A multi-hole air pressure precision sample adding box

Abstract

The present invention discloses a multi-hole air pressure precision sample loading box. The technical key points are as follows: it includes a box body that can be fitted and embedded in a matching sample stage, a microfluidic channel is provided inside the box body, a sample loading slot, a waste liquid slot, and a sample collection slot are recessed on the upper surface of the box body, and the waste liquid slot and the sample collection slot are connected to the microfluidic channel. The waste liquid slot and the sample collection slot are respectively connected to an air pump, and there are multiple groups of sample loading slots; the lower surface of the box body is provided with liquid outlet holes and liquid extraction holes connected to the microfluidic channel, and each group of liquid outlet holes and liquid extraction holes is connected to each group of sample loading slots, waste liquid slots, and sample collection slots on the upper surface of the box body through the microfluidic channel; the lower surface of the box body is provided with a liquid outlet groove that surrounds each group of liquid outlet holes; the lower surface of the box body is provided with a liquid extraction groove located outside the liquid extraction hole, and a flow channel for connection is provided between the liquid extraction groove and the liquid outlet groove. The box body has the functions of automatically and precisely adding reagents, collecting waste liquid, and preparing samples, and has the advantages of no cross contamination and secondary contamination due to the disposable connection method.

Description

Technical Field

[0001] The invention relates to the technical field of micro-droplets, micro-fluidic preparation and reagent mixing detection, and more particularly to a multi-hole air pressure type precision sample adding box. Background Art

[0002] Microfluidics technology is one of the important branches of MEMS technology and one of the most rapidly developing multidisciplinary cross-disciplinary cutting-edge technologies. It has important applications in life sciences, clinical medicine, chemistry and chemical engineering, pharmaceuticals, food hygiene, environmental testing and monitoring, information science and signal detection, and other disciplines.

[0003] Microfluidics technology typically utilizes microanalytical devices as the vehicle for its implementation, and microfluidic chips are the fastest-growing of these devices. Microfluidic chips utilize MEMS technology to fabricate various microstructures, such as microchannels, microreaction cells, and microelectrodes, onto silicon, quartz, glass, or polymer substrates. These microchannels then interconnect components such as micropumps, microvalves, microreservoirs, and microdetectors, enabling fluid transport, control, detection, and monitoring. This allows for the integration of a comprehensive on-chip microanalysis system encompassing dilution, reagent addition, sampling, reaction, separation, dispersion, detection, and monitoring. Microfluidic chips typically range from a few to tens of square centimeters in area, with microchannel dimensions typically in the micrometer or near-millimeter range. When many chemical processes are carried out in microfluidic chips, the tiny volume brings many advantages: if the microchannel size is reduced by one order of magnitude, the reagent usage will be reduced by three orders of magnitude; the diffusion rate of the fluid in the microchannel will be increased by two orders of magnitude, thereby greatly improving the reaction speed; at the same time, microfluidic chips also have the advantages of low cost, batch manufacturing, simple operation, good repeatability, and high reliability.

[0004] However, currently in the field of microfluidic laboratories, most methods are to directly insert a rigid catheter into the sample hole of an elastic polydimethylsiloxane (PDMS) microfluidic chip, or to stick the catheter to the fluid inlet and outlet of a rigid plastic or glass chip. Although the above methods have low material costs, they are cumbersome to operate. Frequent plugging and unplugging of the chip can damage the microfluidic chip interface, PDMS fragments can flow into the channel and cause it to clog, and the sticking process is also prone to clogging. Moreover, due to the small mass and volume of the microfluidic chip, it is easily affected by the connection pipes and can fall over. In addition, in biological experiments, array structures are often used, and the traditional sampling method has scattered pipes and is difficult to manage. Some existing chip fixtures are suitable for rigid microfluidic chips, such as glass chips, PMMA chips, etc. For microfluidic chips made of soft PDMS materials, elastic ring sealing and compression sealing are often used, but excessive compression force can cause microchannel clogging; if the compression force is too small, when a high pressure is introduced into the chip, the liquid will overflow from the sealing surface.

[0005] Therefore, it is of great significance to design a multifunctional sample loading kit that can be used for carrying, fusing, discharging waste liquid, fixing and sealing the liquid inside the microfluidic chip. Summary of the Invention

[0006] In view of the shortcomings of the existing technology, the purpose of the present invention is to provide a multi-porous pneumatic precision sample adding box with the functions of automatic precision adding reagents, collecting waste liquids, and preparing samples. Through a disposable connection method, it has the advantages of no cross contamination and secondary contamination.

[0007] The above-mentioned object of the present invention is achieved through the following technical solutions: a multi-hole air pressure precision loading box, comprising a box body, the box body can be fitted and embedded in a matching sample stage, a microfluidic channel is provided inside the box body, and a loading groove, a waste liquid groove and a sample collection groove are recessed on the upper surface of the box body and connected to the microfluidic channel, the waste liquid groove and the sample collection groove are respectively connected to an air pump, and the loading groove is provided with multiple groups;

[0008] The lower surface of the box body is provided with a liquid outlet and a liquid extraction hole connected to the microfluidic channel. The liquid outlet holes are provided in at least one group and are arranged in a cluster. Each group of the liquid outlet holes and the liquid extraction holes is connected to each group of sample addition slots, waste liquid slots, and sample collection slots on the upper surface of the box body through the microfluidic channel. The lower surface of the box body is extended with a liquid outlet groove surrounding each group of liquid outlet holes;

[0009] A liquid extraction groove located outside the liquid extraction hole is extended from the lower surface of the box body, and a flow channel for connection is provided between the liquid extraction groove and the liquid outlet groove.

[0010] By adopting the above technical solution, the box body is designed to be disposable, which can prevent secondary contamination of the research reagents and simplify the fusion and loading of various reagents. The samples required in the single-cell automatic operation process are injected into the reagent liquid from each loading groove in sequence according to the process, and the reagents are infiltrated through the microfluidic channel inside the box body. The waste liquid tank and the top of the sample collection tank are connected to the air pump, and the positive air pressure applied by the air pump is used to precisely press the reagents in the loading groove to each group of liquid outlet holes. The groups of liquid outlet holes are gathered to confine the reagents in the liquid outlet groove area, preventing various reagent liquids from penetrating to the bottom of the box body, causing cross-contamination and secondary contamination of biosafety. The reagents are introduced into the liquid extraction groove through the flow channel or the external chip in the liquid outlet groove, and then negative air pressure is applied by the air pump. The waste liquid and the final sample generated in the single-cell automatic operation process are drawn back to the waste liquid tank and the sample collection groove on the upper surface of the box body through the liquid extraction groove and the liquid extraction hole according to the negative pressure principle.

[0011] Preferably, the side surface of the liquid outlet groove is cut off with an air hole groove communicating with the outside of the box body.

[0012] By adopting the above technical solution, the reagent inside the microfluidic channel is pressurized directly through the positive pressure of the air pump. There may be a certain bubble segment in the microfluidic channel. Through the air hole grooves added on the side, excess air can be discharged under the action of positive and negative pressure, so that a complete microfluidic column is formed in the channel, further optimizing the structure.

[0013] Preferably, a plurality of groups of supporting feet for supporting the box body are provided on the lower surface of the box body, and the height of the supporting feet is greater than or equal to the depth of the liquid extraction groove and the liquid discharge groove.

[0014] By adopting the above technical solution, the supporting feet cooperate with the sample table below to further play a limiting role, while providing space for the liquid extraction groove and the liquid discharge groove.

[0015] Preferably, the edges of each group of the sample adding slots, waste liquid slots, and sample collecting slots are respectively extended with a sample adding boss, a waste liquid boss, and a sample collecting boss located on the upper surface of the box body.

[0016] By adopting the above technical solution, since the box body needs to be covered with a sealing cover after loading the reagents, the sample loading boss, waste liquid boss and sample collection boss are designed to cooperate with the sealing cover above to achieve a sealed space.

[0017] Preferably, the sample adding groove, waste liquid groove, sample collecting groove, liquid outlet groove and liquid extraction groove can be a microstructure of a cone, a cylinder, a cubic column, a quadrangular column or other polygonal column.

[0018] By adopting the above technical solution, various shapes can be used for this structure.

[0019] Preferably, the diameters of the sample adding tank, waste liquid tank and sample collecting tank are 1-30 mm.

[0020] By adopting the above technical solution, sample adding troughs, waste liquid troughs, and sample collecting troughs of different diameters can carry reagents in different required amounts, and the diversified structure increases the variety of choices.

[0021] Preferably, the heights of the sample adding boss, waste liquid boss and sample collecting boss are 0.1-1 mm.

[0022] By adopting the above technical solution, the micro height is convenient for sealing.

[0023] Preferably, the volume of the sample in each group of sample loading slots pressed into the liquid outlet through the microfluidic channel is 5-1500 μL.

[0024] By adopting the above technical solution, the degree of volume change is relatively large, and precise volume injection can be achieved through precise air pressure injection.

[0025] In summary, the beneficial effects of the present invention are:

[0026] 1. The integrated design of reagent preparation in the single-cell microfluidic channel is not available in the prior art. This application has achieved the precise preparation and sample addition function, waste liquid collection function and sample preparation function through originality from scratch;

[0027] 2. Through the combination of positive and negative pressure and multiple groups of functional slots, the reagent circulation and automated preparation in the microfluidic channel are realized, eliminating the need for manual multi-step operation and intervention, and facilitating the equipment and test process;

[0028] 3. The sample loading groove convex ring, waste liquid convex platform, sample collection convex platform on the box body cooperate with the sealing cover on the top to achieve a good sealing effect during the single-cell automatic operation process. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 A schematic diagram showing the upper surface structure of the box body of this embodiment;

[0030] Figure 2 A schematic diagram showing the lower surface structure of the box body of this embodiment.

[0031] Figure numerals: 1, box body; 2, sample adding slot; 21, sample adding boss; 3, waste liquid slot; 31, waste liquid boss; 4, sample collecting slot; 41, sample collecting boss; 5, liquid outlet; 6, liquid extraction hole; 61, liquid extraction groove; 7, liquid outlet groove; 71, air hole slot; 8, supporting foot. DETAILED DESCRIPTION

[0032] The specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings.

[0033] Example: Figure 1 As shown, a porous air pressure precision sample loading box includes a box body 1. The box body 1 is made of PC, PMMA, COC, PS and other materials with high strength, high elastic coefficient, high impact strength, wide operating temperature range and hydrophobicity greater than 70 degrees. The box body 1 can be fitted and embedded in the sample table matched with it. A microfluidic channel is provided inside the box body 1. A sample loading slot 2, a waste liquid slot 3 and a sample collecting slot 4 connected to the microfluidic channel are recessed on the upper surface of the box body 1. The waste liquid slot 3 and the sample collecting slot 4 are respectively connected to an air pump. The sample loading slot 2 is provided with multiple groups. In this embodiment, the sample loading slot 2 is set to 8 groups, of which 7 groups have smaller diameters and one group has a larger diameter; a liquid outlet 5 and a liquid extraction hole 6 connected to the microfluidic channel are provided on the lower surface of the box body 1. The liquid outlet holes 5 are provided with at least one group and are clustered and arranged. In this embodiment, the number of liquid outlet holes 5 is set to 7. Each group of liquid outlet holes 5 and liquid extraction holes 6 is connected with each group of sample loading slots 2, waste liquid slots 3 and sample collecting slots 4 on the upper surface of the box body 1 through the microfluidic channel. A liquid outlet groove 7 surrounding each group of liquid outlet holes 5 is extended from the lower surface of the box body 1; a liquid extraction groove 61 located outside the liquid extraction hole 6 is extended from the lower surface of the box body 1, and a flow channel for connection is provided between the liquid extraction groove 61 and the liquid outlet groove 7.

[0034] like Figure 1 、 Figure 2 As shown, the side of the liquid outlet groove 7 is interrupted by an air hole groove 71 that connects to the outside of the box body 1. The positive pressure of the air pump is directly used to pressurize the reagent inside the microfluidic channel. There may be a certain amount of air bubbles in the microfluidic channel. The air hole groove 71 added on the side can expel excess air during the positive and negative pressure effects, forming a complete microfluidic column in the channel, further optimizing the structure.

[0035] like Figure 1 、 Figure 2 As shown, the lower surface of the box body 1 is provided with multiple groups of support feet 8 for supporting the box body 1. In this embodiment, the support feet 8 are respectively located at the four corners of the bottom of the box body 1, and the height of the support feet 8 is greater than or equal to the depth of the liquid extraction groove 61 and the liquid outlet groove 7. The support feet 8 cooperate with the sample stage below to further play a limiting role, while providing space for the liquid extraction groove 61 and the liquid outlet groove 7. The edges of each group of sample loading grooves 2, waste liquid grooves 3, and sample receiving grooves 4 are respectively extended with sample loading bosses 21, waste liquid bosses 31, and sample receiving bosses 41 located on the upper surface of the box body 1. The sample loading grooves 2, waste liquid grooves 3, and sample receiving grooves 4 can also be directly a single plane. Because the box body 1 needs to be covered with a sealing cover after loading the reagents, the sample loading bosses 21, waste liquid bosses 31, and sample receiving bosses 41 are designed to cooperate with the sealing cover above to achieve a sealed space.

[0036] like Figure 1 、 Figure 2 As shown, the sample loading groove 2, waste liquid groove 3, sample receiving groove 4, liquid outlet groove 7, and liquid extraction groove 61 can be one of the four microstructures of conical, cylindrical, cubic, and quadrangular shapes. This embodiment adopts a cylindrical structure, and the diameter of the sample loading groove 2, waste liquid groove 3, and sample receiving groove 4 is 1-30mm. Sample loading grooves 2, waste liquid grooves 3, and sample receiving grooves 4 of different diameters can be used to carry reagents with different requirements, and the diversified structure increases the variety of options. The height of the sample loading boss 21, waste liquid boss 31, and sample receiving boss 41 is 0.1-1mm. The volume of each group of sample loading grooves 2 pressed into the liquid outlet hole 5 through the microfluidic channel is 5-1500μL. The degree of volume change is large, and precise volume injection can be achieved through precise air pressure injection.

[0037] A buffer stirring and mixing zone is provided on one of the microfluidic channels in the box body 1 for cooperating with pressure control to achieve stirring and mixing of the liquid / suspension therein.

[0038] Working principle and process: This box body 1 is a disposable design, which can prevent secondary contamination of the research reagents and simplify the fusion and loading of various reagents. The samples required in the single-cell automatic operation process are injected into the reagent liquid from each sample loading groove 2 in sequence according to the process, and the reagents are infiltrated through the microfluidic channel inside the box body 1. The waste liquid tank 3 and the top of the sample collection tank 4 are connected to an air pump. The air pump applies positive air pressure to accurately press the reagents in the sample loading groove 2 to each group of liquid outlet holes 5. The groups of liquid outlet holes 5 are gathered to confine the reagents in the liquid outlet groove 7 area to prevent various reagent liquids from penetrating into the bottom of the box body 1, causing cross-contamination and secondary contamination of biological safety. The reagents are introduced into the liquid extraction groove 61 through the flow channel or the external chip in the liquid outlet groove 7, and then negative air pressure is applied by the air pump. The waste liquid and the final sample generated in the single-cell automatic operation process are drawn back to the waste liquid tank 3 and the sample collection groove 4 on the upper surface of the box body 1 through the liquid extraction groove 61 and the liquid extraction hole 6 through the negative pressure principle.

[0039] This specific embodiment is merely an explanation of the present invention and is not intended to limit the present invention. After reading this specification, those skilled in the art may make non-creative modifications to this embodiment as needed. However, as long as such modifications are within the scope of the claims of the present invention, they are protected by patent law.

Claims

1. A multi-hole air pressure precision sample loading box, characterized by: The invention comprises a box body (1), wherein the box body (1) can be fitted and embedded in a sample stage matched therewith, wherein a microfluidic channel is provided inside the box body (1), wherein a sample adding slot (2), a waste liquid slot (3) and a sample collecting slot (4) communicating with the microfluidic channel are provided in a recessed manner on the upper surface of the box body (1), wherein the waste liquid slot (3) and the sample collecting slot (4) are respectively connected to an air pump, and wherein the sample adding slot (2) is provided with multiple groups; wherein a liquid outlet (5) and a liquid extraction hole (6) communicating with the microfluidic channel are provided on the lower surface of the box body (1), wherein the liquid outlet (5) is provided with a plurality of liquid extraction holes (6) communicating with the microfluidic channel; ... There is at least one group of liquid outlet holes (5) and liquid extraction holes (6) arranged in a cluster, each group of which is connected to each group of sample loading slots (2), waste liquid slots (3), and sample collection slots (4) on the upper surface of the box body (1) through a microfluidic channel, and a liquid outlet groove (7) is extended from the lower surface of the box body (1) to surround each group of liquid outlet holes (5); a liquid extraction groove (61) is extended from the lower surface of the box body (1) and is located outside the liquid extraction holes (6), and a flow channel for connection is provided between the liquid extraction groove (61) and the liquid outlet groove (7); wherein, Positive air pressure is applied by the air pump to precisely press the reagents in the loading tank to each group of liquid outlet holes. The groups of liquid outlet holes are gathered to confine the reagents in the liquid outlet groove area. The reagents are introduced into the liquid extraction groove through the flow channel or external chip in the liquid outlet groove. Then negative air pressure is applied by the air pump. The waste liquid and the final sample generated during the single-cell automatic operation process are drawn back to the waste liquid tank and sample collection tank on the upper surface of the box through the liquid extraction groove and the liquid extraction holes according to the negative pressure principle.

2. The multi-hole air pressure precision sample loading box according to claim 1, characterized in that: The side of the liquid outlet groove (7) is cut off to form an air hole groove (71) communicating with the outside of the box body (1).

3. The multi-hole air pressure precision sample loading box according to claim 1, characterized in that: The lower surface of the box body (1) is provided with a plurality of groups of supporting feet (8) for supporting the box body (1), and the height of the supporting feet (8) is greater than or equal to the depth of the liquid extraction groove (61) and the liquid outlet groove (7).

4. The multi-hole air pressure precision sample loading box according to claim 3, characterized in that: The edges of each group of the sample adding slots (2), waste liquid slots (3), and sample collecting slots (4) are respectively extended with a sample adding boss (21), a waste liquid boss (31), and a sample collecting boss (41) located on the upper surface of the box body (1).

5. The multi-hole air pressure precision sample loading box according to claim 1, characterized in that: The sample adding groove (2), waste liquid groove (3), sample collecting groove (4), liquid outlet groove (7), and liquid extraction groove (61) can be a microstructure of a cone, a cylinder, a cubic column, a quadrangular column, or other polygonal columns.

6. The multi-hole air pressure precision sample loading box according to claim 1, characterized in that: The diameters of the sample adding tank (2), the waste liquid tank (3) and the sample collecting tank (4) are 1-30 mm.

7. The multi-hole air pressure precision sample loading box according to claim 4, characterized in that: The heights of the sample adding boss (21), the waste liquid boss (31), and the sample collecting boss (41) are 0.1-1 mm.

8. The multi-hole air pressure precision sample loading box according to claim 1, characterized in that: The volume of the sample in each group of the sample loading slots (2) pressed into the liquid outlet (5) through the microfluidic channel is 5-1500 μL.

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