Pipetting device suitable for automated workstation and method for using same

Abstract

The present invention relates to the technical field of preparation and transfer of a trace amount of liquid and / or reagent, in particular to a pipetting device suitable for an automated workstation and a method for using same, and particularly to a micropipetting device suitable for automated formulation. Further provided in the present invention are a dispensing method for automated formulation of a liquid, in particular a trace amount of liquid, by means of the pipetting device, and a method for using the pipetting device.

Description

Liquid transfer device suitable for automated workstation and method of use thereof Technical Field

[0001] The present invention relates to the technical field of preparation and transfer of trace samples, liquids and / or reagents, and in particular to a micro-liquid automated configuration pipetting device, and more particularly to a micro-liquid automated configuration pipetting device suitable for an automated workstation and a method for using the same. Background Art

[0002] Liquid handling devices are used in a wide range of industrial and laboratory analyses to transfer liquids. When performing liquid handling operations on automated workstations, pipettes are typically used to aspirate, transfer, pipette, and dispense liquids. To ensure accurate pipetting, precision-engineered liquid handling devices are essential.

[0003] When performing pipetting operations in an automated workstation, reagents are often pre-packed in containers such as reagent troughs, reagent tubes or well plates. The bottom area of ​​these containers is relatively large. In order to ensure that the pipette can accurately detect the liquid level, to ensure that the tip can contact the liquid, and to accurately absorb the liquid, it is necessary to add liquid that is larger than the actual required volume. This extra volume of liquid is the so-called dead volume. In actual operation, there is generally a dead volume of about 10μL, but in some application scenarios, such as in the library construction process, sometimes the amount of certain enzymes or catalysts added is less than 1μL, which results in a dead volume larger than the actual required volume, resulting in a serious waste of reagents. Dead volume is a problem often encountered when working in automated workstations, and there is currently no particularly good solution.

[0004] CN110385153B (Pipette Device, Pipette Tip Connector, and Pipette Tip: Device and Method) describes a Hamilton automated workstation pipette device. While ingeniously designed, most current automated workstation pipette devices struggle to process extremely small amounts of liquid, less than 1 μL. This limits their application in areas such as library construction.

[0005] In order to store reagents more conveniently, CN107557873A proposes a single-person library construction kit, which includes a reagent strip and reagents required for library construction. The reagent strip is provided with multiple storage holes, and the storage holes include a bottom, a lower part and an upper part in sequence. The bottom is closed, the lower parts of the multiple storage holes are independent of each other, and the upper parts of adjacent storage holes are connected to each other. The reagents required for library construction are placed in the storage holes. This invention only processes one sample, reducing the loss of enzyme activity of the tool enzyme due to repeated freezing and thawing; eliminating the risk of inconsistency between the actual amount and the theoretical amount caused by liquid reagents adhering to the wall during storage and transportation; and minimizing the manual operation steps of the operator, reducing the probability of misoperation, reducing the need for operator training, and ensuring the quality and reliability of sample preparation. Although this kit can realize the single-person use of library construction reagents, it still does not propose measures to solve the dead volume problem.

[0006] In the process of automated library construction, insufficient pipetting accuracy will affect the stability and reliability of the results. In particular, when using commercial kits for automated library construction, it is difficult to solve the problem of pipetting accuracy by increasing the reaction system, which will make the results unreliable and cause the number of reactions to be too small. In the process of automated library construction, dead volume will cause serious waste of reagents. In addition, the process of automated library construction involves many steps, which require the use of reaction mixtures. Most reaction mixtures need to be prepared and used immediately and cannot be mixed together in advance, which will require a large number of plate positions and consumables.

[0007] In order to solve the problem of single-dose pre-packaging of reagents that cannot be pre-mixed in advance, CN108641899A discloses a pre-split cover for a centrifuge tube, comprising a accommodating cavity, the accommodating cavity is divided into 2 to 6 mutually unconnected areas, and the size of the accommodating cavity opening is adapted to the caliber of the centrifuge tube. The invention also discloses a centrifuge tube, comprising a tube body and a pre-split cover, the accommodating cavity of the pre-split cover is divided into 2 to 6 mutually unconnected areas. The invention also discloses a reagent pre-split storage and transport device, comprising a pre-split cover and a cover seal, the accommodating cavity of the pre-split cover is divided into 2 to 6 mutually unconnected areas, and the size of the cover seal is adapted to the caliber of the pre-split cover opening. The invention solves the problem of single-dose pre-packaging of reagents that cannot be pre-mixed in advance, and at the same time avoids the problem of large manual operation workload and easy contamination during batch liquid separation before the reaction, ensures that multiple reaction units in the same batch start reactions simultaneously in a short period of time, homogenizes the reaction time of each centrifuge tube and prevents non-specific reactions caused by long operation time. Although this patent solves the problem of single-dose pre-packaging when reagents cannot be pre-mixed in advance, it still cannot solve the previously mentioned problems of pipetting accuracy, dead volume, and the need for automated reagent configuration to save plate space.

[0008] Therefore, there has been a need in the art for a device and method that can simultaneously solve the problems of reagent pipetting accuracy, reducing dead volume, pre-packaging and automated configuration to reduce plate space and consumables consumption.

[0009] Summary of the Invention

[0010] The present invention aims to solve the problems existing in the pipetting operation of the existing automated workstation and provides a newly designed pipetting device and its use method, which solves a series of problems such as insufficient precision of the automated workstation, insufficient operation board space, and excessive dead volume.

[0011] In particular, the present invention solves the following deficiencies of existing automated workstations:

[0012] 1. Current automated workstations have difficulty accurately configuring extremely small amounts of liquid (≤1μL); however, during automated library construction, the amount of enzymes or catalysts added is often ≤1μL.

[0013] 2. When preparing trace liquids, the current automated workstation has a large dead volume to ensure the accuracy of liquid aspiration, which will cause waste of reagents and increase reagent costs.

[0014] 3. The board space of the automated workstation is limited, and the library construction process requires many steps. This may result in the automated workstation being unable to execute the entire library construction process at one time due to the limited board space.

[0015] 4. During the library construction process, many reaction systems (e.g., liquid mixtures containing various reagents) cannot be pre-mixed in advance and need to be prepared on-the-fly.

[0016] The present invention uses a specially designed micropipette device to achieve precise addition of samples to reaction systems (e.g., liquid mixtures containing various reagents), reduce dead volume, store multiple reagents simultaneously, save plate space, and automatically configure and use reagents, allowing more operational processes to be implemented on an automated workstation.

[0017] According to a first aspect of the present invention, a micropipette device suitable for an automated workstation is provided, the device comprising a connector (II), a microstorage tube (I) and a sealing portion (III and / or V). The connector can be connected to a pipette so that the micropipette device can act as a pipette tip, and the pipette can be used to install and separate the micropipette device, and to perform operations such as blowing and pumping the liquid in the microstorage tube. The microstorage tube is used to store liquids, such as pre-packaged liquids, which contain, for example, required reagents. The sealing portion is used to seal the pre-packaged liquid (IV), which contains, for example, required reagents.

[0018] According to a second aspect of the present invention, a micropipetting packaging method suitable for an automated workstation is provided:

[0019] a. Accurately dispensing extremely small amounts of liquids can be achieved by dispensing using external dispensing equipment (e.g., micropumps (including microsyringe pumps, gear pumps, peristaltic pumps, and piston pumps), microdispensers, and automatic filling machines);

[0020] b. Using an alternating packaging method, the reaction solution (including, for example, the required reagents) and the isolation solution are alternately stored in micro-storage tubes to avoid pre-mixing of the reaction solution in advance, so that the same pipetting device can contain multiple reaction solutions for easy storage and use;

[0021] c. By pre-packaging multiple reaction solutions (including, for example, required reagents) in a prescribed volume, it is only necessary to discharge all the reaction solutions from the micro-storage tube with a pipette when using the solution, without having to aspirate each reaction solution from a separate reagent tube, thereby reducing the waste caused by dead volume.

[0022] The third aspect of the present invention provides a method for using a micropipette suitable for an automated workstation:

[0023] a. By pre-packaging multiple reaction solutions (including, for example, required reagents) in alternating volumes according to specified volumes, the required multiple reaction solutions can be stored in the same micro-storage tube. When needed, all reaction solutions are simply discharged from the micro-storage tube using a pipette. The various reagents are then combined to form a mixed solution, thus enabling the automated configuration and use of micro-reaction solutions. This saves plate space and enables more operational processes to be implemented on the automated workstation.

[0024] b. The reaction solution is pre-packaged according to the amount using a high-precision external device, thus solving the problem of insufficient pipette accuracy.

[0025] According to a first aspect of the present invention, a micropipette device for an automated workstation is provided, the device comprising a connector (II), a microstorage tube (I) and a first sealing portion (III), wherein the first sealing portion is connected to a first interface (1) on the microstorage tube to seal the microstorage tube, the connector is connected to a second interface (2) on the microstorage tube, and the connector is used to connect to a pipette of the automated workstation via a third interface (3).

[0026] In one embodiment, the micropipette device is a micropipette device comprising a pre-packaged liquid (IV), preferably, the pre-packaged liquid is contained in the micro storage tube.

[0027] In one embodiment, the connector can be connected to a pipette of an automated workstation. Specifically, the connector (II) can be connected to the pipette (VI), allowing the micropipette device to function as a pipette tip, enabling the micropipette device to be installed and separated, and to perform operations such as aspiration, pipetting, discharge, and transfer of liquid in a microstorage tube.

[0028] In one embodiment, the inner diameter of the micro storage tube is in the range of about 0.2 mm to about 5 mm, preferably, in the range of about 0.5 mm to about 2.5 mm.

[0029] In a preferred embodiment, the inner diameter of the micro storage tube can be, for example, about 0.2 mm, about 0.3 mm, about 0.4 mm, about 0.5 mm, about 0.6 mm, about 0.7 mm, about 0.8 mm, about 0.9 mm, about 1 mm, about 1.1 mm, about 1.2 mm, about 1.3 mm, about 1.4 mm, about 1.5 mm, about 1.6 mm, about 1.7 mm, about 1.8 mm, about 1.9 mm, about 2.0 mm, about 2.1 mm, about 2.2 mm, about 2.3 mm, about 2.4 mm, about 2.5 mm , about 2.6 mm, about 2.7 mm, about 2.8 mm, about 2.9 mm, about 3.0 mm, about 3.1 mm, about 3.2 mm, about 3.3 mm, about 3.4 mm, about 3.5 mm, about 3.6 mm, about 3.7 mm, about 3.8 mm, about 3.9 mm, about 4.0 mm, about 4.1 mm, about 4.2 mm, about 4.4 mm, about 4.4 mm, about 4.5 mm, about 4.6 mm, about 4.7 mm, about 4.8 mm, about 4.9 mm or about 5 mm, or a sub-range consisting of any value within these ranges.

[0030] In one embodiment, the length of the micro storage tube is in the range of about 1 cm to about 20 cm, preferably, in the range of about 2 cm to about 10 cm.

[0031] In a preferred embodiment, the length of the micro storage tube can be, for example, about 1 cm, about 1.1 cm, about 1.2 cm, about 1.3 cm, about 1.4 cm, about 1.5 cm, about 1.6 cm, about 1.7 cm, about 1.8 cm, about 1.9 cm, about 2 cm, about 2.1 cm, about 2.2 cm, about 2.3 cm, about 2.4 cm, about 2.5 cm, about 2.6 cm, about 2.7 cm, about 2.8 cm, about 2.9 cm, about 3 cm, about 3.1 cm, about 3. 2cm, about 3.3cm, about 3.4cm, about 3.5cm, about 3.6cm, about 3.7cm, about 3.8cm, about 3.9cm, about 4cm, about 4.1cm, about 4.2cm, about 4.3cm, about 4.4cm, about 4.5cm, about 4.6cm, about 4.7cm, about 4.8cm, about 4.9cm, about 5cm, about 5.1cm, about 5.2cm, about 5.3cm, about 5.4cm, about 5.5cm, about 5.6cm, about 5.7cm, about 5 .8cm, about 5.9cm, about 6cm, about 6.1cm, about 6.2cm, about 6.3cm, about 6.4cm, about 6.5cm, about 6.6cm, about 6.7cm, about 6.8cm, about 6.9cm, about 7cm, about 7.1cm, about 7.2cm, about 7.3cm, about 7.4cm, about 7.5cm, about 7.6cm, about 7.7cm, about 7.8cm, about 7.9cm, about 8cm, about 8.1cm, about 8.2cm, about 8.3cm, about 8. 4cm, about 8.5cm, about 8.6cm, about 8.7cm, about 8.8cm, about 8.9cm, about 9cm, about 9.1cm, about 9.2cm, about 9.3cm, about 9.4cm, about 9.5cm, about 9.6cm, about 9.7cm, about 9.8cm, about 9.9cm, about 10cm, about 10.1cm, about 10.2cm, about 10.3cm, about 10.4cm, about 10.5cm, about 10.6cm, about 10.7cm, about 10.8cm, About 10.9cm, about 11cm, about 11.1cm, about 11.2cm, about 11.3cm, about 11.4cm, about 11.5cm, about 11.6cm, about 11.7cm, about 11.8cm, about 11.9cm, about 12cm, about 12.1cm, about 12.2cm, about 12.3cm, about 12.4cm, about 12.5cm, about 12.6cm, about 12.7cm m, about 12.8cm, about 12.9cm, about 13cm, about 13.1cm, about 13.2cm, about 13.3cm, about 13.4cm, about 13.5cm, about 13.6cm, about 13.7cm, about 13.8cm, about 13.9cm, about 14cm, about 14.1cm, about 14.2cm, about 14.3cm, about 14.4cm, about 14.5cm, about 14.6cm, about 14.7cm, about 14.8cm, about 14.9cm, about 15cm, about 15.1cm, about 15.2cm, about 15.3cm, about 15.4cm, about 15.5cm, about 15.6cm, about 15.7cm, about 15.8cm, about 15.9cm, about 16cm, about 16.1cm, about 16.2cm, about 16.3cm, about 16.4cm, about 16.5cm, about 16.6cm, about 16.7cm, about 16.8cm, about 16.9cm, about 17cm, about 17.1cm, about 17.2cm, about 17.3cm, about 17.4cm , about 17.5 cm, about 17.6 cm, about 17.7 cm, about 17.8 cm, about 17.9 cm, about 18 cm, about 18.1 cm, about 18.2 cm, about 18.3 cm, about 18.4 cm, about 18.5 cm, about 18.6 cm, about 18.7 cm, about 18.8 cm, about 18.9 cm, about 19 cm, about 19.1 cm, about 19.2 cm, about 19.3 cm, about 19.4 cm, about 19.5 cm, about 19.6 cm, about 19.7 cm, about 19.8 cm, about 19.9 cm or about 20 cm, or sub-ranges consisting of any values ​​within these ranges.

[0032] According to the micro-pipette device provided by the present invention, the liquid stored in the micro-storage tube remains in layers and will not be mixed in the micro-storage tube due to disturbance or density difference.

[0033] In one embodiment, the liquid may be one liquid, or may be a mixture comprising a plurality of liquids (for example, comprising 4-liquid A1, 5-liquid B1, 6-liquid A2, 7-liquid B2, 8-liquid A3, 9-liquid B3, 10-mixed liquid A, 11-mixed liquid B, see FIG4 ).

[0034] In one embodiment, the mixture comprises a water-immiscible liquid and a water-containing liquid. In a preferred embodiment, in the micro-storage tube, the water-immiscible liquid and the water-containing liquid are alternately arranged in the micro-storage tube. In a preferred embodiment, in the micro-storage tube, different water-containing liquids are separated by the water-immiscible liquid, and the different water-containing liquids do not directly contact each other.

[0035] In one embodiment, the density of the water-immiscible liquid is different from the density of the water-containing liquid. After the pre-packaged liquid is discharged from the micro-storage tube, the water-immiscible liquid and the water-containing liquid are separated into layers due to the density difference.

[0036] In one embodiment, after the pre-packaged liquid is discharged from the micro-storage tube, the liquids containing water (e.g., multiple reaction liquids containing reagents) are combined with each other to form a mixed liquid (e.g., multiple reaction liquids containing reagents are mixed to form a reaction mixture, realizing an automated configuration process of the reaction liquid).

[0037] In one embodiment, the liquid immiscible with water is selected from one or more of mineral oil, alkane, silicone oil and liquid paraffin.

[0038] In one embodiment, the water-containing liquid is a liquid with water as a matrix and contains the required reagents. Preferably, the water-containing liquid contains at least one required reagent, such as at least one, two, three, four, five, six, seven, eight, nine or ten required reagents, or even at least 11-20 required reagents, or any sub-range of required reagents consisting of any value in these ranges. Optionally, the water-containing liquid further contains other components.

[0039] In a preferred embodiment, the liquid containing water further contains other components, including one or more of salts, enzymes, polymers, nucleotides, nucleic acids, proteins, and organic substances. In a preferred embodiment, the polymers are one or more of polyethylene glycol, polyethyleneimine, and dextran. In a preferred embodiment, the organic substances are one or more of glycerol, dithiothreitol, tris(hydroxymethyl)aminomethane, ethylenediaminetetraacetic acid, and sodium lauryl sulfate.

[0040] In one embodiment, the micropipette device further comprises a second sealing portion (V) which seals the third interface on the connector.

[0041] In one embodiment, a plurality of the micro-pipette devices can be combined and used together. Preferably, a plurality of the micro-pipette devices can be combined together for use with a pistol gun or a multi-channel pipette.

[0042] According to a second aspect of the present invention, there is provided a method for liquid packaging using the micropipette device of the present invention, comprising the following steps:

[0043] Step S1: removing the sealing portion of the liquid transfer device of the present invention and connecting it to an external dispensing device via a connector;

[0044] Step S2: using an external dispensing device to add a certain volume of liquid into the micro storage tube, wherein the liquid is a liquid immiscible with water or a liquid containing water;

[0045] Step S3: Next, a certain volume of liquid is added to the micro-storage tube, wherein when the liquid in step S2 is a liquid that is immiscible with water, the liquid in step S3 is a liquid containing water; when the liquid in step S2 is a liquid that contains water, the liquid in step S3 is a liquid that is immiscible with water;

[0046] Step S4: repeating steps S2 and S3, so that the water-immiscible liquid and the water-containing liquid are alternately arranged in the micro storage tube;

[0047] Step S5: sealing the micro-storage tube with a sealing portion;

[0048] Step S6: storing the micropipette device according to demand.

[0049] According to the packaging method provided by the present invention, various reagents can be contained in a liquid containing water. Because they are separated by a liquid that is immiscible with water (for example, mineral oil, silicone oil, alkane or liquid paraffin), they do not contact each other and therefore will not react with each other and can be stored for a long time.

[0050] In one embodiment, the external packaging equipment can be a micro pump, a micro dispenser, an automatic filling machine, etc.

[0051] In a preferred embodiment, the micro pump may include a micro syringe pump, a gear pump, a peristaltic pump, a piston pump, and the like.

[0052] In one embodiment, the micropipette device packaged by the above method is stored at room temperature, 2-8°C or frozen, preferably at -15--25°C or ≤-70°C, more preferably at -20°C or -80°C.

[0053] According to a third aspect of the present invention, there is provided a method for performing micropipetting using the micropipetting device of the present invention, comprising the following steps:

[0054] Step S7: If the micropipette device is stored frozen, thaw the liquid in step S6; otherwise, proceed directly to step S8;

[0055] Step S8: opening the sealing portion;

[0056] Step S9: connecting the pipette of the automated workstation to the connector of the micropipette device;

[0057] Step S10: moving the micropipette device to a desired position;

[0058] Step S11: using a pipette on an automated workstation to discharge all the liquid in the micro-storage tube to a desired location, so that the water-containing liquids that were originally not in direct contact with each other merge with each other to form a mixed liquid, thereby realizing an automated configuration process of the reaction system;

[0059] Step S12: Separate the pipette of the automated workstation from the micro-pipetting device.

[0060] In one embodiment, in the method of use, step S9 may be performed first, and then step S8.

[0061] The present invention achieves excellent technical effects by providing the above-mentioned micropipette device and its use method:

[0062] 1. The present invention completes the accurate packaging of extremely small amounts of liquids through an extremely high-precision external packaging device (for example, a micropump (including a microinjection pump, a gear pump, a peristaltic pump and a piston pump, etc.), a microdispenser, an automatic filling machine, etc.). When in use, it is only necessary to discharge all the pre-packaged liquid in the micropipetting device through the pipette, which reduces the demand for pipette accuracy, realizes the automated configuration and use of trace reagents, and avoids the problem of insufficient pipetting accuracy of an automated workstation.

[0063] 2. The present invention pre-packages the required multiple reagents in a prescribed volume in an alternating manner. When in use, all reagents need only be discharged from the micro storage tube using a pipette, without the need to individually aspirate each reagent from a separate reagent tube, thereby reducing the waste caused by dead volume.

[0064] 3. The present invention stores multiple required reagents in the same micropipette device by pre-packaging them alternately according to specified volumes. When needed, all reagents need to be discharged from the micro storage tubes using a pipette, so that various reagents are integrated with each other, realizing the automated configuration and use of trace reagents, saving plate space, and enabling more operation processes to be implemented on the automated workstation. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] In the drawings of the present application, I-micro storage tube, II-connector, III-first sealing part, IV-pre-packaged liquid, V-second sealing part, VI-pipette; 1-first interface, 2-second interface, 3-third interface; 4-liquid A1, 5-liquid B1, 6-liquid A2, 7-liquid B2, 8-liquid A3, 9-liquid B3, 10-mixed liquid A, 11-mixed liquid B.

[0066] FIG1 shows an exemplary structure of a micro-pipette device suitable for an automated workstation according to the present invention.

[0067] FIG. 2 shows an exemplary connection method between a micropipette device and a pipette.

[0068] FIG. 3 shows an exemplary embodiment of a plurality of micro-pipette devices used in combination.

[0069] FIG4 shows the composition structure of pre-packaged liquid in an exemplary micropipette device.

[0070] Figure 5 illustrates the packaging and use of an exemplary micropipette device. A) Initial configuration of the exemplary micropipette device; B) Opening the seal; C) Connecting the automated workstation's pipette to the connector of the micropipette device; D) Discharging all liquid from the micro-storage tube, allowing previously non-contacting compatible liquids to mix to form a mixed liquid, thus automating the reaction system configuration process; E) Separating the automated workstation's pipette from the micropipette device. DETAILED DESCRIPTION

[0071] In order to better understand the present invention, the content of the present invention is further described below in conjunction with the examples, but the content of the present invention is not limited to the following examples. Based on the examples in the present invention, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present invention. The experimental operations described in the following examples, unless otherwise specified, are all routine operations; the reagents and materials, unless otherwise specified, can be obtained from commercial channels.

[0072] Example 1. An automated configuration suitable for an automated workstation, a micropipette device, and a method of use thereof, and its structure and basic usage

[0073] As shown in Figure 1, an exemplary micropipette device suitable for automated configuration in an automated workstation comprises at least the following components: I - micro-storage tube, II - connector, and III - first sealing portion. The micro-storage tube I is connected to the connector II via 2 - second interface. The first sealing portion III seals the micro-storage tube I via 1 - first interface.

[0074] Furthermore, the micro-pipette device may further include a V-second sealing portion, wherein the V-second sealing portion seals the II-connector via a 3-third interface.

[0075] A micropipette device may include an IV pre-dispensed liquid. The IV pre-dispensed liquid is stored in an I micro-storage tube. The IV pre-dispensed liquid is sealed in the I micro-storage tube via a III first seal and, optionally, a V second seal, to prevent leakage of the IV pre-dispensed liquid.

[0076] In some cases, III-first sealing part and V-second sealing part are separate components; in other cases, III-first sealing part and V-second sealing part may not be a separate component, but may be integrated at 1-first interface of I-micro storage tube and 3-third interface of II-connector, and III-first sealing part and V-second sealing part may be opened by the action of external force.

[0077] In the absence of the V-second sealing part to seal the 3-third interface and the III-first sealing part to seal the 1-first interface, the II-connector can be connected to the VI-pipette (as shown in Figure 2), so that the micropipette device can act as the pipette tip, thereby realizing the functions of liquid transfer, absorption, discharge, mixing, blowing, etc. through the pipette.

[0078] In some cases, as shown in FIG. 3 , multiple micropipette devices can be combined for use together, and further combined together for use as a tank gun or multi-channel pipette.

[0079] Example 2. Implementation of liquid pre-packaging and storage

[0080] The accuracy of existing conventional microinjection pumps can reach the pL or nL level. Therefore, with the help of external filling equipment, such as microinjection pumps, micro-liquid filling can be achieved, thereby solving the problem that existing automated workstations are difficult to accurately configure extremely small amounts of liquids (≤1μL).

[0081] In this embodiment, the micropipette device provided by the present invention is used to pre-pack liquid, including the following steps:

[0082] Step S1: The interface is connected to an external packaging device, and with the help of the external packaging device, liquid, including trace liquid, is packaged.

[0083] This method allows liquid (including the required reagents) to be dispensed into a micropipette device via an external dispensing device. When used, the liquid in the I-micro storage tube is completely drained, eliminating the need to draw reagents separately from the reagent tank, thus reducing the generation of dead volume.

[0084] Furthermore, in some cases, pre-packaging of multiple liquids can be achieved, including the following steps:

[0085] Step S2: First, liquid A1 is dispensed into 4 portions with the help of an external dispensing device;

[0086] Step S3: Re-pack 5-liquid B1;

[0087] Step S4: Repeat this process until all required liquids are packaged (IV-pre-packaged liquid after packaging is shown in FIG4 );

[0088] The amount and order of liquids dispensed can be precisely controlled by external dispensing equipment. Liquids A and B are immiscible. In the example composition shown in Figure 4, liquids A1, A2, and A3 are miscible with each other, while liquids B1, B2, and B3 are miscible with each other. B1, B2, and B3 represent different or identical liquids. It should be noted that 1, 2, and 3 are examples only; in practice, any number of liquids can be used as needed. A represents a liquid immiscible with B.

[0089] In some cases, liquid A is a liquid that is immiscible with water, such as mineral oil, silicone oil, alkane, and liquid paraffin; liquid B is a liquid containing water, which contains various components required for the experiment (such as required reagents). Different liquids B (such as liquids B1, B2, and B3) are separated by liquid A to prevent direct contact and mixing between liquids B1, B2, and B3. In other cases, liquid B is a liquid that is immiscible with water, such as mineral oil, silicone oil, alkane, and liquid paraffin; liquid A is a liquid containing water, which contains various components required for the experiment (such as required reagents). Different liquids A (such as liquids A1, A2, and A3) are separated by liquid B to prevent direct contact and mixing between liquids A1, A2, and A3.

[0090] Step S5: sealing the micropipette device with III-first sealing part and, optionally, V-second sealing part;

[0091] This method allows all components required for a reaction mixture (A1, A2, A3, etc. or B1, B2, B3, etc.) to be stored in the same micropipette device, achieving long-term stable storage of the components and avoiding premature mixing of the components, which would shorten the storage time.

[0092] Step S6: storing the micropipette device under suitable conditions.

[0093] Depending on the storage requirements of the various components required for the experiment (e.g., required reagents), the micropipette device can be stored at: room temperature, 2-8°C, or frozen (e.g., -15--25°C or ≤-70°C, e.g., -20°C or -80°C).

[0094] Example 3. Implementation of liquid automation configuration

[0095] This embodiment provides an example method for implementing liquid automation configuration using the micro-pipette device (as shown in FIG5 ), and the steps are as follows:

[0096] Step S7: Thawing the liquid in step S6; (If the micropipette device is not frozen, thawing is not required and step S8 can be directly performed) ( FIG. 5A );

[0097] Step S8: opening III-first sealing portion, and optionally, V-second sealing portion ( FIG. 5B );

[0098] Step S9: connecting the VI-pipette of the automated workstation to the II-connector of the micropipette device ( FIG. 5C );

[0099] Step S10: moving the micropipette device to a desired position driven by the VI-pipette;

[0100] Step S11: All liquids in micro-storage tube 1 (e.g., 4 - liquid A1, 5 - liquid B1, 6 - liquid A2, 7 - liquid B2, 8 - liquid A3, 9 - liquid B3) are discharged to the desired locations, and the miscible liquids that were originally not in direct contact with each other (e.g., A1, A2, A3 and B1, B2, B3) are merged into a mixed liquid (e.g., 10 - mixed liquid A, 11 - mixed liquid B), thereby completing the automated configuration process of the reaction system ( FIG5D );

[0101] Furthermore, 11-mixed liquid B can be drawn to perform library building reaction. Since 11-mixed liquid B is formed by merging 5-liquid B1, 7-liquid B2 and 9-liquid B3 into a mixed liquid, the generation of dead volume can be reduced when the dead volume of the pipette is fixed in a single step. For example, assuming that the dead volume of the pipette is 10 μL, if 11-mixed liquid B is configured separately, 5-liquid B1, 7-liquid B2 and 9-liquid B3 will each leave a dead volume of 10 μL, resulting in a total of 30 μL dead volume. By using the micropipette device of the present invention, 5-liquid B1, 7-liquid B2 and 9-liquid B3 are packaged together in the micropipette device of the present invention in advance, and all are discharged into a reagent tank when used, then the dead volume left by each liquid when configured separately can be reduced, so that the one-time configuration only produces, for example, 10 μL dead volume.

[0102] In Figure 5, 11-mixed liquid B is a reaction system formed after automated configuration. This is only a schematic diagram of a certain case. In other cases, mixed liquid A can also be a reaction system formed after automated configuration.

[0103] Step S12: Separate the VI-pipette of the automated workstation from the micropipette device ( FIG. 5E ).

[0104] The automated configuration of liquid is achieved through the use method described above.

[0105] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention will be further described in detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described in the text are only used to explain the present invention and are not intended to limit the present invention.

Claims

1. A micropipetting device for an automated workstation, the device comprising a connector, a micro storage tube, and a first sealing portion, wherein the first sealing portion is connected to a first interface on the micro storage tube to enclose the micro storage tube, the connector is connected to a second interface on the micro storage tube, and the connector is used to connect to a pipette of the automated workstation through a third interface.

2. The micropipetting device according to claim 1, wherein the micropipetting device is a micropipetting device containing pre - aliquoted liquid, preferably, the pre - aliquoted liquid is contained in the micro storage tube.

3. The micropipetting device according to claim 1, wherein the inner diameter of the tube of the micro storage tube is in the range of 0.2 mm to 5 mm, preferably, in the range of 0.5 mm to 2.5 mm; wherein the tube length of the micro storage tube is in the range of 1 cm to 20 cm, preferably, in the range of 2 cm to 10 cm.

4. The micropipetting device according to claim 1, wherein the connector is connected to the pipette such that the micropipetting device acts as a tip of the pipette.

5. The micropipetting device according to claim 2, wherein the liquid is a single liquid or a mixture containing multiple liquids.

6. The micropipetting device according to claim 5, wherein the mixture contains a liquid immiscible with water and a liquid containing water; preferably, in the micro storage tube, the liquid immiscible with water and the liquid containing water are arranged alternately in the micro storage tube; preferably, in the micro storage tube, different liquids containing water are separated by the liquid immiscible with water, and different liquids containing water do not directly contact each other.

7. The micropipetting device according to claim 6, wherein the density of the liquid immiscible with water is different from the density of the liquid containing water, and when the pre - aliquoted liquid is discharged from the micro storage tube, the liquid immiscible with water and the liquid containing water are layered due to the density difference.

8. The micropipetting device according to claim 6, wherein when the pre - aliquoted liquid is discharged from the micro storage tube, the liquids containing water are fused together to form a mixed liquid.

9. The micropipetting device according to claim 6, wherein the liquid immiscible with water is selected from one or more of: mineral oil, alkane, silicone oil, and liquid paraffin.

10. The micropipetting device according to claim 6, wherein the liquid containing water is a water - based liquid and contains the required reagents, preferably, a liquid containing water contains at least one required reagent; preferably, the liquid containing water also contains other components.

11. The micropipetting device according to claim 10, wherein the liquid containing water also contains other components, and the other components include one or more of acid, base, salt, buffer substance, enzyme, high - molecular polymer, nucleotide, nucleic acid, protein, and organic matter.

12. The micropipetting device according to claim 11, wherein the high - molecular polymer is one or more of polyethylene glycol, polyethyleneimine, and dextran.

13. The micro pipetting device according to claim 11, wherein the organic matter is one or more of glycerol, dithiothreitol, tris(hydroxymethyl)aminomethane, ethylenediaminetetraacetic acid, and sodium dodecyl sulfate.

14. The micro pipetting device according to any one of claims 1-13, wherein the micro pipetting device further comprises a second sealing portion that seals the third interface on the connector.

15. The micro pipetting device according to any one of claims 1-13, wherein a plurality of the micro pipetting devices are combined for use, preferably, for a dispensing gun or a multi-channel pipette.

16. A method for liquid dispensing using the micro pipetting device according to any one of claims 1-15, comprising the following steps: Step S1: Remove the sealing portion on the micro pipetting device and connect to an external dispensing device through the connector; Step S2: Using the external dispensing device, add a certain volume of liquid to the micro storage tube, and the liquid is a liquid immiscible with water or a liquid containing water; Step S3: Immediately thereafter, add a certain volume of liquid to the micro storage tube, wherein when the liquid in Step S2 is a liquid immiscible with water, the liquid in Step S3 is a liquid containing water; when the liquid in Step S2 is a liquid containing water, the liquid in Step S3 is a liquid immiscible with water; Step S4: Repeat Step S2 and Step S3 to alternately arrange the liquid immiscible with water and the liquid containing water in the micro storage tube; Step S5: Seal the micro storage tube with the sealing portion; Step S6: Store the micro pipetting device.

17. The method according to claim 16, wherein the external dispensing device is selected from a micro pump, a micro dispenser, and an automatic filling machine.

18. The method according to claim 17, wherein the micro pump is selected from a micro syringe pump, a gear pump, a peristaltic pump, and a piston pump.

19. The method according to any one of claims 16-18, wherein the dispensed micro pipetting device is stored at room temperature, 2-8 °C, or frozen, preferably the freezing is carried out at -15 to -25 °C or ≤ -70 °C, and more preferably at -20 °C or -80 °C.

20. A method for micro pipetting using the micro pipetting device according to any one of claims 1-15, wherein the micro pipetting device contains pre-dispensed liquid, and the method comprises the following steps: Step S7: If the micro pipetting device is stored frozen, thaw the liquid in the micro pipetting device, otherwise directly proceed to the following Step S8; Step S8: Open the sealing portion; Step S9: Connect the pipette of the automated workstation to the connector of the micro pipetting device; Step S10: Move the micro pipetting device to the required position; Step S11: Drain all the liquid in the micro storage tube to the required position through the pipette of the automated workstation, so that the liquids containing water that were not directly in contact with each other are fused together to form a mixed liquid, realizing the automated configuration process of the reaction system; Step S12: Separate the pipette of the automated workstation from the micro pipetting device.

21. According to the method described in claim 20, in the steps of the method, step S9 can be performed first, and then step S8.

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