CN118122400A - Centrifugal dry biochemical detection micro-fluidic chip and application method thereof - Google Patents
Centrifugal dry biochemical detection micro-fluidic chip and application method thereof Download PDFInfo
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
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- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B—PERFORMING OPERATIONS; TRANSPORTING
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- B—PERFORMING OPERATIONS; TRANSPORTING
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- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
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Abstract
The invention provides a centrifugal dry biochemical detection micro-fluidic chip and a use method thereof, wherein a detection unit is arranged in the micro-fluidic chip, and the detection unit comprises: a shunt cavity for receiving a sample to be detected; each reaction cavity is divided into a sample quantifying cavity and a multilayer dry chemical test paper placing cavity below the sample quantifying cavity by the partition plate part; and a diversion and diversion structure for respectively communicating each sample quantifying cavity to the diversion cavity; the diversion and diversion structure is used for receiving the sample to be detected from the diversion cavity under the action of centrifugal force and sequentially filling the sample quantitative cavities with the diversion and diversion structure; the separator is provided with micropores, and the sample quantifying cavity is communicated with the multilayer dry chemical test paper placing cavity only through the micropores; after the micropores are sequentially filled in the sample quantifying cavities, the to-be-detected samples in the sample quantifying cavities can enter the multilayer dry chemical test paper placing cavity through the micropores by increasing centrifugal force, so that the biochemical reaction is ensured to be carried out in sequence.
Description
Technical Field
The invention relates to a centrifugal micro-fluidic chip, in particular to a centrifugal dry biochemical detection micro-fluidic chip and a use method thereof.
Background
Dry chemical analysis is a technique that involves applying a liquid test sample directly to a dry strip of reagent, and performing a chemical analysis by bringing about a specific chemical reaction with the moisture of the sample being tested as a solvent, and is based on enzymatic methods, also known as dry reagent chemistry or solid phase chemistry.
Common liquid test samples include urine and blood. The qualitative/semi-quantitative detection items of the dry chemical analysis test paper of urine basically comprise the content of routine analysis of urine, and the detection items comprise pH, specific gravity, urine protein, urine sugar, ketone body, bilirubin, urobilinogen, nitrite, leucocytes and erythrocytes. The dry chemical analyzer of blood can quantitatively determine liver function, kidney function, myocardial zymogram, etc.
Although the dry chemical analysis method has the advantages of rapidness, accuracy, convenience, simplicity, good reagent stability and the like, the traditional dry biochemical quantitative detection mode is low in test flux, and generally only one or a plurality of samples and one or a plurality of items can be tested at a time. Moreover, the volume of a sample required by the detection in the traditional dry biochemical quantitative detection mode is the same as that of the sample in the traditional wet chemistry, and for the patients with difficult sample collection such as infants, patients with long-term chemotherapy and small pets, the adoption of the traditional dry biochemical quantitative detection mode for carrying out multi-item biochemical detection is very inconvenient.
Microfluidic chip technology, also known as microfluidic chip labs or lab-on-a-chip, refers to chemical or biological labs built on a few square centimeters of chips that integrate basic operating units involved in the fields of chemistry and biology, etc., such as sample preparation, reaction, separation, detection, cell culture, sorting, lysis, etc., onto a small chip, with microchannels forming a network, through which a controllable fluid is passed, for achieving various functions in the fields of biology, chemistry, medical diagnostics, etc.
The centrifugal biochemical detection microfluidic core based on the dry chemical technology has the advantages of high measurement flux, quick and convenient operation, high detection accuracy and strong repeatability, and can detect a plurality of indexes on a trace sample in parallel.
Disclosure of Invention
The invention aims to: aiming at the defects of the prior art, the invention provides a centrifugal dry biochemical detection micro-fluidic chip which can detect a plurality of indexes on a trace sample in parallel and has the advantages of high measurement flux, quick and convenient operation, high detection accuracy and strong repeatability.
To solve the above technical problem, a first aspect of the present invention discloses a centrifugal dry biochemical detection microfluidic chip, in which a detection unit is disposed, the detection unit includes:
a shunt cavity for receiving a sample to be detected;
Each reaction cavity is divided into a sample quantifying cavity and a multilayer dry chemical test paper placing cavity below the sample quantifying cavity by the corresponding baffle plate part;
The flow diversion and distribution structure is used for enabling each sample quantitative cavity to be communicated with the corresponding flow diversion cavity through the flow diversion and distribution structure; the diversion and diversion structure is used for receiving the sample to be detected from the diversion cavity under the action of centrifugal force and sequentially filling the sample quantitative cavities with the diversion and diversion structure;
the separator is provided with micropores, and the sample quantifying cavity is communicated with the multilayer dry chemical test paper placing cavity only through the micropores; the micropores are formed after the sample quantifying cavities are filled in sequence, and the sample to be detected in each sample quantifying cavity can enter the corresponding multilayer dry chemical test paper placing cavity through the corresponding micropores by increasing centrifugal force.
Specifically, the sample quantifying cavity and the multilayer dry chemical test paper placing cavity are cylindrical cavities, have the same diameter and are arranged opposite to each other along the rotation central axis of the microfluidic chip. The inlet of the micropore is positioned at the center of the bottom of the sample quantifying cavity, and the outlet of the micropore is positioned at the center of the top of the multilayer dry chemical test paper placing cavity. When the multilayer dry chemical test paper is placed in the multilayer dry chemical test paper placing cavity, the multilayer dry chemical test paper is pressed and attached to the lower side of the partition plate part, and the size of the multilayer dry chemical test paper is matched with the multilayer dry chemical test paper placing cavity.
Specifically, at least one of the multi-layer dry chemical test paper placing cavities is preset with a plurality of layers of dry chemical test paper.
Specifically, the multilayer dry chemical test paper comprises a sample dispersion filter paper layer, a dry chemical reagent layer and a color development test paper measuring layer which are sequentially laminated along the longitudinal direction. The sample dispersion filter paper layer is pressed and attached to the lower side of the partition plate portion, so that the multilayer dry chemical test paper is pressed and attached to the lower side of the partition plate portion.
Specifically, the diversion flow distribution structure comprises a radial communication channel, a diversion flow channel and a diversion flow channel, and the diversion cavity, the radial communication channel and the diversion flow channel are sequentially communicated. Each sample quantifying cavity is communicated with the diversion flow channel through a corresponding diversion flow channel. The diversion cavity is positioned at one side of the diversion flow channel, which is close to the rotation central axis of the micro-fluidic chip. The radial communication channel is positioned between the diversion cavity and the diversion flow channel. The diversion flow channel is an arc flow channel with the arc center positioned on the rotation central axis of the micro-fluidic chip. The more than one reaction cavities are positioned at one side of the diversion flow channel far away from the rotation central axis of the microfluidic chip and are distributed at equal angle intervals around the rotation central axis of the microfluidic chip. Each flow dividing channel is respectively arranged between the corresponding reaction cavity and the corresponding flow guiding channel and extends along the radial direction of the microfluidic chip.
Specifically, the inlet of the flow diversion channel is connected with the upper part of the flow diversion channel, and the outlet of the flow diversion channel is connected with the upper part of the sample quantifying cavity.
Specifically, the detection unit comprises a sample adding port, and the sample adding port is positioned at the top of the shunt cavity.
Specifically, the microfluidic chip comprises a first cover layer, a chip substrate and a second cover layer, wherein the chip substrate comprises a first side and a second side which are oppositely arranged, the flow diversion cavity, the flow diversion flow channel and the sample quantifying cavity are respectively formed into grooves in the first side of the chip substrate, and the multilayer dry chemical test paper placing cavity is formed into grooves in the second side of the chip substrate. The first side is covered with a first cover layer, and the second side is covered with a second cover layer to enclose the flow diversion cavity, the flow diversion flow passage and the sample quantifying cavity. The sample adding port is arranged on the first covering layer.
Specifically, the chip substrate and the second cover layer are both formed of a transparent material.
The second aspect of the invention provides a use method of the centrifugal dry biochemical detection microfluidic chip, in a detection unit of the microfluidic chip, at least one multilayer dry chemical test paper placing cavity is preset with a plurality of layers of dry chemical test papers, and the plurality of layers of dry chemical test papers are pressed and attached to the lower side of a partition plate part; the application method comprises the following steps:
Under the action of a certain centrifugal force, the diversion and diversion structure receives a sample to be detected from the diversion cavity and fills the sample to be detected into each sample quantifying cavity in sequence; under the action of the certain centrifugal force, the air pressure in the multi-layer dry chemical test paper placing cavity of the sample to be detected cannot be kept in the corresponding sample quantifying cavity through the corresponding micropores;
Step two, increasing centrifugal force, and enabling the sample to be detected in each sample quantitative cavity to enter a corresponding multilayer dry chemical test paper placing cavity through a corresponding micropore; the sample to be detected enters the sample dispersion filter paper layer of the multilayer dry chemical test paper from the micropores and uniformly diffuses, so that the biochemical reaction in the multilayer dry chemical test paper is ensured to be carried out in sequence.
The beneficial effects are that:
(1) Compared with a dry chemical test paper test card in the prior art, the application can quantify the sample to be detected, can detect a plurality of indexes on the same sample to be detected in parallel, has the advantages of high measurement flux, quick and convenient operation, high detection accuracy, strong repeatability, high flux advantage of the centrifugal micro-fluidic technology, rapidness, accuracy, convenience, simplicity, good reagent stability, single-person independent package, reduction of waste of unsealing and invalidation of liquid reagents, simple structure of a matched analyzer, small volume, easy realization of POCT function and the like.
(2) According to the application, the separator parts with micropores are arranged in each reaction cavity, each separator part divides the corresponding reaction cavity into the sample quantifying cavity and the multilayer dry chemical test paper placing cavity positioned below the sample quantifying cavity, and the multilayer dry chemical test paper placing cavity is relatively airtight and is communicated with the sample quantifying cavity only through the micropores. When the device is used, under the action of a certain centrifugal force, the to-be-detected samples are filled into the sample quantifying cavities, and in the process, the to-be-detected samples cannot enter the multilayer dry chemical test paper placing cavities due to the air pressure in the multilayer dry chemical test paper placing cavities, so that the inflow to-be-detected samples are kept in the sample quantifying cavities and the sample quantifying cavities are filled; then after all sample quantifying cavities are filled in sequence, increasing centrifugal force, and enabling the samples to be detected in each sample quantifying cavity to enter the corresponding multilayer dry chemical test paper placing cavity through corresponding micropores; the sample to be detected enters the sample dispersion filter paper layer of the multi-layer dry chemical test paper from the micropores and is uniformly diffused. Therefore, for parallel multi-project detection, the process of reacting and developing the sample quantity to be detected among the pre-quantitative distribution projects and each distribution project is divided into two steps which are sequentially carried out, the samples in each sample quantitative cavity can only enter the corresponding multilayer dry chemical test paper placing cavity through the micropores, the samples flowing out of the micropores enter the sample dispersing filter paper layer of the multilayer dry chemical test paper and can be uniformly diffused, and therefore the whole biochemical reaction sequence is ensured, and further the high detection accuracy and the high repeatability of the microfluidic chip are ensured. Conversely, if the baffle plate part is not arranged in the reaction cavity, the reaction cavity is used as a quantitative cavity and a multi-layer dry chemical test paper placing cavity, namely, the multi-layer dry chemical test paper is directly placed in the quantitative reaction cavity; because a gap exists between the inner wall of the reaction cavity and the dry chemical test paper, a sample to be detected enters the quantitative reaction cavity under the action of centrifugal force and is flushed to the inner wall of the centrifugal force far end, and the sample liquid to be detected directly enters the bottom layer of the multilayer dry chemical test paper along the gap between the inner wall of the far end and the multilayer dry chemical test paper, so that the whole biochemical reaction sequence is damaged, and the result is uncontrollable and wrong. The present application eliminates the above-described drawbacks.
Drawings
The foregoing and/or other advantages of the invention will become more apparent from the following detailed description of the invention when taken in conjunction with the accompanying drawings and detailed description.
FIG. 1 is an exploded view of a centrifugal dry biochemical detection microfluidic chip according to an embodiment of the present invention;
FIG. 2 is a top view of a chip substrate of the centrifugal dry biochemical detection microfluidic chip shown in FIG. 1;
FIG. 3 is a bottom view of the chip substrate of a centrifugal dry biochemical test microfluidic chip of FIG. 1;
FIG. 4 is a perspective cross-sectional view taken along line A-A of FIG. 2;
Fig. 5 is a cross-sectional view of fig. 4.
The reference numerals are explained as follows:
101. a shunt cavity; 1011. a first sidewall; 1012. a sample adding port; 102. a diversion structure; 1021. a radial communication channel; 1022. a diversion flow passage; 1023. a flow dividing channel; 103. a reaction chamber; 1031. a partition plate portion; 1032. a sample quantification chamber; 1033. a multilayer dry chemical test paper placing cavity; 1034. micropores; 104. a rotation central axis; 106. positioning holes; 107. a ventilation through hole; 110. a first cover layer; 120. a chip substrate; 121. a first side; 122. a second side; 130. and a second cover layer.
Detailed Description
Referring to fig. 1 to 4, the present invention provides a centrifugal dry biochemical detection microfluidic chip, in which a detection unit is disposed, the detection unit includes a diversion cavity 101 for receiving a sample to be detected, one or more reaction cavities 103, and a diversion structure 102.
Referring to fig. 1 and 4, a partition plate portion 1031 is provided in each reaction chamber 103, and each reaction chamber 103 is partitioned by the corresponding partition plate portion 1031 into a sample quantifying chamber 1032 and a multi-layer dry chemical test paper placing chamber 1033 located below the sample quantifying chamber 1032.
Referring to fig. 1 and 4, each sample metering chamber 1032 is in communication with the flow diversion chamber 101 via a flow diversion structure 102, respectively. The flow diversion and diversion structure 102 is used for receiving the sample to be detected from the diversion cavity 101 and filling each sample quantifying cavity 1032 in turn.
Referring to fig. 1 and 4, a separator 1031 is provided with a micropore 1034, and a sample quantification chamber 1032 communicates with a multilayer dry chemical test paper placement chamber 1033 only through the micropore 1034. The microwells 1034 are configured such that after each sample-quantifying chamber 1032 is sequentially filled, the sample in each sample-quantifying chamber 1032 can enter the corresponding multi-layered dry chemical test paper placing chamber 1033 through the corresponding microwells 1034 by increasing the centrifugal force.
In the present application, the sample to be tested may be pre-diluted plasma or urine depending on the biochemical test item. The pre-diluted blood plasma can be used for liver function detection, blood sugar and blood fat detection, kidney function detection and myocardial zymogram detection. Items for liver function detection include total protein, albumin, globulin, albumin-to-ball ratio, total bilirubin, direct, indirect bilirubin, and transaminase. Items for blood glucose and lipid testing include total cholesterol, triglycerides, high and low density lipoproteins, apolipoproteins and fasting blood glucose. Items for renal function testing include creatinine, urea nitrogen, and uric acid; items for myocardial zymogram detection include lactate dehydrogenase and creatine kinase. Urine is used for detecting projects such as urinary creatinine, urinary nitrogen, urinary uric acid, urinary sugar, urinary albumin and the like. For different detection projects, a corresponding multilayer dry chemical test paper should be used.
In the application, by arranging the partition plate part 1031 with the micropores 1034 in each reaction cavity 103, each partition plate part 1031 divides the corresponding reaction cavity 103 into the sample quantifying cavity 1032 and the multi-layer dry chemical test paper placing cavity 1033 positioned below the sample quantifying cavity 1032, and the multi-layer dry chemical test paper placing cavity 1033 is relatively sealed and communicated with the sample quantifying cavity 1032 only through the micropores 1034. When in use, under the action of a certain centrifugal force, the diversion and flow distribution structure 102 receives a sample to be detected from the flow distribution cavity 101 and fills each sample quantitative cavity 1032 with the sample to be detected, and in the process, the sample to be detected cannot enter the multilayer dry chemical test paper placing cavity 1033 due to the air pressure in the multilayer dry chemical test paper placing cavity 1033, so that the inflow sample to be detected is kept in the sample quantitative cavity 1032 and the sample quantitative cavity 1032 is filled. And then after all the sample quantifying chambers 1032 are sequentially filled, increasing the centrifugal force, wherein the pressure applied to the sample to be detected by the centrifugal force is greater than the air pressure in the multilayer dry chemical test paper placing chamber 1033, so that the sample to be detected in each sample quantifying chamber 1032 enters the corresponding multilayer dry chemical test paper placing chamber 1033 through the corresponding micropores 1034. The sample to be detected enters the sample dispersion filter paper layer of the multilayer dry chemical test paper from the micropores 1034 and is uniformly diffused, and the air in the multilayer dry chemical test paper placing cavity 1033 is discharged. Therefore, for parallel multi-project detection, the process of reacting and developing the sample quantity to be detected among the pre-quantitative distribution projects and each distribution project is divided into two steps which are sequentially carried out, and the samples in each sample quantitative cavity 1032 can only enter the corresponding multilayer dry chemical test paper placing cavity 1033 through the micropores 1034, the samples flowing out of the micropores 1034 enter the sample dispersing filter paper layer of the multilayer dry chemical test paper and can be uniformly diffused, and sequentially penetrate into the reagent layers and the developing layers of the multilayer dry chemical test paper, so that the whole biochemical reaction sequence is ensured, and the detection effectiveness, accuracy and repeatability of the microfluidic chip are further ensured.
Conversely, if the baffle plate part is not arranged in the reaction cavity, the reaction cavity is used as a quantitative cavity and a multi-layer dry chemical test paper placing cavity, namely, the multi-layer dry chemical test paper is directly placed in the quantitative reaction cavity. Because a gap exists between the inner wall of the reaction cavity and the dry chemical test paper, a sample to be detected enters the quantitative reaction cavity under the action of centrifugal force and is flushed to the inner wall of the centrifugal force far end, and the sample liquid to be detected directly enters the bottom layer of the multilayer dry chemical test paper along the gap between the inner wall of the far end and the multilayer dry chemical test paper, so that the whole biochemical reaction sequence is damaged, and the result is uncontrollable and wrong. Meanwhile, as the dry test paper absorbs and permeates the sample to be detected at a certain speed, namely, at a certain retardation, the speed of distributing the sample to be detected to different reaction chambers of the diversion and distribution structure 102 can be influenced, namely, the sample to be detected cannot enter the reaction chambers completely in time, the time of the diversion flow channel of the diversion and distribution structure for stay of the sample to be detected is prolonged, and the probability of cross contamination among different reaction chambers is further increased.
Specifically, referring to fig. 2 to 5, the micro holes 1034 penetrate through the partition plate portion 1031 in the direction of the rotation center axis 104 of the microfluidic chip.
Specifically, referring to fig. 2 to 4, the sample quantifying cavity 1032 and the multilayer dry chemical test paper placing cavity 1033 are cylindrical cavities, and have the same diameter and are disposed directly opposite to each other along the rotation central axis 104 of the microfluidic chip. The inlet of the microwell 1034 is located at the center of the bottom of the sample quantifying chamber 1032, and the outlet of the microwell 1034 is located at the center of the top of the multilayer dry chemical test paper placing chamber 1033. When the multilayer dry chemical test paper is placed in the multilayer dry chemical test paper placing cavity 1033, the multilayer dry chemical test paper is pressed and attached to the lower side of the partition plate portion 1031, and the size of the multilayer dry chemical test paper is matched with the size of the multilayer dry chemical test paper placing cavity 1033, namely, the multilayer dry chemical test paper fills the multilayer dry chemical test paper placing cavity 1033.
In this embodiment, the sample to be detected from the outlet of the micropore 1034 is added from the central position of the multilayer dry chemical test paper, so that the situation that the sample to be detected directly permeates into the chromogenic test paper testing layer of the multilayer dry chemical test paper along the gap between the multilayer dry chemical test paper and the cavity side wall of the multilayer dry chemical test paper placing cavity 1033, thereby leading to the reverse of the biochemical reaction sequence and the failure of the chip is avoided. This arrangement is also advantageous in that the sample to be detected is uniformly dispersed, and the result is more accurate.
According to the embodiment, the lower sides of the multi-layer dry chemical test paper and the partition board portion 1031 are pressed and attached, so that the partition board portion 1031 effectively prevents the multi-layer dry chemical test paper from overturning in the transportation process, and the probability of mutual pollution interference in a chip due to falling of biochemical reagent powder of different projects is reduced. Meanwhile, gaps are avoided among layers of the multi-layer dry chemical test paper, and bubbles in the gaps are prevented from blocking a sample to be detected to pass through the dry chemical test paper, so that the sample to be detected is ensured to be uniformly infiltrated from top to bottom.
In the embodiment, the cylindrical cavity and the circular dry chemical test paper are adopted, compared with other shapes such as a cubic cavity and a square dry chemical test paper, the distance from the center to the edge of the circular dry chemical test paper is the same, and the diffusion of the sample to be detected from the center point is uniform, so that the time consistency of biochemical reaction of the dry chemical test paper is ensured, and the detection precision is ensured.
Specifically, in the detection unit, at least one multi-layered dry chemical test strip placement chamber 1033 is pre-set with multi-layered dry chemical test strips.
Specifically, the multi-layer dry chemical test paper comprises a sample dispersion filter paper layer, a dry chemical reagent layer and a color development test paper measuring layer which are sequentially laminated along the longitudinal direction; the sample dispersion filter paper layer is pressed and attached to the lower side of the partition board 1031, so that the multilayer dry chemical test paper is pressed and attached to the lower side of the partition board 1031.
The multilayer dry chemical test paper can be prepared by adopting the same process based on the finished product of the commercially available dry chemical test paper.
Specifically, referring to fig. 2, the flow diversion structure 102 includes a radial communication passage 1021, a flow diversion flow passage 1022, and a flow diversion flow passage 1023, and the flow diversion cavity 101, the radial communication passage 1021, and the flow diversion flow passage 1022 are sequentially communicated. Each sample metering chamber 1032 communicates with a flow diversion flow passage 1022 through a corresponding flow diversion flow passage 1023. The diversion cavity 101 is located on one side of the diversion flow passage 1022 near the rotation central axis 104 of the microfluidic chip. The radial communication passage 1021 is located between the diversion chamber 101 and the diversion flow passage 1022. The flow guide channels 1022 are arc-shaped channels with their arc centers located on the rotational center axis 104 of the microfluidic chip. The more than one reaction cavity 103 is located at one side of the flow guide channel 1022 away from the rotation central axis 104 of the microfluidic chip and is equiangularly distributed around the rotation central axis 104 of the microfluidic chip. Each of the flow dividing channels 1023 is disposed between the corresponding reaction chamber 103 and the flow guiding channel 1022 and extends along the radial direction of the microfluidic chip.
Specifically, referring to fig. 1, the inlet of the flow diversion channel 1023 is connected to the upper portion of the flow diversion channel 1022, and the outlet of the flow diversion channel 1023 is connected to the upper portion of the sample quantifying chamber 1032.
Specifically, referring to fig. 2, the shunt cavity 101 includes a first sidewall 1011 remote from the rotational central axis 104 of the microfluidic chip, the first sidewall 1011 being an arcuate surface having a common rotational central axis 104 with the microfluidic chip.
In this embodiment, the first side wall 1011 of the shunt cavity 101 far away from the rotation central axis 104 of the microfluidic chip is an arc surface, so as to avoid the sample to be detected remaining in the shunt cavity 101 after filling each sample quantifying cavity 1032 with the sample to be detected.
Specifically, referring to fig. 2, the radial communication channels 1021 extend in the radial direction of the microfluidic chip. Referring to fig. 1, an inlet of the radial communication passage 1021 is located at an upper portion of the first side wall 1011 of the flow dividing chamber 101, and an outlet of the radial communication passage 1021 is located at an upper portion of a side of the flow guiding flow passage 1022 near the rotation center axis 104 of the microfluidic chip.
Specifically, referring to fig. 1, the detection unit includes a ventilation through hole 107 directly communicating with the external atmosphere, the ventilation through hole 107 is located at a side of the flow guide channel 1022 near the rotation central axis 104 of the microfluidic chip, and the end of the flow guide channel 1022 is communicated with the ventilation through hole 107 through a curved pipeline.
Specifically, referring to fig. 1, each detection unit includes a sample inlet 1012, and the sample inlet 1012 is located at the top of the flow distribution chamber 101. The sample to be detected, such as pre-diluted blood plasma, can be directly added to the shunt cavity 101 through the sample adding port 1012, and the blood plasma and the diluent can be sequentially added to the shunt cavity 101 to obtain the sample to be detected in the shunt cavity 101.
Specifically, referring to fig. 1, the microfluidic chip includes a first cover layer 110, a chip substrate 120, and a second cover layer 130, the chip substrate 120 includes a first side 121 and a second side 122 disposed opposite to each other, the flow splitting chamber 101, the flow guiding flow channels 1022, the flow splitting flow channels 1023, and the sample quantifying chambers 1032 are respectively formed as grooves in the first side 121 of the chip substrate 120, and the multilayer dry chemical test paper placing chamber 1033 is formed as a groove in the second side 122 of the chip substrate 120; the first side 121 is covered with a first cover layer 110 and the second side 122 is covered with a second cover layer 130 to enclose the flow diversion chamber 101, the flow diversion flow channel 1022, the flow diversion flow channel 1023 and the sample quantifying chamber 1032; the sample inlet 1012 is disposed on the first cover layer 110.
Specifically, referring to fig. 1, the ventilation through-holes 107 penetrate through the first cover layer 110, the chip substrate 120, and the second cover layer 130.
Specifically, the chip substrate 120 and the second cover layer 130 are both formed of a transparent material, such as glass or plastic, to satisfy the need for human eye observation or machine vision.
In this embodiment, the color change of the chromogenic test paper measuring layer is measured by the transparent second cover layer 130, and qualitative detection and quantitative analysis of the analyte concentration can be realized based on the light reflection detection method. Qualitative detection and quantitative analysis of analyte concentration based on light reflection detection are prior art, and have been widely applied to dry chemical analyzers on the market, and therefore are not described here in detail.
In one embodiment, the microfluidic chip is a disk with several detection units arrayed around the central axis of rotation 104 of the microfluidic chip. In another embodiment, referring to fig. 1, the microfluidic chips are fan-shaped discs, each provided with only one detection unit. The fan-shaped discs are spliced into a circular disc shape. Correspondingly, the micro-fluidic chip in the sector disc state is matched with a carrier disc, and the micro-fluidic chip in the sector disc state can be quickly disassembled and assembled with the carrier disc. Specifically, in the microfluidic chip in the fan-shaped disc state, the chip substrate 120, the first cover layer 110, and the second cover layer 130 may all be fan-shaped. The bottoms of the two angles of the outer arc degree of the chip substrate 120 are concave, 2 positioning grooves are respectively formed in two sides of the outer edge of the chip substrate 120, three positioning holes 106 are formed in the chip substrate 120, and through holes are formed in the positions, corresponding to the positioning holes 106, of the first cover layer 110 and the second cover layer 130 so as to avoid shielding the positioning holes 106 during film pasting. The carrier plate is provided with a positioning step matched with the positioning groove and a positioning boss matched with the positioning hole 106, and the carrier plate is clamped with the microfluidic chip in the sector disc state through the positioning step and the positioning boss. In a particular embodiment, six fanned disks are spliced into a circular disk shape.
The second aspect of the present application provides a method for using the centrifugal biochemical detection microfluidic chip, which comprises the following steps:
Under the action of a certain centrifugal force, the diversion and diversion structure 102 receives a sample to be detected from the diversion cavity 101 and sequentially fills each sample quantifying cavity 1032; under the action of the certain centrifugal force, the sample to be detected cannot be held in the corresponding sample quantifying cavity 1032 by the pressure in the multilayer dry chemical test paper placing cavity 1033 through the corresponding micropores 1034;
Step two, increasing centrifugal force, and enabling the sample to be detected in each sample quantitative cavity 1032 to enter a corresponding multilayer dry chemical test paper placing cavity 1033 through a corresponding micropore 1034; the sample to be detected enters the sample dispersion filter paper layer of the multi-layer dry chemical test paper from the micropores 1034 and is uniformly diffused. The biochemical reactions are then carried out sequentially.
In a specific embodiment, the microfluidic chip of the present embodiment is applied to measuring five indicators of liver function on pre-diluted infant heel blood. The five indexes are alanine aminotransferase, aspartic aminotransferase, total bile acid, total bilirubin and direct bilirubin, respectively. Correspondingly, the chip is provided with six reaction chambers 103, and alanine aminotransferase test paper, aspartic acid aminotransferase test paper, total bile acid test paper, total bilirubin test paper, direct bilirubin test paper and blank control test paper are sequentially and respectively placed in each multi-layer dry chemical test paper placing chamber 1033 from one side close to the outlet of the radial communication channel 1021. By arranging the blank control test paper, interference caused by the color background of the sample to be detected can be eliminated, and the accuracy of the result is improved. The multi-layer dry chemical test paper can be prepared by adopting the same process based on the finished product of the commercially available dry chemical test paper, and related companies include ARKRAY Factoy, inc., oson clinical diagnosis (United states) Co., ltd., ortho-Clinical Diagnostics, inc., and Guangzhou Wanfu Biotechnology Co., ltd.
In the microfluidic chip of the present embodiment, the capacity of the shunt chamber 101 is 300ul. The diameter of the loading port 1012 was 1500um. The radial communication passage 1021 has a width dimension of 600um and a depth dimension of 600um. The flow channels 1022 have a width dimension of 600um and a depth dimension of 1500um. The sample quantitative chamber 1032 and the multilayer dry chemical test paper placing chamber 1033 each have a volume of 15ul, and are each cylindrical cavities having a height dimension of 2.1mm and a diameter of 3.0 mm.
The sample to be detected can be prepared before the measurement. The specific process is as follows: collecting 20ul of infant heel blood, adding a trace blood sampling tube, centrifuging at 1500rpm for 5 minutes, and separating serum; diluting the separated serum with purified water for 30 times, if the separated serum is 10ul, adding purified water 290ul, and mixing. The diluted sample obtained by mixing is used as a sample to be detected.
The measuring process comprises the following steps:
Step one, 200ul of diluted sample is taken and added into a diversion cavity 101 of the chip, the chip is put into a matched analyzer, the analyzer automatically executes a centrifugal program, and under the action of a certain centrifugal force, for example, 1500 rpm-2000 rpm, the sample to be detected is automatically and quantitatively distributed into 6 sample quantitative cavities 1032 through a diversion and diversion structure 102;
Step two, increasing the centrifugal force to 2500 rpm-3500 rpm, enabling the sample to be detected in the sample quantitative cavity 1032 to enter the multilayer dry chemical test paper placing cavity 1033 through the micropores 1034 in the middle of the partition plate portion 1031, sequentially passing through the sample dispersing filter paper layer and the dry chemical reagent layer to the color development test paper measuring layer, performing biochemical reaction on the dry chemical reagent layer, performing color development reaction on the color development test paper measuring layer, and finally analyzing and calculating the activity value of biochemical enzyme and the content of substances in the sample through measuring the color variation of the color development test paper measuring layer.
The invention provides a centrifugal dry biochemical detection micro-fluidic chip, a thought of using method thereof and a method thereof, and a method and a way for realizing the technical scheme are numerous, the above is only a preferred embodiment of the invention, and it should be pointed out that a plurality of improvements and modifications can be made to those skilled in the art without departing from the principle of the invention, and the improvements and modifications are also regarded as the protection scope of the invention. The components not explicitly described in this embodiment can be implemented by using the prior art.
Claims (10)
1. A centrifugal dry biochemical detection micro-fluidic chip, characterized in that a detection unit is arranged in the centrifugal dry biochemical detection micro-fluidic chip, and the detection unit comprises:
a shunt chamber (101) for receiving a sample to be detected;
More than one reaction cavity (103), each reaction cavity (103) is internally provided with a baffle plate part (1031), and each reaction cavity (103) is divided into a sample quantifying cavity (1032) and a multi-layer dry chemical test paper placing cavity (1033) positioned below the sample quantifying cavity (1032) by the corresponding baffle plate part (1031);
And a flow diversion structure (102), each sample quantifying cavity (1032) is respectively communicated with the flow diversion cavity (101) through the flow diversion structure (102); the diversion and diversion structure (102) is used for receiving the sample to be detected from the diversion cavity (101) under the action of centrifugal force and sequentially filling each sample quantifying cavity (1032);
The separator part (1031) is provided with micropores (1034), and the sample quantifying cavity (1032) is communicated with the multilayer dry chemical test paper placing cavity (1033) only through the micropores (1034); the micropores (1034) are configured such that after each of the sample quantification chambers (1032) is filled in sequence, a sample to be detected in each of the sample quantification chambers (1032) can enter the corresponding multi-layer dry chemical test paper placement chamber (1033) through the corresponding micropores (1034) by increasing centrifugal force.
2. The centrifugal dry biochemical detection micro-fluidic chip according to claim 1, wherein the sample quantifying cavity (1032) and the multilayer dry chemical test paper placing cavity (1033) are cylindrical cavities, have the same diameter and are arranged opposite to each other along a rotation central axis (104) of the micro-fluidic chip; the inlet of the micropore (1034) is positioned at the center of the bottom of the sample quantifying cavity (1032), and the outlet of the micropore (1034) is positioned at the center of the top of the multilayer dry chemical test paper placing cavity (1033); when the multilayer dry chemical test paper is placed in the multilayer dry chemical test paper placing cavity (1033), the multilayer dry chemical test paper is pressed and attached to the lower side of the partition plate portion (1031), and the size of the multilayer dry chemical test paper is matched with the multilayer dry chemical test paper placing cavity (1033).
3. A centrifugal dry biochemical detection micro-fluidic chip according to claim 2, wherein at least one of said multi-layer dry chemical test strip placement chambers (1033) is pre-loaded with multi-layer dry chemical test strips.
4. A centrifugal dry biochemical test microfluidic chip according to claim 3, wherein the multilayer dry chemical test paper comprises a sample dispersion filter paper layer, a dry chemical reagent layer and a chromogenic test paper measuring layer which are sequentially laminated in a longitudinal direction; the sample dispersion filter paper layer is pressed and attached to the lower side of the partition board portion (1031), so that the multilayer dry chemical test paper is pressed and attached to the lower side of the partition board portion (1031).
5. The centrifugal dry biochemical detection micro-fluidic chip according to claim 4, wherein the diversion flow distribution structure (102) comprises a radial communication channel (1021), a diversion flow channel (1022) and a diversion flow channel (1023), and the diversion cavity (101), the radial communication channel (1021) and the diversion flow channel (1022) are sequentially communicated; each sample quantifying cavity (1032) is respectively communicated with the diversion flow channel (1022) through a corresponding diversion flow channel (1023); the diversion cavity (101) is positioned at one side of the diversion flow channel (1022) close to the rotation central axis (104) of the microfluidic chip; the radial communication channel (1021) is positioned between the diversion cavity (101) and the diversion flow channel (1022); the diversion flow channel (1022) is an arc flow channel with the arc center positioned on the rotation central axis (104) of the micro-fluidic chip; the more than one reaction cavities (103) are positioned at one side of the diversion flow channel (1022) far away from the rotation central axis (104) of the micro-fluidic chip and are distributed at equal angular intervals around the rotation central axis (104) of the micro-fluidic chip; each flow dividing flow passage (1023) is respectively arranged between the corresponding reaction cavity (103) and the corresponding flow guiding flow passage (1022) and extends along the radial direction of the microfluidic chip.
6. The centrifugal dry biochemical detection micro-fluidic chip according to claim 5, wherein an inlet of the flow diversion channel (1023) is connected to an upper portion of the flow diversion channel (1022), and an outlet of the flow diversion channel (1023) is connected to an upper portion of the sample quantifying cavity (1032).
7. The centrifugal dry biochemical detection micro-fluidic chip according to claim 6, wherein the detection unit comprises a sample inlet (1012), and the sample inlet (1012) is positioned at the top of the shunt cavity (101).
8. The centrifugal dry biochemical detection microfluidic chip according to claim 7, wherein the microfluidic chip comprises a first cover layer (110), a chip substrate (120) and a second cover layer (130), the chip substrate (120) comprises a first side (121) and a second side (122) which are oppositely arranged, the flow diversion chamber (101), the flow diversion flow channel (1022), the flow diversion flow channel (1023) and the sample quantification chamber (1032) are respectively formed as grooves in the first side (121) of the chip substrate (120), and the multi-layer dry chemical test paper placement chamber (1033) is formed as grooves in the second side (122) of the chip substrate (120); -the first side (121) is covered with a first cover layer (110) and the second side (122) is covered with a second cover layer (130) to enclose the flow diversion chamber (101), the flow diversion flow channel (1022), the flow diversion flow channel (1023) and the sample quantification chamber (1032); the sample adding port (1012) is arranged on the first covering layer (110).
9. The centrifugal dry biochemical detection micro-fluidic chip according to claim 8, wherein the chip substrate (120) and the second cover layer (130) are both formed of a transparent material.
10. The method for using a centrifugal dry biochemical detection micro-fluidic chip according to claim 1, wherein in the detection unit of the micro-fluidic chip, at least one multi-layer dry chemical test paper placing cavity (1033) is preset with a plurality of layers of dry chemical test papers, and the multi-layer dry chemical test papers are pressed and attached to the lower side of a partition plate part (1031); the application method comprises the following steps:
Under the action of a certain centrifugal force, the diversion and diversion structure (102) receives a sample to be detected from the diversion cavity (101) and sequentially fills each sample quantifying cavity (1032); under the action of the certain centrifugal force, the sample to be detected cannot be kept in the corresponding sample quantifying cavity (1032) through the corresponding micropores (1034) due to the air pressure in the multilayer dry chemical test paper placing cavity (1033);
Step two, increasing centrifugal force, and enabling the sample to be detected in each sample quantifying cavity (1032) to enter a corresponding multilayer dry chemical test paper placing cavity (1033) through a corresponding micropore (1034); the sample to be detected enters the sample dispersion filter paper layer of the multi-layer dry chemical test paper from the micropores (1034) and is uniformly diffused.
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