CN110586209A - Micro-fluidic chip and in-vitro detection device comprising same - Google Patents
Micro-fluidic chip and in-vitro detection device comprising same Download PDFInfo
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- CN110586209A CN110586209A CN201910803534.0A CN201910803534A CN110586209A CN 110586209 A CN110586209 A CN 110586209A CN 201910803534 A CN201910803534 A CN 201910803534A CN 110586209 A CN110586209 A CN 110586209A
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Classifications
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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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- 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
- B01L3/502753—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 bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces
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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
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N2021/7756—Sensor type
- G01N2021/7759—Dipstick; Test strip
Abstract
The invention relates to a micro-fluidic chip and an in-vitro detection device comprising the same. The micro-fluidic chip is provided with a sample adding cavity, a first micro-channel, a precipitation cavity, a second micro-channel, a first capillary channel, a third micro-channel and a quantitative detection cavity. The micro-fluidic chip is provided with a sample adding cavity, a precipitation cavity and a quantitative detection cavity, a sample to be detected can be added into the sample adding cavity through a sample adding hole, solid precipitates can be separated from liquid through centrifugal separation, a sample solution to be detected containing a target substance is obtained, the sample solution to be detected in the sample adding cavity and the first micro-channel and the second micro-channel can drive liquid to flow through the capillary force of the first capillary channel, and finally, siphonage and external centrifugation are formed and flow into the quantitative detection cavity to realize the quantification of the sample solution to be detected. The micro-fluidic chip has a relatively simple structure, is easy to manufacture and mold, and can be widely popularized and used.
Description
Technical Field
The invention relates to the technical field of in-vitro diagnosis, in particular to a micro-fluidic chip and an in-vitro detection device comprising the same.
Background
In Vitro Diagnosis (IVD) refers to a technique for diagnosing diseases by taking samples (blood, body fluid, tissue, etc.) from a human body and performing detection analysis, and corresponding instruments and reagents are required In the detection process, and the instruments and the reagents constitute an In Vitro Diagnosis system. In vitro diagnostic systems are broadly divided into two categories: one is represented by a detection center laboratory, has the characteristics of system modularization and automation, and is used for carrying out pipelined detection on a sample, so that the system also has the advantages of high flux, high efficiency and high sensitivity, but the whole system also has the defects of high cost, large occupied volume, requirement of professional operation and the like, and is mainly applied to large hospitals; in addition, the other method is represented by point-of-care testing (POCT), and the system has the characteristics of integration and miniaturization, can carry out sample testing at any time and any place, and further has the advantages of low price, simplicity in operation and timely result report. The microfluidic chip technology has the advantages of high functional integration, miniaturization and automation, so that the microfluidic chip technology is widely applied to the field of POCT. However, the conventional microfluidic chip for POCT generally has a complicated structure, so that the conventional microfluidic chip is relatively expensive, and further application of the conventional microfluidic chip in the POCT field is limited.
Disclosure of Invention
In view of the above, there is a need for a microfluidic chip with a relatively simple structure and capable of separating and quantifying a sample, and an in vitro detection apparatus including the microfluidic chip.
A micro-fluidic chip is provided with a sample adding cavity, a first micro-channel, a precipitation cavity, a second micro-channel, a first capillary channel, a third micro-channel and a quantitative detection cavity; the sample adding cavity is provided with a sample adding hole, the sample adding cavity is communicated with the precipitation cavity through the first micro-channel, two ends of the second micro-channel are respectively communicated with the first micro-channel and the first capillary channel, and the first capillary channel is communicated with the quantitative detection cavity through the third micro-channel;
the micro-fluidic chip is provided with a rotation center, the sample adding cavity is closer to the rotation center than the precipitation cavity, the first capillary channel is gradually extended from one end communicated with the second micro-channel to the direction close to the rotation center and is bent to be extended to the direction far away from the rotation center so as to be communicated with the third micro-channel, and the quantitative detection cavity is closer to the first capillary channel and is far away from the rotation center.
In one embodiment, the sample-adding cavity is provided with a first air vent.
In one embodiment, the sample-adding cavity partially surrounds the rotation center, and the width of the sample-adding cavity gradually increases from one end provided with the sample-adding hole to the other end, and the connection position of the first micro-channel and the sample-adding cavity is far away from the rotation center than the first air hole.
In one embodiment, the second microchannel extends from a connection position with the first microchannel toward a direction closer to the rotation center.
In one embodiment, the quantitative detection cavity is provided with a plurality of cavities, and the plurality of cavities are distributed at intervals along the third microchannel.
In one embodiment, the third microchannel is disposed around the center of rotation.
In one embodiment, the quantitative detection cavity is communicated with the third micro-channel through a fourth micro-channel, and an included angle of 30 to 90 degrees is formed between a downstream tangential direction of a connecting position of the third micro-channel and the fourth micro-channel.
In one embodiment, the end of the third micro flow channel is provided with a second air vent, and the second air vent is closer to the rotation center than the quantitative detection cavity.
In one embodiment, the microfluidic chip further has a first waste liquid cavity, the first waste liquid cavity is communicated with the third microchannel, and the first waste liquid cavity is closer to the end of the third microchannel than the quantitative detection cavity.
In one embodiment, the microfluidic chip further comprises a predetermined volume cavity, a second capillary channel, a fifth microchannel and a sixth microchannel;
one end of the second capillary channel is communicated with the second micro channel, the other end of the second capillary channel is communicated with the fifth micro channel, the preset amount cavity is communicated with one end of the first capillary channel through the sixth micro channel, and compared with the communication position of the fifth micro channel, the communication position of the preset amount cavity and the sixth micro channel is closer to the rotation center;
the second capillary channel gradually extends from one end communicated with the second micro-channel to the direction close to the rotation center, bends and then extends to the direction far away from the rotation center so as to be communicated with the fifth micro-channel; the predetermined amount cavity is far away from the rotation center than the second capillary flow channel;
the quantitative detection cavity and the third micro flow channel are far away from the rotation center than the preset amount cavity.
In one embodiment, the microfluidic chip further has a second waste liquid chamber, the second waste liquid chamber communicates with the fifth microchannel, and a position where the second waste liquid chamber communicates with the fifth microchannel is closer to an end of the fifth microchannel than a position where the predetermined amount chamber communicates with the fifth microchannel.
In one embodiment, the tail of the fifth microchannel extends gradually toward the center of rotation; and/or
And a third air hole is formed at the tail end of the fifth micro flow channel, and the third air hole is closer to the rotating center than the preset amount cavity.
In one embodiment, the width of the first capillary flow channel and/or the second capillary flow channel is 0.1mm to 0.2mm, and the depth is 0.1mm to 0.2 mm; or
The width of the first capillary flow channel and/or the second capillary flow channel is 0.2 mm-0.5 mm, the depth of the first capillary flow channel and/or the second capillary flow channel is 0.2 mm-0.5 mm, and the channel wall of the first capillary flow channel and/or the second capillary flow channel is subjected to surface treatment by PEG 4000.
In one embodiment, the microfluidic chip is disc-shaped, and a rotation mounting part is arranged in the middle of the microfluidic chip, and the center of the rotation mounting part is the rotation center.
In one embodiment, the microfluidic chip comprises a chip body and a transparent cover film covering the chip body, wherein the chip body and the transparent cover film are matched to form cavity structures and flow channel structures.
In one embodiment, the transparent cover film is a transparent pressure sensitive adhesive tape.
An in vitro detection device comprises the microfluidic chip and a detection unit in any embodiment, wherein the detection unit is used for detecting a sample in the quantitative detection cavity.
In one embodiment, the detection unit is a lyophilized reagent disposed in the quantitative detection cavity.
In one embodiment, the quantitative detection cavity is provided with a penetration hole, a water-soluble film covers the penetration hole, and the sample inlet of the detection unit is butted with the penetration hole and separated by the water-soluble film.
In one embodiment, the detection unit is a dry chemical strip.
In one embodiment, the dry chemical test paper comprises a support layer, and a reaction indicating layer and a diffusion layer which are sequentially stacked on the support layer, wherein the reaction indicating layer contains a reaction reagent and an indicating reagent which can react with a target substance in a sample to be tested, and the diffusion layer faces the water-soluble film through the sample inlet.
In one embodiment, the microfluidic chip is provided with a mounting groove around the permeation hole, and the detection unit is embedded in the mounting groove.
The micro-fluidic chip is provided with a sample adding cavity, a precipitation cavity and a quantitative detection cavity, a sample to be detected can be added into the sample adding cavity through the sample adding hole, solid precipitates can be separated from liquid through centrifugal separation, a sample solution to be detected containing a target substance is obtained, the sample solution to be detected in the sample adding cavity and the first micro-channel and the second micro-channel can drive liquid to flow through the capillary force of the first capillary channel, and finally, a siphoning effect is formed, and the sample solution to be detected can flow into the quantitative detection cavity by combining with the external centrifugal effect to realize the quantification of the sample solution to be detected. The micro-fluidic chip has a relatively simple structure, is easy to manufacture and mold, and can be widely popularized and used.
The in vitro detection device is provided with the detection unit, can directly carry out quantitative detection on quantitative sample solution to be detected in the quantitative detection cavity, and is simple to operate and high in detection efficiency.
Drawings
Fig. 1 is a schematic front structure diagram of a microfluidic chip according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a back side structure of the microfluidic chip shown in FIG. 1;
FIG. 3 is a side view of the microfluidic chip fit detection unit shown in FIG. 1;
FIGS. 4-1, 4-2, 4-3, 4-4, 4-5, 4-6 and 4-7 are schematic diagrams illustrating a process of separating and quantifying plasma (or serum) from a whole blood sample by the microfluidic chip shown in FIG. 1, and FIGS. 4-2-1, 4-3-1, 4-4-1, 4-5-1 and 4-6-1, and stage 4-7-1 are corresponding enlarged partial schematic diagrams;
FIGS. 5-1, 5-2 and 5-3 are schematic diagrams illustrating a process of dissolving a water-soluble film into plasma (or serum) of an in vitro test device according to an embodiment of the present invention;
FIG. 6 is a schematic diagram of a dry chemical indicator paper according to an embodiment.
The reference numerals are explained below:
10: a microfluidic chip; 11: a chip body; 12: a transparent cover film; 110: a sample adding cavity; 111: a sample application hole; 112: a first air vent; 120: a first microchannel; 130: a settling chamber; 140: a second microchannel; 150: a first capillary flow passage; 160: a third microchannel; 161: a second air hole; 170: a quantitative detection cavity; 171: a penetration hole; 172: a water-soluble film; 180: a fourth microchannel; 190: a first waste liquid chamber; 210: a predetermined amount of cavities; 220: a second capillary flow passage; 230: a fifth microchannel; 231: a third air hole; 240: a sixth microchannel; 250: a second waste liquid chamber; 260: a rotation mounting part; 18: a center of rotation; 270: mounting grooves; 220a, 220b, 220c are different points on the second capillary flow passage 220; 150a, 150b, and 150c are different points on the first capillary flow passage 150;
20: a detection unit; 21: a sample inlet; 22: a support layer; 23: a reaction indicating layer; 232: an indicator layer; 233: a reagent layer; 24: a diffusion layer.
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 "on" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" or "in communication with" another element, it can be directly connected or intervening elements may also be present.
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.
Referring to fig. 1 and fig. 2, an embodiment of the invention provides a microfluidic chip 10, which has a sample application cavity 110, a first microchannel 120, a precipitation cavity 130, a second microchannel 140, a first capillary channel 150, a third microchannel 160, and a quantitative detection cavity 170. The sample adding cavity 110 is used for adding a sample to be tested and is provided with a sample adding hole 111. The sedimentation chamber 130 is mainly used for collecting useless matters in the solid sample to be detected with high density, such as various cells in blood plasma. The sedimentation chamber 130 is in communication with the sample application chamber 110 via the first microchannel 120. The two ends of the second micro flow channel 140 are respectively connected to the first micro flow channel 120 and the first capillary flow channel 150, and the first capillary flow channel 150 is connected to the quantitative detection cavity 170 through the third micro flow channel 160. The quantitative detection cavity 170 is used for quantifying the sample solution to be detected.
The microfluidic chip 10 has a center of rotation 18. Sample application chamber 110 is closer to center of rotation 18 than is deposition chamber 130. The first capillary channel 150 extends from the end communicating with the second microchannel 140 to a direction gradually approaching the rotation center 18 (may be a direction gradually approaching the rotation center 18, and may be, for example, but not limited to, a radial direction) and bends to extend to a direction away from the rotation center 18 (may be a direction gradually departing from the rotation center 18, and may be, for example, but not limited to, a radial direction) to communicate with the third microchannel 160, and the entire first capillary channel 150 has a U-shape. The quantitative detection cavity 170 is farther from the rotation center 18 than the first capillary channel 150.
In the illustrated embodiment, the sample application cavity 110 is provided with a first vent 112. The first air hole 112 and the sample adding hole 111 are respectively located at two ends of the sample adding cavity 110. The first air vent 112 is used for timely exhausting air in the sample adding cavity 110 during sample adding, so that influence on sample adding due to increase of internal air pressure is avoided.
Further, the sample application chamber 110 is partially disposed around the center of rotation 18. Preferably, the width of the sample application cavity 110 gradually increases from the end having the sample application hole 111 to the other end, the connection position of the first microchannel 120 and the sample application cavity 110 is located at the end of the sample application cavity 110 opposite to the end where the sample application hole 111 is located, and the connection position is far away from the rotation center 18 than the first air vent 112, and preferably, the connection position of the first microchannel 120 and the sample application cavity 110 is located at the farthest position of the sample application cavity 110 from the rotation center. The side wall of the whole sample-adding cavity 110 is arc-shaped to facilitate the flow of the added sample to be tested (such as a whole blood sample).
The first microchannel 120 is required to have a size enough to allow the passage of unnecessary substances having a large density or size in the sample to be measured, and for example, in the case of a whole blood sample, when separating plasma or serum, the first microchannel 120 is required to allow the passage of blood cells in the whole blood.
The sedimentation chamber 130 is preferably funnel-shaped, and one end with a smaller size is connected to the first micro flow channel 120, so that the unwanted materials such as cells are not easy to flow out after entering the sedimentation chamber 130.
In the particular example illustrated, the second fluidic channel 140 is directly connected to the first fluidic channel 120. More specifically, the second microchannel 140 extends from the connection position with the first microchannel 120 gradually toward the direction near the rotation center 18 to connect with other channel structures.
Preferably, for example, in the illustrated specific example, there are a plurality of quantitative detection chambers 170, and the plurality of quantitative detection chambers 170 are distributed at intervals along the third micro flow channel 160. By arranging the plurality of quantitative detection cavities 170, repeated detection of one sample for multiple times or multi-item detection of one sample can be realized, the consistency is good, the integration level is high, and the flux of single detection can be obviously improved.
Further, a third microchannel 160 is disposed around the center of rotation 18. More preferably, the plurality of quantitative detection chambers 170 are equally spaced apart on the third microchannel 160. The dimensions of the quantitative detection chambers 170 may be uniform or non-uniform to facilitate repeated detection of the same item or multi-item, multi-factor (e.g., different sample amounts) detection of the same sample.
In the particular example illustrated, the quantitative detection chamber 170 is in communication with the third microchannel 160 via a fourth microchannel 180. Preferably, the downstream tangential direction of the third fluidic channel 160 at the connection location with the fourth fluidic channel 180 (the side of the tangent of the third fluidic channel 160 at the connection location downstream of the connection location) is at an angle of 30 ° to 90 ° with the fourth fluidic channel 180 (i.e., angle a in fig. 4-7-1). Further preferably, an included angle of less than 60 ° is formed between the downstream tangential direction of the connection position of the third microchannel 160 and the fourth microchannel 180, that is, along the liquid flow direction of the third microchannel 160, the whole fourth microchannel 180 is inclined towards the direction close to the end of the third microchannel 160, so that the sample solution can enter the quantitative detection cavity 170 more smoothly through the fourth microchannel 180.
Further, a second air hole 161 is formed at the end of the third micro flow channel 160. The second vent 161 is closer to the rotation center 18 than the quantitative detection chamber 170. Preferably, the tail of the third microchannel 160 is bent and extended toward the rotation center 18, and the second vent 161 is located at the end of the extended portion. Through the second air hole 161, the air in the third micro flow channel 160 can be discharged in time, and the influence of the air pressure increase caused by the air remaining in the third micro flow channel 160 on the sample solution entering the third micro flow channel 160 and further entering the quantitative detection cavity 170 is avoided.
Further, preferably, the microfluidic chip 10 further has a first waste liquid chamber 190. The first waste chamber 190 is in communication with the third microchannel 160, and the first waste chamber 190 is closer to the end of the third microchannel 160 than the quantitative detection chamber 170. The first waste liquid chamber 190 is farther from the rotation center 18 with respect to the second vent 161. The first waste liquid cavity 190 is located at the downstream of the plurality of quantitative detection cavities 170, so that it can be fully ensured that each quantitative detection cavity 170 is filled with the sample solution to be detected, and when the sample solution appears in the first waste liquid cavity 190, it can be described that each quantitative detection cavity 170 located at the upstream is inevitably filled with the sample solution to be detected, and therefore, by observing the liquid state in the first waste liquid cavity 190, the liquid containing condition of each quantitative detection cavity 170 can be judged.
In the illustrated preferred example, the microfluidic chip 10 further has a predetermined volume chamber 210, a second capillary flow channel 220, a fifth micro flow channel 230, and a sixth micro flow channel 240.
One end of the second capillary flow channel 220 communicates with the second microchannel 140, and the other end communicates with the fifth microchannel 230. The predetermined volume 210 is in communication with the fifth microchannel 230, so that the sample solution in the sample application chamber 110 and the second microchannel 140 can enter the predetermined volume 210 via the second capillary channel 220 and the fifth microchannel 230. The predetermined amount chamber 210 is used to perform a predetermined amount of the sample solution to be measured, and may further precipitate and separate solid useless materials and the like that may remain in the sample solution. The predetermined amount chamber 210 communicates with one end of the first capillary flow channel 150 through the sixth micro flow channel 240, and the predetermined amount chamber 210 communicates with the fifth micro flow channel 230 at a position closer to the rotation center 18 than the position where it communicates with the sixth micro flow channel 240. Preferably, the tail of the sixth fluidic channel 240 also extends in a curved manner toward the center of rotation 18 and terminates directly in an end of the first capillary channel 150.
The second capillary channel 220 gradually extends from the end communicating with the second microchannel 140 toward the rotation center 18, bends and extends away from the rotation center 18 to communicate with the fifth microchannel 230. The predetermined amount cavity 210 is farther from the rotation center 18 than the second capillary flow passage 220. The quantitative detection chamber 170 and the third microchannel 160 are further from the rotation center 18 than the predetermined volume 210.
Further preferably, the microfluidic chip 10 further has a second waste liquid chamber 250. The second waste chamber 250 communicates with the fifth microchannel 230 and the position at which the second waste chamber 250 communicates with the fifth microchannel 230 is closer to the end of the fifth microchannel 230 than the position at which the predetermined volume chamber 210 communicates with the fifth microchannel 230. The second waste chamber 250 is used to contain the excess sample solution that cannot be contained in the predetermined volume 210, and the second waste chamber 250 is located downstream of the predetermined volume 210, so that, like the first waste chamber 190, the second waste chamber 250 can be used to indicate whether the predetermined volume 210 is filled with the sample solution, and when the second waste chamber 250 contains the sample solution, the predetermined volume 210 is necessarily filled with the sample solution.
Further, in the illustrated specific example, the tail of the fifth microchannel 230 gradually extends toward the rotation center 18. The end of the fifth micro flow channel 230 is provided with a third air hole 231. The third airing hole 231 is closer to the rotation center 18 than the predetermined amount cavity. The third vent 231 serves to discharge air in time when the sample solution flows into the predetermined amount chamber 210, so that the sample solution enters the predetermined amount chamber 210 and the second waste liquid chamber 250.
The first capillary flow passage 150 and the second capillary flow passage 220 are both U-shaped, and the bent portions thereof are both close to the rotation center 18. In one specific example, the first capillary flow passage 150 and/or the second capillary flow passage 220 has a width of 0.1mm to 0.2mm and a depth of 0.1mm to 0.2 mm; or the width of the first capillary flow channel 150 and/or the second capillary flow channel 220 is 0.2mm to 0.5mm, and the depth is 0.2mm to 0.5 mm. When the width of the first capillary flow channel 150 and/or the second capillary flow channel 220 is 0.1mm to 0.2mm and the depth is 0.1mm to 0.2mm, the channel walls of the first capillary flow channel 150 and/or the second capillary flow channel 220 do not need to be surface-treated; when the width of the first capillary flow channel 150 and/or the second capillary flow channel 220 is 0.2mm to 0.5mm and the depth is 0.2mm to 0.5mm, the channel wall of the first capillary flow channel 150 and/or the second capillary flow channel 220 is preferably surface-treated with PEG 4000. Further preferably, the width of the first capillary flow passage 150 and/or the second capillary flow passage 220 is 0.2mm, and the depth is also 0.2 mm. The first capillary channel 150 and/or the second capillary channel 220 allow the sample solution to flow to the other end thereof by capillary action after the sample solution enters, and finally form a siphon action between the micro channels connected to the two ends thereof. The PEG4000 surface treatment can be, but is not limited to, adding 1% (g/v, i.e. adding 1g of PEG4000 to 100ml of water) of PEG4000 solution into the capillary channel, and naturally drying to form the PEG4000 surface treatment. The PEG4000 surface treatment is beneficial to increasing the capillary force of the capillary flow channel, and the PEG4000 belongs to an inert substance in a reaction system and generally does not react with a sample, a detection reagent and the like, so that the detection result is not influenced.
It is understood that, in other embodiments, the microfluidic chip 10 may not include the predetermined volume cavity 210 and the channel structure (including the second capillary channel 220, the fifth microchannel 230, and the sixth microchannel 240) attached thereto, and in this case, the second microchannel 140 may be directly connected to the first capillary channel 150.
In the illustrated embodiment, the microfluidic chip 10 has a disk shape, and the central portion of the microfluidic chip 10 has a rotation mounting portion 260. The center of the rotation mounting portion 260 is the rotation center 18. The rotary mounting portion 260 may be various types of slots or posts.
As shown in fig. 3, in a specific example, the microfluidic chip 10 includes a chip body 11 and a transparent cover film 12 covering the chip body 11. The chip body 11 and the transparent cover film 12 are matched to form cavity structures and flow passage structures. Specifically, the grooves of the cavity structures and the flow channel structures are preformed on the chip body 11, as shown in fig. 2, the holes may be opened on the back of the chip body 11, and the cavity structures and the flow channel structures may be encapsulated by covering and sealing the transparent cover film 12 on the front of the chip body 11, so as to form complete cavity structures and flow channel structures.
The transparent cover film 12 can be but is not limited to transparent pressure-sensitive adhesive and the like, is matched with the chip body 11 to form the whole microfluidic chip 10, is simple to assemble, does not need to use a complex and expensive ultrasonic welding technology, can be directly bonded, and can obviously reduce the manufacturing cost. It is understood that in other specific examples, the microfluidic chip 10 may be formed by welding using a costly ultrasonic welding technique or integrally formed by using a 3D printing technique.
The microfluidic chip is provided with a sample adding cavity 110, a precipitation cavity 130 and a quantitative detection cavity 170, a sample to be detected can be added into the sample adding cavity 110 through the sample adding hole 111, solid precipitates can be separated from liquid through centrifugal separation, a sample solution to be detected containing a target substance is obtained, the sample solution to be detected in the sample adding cavity 110 and the first micro flow channel 120 and the second micro flow channel 140 can drive liquid to flow through the capillary force of the first capillary flow channel 150, and finally, siphonage and external centrifugation are formed to flow into the quantitative detection cavity 170 so as to realize the quantification of the sample solution to be detected. The micro-fluidic chip has a relatively simple structure, is easy to manufacture and mold, and can be widely popularized and used.
Referring to fig. 2, fig. 5-1, fig. 5-2, and fig. 5-3, the present invention further provides an in vitro detection apparatus, which includes the above microfluidic chip 10 and the detection unit 20, wherein the detection unit 20 is used for detecting a sample in the quantitative detection cavity 170.
In one particular example, the detection unit 20 is a lyophilized reagent disposed within the quantitative detection cavity 170. The lyophilized reagent is placed in the quantitative detection cavity 170, and when the sample solution flows into the quantitative detection cavity 170, the lyophilized reagent can be dissolved and react with the sample solution, and the reaction result can be detected. The multiple quantitative detection chambers 170 may contain the same or different lyophilized reagents, so as to facilitate repeated detection of the same item or detection of different items.
Due to the complicated manufacturing process and high cost of the lyophilized reagent, in another specific example, the detecting unit 20 is externally disposed. Specifically, the quantitative detection chamber 170 has a penetration hole 171, and the penetration hole 171 is covered with a water-soluble film 172. The sample inlet 21 of the detection unit 20 is interfaced with the permeation pore 171 and separated by a water-soluble film 172.
Further, in one particular example, the detection unit 20 is a dry chemical strip. As shown in fig. 6, the dry chemical test paper 20 may include a support layer 22, and a reaction indicating layer 23 and a diffusion layer 24 sequentially stacked on the support layer 22. The reaction indicator layer 23 contains a reaction reagent and an indicator reagent that can react with a target substance in a sample to be measured. The reaction indicating layer 23 may be one layer or a plurality of layers, for example, in the specific example shown in fig. 6, the reaction indicating layer 23 includes two layers, i.e., an indicating layer 232 and a reagent layer 233, the indicating layer 232 is adjacent to the support layer 22 and contains a color-developing indicating reagent, and the reagent layer 233 is adjacent to the diffusion layer 24 and contains a reaction reagent capable of reacting with the target substance; further, the reagents contained in the indicator layer 232 and the reagent layer 233 may be exchanged or appropriately mixed. The diffusion layer 24 faces the water-soluble film 172 through the injection port 21.
Further, the microfluidic chip 10 is provided with a mounting groove 270 around the penetration hole 171. The external detection unit 20 is embedded in the mounting groove 270.
The in vitro detection device can directly detect quantitative sample solution to be detected in the quantitative detection cavity 170, and is simple to operate and high in detection efficiency.
Taking the whole blood sample loading detection as an example, the whole blood sample dry chemical detection process is mainly divided into four processes, namely, the whole blood is subjected to a plasma (serum) separation process; plasma (serum) quantification; plasma (serum) distribution and water-soluble membrane dissolution; plasma (serum) was reacted with dry chemical paper. Specifically, the detection process using the in vitro detection apparatus having the structure of the microfluidic chip shown in fig. 1 can be referred to as follows.
Referring to FIG. 4-1, a predetermined amount of whole blood is added to the sample addition chamber 110 through the sample addition hole 111. After the sample is added, the rotary mounting part 180 of the microfluidic chip 10 (hereinafter referred to as chip) is mounted in a matched rotary centrifugal instrument, the instrument is started to rotate, red blood cells and the like are deposited at the bottom of the precipitation cavity 130 under the action of centrifugal force, a small amount of partial plasma (serum) generally exists at the upper part of the precipitation cavity 130, meanwhile, the first micro-channel 120 and the second micro-channel 140 are full of plasma (serum), and plasma (serum) also exists at one side of the sample adding cavity 110, which is far away from the rotation center 18.
As shown in fig. 4-2, the plasma (serum) only partially fills the second capillary flow channel 220 during the rotation of the chip, and as shown in fig. 4-2-1, the plasma (serum) only can pass 220a, but not 220 b.
When the rotation is stopped, as shown in fig. 4-3 and 4-3-1, the second capillary flow path 220 passes the plasma (serum) over 220b by capillary force and stops after reaching 220c, and a siphon action is formed because the point of 220c is lower than the upper liquid level of the plasma (serum).
As shown in fig. 4-4 and 4-4-1, when the second capillary channel 220 is completely filled with plasma (serum), the control chip starts to rotate again. Due to the siphon action, the plasma (serum) enters the predetermined amount chamber 210 through the fifth micro flow channel 230. Since the third ventilation hole 231 is closer to the rotation center 18 than the second capillary flow path 220, the plasma (serum) does not flow out from the second capillary flow path 220. Under centrifugal force and siphon action, the plasma (serum) enters the predetermined amount chamber 210, completing a predetermined amount process of the plasma (serum).
In the particular example shown, excess plasma (serum) enters the second waste chamber 250, the instrument will detect the presence of liquid in the second waste chamber 250, and if the instrument detects the presence of liquid in the second waste chamber 250, the predetermined volume 210 is filled with plasma (serum); if the presence of liquid is not detected in the second waste chamber 250, indicating that the predetermined volume 210 is likely not filled with liquid, or that the capillary is blocked, etc., in which case the instrument indicates an error and stops further testing. The down test process will continue only when the second waste chamber 250 has liquid present. It is understood that, in other specific examples, whether the predetermined volume 210 is filled with liquid can be determined according to whether liquid remains in the fifth micro flow channel 230 downstream of the predetermined volume 210, which is substantially the same as the principle according to whether liquid remains in the second waste volume 250.
As shown in fig. 4-5 and 4-5-1, during the rotation of the chip, a part of the liquid exists in the first capillary channel 150, and similarly, the liquid passes through 150a but does not pass through 150 b.
As shown in fig. 4-6 and 4-6-1, when the rotation is stopped, the liquid in the first capillary channel 150 will cross 150b under the capillary action, and will stop after reaching 150c, and will fill the whole first capillary channel 150, so as to form the siphon action. When the chip rotates again, under the action of centrifugal force and siphon, the plasma (serum) liquid enters the third micro flow channel 160, and since the second air vent 161 is higher than the upper liquid level of the quantitative detection cavity 170, the liquid does not flow out from the second air vent 161, and the liquid is convenient to flow into the third micro flow channel 160, and air in the quantitative detection cavity 170 is discharged.
As shown in fig. 4-7 and 4-7-1, the fourth micro flow channel 180 between the third micro flow channel 160 and the quantitative detection cavity 170 is preferably not perpendicular to the third micro flow channel 160, but is inclined downstream along the flow direction of the liquid, and the inclined angle a is preferably less than 60 °, so that the liquid can enter the quantitative detection cavity 170 in a shorter time, and bubbles can be sufficiently discharged.
When the plasma (serum) completely fills all of the quantitative detection chambers 170, as shown in fig. 4-7, the excess fluid enters the first waste chamber 190. When the matched instrument detects that the first waste liquid cavity 190 is filled with liquid, the quantitative detection cavities 170 are all filled, and the instrument can smoothly perform the next step; the volume of the quantitative detection cavity 170 is the sample loading amount required by the sample, so that the volume of the quantitative detection cavity 170 can play a role in quantification.
The time required for the plasma (serum) liquid to be dispensed from the predetermined volume 210 into each of the quantitative detection chambers 170 is short, typically within 1 min. When all the quantitative determination chamber 170 is filled with the liquid, the control chip stops rotating, as shown in fig. 5-1, the penetration hole 171 is sealed by the water-soluble film 172, so that the plasma (serum) liquid is temporarily sealed in the quantitative determination chamber 170, and after standing for 1min, the water-soluble film 172 is dissolved by the plasma (serum), as shown in fig. 5-2.
As shown in fig. 5-2, after the water-soluble film 172 is dissolved by the plasma (serum) liquid, the liquid does not flow into the detection unit (dry chemical test strip) 20 by gravity. At this time, the chip can be controlled to rotate at the low speed of 1800-;
the sample solution is diffused to the reaction indication layer 23 through the diffusion layer 24 in sequence to carry out color reaction, the concentration of the detection object can be reflected by the depth of color development, signal acquisition can be carried out through the detection hole 25 of the detection unit, and finally the concentration data of the detection object is converted. The in vitro detection device can solve the problems of low sensitivity and stability of the dry chemical test paper whole blood, and has the advantages of high detection flux, low cost and the like.
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 (22)
1. A micro-fluidic chip is characterized in that the micro-fluidic chip is provided with a sample adding cavity, a first micro-channel, a precipitation cavity, a second micro-channel, a first capillary channel, a third micro-channel and a quantitative detection cavity; the sample adding cavity is provided with a sample adding hole, the sample adding cavity is communicated with the precipitation cavity through the first micro-channel, two ends of the second micro-channel are respectively communicated with the first micro-channel and the first capillary channel, and the first capillary channel is communicated with the quantitative detection cavity through the third micro-channel;
the micro-fluidic chip is provided with a rotation center, the sample adding cavity is closer to the rotation center than the precipitation cavity, the first capillary channel is gradually extended from one end communicated with the second micro-channel to the direction close to the rotation center and is bent to be extended to the direction far away from the rotation center so as to be communicated with the third micro-channel, and the quantitative detection cavity is closer to the first capillary channel and is far away from the rotation center.
2. The microfluidic chip according to claim 1, wherein the sample application chamber is provided with a first vent.
3. The microfluidic chip according to claim 2, wherein the sample loading cavity is partially disposed around the rotation center, and has a width gradually increasing from one end having the sample loading hole to the other end, and the connection position of the first microchannel and the sample loading cavity is farther from the rotation center than the first vent.
4. The microfluidic chip according to claim 1, wherein the second microchannel extends from a connection position with the first microchannel in a direction gradually closer to the rotation center.
5. The microfluidic chip according to claim 1, wherein the plurality of quantitative detection chambers are spaced apart along the third microchannel.
6. The microfluidic chip of claim 5, wherein the third microchannel is disposed around the center of rotation.
7. The microfluidic chip according to claim 6, wherein the quantitative detection cavity is connected to the third microchannel through a fourth microchannel, and a downstream tangential direction at a connection position of the third microchannel and the fourth microchannel forms an included angle of 30 ° to 90 ° with the fourth microchannel.
8. The microfluidic chip according to claim 1, wherein the end of the third microchannel has a second vent, and the second vent is closer to the rotation center than the quantitative detection cavity.
9. The microfluidic chip according to claim 1, wherein the microfluidic chip further comprises a first waste chamber, the first waste chamber is in communication with the third microchannel, and the first waste chamber is closer to an end of the third microchannel than the quantitative detection chamber.
10. The microfluidic chip according to any one of claims 1 to 9, wherein the microfluidic chip further comprises a predetermined volume chamber, a second capillary channel, a fifth microchannel, and a sixth microchannel;
one end of the second capillary channel is communicated with the second micro channel, the other end of the second capillary channel is communicated with the fifth micro channel, the preset amount cavity is communicated with one end of the first capillary channel through the sixth micro channel, and compared with the communication position of the fifth micro channel, the communication position of the preset amount cavity and the sixth micro channel is closer to the rotation center;
the second capillary channel gradually extends from one end communicated with the second micro-channel to the direction close to the rotation center, bends and then extends to the direction far away from the rotation center so as to be communicated with the fifth micro-channel; the predetermined amount cavity is far away from the rotation center than the second capillary flow channel;
the quantitative detection cavity and the third micro flow channel are far away from the rotation center than the preset amount cavity.
11. The microfluidic chip according to claim 10, wherein the microfluidic chip further comprises a second waste chamber, wherein the second waste chamber is in communication with the fifth microchannel, and wherein the position where the second waste chamber is in communication with the fifth microchannel is closer to the end of the fifth microchannel than the position where the predetermined-amount chamber is in communication with the fifth microchannel.
12. The microfluidic chip according to claim 10, wherein the tail of the fifth microchannel extends gradually toward the center of rotation; and/or
And a third air hole is formed at the tail end of the fifth micro flow channel, and the third air hole is closer to the rotating center than the preset amount cavity.
13. The microfluidic chip according to claim 10, wherein the first capillary flow channel and/or the second capillary flow channel has a width of 0.1mm to 0.2mm and a depth of 0.1mm to 0.2 mm; or
The width of the first capillary flow channel and/or the second capillary flow channel is 0.2 mm-0.5 mm, the depth of the first capillary flow channel and/or the second capillary flow channel is 0.2 mm-0.5 mm, and the channel wall of the first capillary flow channel and/or the second capillary flow channel is subjected to surface treatment by PEG 4000.
14. The microfluidic chip according to any one of claims 1 to 9 and 11 to 13, wherein the microfluidic chip has a disk shape, and a rotational mounting portion is provided in a central portion of the microfluidic chip, and a center of the rotational mounting portion is the rotational center.
15. The microfluidic chip according to any one of claims 1 to 9 and 11 to 13, wherein the microfluidic chip comprises a chip body and a transparent cover film covering the chip body, and the chip body and the transparent cover film cooperate to form each cavity structure and each flow channel structure.
16. The microfluidic chip of claim 15, wherein the transparent cover film is a transparent pressure sensitive adhesive tape.
17. An in vitro detection device, comprising the microfluidic chip according to any one of claims 1 to 16 and a detection unit for detecting a sample in the quantitative detection cavity.
18. The in vitro assay device of claim 17, wherein the assay unit is a lyophilized reagent disposed within the quantitative detection chamber.
19. The in-vitro detection device according to claim 17, wherein the quantitative detection cavity has a penetration hole covered with a water-soluble film, and the sample inlet of the detection unit is butted with the penetration hole and separated by the water-soluble film.
20. The in vitro test device of claim 19, wherein the test element is a dry chemical strip.
21. The in-vitro detection device according to claim 20, wherein the dry chemical test paper comprises a support layer, and a reaction indication layer and a diffusion layer which are sequentially stacked on the support layer, the reaction indication layer contains a reaction reagent and an indication reagent which can react with a target substance in a sample to be detected, and the diffusion layer faces the water-soluble film through the sample inlet.
22. The in vitro detection device according to any one of claims 19 to 21, wherein the microfluidic chip is provided with a mounting groove around the permeation hole, and the detection unit is embedded in the mounting groove.
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| PCT/CN2019/122796 WO2021036074A1 (en) | 2019-08-28 | 2019-12-03 | Microfluidic chip, and in vitro test device including same |
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