extending away from the center of rotation after exiting the common reagent reservoir, the first centrifugal force channel having a first exit microfluidic valve.

8. The microfluidic chip according to claim 7, wherein the reagent output channel corresponding to at least one of the plurality of sub-reservoirs further comprises a delay unit, the delay unit comprises a first diverting flow channel, a capillary flow channel, a second diverting flow channel, and a second centrifugal flow channel, the first centrifugal flow channel extends from the common reagent reservoir chamber to be far from the rotation center and is connected to one end of the first diverting flow channel, the other end of the first diverting flow channel is connected to one end of the capillary flow channel, the capillary flow channel extends from the first diverting flow channel to be close to the rotation center, the other end of the capillary flow channel is connected to one end of the second diverting flow channel, the other end of the second diverting flow channel is connected to the second centrifugal flow channel, and the second centrifugal flow channel extends from the second diverting flow channel to be far from the rotation center, the second centrifugal force flow channel is provided with a second discharging micro-flow valve.

9. The microfluidic chip according to claim 8, wherein the reagent output channel corresponding to at least one of the sub-reservoirs has a plurality of the delay units, and the plurality of delay units are sequentially connected.

10. The microfluidic chip according to any one of claims 4, 5, and 7 to 9, further comprising a sample solvent addition layer, wherein the sample solvent addition layer, the common reagent layer, and the reaction layer are sequentially stacked;