CN110045102B - Reagent sequential loading device, centrifugal microfluidic device and analysis system - Google Patents
Reagent sequential loading device, centrifugal microfluidic device and analysis system Download PDFInfo
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- CN110045102B CN110045102B CN201910275523.XA CN201910275523A CN110045102B CN 110045102 B CN110045102 B CN 110045102B CN 201910275523 A CN201910275523 A CN 201910275523A CN 110045102 B CN110045102 B CN 110045102B
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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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
Abstract
The application relates to a reagent sequential loading device, a centrifugal microfluidic device and an analysis system, wherein the reagent sequential loading device comprises a liquid outlet pipeline, a hydraulic control chamber, a collection chamber and at least two loading chambers; the liquid outlet pipeline is communicated with the hydraulic control chamber and the collecting chamber and is provided with a resistance increasing structure; the loading cavity is provided with a hydraulic control pipeline and a loading pipeline; the collecting cavity is provided with a collecting cavity hole, the hydraulic control cavity is provided with a liquid injection hole, and the distances between the communication ends and the liquid injection hole are different; the distances from the rotation center are arranged from small to large: the loading chamber is communicated with the hydraulic control pipeline, and the loading chamber is communicated with the loading pipeline. The method is simple to implement, the design difficulty is reduced, and the processing cost is saved; the time interval for loading different reagents can be controlled by controlling the centrifugal rotating speed, and the time is reserved for reagent reaction or reagent treatment; sequential loading of any number of reagents can be achieved by adjusting the different reagents in the different loading chambers.
Description
Technical Field
The present application relates to the field of centrifugal microfluidics, and in particular to a reagent sequential loading device, a centrifugal microfluidic device and an analysis system.
Background
Microfluidics (Microfluidics) refers to the manipulation of liquids on a sub-millimeter scale, which is typically several microns to several hundred microns. The microfluidic technology integrates the basic operation units related to the biological and chemical fields, even the functions of the whole laboratory, including sampling, dilution, reaction, separation, detection, etc., on a small Chip, so it is also called Lab-on-a-Chip. The chip generally comprises various liquid storage tanks and a micro-channel network which is connected with each other, can greatly shorten the sample processing time, and realizes the maximum utilization efficiency of reagent consumables by precisely controlling the liquid flow. The micro-fluidic provides a very wide prospect for the application in numerous fields such as biomedical research, drug synthesis screening, environmental monitoring and protection, health quarantine, judicial identification, biological reagent detection and the like. Microfluidic systems refer to devices that manipulate liquids on a sub-millimeter scale. In the industry, microfluidics is generally classified into the following types: pressure (pneumatic or hydraulic) driven microfluidics, centrifugal microfluidics, droplet microfluidics, digital microfluidics, paper microfluidics, and the like. Centrifugal microfluidics belongs to a branch of microfluidics, and particularly relates to a method for driving liquid to flow by rotating a centrifugal microfluidic chip, so that the liquid is controlled on a sub-millimeter scale by using centrifugal force. Centrifugal microfluidics integrates basic operational units involved in biological and chemical fields into a small disc-shaped (disc-shaped) chip. In addition to the advantages specific to microfluidics, the overall device is more compact since only one motor is required for centrifugal microfluidics to provide the force required for liquid manipulation. And the ubiquitous centrifugal field on the disc chip can not only make liquid drive more effective and ensure that no liquid remains in the pipeline, but also can effectively realize sample separation based on density difference and make parallel processing simpler. Microfluidics can well meet the demand of Point-of-care testing (POCT) miniaturized instruments, and therefore, centrifugal microfluidics is also increasingly applied to Point-of-care testing (POCT).
When the microfluidic is applied to the field of in vitro diagnosis, the very important and key operation is to make reagents react according to a certain sequence, and finally obtain a diagnosis result. In order to realize the reaction of a plurality of reagents according to a certain sequence, the precondition is to realize the loading of the plurality of reagents into the appointed reaction chamber according to a certain sequence. In centrifugal microfluidics, sequential loading of reagents is mainly achieved by means of valves, including capillary valves, siphon valves, paraffin valves, etc. When the micro-fluidic chip rotates according to the set rotating speed time sequence, different reagents can break through different valves in sequence, and therefore sequential loading of the reagents is achieved. However, the use of valves tends to increase the processing cost of the entire microfluidic chip, and the sequential loading achieved with valves tends to be difficult to ensure with repeatability and reliability. Also, in centrifugal microfluidics, these valves are not easy to implement. The capillary valve has high requirements on the processing precision of the pipeline, the capillary valve is related to the contact angle of the liquid reagent on the surface of the material, and different reagents need different pipeline sizes to realize the effect of the capillary valve; the siphon valve needs to perform hydrophilic treatment on the siphon pipeline, and the process requirement of the treatment is high, so that the processing cost of the chip is often greatly increased; the melting of paraffin in the paraffin valve usually needs the instrument to carry out corresponding temperature control, and the design difficulty of the instrument is increased.
Disclosure of Invention
In view of the above, there is a need for a sequential reagent loading device, a centrifugal microfluidic device, and an analytical system.
A reagent sequential loading device is provided with a rotation center and comprises a liquid outlet pipeline, a hydraulic control chamber, a collection chamber and at least two loading chambers; the liquid inlet end of the liquid outlet pipeline is communicated with the hydraulic control chamber, the liquid outlet end of the liquid outlet pipeline is communicated with the collecting chamber, and the liquid outlet pipeline is provided with a resistance increasing structure; the reagent sequential loading device is provided with a hydraulic control pipeline and a loading pipeline in each loading chamber respectively and correspondingly; the collection cavity is equipped with collects the accent, the liquid accuse cavity is equipped with annotates the liquid hole, annotate the liquid hole with the feed liquor end of liquid outlet pipe way is located respectively the both ends of liquid accuse cavity, and with the distance of rotation center is arranged according to order from little to big: the liquid injection hole, the liquid inlet end of the liquid outlet pipeline and the liquid outlet end of the liquid outlet pipeline are arranged; each loading chamber is communicated with the hydraulic control chamber through the corresponding hydraulic control pipeline, each hydraulic control pipeline is provided with a communication end which is communicated with the hydraulic control chamber and is positioned between the liquid inlet end of the liquid outlet pipeline and the liquid injection hole, and the distances between the communication ends and the liquid injection hole are different; each loading chamber is also communicated with the collection chamber through the corresponding loading pipeline, and the distances between the loading chambers and the rotation center are arranged from small to large in sequence: the loading chamber is communicated with the hydraulic control pipeline, and the loading chamber is communicated with the loading pipeline.
The reagent sequential loading device is suitable for a centrifugal microfluidic device, is ingenious in design, can realize sequential loading of various reagents only in a centrifugal rotation environment without using various valves, is simple to realize, reduces the design difficulty and saves the processing cost; particularly, the time interval of loading different reagents can be controlled by controlling the centrifugal rotating speed, and the time is reserved for reagent reaction or reagent treatment; further, sequential loading of any number of reagents can be achieved by adjusting the different reagents in the different loading chambers.
In one embodiment, the liquid outlet pipeline is provided with a resistance increasing structure, and the resistance increasing structure is used for matching with a rotating speed setting so as to control the passing speed of the liquid outlet pipeline during rotation.
In one embodiment, the resistance-increasing structure comprises at least two communicated bends.
In one embodiment, the resistance increasing structure comprises a through area which is arranged in a reduced mode.
In one embodiment, the reagent sequential loading device has a hydrophobic material body, and the liquid outlet pipeline, the liquid control chamber, the collection chamber and each loading chamber are all opened in the hydrophobic material body.
In one embodiment, the reagent sequential loading device includes three loading chambers, the three loading chambers are respectively a first reagent loading chamber, a second reagent loading chamber and a third reagent loading chamber, and the distances from the liquid injection hole are arranged in the order from small to large: a communication end of the first reagent loading chamber, a communication end of the second reagent loading chamber, and a communication end of the third reagent loading chamber.
In one embodiment, the collection chamber comprises a collection cavity, an acquisition cavity and a waste liquid cavity, and a connecting line of the center of the collection cavity and the rotation center is taken as a reference line, and the acquisition cavity and the waste liquid cavity are respectively positioned on two sides of the reference line; each loading chamber is communicated with the collecting cavity through the corresponding loading pipeline, and the liquid outlet end of the liquid outlet pipeline is communicated with the waste liquid cavity; the collecting cavity is communicated with the acquiring cavity through an acquiring pipeline and is also communicated with the waste liquid cavity through a waste liquid pipeline; the acquisition cavity is provided with an acquisition cavity hole, and the acquisition cavity hole is positioned at the position of the acquisition cavity close to the rotation center and is arranged at an interval with the acquisition pipeline; the waste liquid cavity is provided with a waste liquid cavity hole, and the waste liquid cavity hole is located at the position, closest to the hydraulic control cavity, of the waste liquid cavity and is arranged at an interval with the liquid outlet end of the liquid outlet pipeline.
In one embodiment, the collection cavity is provided with an outlet, and the collection cavity is communicated with the acquisition cavity and the waste liquid cavity through the outlet respectively; and the outlet is provided with a filter membrane.
A centrifugal microfluidic device comprising any one of the reagent sequential loading devices.
The centrifugal microfluidic device is very ingenious in design, various valves are not needed, and multiple reagents can be loaded sequentially only in a centrifugal rotation environment, so that the centrifugal microfluidic device is simple to realize, the design difficulty of sequential reagent loading is reduced, and the processing cost is saved; particularly, the time interval of loading different reagents can be controlled by controlling the centrifugal rotating speed, so that the time for the reaction of the reagents to occur is reserved; further, sequential loading of any number of reagents can be achieved by adjusting the different reagents in the different loading chambers.
An analytical system comprising the centrifugal microfluidic device.
Drawings
FIG. 1 is a schematic structural diagram of an embodiment of the present application; FIG. 2 is an enlarged schematic view of the embodiment of FIG. 1 at A; FIG. 3 is another schematic view of the embodiment of FIG. 1; FIG. 4 is a schematic view of another direction of the embodiment shown in FIG. 1; FIG. 5 is another schematic view of the embodiment of FIG. 1; FIG. 6 is an enlarged view of the embodiment of FIG. 5 at B; FIG. 7 is another schematic directional diagram of the embodiment of FIG. 1; FIG. 8 is an enlarged schematic view of the embodiment of FIG. 7 at C; FIG. 9 is a schematic structural diagram of another embodiment of the present application; FIG. 10 is a schematic view of another alternative embodiment of the embodiment of FIG. 9; FIG. 11 is another schematic directional diagram of the embodiment of FIG. 9; FIG. 12 is another schematic directional view of the embodiment of FIG. 9; FIG. 13 is an enlarged schematic view of the embodiment of FIG. 12 at D; FIG. 14 is an enlarged schematic view at F of the embodiment shown in FIG. 13; FIG. 15 is an enlarged schematic view at E of the embodiment shown in FIG. 12; FIG. 16 is another schematic directional view of the embodiment of FIG. 9; FIG. 17 is an enlarged schematic view at G of the embodiment of FIG. 16; fig. 18 is an enlarged view at H of the embodiment shown in fig. 16.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present application more comprehensible, embodiments accompanying the present application are described in detail below with reference to the accompanying drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. This application is capable of embodiments in many different forms than those described herein and that modifications may be made by one skilled in the art without departing from the spirit and scope of the application and it is therefore not intended to be limited to the specific embodiments disclosed below. In one embodiment of the present application, a sequential reagent loading device has a rotation center, and the sequential reagent loading device includes a liquid outlet pipe, a hydraulic control chamber, a collection chamber and at least two loading chambers; the liquid inlet end of the liquid outlet pipeline is communicated with the hydraulic control chamber, and the liquid outlet end of the liquid outlet pipeline is communicated with the collection chamber; the reagent sequential loading device is provided with a hydraulic control pipeline and a loading pipeline in each loading chamber respectively; the collection cavity is equipped with collects the accent, and the liquid accuse cavity is equipped with annotates the liquid hole, annotates the liquid hole and holds the both ends that are located the liquid accuse cavity respectively with the feed liquor that goes out the liquid pipeline, and arranges according to from little to big order with rotation center's distance: the liquid inlet end of the liquid outlet pipeline is connected with the liquid inlet hole; each loading chamber is communicated with the hydraulic control chamber through the corresponding hydraulic control pipeline, each hydraulic control pipeline is provided with a communication end which is communicated with the hydraulic control chamber and is positioned between the liquid inlet end of the liquid outlet pipeline and the liquid injection hole, and the distances between each communication end and the liquid injection hole are different; each loading chamber is also communicated with the collecting chambers through the corresponding loading pipelines, and the distances between the loading chambers and the rotating center are arranged from small to large in sequence: the loading chamber is communicated with the hydraulic control pipeline, and the loading chamber is communicated with the loading pipeline. The reagent sequential loading device is suitable for a centrifugal microfluidic device, is ingenious in design, can realize sequential loading of various reagents only in a centrifugal rotation environment without using various valves, is simple to realize, reduces the design difficulty and saves the processing cost; particularly, the time interval of loading different reagents can be controlled by controlling the centrifugal rotating speed, and the time is reserved for reagent reaction or reagent treatment; further, sequential loading of any number of reagents can be achieved by adjusting the different reagents in the different loading chambers. In one embodiment, the reagent sequence loading device comprises a part of or the whole structure of the following embodiments; namely, the reagent sequence loading device comprises the following technical characteristics in part or all. In one embodiment, as shown in fig. 1, a reagent sequential loading device for placement in a centrifugal microfluidic device having a center of rotation, i.e., a reagent sequential loading device having a center of rotation 999 or rotatable about the center of rotation 999. In one of the embodiments, the reagent sequential loading device has a cylindrical body 100, and in one of the embodiments, the reagent sequential loading device has a cylindrical body of a flat shape. In one embodiment, the reagent sequential loading device has a rectangular body or a centrally symmetric structural body such as a regular polygonal body. In one embodiment, the reagent sequential loading device comprises a liquid outlet pipeline, a hydraulic control chamber, a collection chamber and at least two loading chambers, wherein the hydraulic control chamber is used for realizing reagent sequential loading control by filling liquid and liquid level positions of the liquid during rotation, the collection chamber is used for collecting the liquid in the hydraulic control chamber through the liquid outlet pipeline, the loading chambers are used for accommodating reagents to be loaded, different loading chambers are used for accommodating different reagents, and in each embodiment, the reagents are liquid; in one embodiment, as shown in fig. 1, 3 and 5, the sequential reagent loading device includes a liquid outlet pipe 220, a hydraulic control chamber 200, a collection chamber 600 and three loading chambers, in one embodiment, the three loading chambers are a first reagent loading chamber 300, a second reagent loading chamber 400 and a third reagent loading chamber 500 respectively according to the reagent loading sequence; and the distances from the liquid injection hole 210 are arranged from small to large: a communication end of the first reagent loading chamber, a communication end of the second reagent loading chamber, and a communication end of the third reagent loading chamber; as shown in fig. 1-3 and 5-8, the distance between the communication end 315 of the first reagent loading chamber 300 and the injection hole 210 is smaller than the distance between the communication end 415 of the second reagent loading chamber 400 and the injection hole 210, and the distance between the communication end 415 of the second reagent loading chamber 400 and the injection hole 210 is smaller than the distance between the communication end 515 of the third reagent loading chamber 500 and the injection hole 210, so as to ensure the reagent loading sequence, the first reagent, such as the sample, in the first reagent loading chamber, the second reagent, such as the cleaning solution, in the second reagent loading chamber, and the third reagent, such as the cleaning solution, in the third reagent loading chamber are added first. It can be understood that when the reagent loading sequence needs to be adjusted, only the reagent position in the loading chamber needs to be adjusted, or the loading chamber needs to be adjusted, and the application is simple and convenient.
In one embodiment, the reagent sequential loading device has a hydrophobic material body, and the liquid outlet pipe 220, the liquid control chamber 200, the collection chamber 600 and each loading chamber are all opened in the hydrophobic material body. In this embodiment, the cylindrical body 100 is a body of hydrophobic material. In one embodiment, the reagent sequential loading device has a hydrophobic plastic body, that is, the cylindrical body 100 is a hydrophobic plastic body, and the liquid outlet pipe 220, the hydraulic control chamber 200, the collection chamber 600 and each loading chamber are all opened in the hydrophobic plastic body; the hydrophobic plastic body includes a body structure of a hydrophobic plastic, or a body structure of a plastic subjected to surface hydrophobization treatment. Further, in one embodiment, the hydrophobic material body or the hydrophobic plastic body is a PMMA (polymethyl methacrylate), PDMS (polydimethylsiloxane), PC (Polycarbonate), ABS (Acrylonitrile-Butadiene-Styrene copolymer), COC (copolymers of Cyclo Olefin copolymer) or COP (Cyclo Olefin Polymer) part. The design of the hydrophobic material body such as the hydrophobic plastic body is beneficial to enhancing the hydrophobicity of the liquid outlet pipeline and the hydraulic control pipeline, so that liquid, especially water, in the hydraulic control cavity is prevented from entering or smoothly entering the liquid outlet pipeline and/or the hydraulic control pipeline, the speed of the liquid in the hydraulic control cavity entering the collection cavity or the waste liquid cavity is controlled on one hand, the time of sample adding reaction or sample adding treatment is controlled, and on the other hand, the liquid in the hydraulic control cavity is prevented from entering the loading cavity through the hydraulic control pipeline. The hydrophobic plastic body is a resistance-increasing structure.
In one embodiment, referring to fig. 6, the liquid inlet 221 of the liquid outlet pipe 220 is connected to the liquid control chamber 200, and the liquid outlet 222 of the liquid outlet pipe is connected to the collection chamber 600; referring to fig. 9, the area of the inlet end 221 of the outlet pipe 220 is larger than the area of any one of the first, second and third communication ends 315, 415 and 515. By the design, the liquid in the hydraulic control chamber 200 can flow out from the liquid inlet end 221 of the liquid outlet pipe 220.
In one embodiment, the reagent sequential loading device is provided with a hydraulic control pipeline and a loading pipeline in each loading chamber respectively and correspondingly; as shown in fig. 1, the reagent sequential loading apparatus is provided with a first hydraulic control pipeline 310 and a first loading pipeline 320 communicating with the first reagent loading chamber 300 in the first reagent loading chamber 300, a second hydraulic control pipeline 410 and a second loading pipeline 420 communicating with the second reagent loading chamber 400 in the reagent sequential loading apparatus, and a third hydraulic control pipeline 510 and a third loading pipeline 520 communicating with the third reagent loading chamber 500 in the reagent sequential loading apparatus. In one embodiment, each loading chamber is communicated with the hydraulic control chamber through its corresponding hydraulic control pipeline, and each hydraulic control pipeline has a communication end which is communicated with the hydraulic control chamber 200 and is located between the liquid inlet end 221 of the liquid outlet pipeline 220 and the liquid injection hole 210, and the distance between each communication end and the liquid injection hole 210 is different; as shown in fig. 1 to 3 and 5 to 8, the first reagent loading chamber 300 is communicated with the pilot-controlled chamber 200 through the first pilot-controlled conduit 310, and is communicated with the collection chamber 600 through the first loading conduit 320; the first pilot-controlled pipeline 310 has a first connection pipe 311, a first second connection pipe 312, a first third connection pipe 313 and a first fourth connection pipe 314 which are sequentially communicated, and the end of the first connection pipe 311 has a first communication end 315, the first communication end 315 is communicated with the pilot-controlled chamber 200, and the first fourth connection pipe 314 is communicated with the first reagent loading chamber 300; the second reagent loading chamber 400 is communicated with the pilot-controlled chamber 200 through a second pilot-controlled conduit 410, and is communicated with the collection chamber 600 through a second loading conduit 420; the second hydraulic control pipeline 410 has a second connection pipe 411, a second connection pipe 412, a second third connection pipe 414 and a second fourth connection pipe 414 which are sequentially communicated, and a second communication end 415 is provided at an end of the second connection pipe 411, the second communication end 415 is communicated with the hydraulic control chamber 200, and the second fourth connection pipe 414 is communicated with the second reagent loading chamber 400; the third reagent loading chamber 500 is communicated with the hydraulic control chamber 200 through a third hydraulic control pipeline 510, and is communicated with the collection chamber 600 through a third loading pipeline 520; the third pilot-controlled pipeline 510 has a third connection pipe 511, a third connection pipe 512, a third connection pipe 513 and a third fourth connection pipe 514, which are sequentially connected, and a third connection end 515 is provided at an end of the third connection pipe 511, the third connection end 515 is connected to the pilot-controlled chamber 200, and the third fourth connection pipe 514 is connected to the third reagent loading chamber 500.
Referring to fig. 6, the first hydraulic control pipeline 310 is communicated with the hydraulic control chamber 200 and has a first communication end 315, the second hydraulic control pipeline 410 is communicated with the hydraulic control chamber 200 and has a second communication end 415, the third hydraulic control pipeline 510 is communicated with the hydraulic control chamber 200 and has a third communication end 515, and the first communication end 315, the second communication end 415 and the third communication end 515 are arranged at different distances from the injection hole 210, so that when the reagent sequence loading device rotates, the liquid level of the hydraulic control chamber 200 slowly descends from the injection hole 210 to the liquid inlet 221 of the liquid outlet pipeline 220, and then descends to the first communication end 315, the second communication end 415 and the third communication end 515 in sequence; when the liquid level in the pilot-controlled chamber 200 drops below the first communication end 315, the first pilot-controlled conduit 310 communicates with the external environment through the first communication end 315, the pilot-controlled chamber 200 and the injection hole 210, and under the combined action of the atmospheric pressure and the centrifugal force, the first reagent in the first reagent loading chamber 300 enters the collection chamber 600 from the first loading conduit 320; when the liquid level in the pilot-controlled chamber 200 drops below the second communication end 415, the second pilot-controlled pipeline 410 is communicated with the external environment through the second communication end 415, the pilot-controlled chamber 200 and the liquid injection hole 210, and under the combined action of atmospheric pressure and centrifugal force, the second reagent in the second reagent loading chamber 400 enters the collection chamber 600 from the second loading pipeline 420; when the liquid level in the hydraulic control chamber 200 drops below the third communication end 515, the third hydraulic control pipeline 510 communicates with the external environment through the third communication end 515, the hydraulic control chamber 200 and the liquid injection hole 210, and under the combined action of atmospheric pressure and centrifugal force, the third reagent in the third reagent loading chamber 500 enters the collection chamber 600 from the third loading pipeline 520; this achieves the sequential loading of reagents without valve control or so-called water-lock control, and it is understood that the present embodiment is exemplified by three loading chambers, and in practical applications, the number of loading chambers may be 2, 4, 5, 6 or more, and the number of loading chambers in the present embodiment should not be construed as limiting the scope of the present application.
In one embodiment, referring to fig. 4, the collection chamber 600 has a collection chamber hole 610, and the pilot-controlled chamber 200 has a liquid injection hole 210, in one embodiment, the relative positions of the collection chamber hole 610 and the liquid injection hole 210 are set according to the rotation direction of the reagent sequence loading device, or the rotation direction of the reagent sequence loading device determines the relative positions of the collection chamber hole 610 and the liquid injection hole 210, as shown in fig. 1, the reagent sequence loading device or the cylindrical body 100 rotates clockwise, and the liquid in the pilot-controlled chamber 200 slowly flows into the collection chamber 600 along the liquid outlet pipe 220. In one embodiment, as shown in fig. 1, 3 and 7, the collecting cavity 610 is located at a position where the collecting cavity 600 is closest to the pilot-controlled cavity 200, and the collecting cavity 610 is spaced from the outlet end 222 of the outlet pipe 220.
In one embodiment, the liquid inlet port 210 and the liquid inlet end 221 of the liquid outlet pipe 220 are respectively located at two ends of the hydraulic control chamber 200, and the distances from the rotation center are arranged from small to large: the liquid inlet hole 210, the liquid inlet end 221 of the liquid outlet pipeline and the liquid outlet end 222 of the liquid outlet pipeline; that is, the distance between the liquid injection hole 210 and the rotation center is smaller than the distance between the liquid inlet end 221 of the liquid outlet pipe and the rotation center, and the distance between the liquid inlet end 221 of the liquid outlet pipe and the rotation center is smaller than the distance between the liquid outlet end 222 of the liquid outlet pipe and the rotation center; that is, the torque of the liquid inlet 210 is smaller than that of the liquid inlet 221 of the liquid outlet pipe, the torque of the liquid inlet 221 of the liquid outlet pipe is smaller than that of the liquid outlet 222 of the liquid outlet pipe, and so on in other embodiments. In this way, it is ensured that under normal centrifugation conditions, which for the embodiment shown in fig. 1 are clockwise rotations; at this time, the liquid in the pilot control chamber 200 can flow from the liquid inlet end 221 to the liquid outlet end 222 of the liquid outlet pipe 220, rather than overflowing from the liquid inlet hole 210. As shown in fig. 1 to 8, the shape of the liquid outlet pipe 220 is a resistance increasing structure.
In one embodiment, each loading chamber is further communicated with the collection chamber through a corresponding loading pipeline, and the distances from the rotation center to the rotation center are arranged in the order from small to large: the loading chamber is communicated with the hydraulic control pipeline, and the loading chamber is communicated with the loading pipeline. As shown in fig. 1, the first reagent loading chamber 300 communicates with the collection chamber 600 through the first loading duct 320, the second reagent loading chamber 400 communicates with the collection chamber 600 through the second loading duct 420, the third reagent loading chamber 500 communicates with the collection chamber 600 through the third loading duct 520, the position where the first pilot-controlled duct 310 communicates with the first reagent loading chamber 300 is closest to the rotation center, the position where the first loading duct 320 communicates with the first reagent loading chamber 300 is farthest from the rotation center, the first reagent loading chamber 300 is centered from the rotation center, and so on in the other embodiments. The reagent sequential loading device is suitable for a centrifugal micro-fluidic device, various valves are not needed, sequential loading of various reagents can be realized only in a centrifugal rotating environment, sequential loading of any reagents can be realized by adjusting different reagents in different loading chambers, the realization is simple, the design difficulty is reduced, and the processing cost is saved; in particular, the time interval between the different reagent loadings can be controlled by controlling the centrifugation speed, thereby allowing time for the reagent reaction or reagent treatment.
In one embodiment, the loading chamber is also provided with a liquid injection hole, so that liquid, such as sample liquid, cleaning liquid or eluent, can be injected into the loading chamber directly through the liquid injection hole. In one embodiment, a liquid storage container is arranged in the loading chamber, and the liquid storage container is provided with a liquid releasing structure. In one embodiment, the liquid releasing structure is a rotary liquid releasing structure or a hot melt liquid releasing structure. The rotary liquid releasing structure is used for releasing liquid in the liquid storage container in a rotating state. The hot melt liquid releasing structure is used for releasing liquid in the liquid storage container at a certain temperature. Further, in one embodiment, the liquid storage container has an opening and a paraffin sealing portion covering the opening, and the paraffin sealing portion is used for melting at a certain temperature and releasing liquid in the liquid storage container. By the design, the liquid in the liquid storage container can be conveniently released in a heating mode or a rotating mode. In one embodiment, the rotary liquid release structure comprises a membrane layer and a rotary rupture member thereof. The rotating rupture member is adapted to rupture the membrane layer to release fluid from the reservoir when rotated, i.e., rotated, to a rate. In one embodiment, the liquid storage container is provided with an opening area, a puncturing member, an elastic member and a film layer, wherein the film layer is used for sealing the opening area, one end of the elastic member is connected with the puncturing member, the other end of the elastic member is fixed in the loading chamber, and the puncturing member is used for cooperating with the elastic member to generate displacement to puncture the film layer during centrifugation. Further, in one embodiment, the loading chamber is provided with a liquid reagent. In one embodiment, the liquid reagent is disposed in a heat-fusible wrapping layer disposed in the loading chamber. In one embodiment, the liquid reagent is disposed in a casing disposed in the loading chamber and the casing is provided with an open area closed with a hot melt layer.
In one embodiment, the loading chamber is provided with an opening and a cover part covering the opening. By adopting the design, the sealing cover part can be directly opened, a sample or other reagents needing to be loaded are placed into the loading chamber through the opening, and then the sealing cover part is used for sealing. With the design, the reagent can be conveniently placed in the loading chamber.
In one embodiment, the passing area of the hydraulic control pipeline is 0.001 to 0.03 square millimeter; further, in one embodiment, the pilot operated conduit has a circular cross-section, which may also be referred to as a cross-section, with a radius of 0.01784 mm to 0.09772 mm; or the hydraulic control pipeline has a rectangular passing section, the length of the hydraulic control pipeline is 0.05mm to 0.3mm, and the width of the hydraulic control pipeline is 0.02mm to 0.1 mm; further, in one embodiment, the pilot operated conduit has a rectangular cross-section with a length of 0.2mm and a width of 0.1 mm. The limitation on the passing area is beneficial to avoiding the liquid in the hydraulic control chamber from flowing into the loading chamber along the hydraulic control pipeline, namely, when the passing area of the hydraulic control pipeline is smaller, the hydraulic control pipeline has certain resistance to the liquid flowing in the hydraulic control chamber; particularly, by adopting the hydrophobic material body such as the hydrophobic plastic body, the liquid in the hydraulic control chamber can be still prevented from flowing into the loading chamber along the hydraulic control pipeline under the condition that the hydraulic control chamber is filled with the liquid such as water, so that the technical effect that the loading chamber has a valve function without a valve is achieved. In one embodiment, as shown in fig. 1 and fig. 6, the distances from the first connection end 315, the second connection end 415, and the third connection end 515 to the rotation center 999 are all smaller than the distance from the liquid inlet end 221 of the liquid outlet pipe 220 to the rotation center 999, and are also smaller than the distance from the liquid outlet end 222 of the liquid outlet pipe to the rotation center 999, such a design is also to ensure that the liquid in the hydraulic control chamber 200 flows out through the liquid outlet pipe 220 during rotation, and does not flow into the first hydraulic control pipe 310, the second hydraulic control pipe 410, and the third hydraulic control pipe 510.
Further, in one embodiment, the passing area of the liquid outlet pipe is larger than that of each hydraulic control pipe, and the design is beneficial to ensuring that liquid in the hydraulic control chamber flows out through the liquid outlet pipe and does not flow into each hydraulic control pipe when the hydraulic control chamber rotates. In one embodiment, the liquid outlet pipeline is provided with a resistance-increasing structure; the resistance increasing structure is used for increasing the liquid flow resistance inside the liquid outlet pipeline, so that the liquid in the liquid control cavity cannot flow out of the liquid outlet pipeline to the collection cavity or the waste liquid cavity of the collection cavity too fast. In one embodiment, the resistance-increasing structure is used for matching with the rotating speed setting so as to control the passing speed of the liquid outlet pipeline during rotation; further, in one embodiment, the resistance-increasing structure is arranged in an extended manner. In one embodiment, the resistance-increasing structure comprises at least two communicated bent parts or a reduced passing area; in one embodiment, the resistance-increasing structure comprises at least two communicated bending parts and a reduced passing area; further, in one embodiment, as shown in fig. 1-8, the resistance-enhancing structure has a single Z-shaped or S-shaped structure; or the resistance-increasing structure is provided with a plurality of Z-shaped or S-shaped structures which are connected end to end, namely the resistance-increasing structure comprises a plurality of communicated bent parts. In one embodiment, the resistive enhancing structures have a pass through area of 0.01 to 0.1 square millimeters, such as 0.0625 square millimeters, and the like.
In one embodiment, as shown in fig. 9 and 10, the collection chamber includes a collection chamber 700, an acquisition chamber 800 and a waste liquid chamber 900, and the acquisition chamber 800 and the waste liquid chamber 900 are respectively located at two sides of a reference line by taking a connecting line of the center of the collection chamber 700 and the rotation center as the reference line; each loading chamber is communicated with the collection chamber 700 through its corresponding loading pipe, please refer to fig. 11 and 12, the liquid outlet end 222 of the liquid outlet pipe is communicated with the waste liquid chamber 900; the collection cavity 700 is communicated with the acquisition cavity 800 through an acquisition pipeline 720, and the collection cavity 700 is also communicated with the waste liquid cavity 900 through a waste liquid pipeline 730; the acquisition cavity 800 is provided with an acquisition cavity hole 810, and the acquisition cavity hole 810 is positioned at a position of the acquisition cavity 800 close to the rotation center and is arranged at an interval with the acquisition pipeline 720; the waste liquid cavity 900 is provided with a waste liquid cavity hole 910, and the waste liquid cavity hole 910 is located at a position of the waste liquid cavity 900 closest to the hydraulic control cavity 200 and is arranged at an interval with the liquid outlet end 222 of the liquid outlet pipeline; with this arrangement, the liquid in the pilot-controlled chamber 200 flows into the waste liquid chamber 900 from the liquid outlet pipe 220 and the liquid in the collection chamber 700 flows into the waste liquid chamber 900 from the waste liquid pipe 730 when rotating clockwise, and the liquid in the collection chamber 700 flows into the taking chamber 800 from the taking pipe 720 when rotating counterclockwise. In one embodiment, referring to fig. 15 and 16, the collection cavity 700 has an outlet 710, and the collection cavity 700 is connected to the access cavity 800 and the waste liquid cavity 900 through the outlet 710; in one embodiment, referring to fig. 15 and 17, the outlet 710 is provided with a filter 740; further, the outlet 710 is provided with a pair of mounting holes, the structure and the sealing of the mounting holes are similar to those of the liquid injection holes and are arranged in a manner of matching the shape of the filter membrane, and the outlet is used for opening the pair of mounting holes, filling the filter membrane and then sealing the pair of mounting holes when the filter membrane is mounted; when the filter membrane is replaced, the pair of mounting holes is opened, the used filter membrane is poked out, and then a new filter membrane is mounted to close the pair of mounting holes. In each embodiment, the specific type and model of the filter membrane is not limited, and the filter membrane is suitable for a target object of centrifugal microfluidics. In one embodiment, referring to fig. 9 and 18, the distance between the first communicating end 315 and the liquid injecting hole 210 is smaller than the distance between the second communicating end 415 and the liquid injecting hole 210, the distance between the second communicating end 415 and the liquid injecting hole 210 is smaller than the distance between the third communicating end 515 and the liquid injecting hole 210, and the distances from the first communicating end 315, the second communicating end 415 and the third communicating end 515 to the rotation center 999 are smaller than the distance from the liquid inlet end 221 of the liquid outlet pipe 220 to the rotation center 999 and smaller than the distance from the liquid outlet end 222 of the liquid outlet pipe to the rotation center 999.
In one embodiment, the passing area of the hydraulic control pipeline is 0.001 to 0.03 square millimeter; the liquid outlet pipeline is provided with a resistance increasing structure; the resistance-increasing structure comprises at least two communicated bending parts and/or a reduced passing area; the reagent sequential loading device is provided with a hydrophobic plastic body, and the liquid outlet pipeline 220, the hydraulic control chamber 200, the collecting chamber 600 and each loading chamber are all arranged in the hydrophobic plastic body; the collection chamber comprises a collection cavity 700, an acquisition cavity 800 and a waste liquid cavity 900, and the acquisition cavity 800 and the waste liquid cavity 900 are respectively positioned at two sides of a reference line by taking a connecting line of the center of the collection cavity 700 and the rotation center as the reference line; each loading chamber is communicated with the collecting cavity 700 through a corresponding loading pipeline, and the liquid outlet end 222 of the liquid outlet pipeline is communicated with the waste liquid cavity 900; the collection cavity 700 is communicated with the acquisition cavity 800 through an acquisition pipeline 720, and the collection cavity 700 is also communicated with the waste liquid cavity 900 through a waste liquid pipeline 730; the acquisition cavity 800 is provided with an acquisition cavity hole 810, and the acquisition cavity hole 810 is positioned at a position of the acquisition cavity 800 close to the rotation center and is arranged at an interval with the acquisition pipeline 720; the waste liquid cavity 900 is provided with a waste liquid cavity hole 910, and the waste liquid cavity hole 910 is located at a position of the waste liquid cavity 900 closest to the hydraulic control cavity 200 and is arranged at an interval with the liquid outlet end 222 of the liquid outlet pipeline; the collection cavity 700 is provided with an outlet 710, and the collection cavity 700 is respectively communicated with the acquisition cavity 800 and the waste liquid cavity 900 through the outlet 710; the outlet 710 is provided with a filter membrane 740; the reagent sequential loading device comprises three loading chambers, wherein the three loading chambers are respectively a sample chamber for containing sample liquid, a cleaning liquid chamber for containing cleaning liquid and an eluent chamber for containing eluent, and the distances between the three loading chambers and the liquid injection hole 210 are arranged from small to large in sequence: the communication end of the sample chamber, the communication end of the cleaning liquid chamber and the communication end of the eluent chamber; that is, the sequential reagent loading device includes three loading chambers, which are a first reagent loading chamber 300, a second reagent loading chamber 400, and a third reagent loading chamber 500, respectively, the first reagent loading chamber 300 is a sample chamber, the second reagent loading chamber 400 is a cleaning solution chamber, the third reagent loading chamber is an eluent chamber, a communication end 315 of the first reagent loading chamber is a communication end of the sample chamber, a communication end 415 of the second reagent loading chamber is a communication end of the cleaning solution chamber, and a communication end 515 of the third reagent loading chamber is a communication end of the eluent chamber.
Continuing with fig. 1 to 8, the reagent sequential loading device and its specific application are described, wherein a liquid, such as water, is injected into the pilot-controlled chamber 200 through the injection hole 210, and the liquid level of the injected liquid is higher than the first connection end 315, which is the connection port between the first pilot-controlled pipeline 310 and the pilot-controlled chamber 200. Liquid reagents are preset in the first reagent loading chamber 300, the second reagent loading chamber 400 and the third reagent loading chamber 500, or in other embodiments, the reagent sequential loading device is provided with liquid injection holes in the first reagent loading chamber 300, the second reagent loading chamber 400 and the third reagent loading chamber 500 respectively; and sealing the corresponding injection hole after the injection is finished, wherein the sealing mode comprises but is not limited to paraffin sealing, glue sealing, adhesive tape sealing and the like. In this embodiment, the first reagent loading chamber 300, the second reagent loading chamber 400, and the third reagent loading chamber 500 are all pre-loaded with liquid storage containers, and the reagents are pre-loaded in the liquid storage containers. It is understood that the reagent sequential loading device of the present embodiment can realize sequential loading of 3 reagents, and in other embodiments, the reagent sequential loading device of the present application can realize sequential loading of a plurality of reagents, including but not limited to 3. In one embodiment, taking the dilution step as an example, one of the loading chambers is a dilution chamber, i.e., a diluted reagent loading chamber, the diluent is disposed in the thermal melting packaging layer, and the thermal melting packaging layer is disposed in the dilution chamber. In one embodiment, the diluent is disposed in a casing, the casing is disposed in the dilution chamber and the casing is provided with an opening closed with a hot melt layer. In one embodiment, the liquid storage container is adhered to the dilution cavity. In one embodiment, a liquid reservoir is provided in each loading chamber and the liquid reservoir has a liquid release structure/in one embodiment, the liquid reservoir has an aluminum foil layer. In one embodiment, the liquid storage container is provided with an opening, a puncturing piece, an elastic piece and a sealing film, wherein the sealing film is used for closing the opening, one end of the elastic piece is connected with the puncturing piece, the other end of the elastic piece is fixed in a loading chamber such as a dilution cavity, and the puncturing piece is used for being matched with the elastic piece to generate displacement to puncture the sealing film during centrifugation.
In application, first, for a liquid storage container having a hot-melt liquid releasing structure, the liquids in the liquid storage containers in the first reagent loading chamber 300, the second reagent loading chamber 400 and the third reagent loading chamber 500 are released into the corresponding chambers by heating. For reservoirs having a rotating release structure, the liquid reagents in the reservoirs in the first, second and third reagent loading chambers 300, 400, 500 are released into the corresponding chambers by centrifugation, e.g., at higher speed.
Secondly, since the first communication end 315, the second communication end 415, and the third communication end 515 between the first hydraulic control pipeline 310, the second hydraulic control pipeline 410, and the third hydraulic control pipeline 510 and the hydraulic control chamber 200 are respectively sealed by the liquid in the hydraulic control chamber 200, the liquid reagents in the first reagent loading chamber 300, the second reagent loading chamber 400, and the third reagent loading chamber 500 do not flow into the collection chamber 600 due to the action of the atmospheric pressure, especially in cooperation with the blocking action of the minute pipelines, such as the hydrophobic plastic body.
Thirdly, since the distance between the liquid injection hole 210 and the rotation center is smaller than the distance between the communication end of any one of the loading chambers and the rotation center, the distance between the communication end of any one of the loading chambers and the rotation center is smaller than the distance between the liquid inlet end 221 of the liquid outlet pipe and the rotation center, and the distance between the liquid inlet end 221 of the liquid outlet pipe and the rotation center is smaller than the distance between the liquid outlet end 222 of the liquid outlet pipe and the rotation center, under the medium-low speed centrifugal action, the liquid in the hydraulic control chamber 200 flows into the collection chamber 600 through the liquid outlet pipe 220. In one embodiment, the tapping line 220 is designed as a serpentine line that is continuously curved. Further, in one embodiment, the design of the outlet pipe 220 is required to increase the liquid flow resistance, including but not limited to the design of small-sized pipes or serpentine pipes, to increase the liquid flow resistance.
Finally, the liquid level of the hydraulic control chamber 200 slowly descends from the liquid injection hole 210 to the liquid inlet end 221 of the liquid outlet pipeline 220, and sequentially descends to the first communication end 315, the second communication end 415 and the third communication end 515; when the liquid level in the pilot-controlled chamber 200 drops below the first communication end 315, the first pilot-controlled conduit 310 communicates with the external environment through the first communication end 315, the pilot-controlled chamber 200 and the injection hole 210, and under the combined action of the atmospheric pressure and the centrifugal force, the first reagent in the first reagent loading chamber 300 enters the collection chamber 600 from the first loading conduit 320; when the liquid level in the pilot-controlled chamber 200 drops below the second communication end 415, the second pilot-controlled pipeline 410 is communicated with the external environment through the second communication end 415, the pilot-controlled chamber 200 and the liquid injection hole 210, and under the combined action of atmospheric pressure and centrifugal force, the second reagent in the second reagent loading chamber 400 enters the collection chamber 600 from the second loading pipeline 420; when the liquid level in the hydraulic control chamber 200 drops below the third communication end 515, the third hydraulic control pipeline 510 communicates with the external environment through the third communication end 515, the hydraulic control chamber 200 and the injection hole 210, and under the combined action of atmospheric pressure and centrifugal force, the third reagent in the third reagent loading chamber 500 enters the collection chamber 600 from the third loading pipeline 520, so that the sequential loading of the reagents is realized. Such a design may ensure a reagent loading sequence, first adding a first reagent, e.g., a sample, then a second reagent, e.g., a wash solution, and finally a third reagent, e.g., an eluent. During centrifugation, the centrifugal rotation speed may be controlled to control the flow speed of the liquid in the pilot-controlled chamber 200, so as to control the time interval of releasing the liquid reagent in the first, second, and third reagent loading chambers 300, 400, and 500.
Taking nucleic acid purification as an example, the reagent sequential loading device and its specific application will be described with reference to fig. 9 to 18, in the molecular diagnostic process, the purification of nucleic acid in the nucleic acid extraction step is very critical, and since the nucleic acid purification involves sequential loading of several liquid reagents, it is often difficult to integrate into a microfluidic chip or other centrifugal microfluidic devices, and it can be conveniently realized by using the embodiments related to the present application.
The lysed sample, the cleaning solution, and the eluent are respectively added into the first reagent loading chamber 300, the second reagent loading chamber 400, and the third reagent loading chamber 500 through injection holes (not shown in the injection hole diagram), and then the injection holes are sealed. Or by adopting a reagent presetting method, the cracked sample, the cleaning solution and the eluent are respectively preset in three liquid storage containers, the three liquid storage containers are respectively preset in the first reagent loading chamber 300, the second reagent loading chamber 400 and the third reagent loading chamber 500, the first reagent loading chamber 300 is used as a sample chamber for containing the sample solution, the second reagent loading chamber 400 is used as a cleaning solution chamber for containing the cleaning solution, and the third reagent loading chamber 500 is used as an eluent chamber for containing the eluent, and specific reagent presetting and releasing methods are as before and are not repeated.
Water is then injected into the pilot chamber 200 through the injection hole 210 until the liquid level is above the interface between the first pilot conduit 310 and the pilot chamber 200, i.e., the first communication port 315.
Fixing the reagent sequence loading device or the rotation center thereof in a rotation system, for example, fixing the position of the rotation center of the reagent sequence loading device on the rotation shaft of the motor; or fixing the centrifugal microfluidic device in which the reagent sequential loading device is positioned in a rotating system, for example, fixing the position of the rotating center of the centrifugal microfluidic device on the rotating shaft of the motor; the order of the steps can be interchanged with the previous steps.
Rotating clockwise, for example at 1500rpm, rotates the reagent sequential loading device or centrifugal microfluidic device clockwise, and the liquid in the pilot-controlled chamber 200 begins to flow into the waste chamber 900 through the serpentine outlet conduit 220. When the liquid level in the hydraulic control chamber 200 is lower than the interface between the first hydraulic control pipeline 310 and the hydraulic control chamber 200, i.e. the first communication end 315, the first reagent loading chamber 300 is communicated with the atmosphere through the liquid injection hole 210, the cracked sample in the first reagent loading chamber 300 flows out to the collection chamber 700 through the first loading pipeline 320, and when the sample passes through the filter membrane 740 of the outlet 710, nucleic acid such as DNA or RNA in the cracked sample remains on the silica gel membrane, and the waste liquid completely enters the waste liquid chamber 900 through the waste liquid pipeline 730 under the action of Coriolis force (Coriolis force). In this embodiment, the filter membrane 740 is a silica gel membrane, and the specific type and specification can be selected according to the requirement. Coriolis forces are a description of the deflection of a particle undergoing linear motion in a rotating system relative to linear motion produced by the rotating system due to inertia. The coriolis force is derived from the inertia of the motion of the object. When a mass point moves linearly relative to an inertial system, the track of the mass point is a curve relative to a rotating system, and the rotating system is used as a reference system according to the theory of Newton mechanics, and the tendency of the linear motion of the mass point to deviate from the original direction is attributed to an external force action, namely Coriolis force.
The liquid in the hydraulic control chamber 200 continuously flows into the waste liquid chamber 900 through the serpentine liquid outlet pipe 220, when the liquid level in the hydraulic control chamber 200 is lower than the interface between the second hydraulic control pipe 410 and the hydraulic control chamber 200, i.e. the second communication end 415, the second reagent loading chamber 400 is communicated with the atmosphere through the liquid injection hole 210, the cleaning liquid in the second reagent loading chamber 400 flows out into the collection chamber 700 through the second loading pipe 420, and when passing through the filter membrane 740 of the outlet 710, the nucleic acid (DNA or RNA) on the filter membrane 740 is cleaned, and the waste liquid completely enters the waste liquid chamber 900 through the waste liquid pipe 730 under the action of coriolis force.
When the liquid level in the hydraulic control chamber 200 is lower than the interface between the third hydraulic control pipeline 510 and the hydraulic control chamber 200, i.e., the third communication end 515, the third reagent loading chamber 500 is communicated with the atmosphere through the liquid injection hole 210, the eluent in the third reagent loading chamber 500 flows out through the third loading pipeline 520 to the collection chamber 700, and passes through the filter membrane 740 of the outlet 710, the nucleic acid (DNA or RNA) on the filter membrane 740 is eluted, and the eluted nucleic acid solution completely enters the acquisition chamber 800 through the acquisition pipeline 720 under the action of coriolis force.
Therefore, the simple reagent sequential loading device for sequentially loading various reagents in the centrifugal microfluidic is realized, the sequential loading mode of the various reagents does not need an additional valve device, the realization is simple, additional processing technology and cost are not increased, the sequential loading of the reagents can be realized only by the microfluidic chip at a constant centrifugal rotating speed, the sequential loading of any kind of reagents can be realized only by simply changing the positions of the reagents or the number of loading chambers, the centrifugal rotating speed can be controlled, the loading time intervals of different reagents are controlled, and the time is reserved for reagent reaction or reagent treatment. In other embodiments, the purification of nucleic acid requires two washing processes, and the reagent sequential loading device is designed with four loading chambers, two of which are respectively used as washing solution chambers, and only washing solution is contained therein; thus, the sample, the first cleaning solution, the second cleaning solution and the eluent after the cracking can be loaded sequentially, and the rest of the embodiments can be analogized.
In one embodiment, a centrifugal microfluidic device comprises a sequential reagent loading device as described in any of the embodiments. The centrifugal microfluidic device is very ingenious in design, various valves are not needed, and multiple reagents can be loaded sequentially only in a centrifugal rotation environment, so that the centrifugal microfluidic device is simple to realize, the design difficulty of sequential reagent loading is reduced, and the processing cost is saved; particularly, the time interval of loading different reagents can be controlled by controlling the centrifugal rotating speed, and the time is reserved for reagent reaction or reagent treatment; further, sequential loading of any number of reagents can be achieved by adjusting the different reagents in the different loading chambers.
In one embodiment, an analytical system includes the centrifugal microfluidic device of any of the embodiments. The analysis system can be used for analyzing nucleic acid, protein, cells, tissues, pathogens, viruses and the like, various valves are not needed, and various reagents can be loaded sequentially only in a centrifugal rotation environment, so that the analysis system is simple to realize, the design difficulty of sequential reagent loading is reduced, and the processing cost is saved. In one embodiment, the assay system is a nucleic acid assay system comprising the centrifugal microfluidic device of any of the embodiments.
Other embodiments of the present application include a reagent sequential loading device, a centrifugal microfluidic device, and an analysis system, which are capable of being implemented by combining technical features of the above embodiments. The technical features of the above embodiments can be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the above embodiments are not described, but should be considered as the scope of the present specification as long as there is no contradiction between the combinations of the technical features. The above examples only express several embodiments of the present application, and the description thereof is more specific and detailed, but not construed as limiting the claims. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the concept of the present application, which falls within the scope of protection of the present application. Therefore, the protection scope of the present application shall be subject to the appended claims.
Claims (10)
1. A reagent sequential loading device is characterized in that the reagent sequential loading device is provided with a rotation center and comprises a liquid outlet pipeline, a hydraulic control chamber, a collection chamber and at least two loading chambers;
the liquid inlet end of the liquid outlet pipeline is communicated with the liquid control chamber, the liquid outlet end of the liquid outlet pipeline is communicated with the collection chamber,
the reagent sequential loading device is provided with a hydraulic control pipeline and a loading pipeline in each loading chamber respectively and correspondingly;
the collection cavity is equipped with collects the accent, the liquid accuse cavity is equipped with annotates the liquid hole, annotate the liquid hole with the feed liquor end of liquid outlet pipe way is located respectively the both ends of liquid accuse cavity, and with the distance of rotation center is arranged according to order from little to big: the liquid injection hole, the liquid inlet end of the liquid outlet pipeline and the liquid outlet end of the liquid outlet pipeline are arranged;
each loading chamber is communicated with the hydraulic control chamber through the corresponding hydraulic control pipeline, each hydraulic control pipeline is provided with a communication end which is communicated with the hydraulic control chamber and is positioned between the liquid inlet end of the liquid outlet pipeline and the liquid injection hole, and the distances between the communication ends and the liquid injection hole are different;
each loading chamber is also communicated with the collection chamber through the corresponding loading pipeline, and the distances between the loading chambers and the rotation center are arranged from small to large in sequence: the loading chamber is communicated with the hydraulic control pipeline, and the loading chamber is communicated with the loading pipeline.
2. The sequential reagent loading device of claim 1, wherein the outlet pipeline has a resistance increasing structure, and the resistance increasing structure is used for matching with a rotating speed setting so as to control the passing speed of the outlet pipeline during rotation.
3. The sequential reagent loading device of claim 2, wherein the resistance-enhancing structure comprises at least two bends in communication.
4. The sequential reagent loading device of claim 2, wherein the resistance-enhancing structures comprise a reduced area of passage.
5. The sequential reagent loading device of claim 1, wherein the sequential reagent loading device has a hydrophobic body, and the outlet conduit, the pilot-controlled chamber, the collection chamber and each loading chamber are all opened in the hydrophobic body.
6. The sequential reagent loading device according to claim 1, wherein the sequential reagent loading device comprises three loading chambers, the three loading chambers are a first reagent loading chamber, a second reagent loading chamber and a third reagent loading chamber, and the distances from the liquid injection hole are arranged from small to large in sequence: a communication end of the first reagent loading chamber, a communication end of the second reagent loading chamber, and a communication end of the third reagent loading chamber.
7. The sequential reagent loading device according to any one of claims 1 to 6, wherein the collection chamber comprises a collection chamber, an acquisition chamber and a waste liquid chamber, and a connecting line of the center of the collection chamber and the rotation center is taken as a reference line, and the acquisition chamber and the waste liquid chamber are respectively positioned at two sides of the reference line;
each loading chamber is communicated with the collecting cavity through the corresponding loading pipeline, and the liquid outlet end of the liquid outlet pipeline is communicated with the waste liquid cavity;
the collecting cavity is communicated with the acquiring cavity through an acquiring pipeline and is also communicated with the waste liquid cavity through a waste liquid pipeline;
the acquisition cavity is provided with an acquisition cavity hole, and the acquisition cavity hole is positioned at the position of the acquisition cavity close to the rotation center and is arranged at an interval with the acquisition pipeline;
the waste liquid cavity is provided with a waste liquid cavity hole, and the waste liquid cavity hole is located at the position, closest to the hydraulic control cavity, of the waste liquid cavity and is arranged at an interval with the liquid outlet end of the liquid outlet pipeline.
8. The sequential reagent loading device of claim 7, wherein the collection chamber is provided with an outlet, and the collection chamber is communicated with the acquisition chamber and the waste liquid chamber through the outlet respectively; and the outlet is provided with a filter membrane.
9. A centrifugal microfluidic device comprising a sequential reagent loading device according to any one of claims 1 to 8.
10. An analytical system comprising a centrifugal microfluidic device according to claim 9.
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| CN112808336A (en) * | 2021-02-09 | 2021-05-18 | 深圳市亚辉龙生物科技股份有限公司 | Micro-fluidic chip |
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