CN112619719A - Droplet generation microdevice for digital PCR - Google Patents
Droplet generation microdevice for digital PCR Download PDFInfo
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- CN112619719A CN112619719A CN202011415287.6A CN202011415287A CN112619719A CN 112619719 A CN112619719 A CN 112619719A CN 202011415287 A CN202011415287 A CN 202011415287A CN 112619719 A CN112619719 A CN 112619719A
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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
- B01L3/02—Burettes; Pipettes
- B01L3/0241—Drop counters; Drop formers
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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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- 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
- B01L3/502769—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 multiphase flow arrangements
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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
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/0864—Configuration of multiple channels and/or chambers in a single devices comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
Abstract
The invention provides a micro device for generating liquid drops for digital PCR (polymerase chain reaction), which comprises an outer container, an inner container and a micro liquid drop generating structure, wherein the outer container is provided with a first accommodating cavity, stable liquid is accommodated in the first accommodating cavity, the micro liquid drop generating structure is immersed in the stable liquid, and the inner container is accommodated in the first accommodating cavity; the inner container is provided with a second containing cavity for containing the liquid to be detected, the micro-droplet generation structure comprises a micro-droplet generation flow channel, the inner container is connected with the micro-droplet generation structure, and the second containing cavity is communicated with the micro-droplet generation flow channel; and the inner container is also provided with an air inlet pipeline to communicate the second accommodating cavity with the outside, outside air enters the second accommodating cavity through the air inlet pipeline to push the liquid to be detected to enter the micro-droplet generating structure and form micro-droplets through the micro-droplet generating flow channel, and the micro-droplets are discharged into the stabilizing liquid through the micro-droplet generating flow channel. The liquid drop generation micro device for the digital PCR is simple in structure and convenient to operate.
Description
Technical Field
The field belongs to the technical field of microfluidics, and particularly relates to a droplet generation micro device for digital PCR.
Background
With the development of digital PCR technology, digital PCR technology is applied to a plurality of technical fields such as biology, medicine, chemistry, etc., and in the process of absolute quantitative analysis of nucleic acid molecules using digital PCR technology, a liquid to be detected needs to be divided into a plurality of micro-droplets. However, the conventional droplet generating device for digital PCR has a complicated structure and high operation difficulty, which is not favorable for improving the generation efficiency of micro-droplets.
Disclosure of Invention
The invention aims to provide a droplet generation micro device for digital Polymerase Chain Reaction (dPCR), which has a simpler structure and lower operation difficulty and is beneficial to improving the generation efficiency of micro droplets.
In order to realize the purpose of the invention, the invention provides the following technical scheme:
the invention provides a micro-device for generating liquid drops for digital PCR, which comprises an outer container, an inner container and a micro-liquid-drop generating structure, wherein the outer container is provided with a first accommodating cavity, a stable liquid is accommodated in the first accommodating cavity, the micro-liquid-drop generating structure is immersed in the stable liquid, and the inner container is accommodated in the first accommodating cavity; the inner container is provided with a second containing cavity for containing liquid to be detected, the micro-droplet generation structure comprises a micro-droplet generation flow channel, the inner container is connected with the micro-droplet generation structure, and the second containing cavity is communicated with the micro-droplet generation flow channel; and the inner container is also provided with an air inlet pipeline so as to communicate the second accommodating cavity with the outside, outside air enters the second accommodating cavity through the air inlet pipeline so as to push the liquid to be detected to enter the micro-droplet generating structure and pass through the micro-droplet generating flow channel to form micro-droplets, and the micro-droplets are discharged into the stabilizing liquid through the micro-droplet generating flow channel.
In one embodiment, a liquid inlet pipeline is connected between the inner container and the micro-droplet generating structure, the micro-droplet generating structure further comprises a diffusion cavity, a liquid inlet end of the diffusion cavity is communicated with the second accommodating cavity through the liquid inlet pipeline, a liquid outlet end of the diffusion cavity is communicated with the micro-droplet generating flow channel, and the cross-sectional area of the liquid outlet end of the diffusion cavity is larger than that of the liquid inlet end.
In one embodiment, the number of the diffusion cavities is a plurality of, and each of the diffusion cavities has the liquid inlet end communicated with the liquid inlet pipeline, and each of the liquid outlet ends of the diffusion cavities is connected with the micro-droplet generation flow channel.
In one embodiment, the number of the droplet generating channels is multiple, multiple droplet generating channels are arranged side by side, and the multiple droplet generating channels are all communicated with the diffusion chamber.
In one embodiment, the droplet generation structure further includes a plurality of partitions arranged side by side and at intervals, a gap between every two adjacent partitions forms one droplet generation channel, the droplet generation channel includes a liquid inlet channel and a liquid outlet channel, which are communicated with each other, and the cross-sectional area of the liquid inlet channel is larger than that of the liquid outlet channel.
In one embodiment, the cross section of the liquid outlet flow passage is circular, and the diameter of the circle ranges from 3 μm to 90 μm; and/or the cross section of the liquid outlet flow channel is rectangular, the size range of the long side of the rectangle is 9-90 μm, and the size range of the wide side of the rectangle is 3-30 μm.
In one embodiment, the length of the liquid outlet flow passage ranges from 100 μm to 300 μm.
In one embodiment, the droplet generation structure includes a bearing portion and a covering portion, the covering portion and the bearing portion enclose a diffusion chamber, a support is further disposed in the diffusion chamber, one end of the support is connected to an inner surface of the covering portion, and the other end of the support is connected to an inner surface of the bearing portion.
In one embodiment, the outer container is further provided with a vent hole, and the vent hole is communicated with the first accommodating cavity and the outside of the outer container for discharging the gas in the first accommodating cavity.
In one embodiment, the test solution is poorly soluble or insoluble in the stabilizing solution and the density of the test solution is greater than or less than the density of the stabilizing solution.
The invention provides a micro-device for generating liquid drops for digital PCR, which is provided with an inner container, an outer container and a micro-liquid drop generating structure with a micro-liquid drop generating flow channel. The liquid drop generation micro device for the digital PCR is simple in structure and convenient to operate.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic diagram of a droplet generation micro device for digital PCR according to an embodiment;
FIG. 2 is a schematic cross-sectional view of the droplet generation microdevice for digital PCR shown in FIG. 1 along the A-A direction;
FIG. 3 is a schematic structural diagram of a microdroplet generating structure according to an embodiment;
FIG. 4 is an enlarged schematic view of region I of the microdroplet generation structure of FIG. 3;
FIG. 5 is a schematic right-side view of the micro-droplet generation structure of FIG. 3 in one embodiment;
fig. 6 is a schematic right-side view of the microdroplet generating structure of fig. 3 in another embodiment.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
Referring to fig. 1 and 2 together, fig. 1 is a schematic structural diagram of a droplet generation micro device 1000 for digital PCR according to an embodiment;
FIG. 2 is a schematic sectional view of the droplet-generating microdevice 1000 for digital PCR shown in FIG. 1, taken along the A-A direction.
The embodiment of the application provides a micro device 1000 for generating liquid drops for digital PCR, the micro device 1000 for generating liquid drops comprises an outer container 100, an inner container 200 and a micro liquid drop generating structure 300, wherein the outer container 100 is provided with a first containing cavity 101, a stabilizing liquid 500 is contained in the first containing cavity 101, the micro liquid drop generating structure 300 is immersed in the stabilizing liquid 500, and the inner container 200 is contained in the first containing cavity 101; the inner container 200 is provided with a second containing cavity 201 for containing the liquid 400 to be detected, the micro-droplet generation structure 300 comprises a micro-droplet generation flow channel 301, the inner container 200 is connected with the micro-droplet generation structure 300, and the second containing cavity 201 is communicated with the micro-droplet generation flow channel 301; the inner container 200 is further provided with an air inlet pipe 600 to communicate the second accommodating cavity 201 with the outside, and outside air enters the second accommodating cavity 201 through the air inlet pipe 600 to push the liquid to be tested 400 to enter the micro-droplet generating structure 300 and form micro-droplets through the micro-droplet generating flow channel 301, and the micro-droplets are discharged into the stabilizing liquid 500 through the micro-droplet generating flow channel 301.
It can be understood that the application fields of the droplet generation micro device 1000 provided in the embodiments of the present application include, but are not limited to, the field of digital PCR, and can also be applied to other fields requiring generation of micro droplets, and details are not repeated herein.
The outer container 100 may have various shapes and structures, as long as the first receiving chamber 101 is provided for receiving the inner container 200, the micro-droplet generating structure 300, and the stabilizing liquid 500, and the shapes and structures of the outer container 100 are not specifically limited herein. It can be understood that, when the droplet generation micro device 1000 provided by the present application is applied to a digital Polymerase Chain Reaction (dPCR) technology, the outer container 100 may be a centrifuge tube, so that after the droplets are generated, the droplet generation micro device 1000 may be directly placed into a dPCR detection apparatus for detection, without taking the droplets out of the droplet generation micro device 1000 first, and then transferring the droplets into the dPCR detection apparatus, thereby reducing the number of intermediate transfer steps, reducing the operation difficulty, effectively avoiding the problems of droplet fracture and fusion and the like caused in the transfer process, and improving the success rate of detection.
The inner container 200 is accommodated in the first accommodating chamber 101, the size of the inner container 200 is matched with that of the first accommodating chamber 101, so that the inner container 200 can be accommodated in the first accommodating chamber 101, and the first accommodating chamber 101 has enough space to accommodate the micro-droplet generating structure 300 and the stabilizing liquid 500 besides the space occupied by the inner container 200. The inner container 200 is provided with a second containing cavity 201 for containing the liquid 400 to be detected, so that the liquid 400 to be detected is separated from the stabilizing liquid 500, and the subsequent generation process of micro-droplets is facilitated. It will be appreciated that in one particular embodiment, the inner container 200 is removably connected to the outer container 100, thereby increasing the structural stability and flexibility of the droplet-generating microdevice 1000 for digital PCR.
The liquid 400 to be measured is contained in the second containing cavity 201, the liquid 400 to be measured is a sample that needs to be divided into a plurality of micro droplets, and when the droplet generation micro device 1000 is applied to different fields, the liquid 400 to be measured can be different types of samples. In one embodiment, the test solution 400 is an aqueous solution containing a plurality of nucleic acid molecules for performing a dPCR test.
The stabilizing solution 500 is accommodated in the first accommodating chamber 101, the stabilizing solution 500 is a solution immiscible with the solution 400 to be tested, and when the solution 400 to be tested is of different types, the stabilizing solution 500 should be of a corresponding immiscible type, and the type of the stabilizing solution 500 is not specifically limited herein. In one embodiment, the test solution 400 is an aqueous solution containing a plurality of nucleic acid molecules, and the stabilizing solution 500 is an oil solution immiscible with the test solution 400.
It is understood that there should be a difference in density between the stabilizing liquid 500 and the liquid 400 to be tested, i.e. the stabilizing liquid 500 floats above the liquid 400 to be tested, or the liquid 400 floats above the stabilizing liquid 500. So that the micro-droplets formed by the liquid 400 to be tested are suspended on the surface of the stabilizing liquid 500 after entering the stabilizing liquid 500, or sink to the bottom of the stabilizing liquid 500. So as to collect micro-droplets suspended on the surface of the stabilizing solution 500, or directly test micro-droplets settled on the bottom of the stabilizing solution 500.
The droplet generating structure 300 is disposed in the first accommodating chamber 101, and the droplet generating structure 300 includes a droplet generating channel 301. After entering the droplet generation structure 300 from the second accommodating chamber 201, the liquid 400 to be tested flows through the droplet generation channel 301, and the liquid 400 to be tested is discharged from the droplet generation channel 301 to the stabilizing liquid 500 to form a stable droplet. It can be understood that the micro-droplet generating structure 300 should be immersed in the stabilizing liquid 500, that is, the liquid 400 to be tested will directly enter the stabilizing liquid 500 after being discharged from the micro-droplet generating flow channel 301, so as to form micro-droplets in which the liquid 400 to be tested is wrapped by the stabilizing liquid 500.
Wherein, an air inlet pipe 600 is further provided on the inner container 200 to communicate the second receiving cavity 201 with the outside. It can be understood that, by introducing gas into the second accommodating chamber 201 from the gas inlet pipe 600, the liquid to be measured 400 can be pushed into the droplet generation flow channel 301 by the gas inlet pressure, so that the droplets are formed. With the above structure, the operation difficulty of forming micro droplets is low, and the size of the generated micro droplets can be adjusted by controlling the magnitude of the intake pressure, thereby improving the controllability of the droplet generation micro device 1000. In one embodiment, a small syringe may be used in conjunction with inlet conduit 600 to produce a large number of uniform micro-droplets by pushing the syringe very slowly. In another embodiment, a precision air pressure control device can be used to connect the air inlet pipe 600, and when the precision air pressure control device is opened, a large amount of micro-droplets with good uniformity can be generated. It is understood that the manner of introducing the gas into the second accommodating chamber 201 through the gas inlet duct 600 includes, but is not limited to, the above two manners, and any structure or method that satisfies the corresponding functional requirements may be adopted, and is not specifically limited herein.
The micro-device 1000 for droplet generation for digital PCR provided in the embodiment of the present application is provided with an inner container 200, an outer container 100, and a micro-droplet generation structure 300 having a micro-droplet generation flow channel 301, and can form stable and uniform micro-droplets by introducing the liquid 400 to be detected in the first accommodating chamber 101 into the micro-droplet generation structure 300, passing through the micro-droplet generation flow channel 301, and then discharging the liquid into the stabilizing liquid 500 in the second accommodating chamber 201. In addition, the droplet generation micro device 1000 for digital PCR provided by the embodiment of the present application has a simple structure and is convenient to operate.
In one embodiment, a liquid inlet pipe 700 is connected between the inner container 200 and the micro-droplet generating structure 300, the micro-droplet generating structure 300 further includes a diffusion cavity 302, one end of the diffusion cavity 302 is communicated with the second accommodating cavity 201 through the liquid inlet pipe 700, and the other end of the diffusion cavity 302 is communicated with the micro-droplet generating flow channel 301. The presence of the inlet conduit 700 allows the inner container 200 to communicate with the droplet generating structure 300, and the liquid under test 400 may enter the droplet generating structure 300 through the inlet conduit 700. The diffusion cavity 302 in the micro-droplet generation structure 300 is used for providing a space for diffusing the liquid 400 to be detected, the liquid 400 to be detected enters the diffusion cavity 302 through the liquid inlet pipeline 700 and is diffused in the diffusion cavity 302, so that the liquid 400 to be detected is uniformly distributed in the diffusion cavity 302 and is uniformly discharged from each micro-droplet generation flow channel 301, the micro-droplet generation efficiency is improved, and the uniformity and stability of micro-droplet generation are ensured.
In a specific embodiment, a main channel 303 is further connected between the diffusion chamber 302 and the liquid inlet pipe 700, and the liquid 400 to be measured in the liquid inlet pipe 700 can enter the diffusion chamber 302 through the main channel 303. It should be noted that the main channel 303 should be designed to have a smaller size to reduce the flow rate of the liquid 400 to be detected entering the diffusion chamber 302 per unit time, so that the detection personnel can control the amount and size of the generated micro-droplets more accurately.
In one embodiment, the number of the diffusion chambers 302 is multiple, each diffusion chamber 302 is communicated with the liquid inlet pipe 700, and each diffusion chamber 302 is connected with a micro-droplet generation flow channel 301. It can be understood that the number of the diffusion cavities 302 may be multiple, and the multiple diffusion cavities 302 are all communicated with the second accommodating cavity 201 through the liquid inlet pipe 700, so that the liquid 400 to be detected in the second accommodating cavity 201 can enter the multiple diffusion cavities 302 and simultaneously flow through the micro-droplet generation flow channel 301 to form micro-droplets, thereby further improving the micro-droplet generation efficiency of the droplet generation micro-device 1000. In a specific embodiment, the number of the diffusion chambers 302 is two (as shown in fig. 2), the two diffusion chambers 302 share a main flow channel 303 to communicate with the liquid inlet pipe 700, and the two diffusion chambers 302 are symmetrically distributed about the main flow channel 303.
In one embodiment, the outer container 100 is further provided with a vent 102, the vent 102 communicating the first receiving chamber 101 with the exterior of the outer container 100 for venting gas from the first receiving chamber 101. It can be understood that, after the liquid to be measured 400 in the inner container 200 enters the first accommodating cavity 101 through the micro-droplet generation flow channel 301, the space occupied by the gas in the first accommodating cavity 101 is squeezed, so that the gas pressure is increased, and the structural damage of the outer container 100 is easily caused, and the gas in the first accommodating cavity 101 can be effectively discharged due to the existence of the vent hole 102, so that the structural damage of the outer container 100 caused by the internal gas pressure is avoided.
Referring to fig. 3 and 4 together, fig. 3 is a schematic structural diagram of an exemplary droplet generation structure 300;
fig. 4 is an enlarged schematic view of region I of the microdroplet generation structure 300 shown in fig. 3.
In one embodiment, the droplet generating structure 300 includes a bearing portion 31 and a covering portion 32, and the covering portion 32 and the bearing portion 31 enclose to form a diffusion chamber 302. In a specific embodiment, a micro-nano structure template composed of a main channel 303, a diffusion cavity 302 and a plurality of droplet generation channels can be processed and prepared on a silicon wafer by adopting a micro-fluidic technology. Then, performing silanization treatment on the prepared micro-nano structure template; pouring Polydimethylsiloxane (PDMS) prepolymer on the surface of the template, and then placing the template covered with PDMS in a vacuum drying oven to cure and shape the PDMS liquid; finally, the template and the PDMS are separated, a punch hole is punched at the position where the PDMS is marked with the punch hole by the puncher, and the portion to be cut is cut by the utility knife, so that the covering portion 32 of the droplet generating structure 300 is obtained. Another blank silicon wafer is taken and a blank PDMS is fabricated by the same method to be used as the carrying part 31 of the micro-droplet generating structure 300. Then, the bonding surfaces of the covering part 32 and the bearing part 31 are upward, and the covering part and the bearing part are cleaned in an oxygen plasma cleaning machine for 30 seconds and then taken out for bonding, so that the micro-droplet generation structure 300 is obtained. It can be understood that the manufacturing manners of the covering portion 32 and the carrying portion 31 of the droplet generating structure 300 include, but are not limited to, the above, and any manner that meets the corresponding functional requirements may also be used, which is not described in detail herein.
In one embodiment, the diffusion chamber 302 includes a liquid inlet end 3021 and a liquid outlet end 3022, the liquid inlet end 3021 of the diffusion chamber 302 is in communication with the liquid inlet conduit 700, the liquid outlet end 3022 of the diffusion chamber 302 is in communication with the droplet generation channels 301, and the cross-sectional area of the diffusion chamber 302 at the liquid outlet end 3022 is greater than the cross-sectional area at the liquid inlet end 3021. It can be understood that the liquid to be measured 400 enters the diffusion cavity 302 from the liquid inlet end 3021 through the liquid inlet pipe 700, and then enters the droplet generation flow channel 301 from the liquid outlet end 3022 after being diffused in the diffusion cavity 302, and in the above structure, when the cross-sectional area of the diffusion cavity 302 at the liquid outlet end 3022 is large, the liquid to be measured 400 entering the diffusion cavity 302 can be effectively diffused when flowing to the liquid outlet end 3022 of the diffusion cavity 302, and thus is uniformly distributed at the liquid outlet end 3022 of the diffusion cavity 302 and enters the droplet generation flow channel 301 communicated with the liquid outlet end 3022. It can be understood that when the solution to be detected 400 is uniformly diffused and then enters the micro-droplet generation flow channel 301, uniform and stable micro-droplets can be formed more favorably, and the micro-droplet generation effect of the micro-droplet generation device 1000 for digital PCR can be further optimized.
In one embodiment, the droplet generating structure 300 further includes a plurality of separating portions 33 arranged side by side and at intervals, a gap between every two adjacent separating portions 33 forms a droplet generating flow channel 301, the droplet generating flow channel 301 includes a liquid inlet flow channel 3011 and a liquid outlet flow channel 3012, which are communicated, and a cross-sectional area of the liquid inlet flow channel 3011 is larger than a cross-sectional area of the liquid outlet flow channel 3012. Under the structure, the shape of the micro-droplet generation flow channel 301 can be changed by changing the shape of the partition part 33, and the size of the cross section of the micro-droplet generation flow channel 301 can be changed by changing the spacing distance between two adjacent partition parts 33, so that the shape and the size of the micro-droplet generation flow channel 301 can be more conveniently and effectively adjusted. It can be understood that the liquid inlet channel 3011 is communicated with the diffusion cavity 302, the liquid outlet channel 3012 is communicated to the stabilizing liquid 500, the liquid 400 to be detected in the diffusion cavity 302 enters the micro-droplet generating channel 301 through the liquid inlet channel 3011, and is discharged into the stabilizing liquid 500 through the liquid outlet channel 3012 to form micro-droplets. It should be noted that the size of the liquid outlet channel 3012 determines the size of the generated micro-droplets to a certain extent, therefore, the cross-sectional area of the liquid outlet channel 3012 should be smaller to meet the size requirement of the generated micro-droplets, and the liquid outlet channel 3012 with a smaller cross-sectional area can effectively prevent the stabilizing liquid 500 from flowing back into the diffusion cavity 302 through the liquid outlet pipeline; the cross section of the liquid inlet channel 3011 should be larger than that of the liquid outlet channel 3012, so that the liquid 400 to be measured can smoothly enter the micro-droplet generation channel 301 through the liquid inlet channel 3011, and the use performance of the micro-droplet generation device 1000 is further optimized.
In one embodiment, the number of the droplet generating channels 301 is multiple, multiple droplet generating channels 301 are arranged side by side, and the multiple droplet generating channels 301 are all communicated with the diffusion chamber 302. It can be understood that the existence of the plurality of micro-droplet generation flow channels 301 enables the micro-droplet generation structure 300 to generate micro-droplets in a plurality of paths at the same time, further improving the generation efficiency of micro-droplets, and even if one micro-droplet generation flow channel 301 is blocked and has a functional failure, the other micro-droplet generation flow channels 301 can still meet the corresponding functional requirements, thereby improving the fault tolerance of the micro-droplet generation device 1000 for digital PCR. It should be noted that the plurality of droplet generation channels 301 are arranged side by side, so that a certain distance is maintained between the droplets generated by the droplet generation channels 301, thereby effectively avoiding the problem of fusion or breakage caused by collision between the generated droplets.
Referring to fig. 5 and 6 together, fig. 5 is a schematic right-side view of the droplet generation structure 300 shown in fig. 3 in one embodiment; fig. 6 is a schematic right-side view of the microdroplet generating structure 300 of fig. 3 in another embodiment.
In one embodiment, the cross section of the liquid outlet channel 3012 is circular, and the diameter of the circle ranges from 3 μm to 90 μm. It can be understood that when the liquid under test 400 is discharged from the micro-droplet generation flow channel 301 into the stabilizing liquid 500, the liquid under test 400 is subjected to the shearing force of the stabilizing liquid 500 at the opening of the micro-droplet generation flow channel 301. When the shearing force applied to the liquid 400 to be tested is the same in all directions, the cross section of the liquid outlet channel 3012 can be designed to be circular, so that when the liquid 400 to be tested is discharged from the liquid outlet channel 3012, stable spherical micro-droplets can be formed even under the action of the shearing force of the stabilizing liquid 500. In a specific embodiment, the diameter of the circular cross section of the outlet flow channel 3012 is in the range of 3 μm to 90 μm, and the size of the formed micro-droplets in the above size range can satisfy the corresponding test requirements.
In one embodiment, the cross section of the liquid outlet channel 3012 is rectangular, the size of the long side of the rectangle is in the range of 9 μm to 90 μm, and the size of the wide side of the rectangle is in the range of 3 μm to 30 μm. When the liquid 400 to be tested is discharged from the droplet generation channel 301 to the stabilizing liquid 500, the liquid 400 to be tested is subjected to the shearing force of the stabilizing liquid 500 at the opening of the droplet generation channel 301. When the liquid 400 to be measured is subjected to only the shearing force of the stabilizing liquid 500 in one direction, the cross section of the liquid outlet channel 3012 may be designed to be rectangular, the long side direction of the rectangle corresponds to the direction of the shearing force applied to the liquid 400 to be measured, and when the liquid 400 to be measured is discharged from the liquid outlet channel 3012, the size of the liquid 400 to be measured in the long side direction is reduced by the action of the shearing force to be close to the size of the liquid 400 in the short side direction, so as to form approximately spherical stable micro-droplets. It can be understood that, according to the magnitude of the shearing force provided by the stabilizing liquid 500, the long side dimension of the rectangular cross section of the liquid outlet channel 3012 should be greater than or equal to three times of the short side dimension, so that the dimension of the liquid 400 to be measured discharged from the liquid outlet channel 3012 in the long side direction is close to the dimension in the short side direction under the action of the shearing force in the long side direction. In a specific embodiment, the size of the long side of the rectangular cross section of the outlet liquid flow channel 3012 is in the range of 9 μm to 90 μm, and the size of the wide side of the rectangle is in the range of 3 μm to 30 μm. Within the above size range, the size of the formed micro-droplets can meet the corresponding test requirements.
In one embodiment, the length of the liquid outlet channel 3012 is in the range of 100 μm to 300 μm. It can be understood that when the length of the liquid outlet channel 3012 is small, it is not beneficial to the formation of micro-droplets, and when the size of the liquid outlet channel 3012 is large, it is not beneficial to the miniaturization of the micro-droplet generation structure 300. In a specific embodiment, the length of the liquid outlet channel 3012 is in a range of 100 μm to 300 μm, and in the above length range, the requirement of miniaturization of the droplet generation structure 300 can be ensured while the droplets can be generated effectively.
In one embodiment, a support 34 is further disposed in the diffusion chamber 302, one end of the support 34 is connected to the inner surface of the covering portion 32, and the other end of the support 34 is connected to the inner surface of the carrying portion 31. It can be understood that the supporting body 34 is supported between the covering portion 32 and the carrying portion 31, so that the inner surfaces of the covering portion 32 and the carrying portion 31 are not attached together, and the diffusion chamber 302 is ensured to have a sufficient diffusion space, so that the liquid 400 to be measured can be effectively diffused after entering the diffusion chamber 302.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. A micro-device for generating liquid drops for digital PCR is characterized by comprising an outer container, an inner container and a micro-liquid drop generating structure, wherein the outer container is provided with a first containing cavity, a stabilizing liquid is contained in the first containing cavity, the micro-liquid drop generating structure is immersed in the stabilizing liquid, and the inner container is contained in the first containing cavity;
the inner container is provided with a second containing cavity for containing liquid to be detected, the micro-droplet generation structure comprises a micro-droplet generation flow channel, the inner container is connected with the micro-droplet generation structure, and the second containing cavity is communicated with the micro-droplet generation flow channel;
and the inner container is also provided with an air inlet pipeline so as to communicate the second accommodating cavity with the outside, outside air enters the second accommodating cavity through the air inlet pipeline so as to push the liquid to be detected to enter the micro-droplet generating structure and pass through the micro-droplet generating flow channel to form micro-droplets, and the micro-droplets are discharged into the stabilizing liquid through the micro-droplet generating flow channel.
2. The droplet generation micro-device according to claim 1, wherein a liquid inlet pipe is connected between the inner container and the micro-droplet generation structure, the micro-droplet generation structure further comprises a diffusion chamber, a liquid inlet end of the diffusion chamber is communicated with the second accommodating chamber through the liquid inlet pipe, a liquid outlet end of the diffusion chamber is communicated with the micro-droplet generation flow passage, and a cross-sectional area of the diffusion chamber at the liquid outlet end is larger than a cross-sectional area of the diffusion chamber at the liquid inlet end.
3. The droplet generation micro-device according to claim 2, wherein the number of the diffusion chambers is plural, the liquid inlet end of each diffusion chamber is communicated with the liquid inlet pipeline, and the liquid outlet end of each diffusion chamber is connected with the micro-droplet generation flow channel.
4. The droplet generation microdevice of claim 2, wherein the number of the droplet generation channels is plural, the plural droplet generation channels are arranged side by side, and the plural droplet generation channels are all communicated with the diffusion chamber.
5. The droplet generation micro-device of claim 4, wherein the droplet generation structure further comprises a plurality of partitions arranged side by side and at intervals, a gap between every two adjacent partitions forms one droplet generation channel, the droplet generation channel comprises a liquid inlet channel and a liquid outlet channel which are communicated, and the cross-sectional area of the liquid inlet channel is larger than that of the liquid outlet channel.
6. The droplet generation micro-device of claim 5, wherein the cross section of the liquid outlet channel is a circle, and the diameter of the circle is in the range of 3 μm to 90 μm; and/or the cross section of the liquid outlet flow channel is rectangular, the size range of the long side of the rectangle is 9-90 μm, and the size range of the wide side of the rectangle is 3-30 μm.
7. The droplet generation micro-device of claim 6, wherein the length of the outlet channel is in the range of 100 μm-300 μm.
8. The droplet generation micro-device according to claim 2, wherein the micro-droplet generation structure comprises a bearing portion and a covering portion, the covering portion and the bearing portion enclose a diffusion chamber, a support is further disposed in the diffusion chamber, one end of the support is connected to an inner surface of the covering portion, and the other end of the support is connected to an inner surface of the bearing portion.
9. The droplet-generating microdevice of claim 1, wherein the outer container further comprises a vent communicating the first chamber with the exterior of the outer container for venting gas from the first chamber.
10. The droplet generation microdevice of claim 1, wherein the solution to be measured is poorly soluble or insoluble in the stabilizing solution, and the density of the solution to be measured is greater than or less than the density of the stabilizing solution.
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