CN117816268A - Pipetting device suitable for automatic workstation and application method thereof - Google Patents

Abstract

The invention relates to the technical field of preparation and transfer of trace liquid and/or reagent, in particular to a pipetting device suitable for an automatic workstation and a using method thereof, and is particularly suitable for an automatically configured trace pipetting device. The invention also provides an automatic configuration of the liquid through the liquid transfer device, in particular a split charging method of a trace amount of liquid and a using method.

Description

Pipetting device suitable for automatic workstation and application method thereof

Technical Field

The invention relates to the technical field of preparation and transfer of micro-samples, liquids and/or reagents, in particular to a pipetting device for automatic configuration of micro-liquids, and particularly relates to a micro-pipetting device suitable for an automatic workstation and a use method thereof.

Background

Pipetting devices are used in a wide range of industrial and laboratory analyses for transferring liquids. In the case of an automated workstation, a pipette is generally used to aspirate, transfer, blow, discharge, and the like, of a liquid. A precisely designed pipetting device is required in order to ensure the accuracy of pipetting.

In pipetting operations in automated workstations, reagents are often pre-packaged in containers such as reagent tanks, reagent tubes or well plates. The bottom area of these containers is relatively large, and in order to ensure that the pipette accurately ascertains the liquid level, to ensure that the pipette tip can contact the liquid, and to accurately aspirate the liquid, it is necessary to add a volume of liquid that is greater than actually required. This part of the added liquid volume is the so-called dead volume. In the practical operation process, a dead volume of about 10 mu L is generally generated, but in some application scenarios, such as in the warehouse building process, the addition amount of some enzymes or catalysts is sometimes less than 1 mu L, which leads to a dead volume larger than the actually required volume, resulting in serious reagent waste. Dead volume is a problem that is often encountered when working in current automated workstations, and no particularly good solution is available at present.

CN110385153B (pipetting devices, pipette tip couplings and pipette tips: devices and methods) describes pipetting devices for Hamilton automated workstations. Although very skillful in design, the pipetting devices of most of the automated workstations currently have difficulty handling very small amounts of liquid below 1 μl, which limits the application of the automated workstations in the fields of warehouse construction and the like as described above.

In order to store reagents more conveniently, CN107557873a proposes a single-person library construction kit, which comprises a reagent strip and reagents required for library construction, wherein the reagent strip is provided with a plurality of storage holes, the storage holes sequentially comprise a bottom, a lower part and an upper part, the bottom is closed, the lower parts of the plurality of storage holes are independent, the upper parts of adjacent storage holes are connected with each other, and the reagents required for library construction are placed in the storage holes. The invention only processes one sample, so that the enzyme activity loss of tool enzyme caused by repeated freezing and thawing is reduced; eliminating the risk of inconsistent actual consumption and theoretical consumption caused by adherence of liquid reagents in the storage, transportation and transportation processes; manual operation steps of operators are reduced to the greatest extent, probability of misoperation is reduced, training requirements of the operators are reduced, and sample preparation quality and reliability are guaranteed. Although such a kit may be used by a single person for the purpose of creating a depot, no means for resolving dead volumes have been proposed.

In the automatic library construction process, the pipetting accuracy is insufficient to influence the stability and reliability of the result, and particularly under the condition that the automatic library construction is performed by using a commercial kit, the pipetting accuracy is difficult to solve by increasing a reaction system, so that the result is unreliable and the reaction number is less. In the automated warehouse-building process, dead volumes can cause serious reagent waste. In addition, the automated warehouse-building process involves a number of steps that require the use of reaction mixtures, most of which are ready to use and cannot be mixed together in advance, which can require a significant amount of board and consumable consumption.

In order to solve the problem of single-dose pre-filling of a reagent which cannot be pre-mixed in advance, CN108641899A discloses a pre-filling cover of a centrifuge tube, which comprises a containing cavity, wherein the containing cavity is divided into 2-6 areas which are not communicated with each other, and the size of an opening of the containing cavity is matched with the caliber of the centrifuge tube. The invention also discloses a centrifuge tube, which comprises a tube body and a pre-cover, wherein the accommodating cavity of the pre-cover is divided into 2-6 areas which are not communicated with each other. The invention also discloses a reagent pre-dispensing and storing device, which comprises a pre-dispensing cover and a cover seal, wherein the accommodating cavity of the pre-dispensing cover is divided into 2-6 areas which are not communicated with each other, and the size of the cover seal is matched with the caliber of an opening of the pre-dispensing cover. The invention solves the problem that single-dose pre-split charging of the pre-mixed reagent can not be carried out in advance, can simultaneously avoid the problems of large manual operation workload and easy pollution in the batch liquid separation process before reaction, ensures that a plurality of reaction units in the same batch start reaction simultaneously in a short time, homogenizes the reaction time of each centrifuge tube and prevents non-specific reaction caused by long operation time. Although this patent solves the problem of single-dose pre-loading of pre-mixed reagents in advance, it still fails to address the previously proposed pipetting accuracy, dead volume issues and the need for automated reagent placement to save plate sites.

Accordingly, there is a continuing need in the art for an apparatus and method that simultaneously addresses the problems of pipetting accuracy, reduced dead volume, pre-dispensing, and automated configuration of reagents to reduce plate and consumable consumption.

Disclosure of Invention

The invention provides a completely new design of pipetting device and a using method thereof, aiming at the problems of the existing automatic workstation in pipetting operation, and solving a series of problems of insufficient precision, insufficient operation plate position, overlarge dead volume and the like of the automatic workstation.

In particular, the present invention addresses the following drawbacks of existing automated workstations:

1. the current automatic workstation has difficulty in performing accurate configuration operation on very small amounts (less than or equal to 1 mu L) of liquid; however, in the automated warehouse-building process, the addition amount of a plurality of enzymes or catalysts is often less than or equal to 1 mu L.

2. When carrying out trace liquid configuration, current automated workstation in order to guarantee the accuracy of imbibition, can have great dead volume, can cause the waste of reagent, has increased reagent cost.

3. The board position of the automatic workstation is limited, and the library construction process requires a plurality of operation steps, which may cause that the automatic workstation cannot execute the whole library construction process at one time due to the limitation of the board position in the library construction process.

4. In the warehouse building process, many reaction systems (e.g., liquid mixtures containing various reagents) have no way to pre-mix in advance and need to be prepared on site.

According to the invention, through the special-designed micropipette, the accurate sample feeding of a reaction system (such as a liquid mixture containing various reagents) is realized, the dead volume is reduced, various reagents are stored simultaneously, the plate positions are saved, and the reagents are automatically configured and used, so that more operation flows can be realized on an automatic workstation.

According to a first aspect of the present invention, there is provided a micropipette adapted for use in an automated workstation, the apparatus comprising a joint (II), a micropipette (I) and a seal (III and/or V). The connector can be connected with the liquid dispenser, so that the micro-liquid dispenser can serve as a suction head of the liquid dispenser, and the micro-liquid dispenser can be installed and separated through the liquid dispenser to blow and beat liquid in the micro-storage tube. The micro-reservoir is used for storing a liquid, e.g. a pre-dispensed liquid, containing e.g. a desired reagent. The seal is used to seal the pre-packaged liquid (IV) containing, for example, the desired reagents.

According to a second aspect of the present invention, there is provided a method of sub-packaging micropipettes suitable for use in an automated workstation:

a. the accurate split charging of the trace liquid is realized by split charging of external split charging equipment (such as a trace pump (including a trace injection pump, a gear pump, a peristaltic pump, a piston pump and the like), a trace liquid dispenser, an automatic filling machine and the like);

b. by adopting an alternate split charging mode, the reaction liquid (containing the required reagent for example) and the isolation liquid are alternately stored in the micro-storage tube, so that the reaction liquid is prevented from being premixed in advance, the same pipetting device can contain multiple reaction liquids, and the storage and the use are convenient;

c. by alternately pre-dispensing a plurality of reaction solutions (containing, for example, required reagents) in prescribed volumes, all the reaction solutions need to be discharged from the micro-storage tube by a pipette when in use, and each reaction solution does not need to be sucked out of a separate reagent tube, thereby reducing the waste caused by dead volumes.

In a third aspect of the invention, a method of use for micropipetting in an automated workstation is provided:

a. the method has the advantages that the multiple reaction liquids (containing the required reagents for example) are stored in the same micro-storage tube in an alternating and pre-split mode according to the specified volume, all the reaction liquids are discharged from the micro-storage tube by using a liquid shifter when the method is needed, the various reagents are fused with each other to form a mixed solution, the automatic configuration and the use of the micro-reaction liquids are realized, the plate positions are saved, and more operation flows can be realized on an automatic workstation.

b. The reaction solution is packaged in advance by using a high-precision external device, so that the problem of insufficient precision of the pipettor can be solved.

According to a first aspect of the present invention, a micropipette for an automated workstation is provided, the device comprising a joint (II), a micropipette (I) and a first seal (III), wherein the first seal is connected to a first port (1) on the micropipette to enclose the micropipette, the joint is connected to a second port (2) on the micropipette, and the joint is for connection to a pipette of an automated workstation through a third port (3).

In one embodiment, the micropipette is a micropipette containing a pre-filled liquid (IV), preferably contained in the micropipette.

In one embodiment, the adapter may be connected to a pipette of an automated workstation. Specifically, the joint (II) can be connected with the pipette (VI) so that the micropipette serves as a suction head of the pipette, and the micropipette is installed and separated through the pipette to suck, blow, discharge, transfer and other operations of liquid in the micropipette.

In one embodiment, the micro-storage tube has a tube inner diameter in the range of about 0.2mm to about 5mm, preferably in the range of about 0.5mm to about 2.5 mm.

In preferred embodiments, the micro-storage tube may have a tube inner diameter of, for example, about 0.2mm, about 0.3mm, about 0.4mm, about 0.5mm, about 0.6mm, about 0.7mm, about 0.8mm, about 0.9mm, about 1mm, about 1.1mm, about 1.2mm, about 1.3mm, about 1.4mm, about 1.5mm, about 1.6mm, about 1.7mm, about 1.8mm, about 1.9mm, about 2.0mm, about 2.1mm, about 2.2mm, about 2.3mm, about 2.4mm, about 2.5mm, about 2.6mm, about 2.7mm, about 2.8mm, about 2.9mm, about 3.0mm, about 3.1mm, about 3.3mm, about 3.4mm, about 3.5mm, about 3.6mm, about 3.7mm, about 3.8mm, about 3.4.4 mm, about 4.4mm, or any of these values.

In one embodiment, the micro-storage tube has a tube length in the range of about 1cm to about 20cm, preferably in the range of about 2cm to about 10 cm.

In a preferred embodiment of the present invention, the micro-storage tube may have a tube length of, for example, about 1cm, about 1.1cm, about 1.2cm, about 1.3cm, about 1.4cm, about 1.5cm, about 1.6cm, about 1.7cm, about 1.8cm, about 1.9cm, about 2cm, about 2.1cm, about 2.2cm, about 2.3cm, about 2.4cm, about 2.5cm, about 2.6cm, about 2.7cm, about 2.8cm, about 2.9cm, about 3cm, about 3.1cm, 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 3.8cm 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 8.8.1 cm, about 8.3cm, about 9.9cm, about 9.1cm, about 7.9cm, about 7.4cm, about 7.8.4 cm, about 7.5cm 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 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 17.3cm, about 17.4cm, about 17.5cm, about 17.6cm, about 17.7cm, about 17.8cm, about 17.9cm, about 18cm, about 18.1cm, about 18.2cm, about 18.3cm, about 18.4cm, about 18.5cm, about 18.6cm, about 18.7cm, about 18.8cm, about 18.9cm, about 19cm, about 19.1cm, about 19.2cm, about 19.3cm, about 19.4cm, about 19.5cm, about 19.6cm, about 19.7cm, about 19.8cm, about 19.9cm, or about 20cm, or a sub-range consisting of any value in these ranges.

According to the micropipette provided by the present invention, the liquid stored in the micropipette remains stratified and does not mix in the micropipette due to turbulence or due to density differences.

In one embodiment, the liquid may be one liquid, or may be a mixture containing a plurality of liquids (for example, containing 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 fig. 4).

In one embodiment, the mixture comprises a water-immiscible liquid and a liquid comprising water. 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-reservoir tube, different water-containing liquids are separated by a water-immiscible liquid, the different water-containing liquids not being in direct contact with each other.

In one embodiment, the water-immiscible liquid has a density that is different from the density of the water-containing liquid, and the water-immiscible liquid and the water-containing liquid delaminate due to the density difference after the pre-packaged liquid exits the micro-reservoir.

In one embodiment, after the pre-dispensed liquid exits the micro-reservoir, the water-containing liquids (e.g., the plurality of reagent-containing reaction fluids) are combined with one another to form a mixed liquid (e.g., the plurality of reagent-containing reaction fluids are mixed to form a reaction mixture, allowing for an automated configuration process of the reaction fluids).

In one embodiment, the water-immiscible liquid is selected from the group consisting of: mineral oil, alkane, silicone oil and liquid paraffin.

In one embodiment, the water-containing liquid is a water-based liquid and comprises the desired agents, preferably one water-containing liquid comprises at least one desired agent, such as at least one, two, three, four, five, six, seven, eight, nine or ten desired agents, even such as at least 11-20 desired agents, or a sub-range of desired agents consisting of any of these ranges. Optionally, the water-containing liquid also contains other ingredients.

In a preferred embodiment, the aqueous liquid further comprises other components, including one or more of salts, enzymes, polymers, nucleotides, nucleic acids, proteins, and organics. In a preferred embodiment, the high molecular polymer is one or more of polyethylene glycol, polyethylenimine, dextran. In a preferred embodiment, the organic substance is one or more of glycerol, dithiothreitol, tris, ethylenediamine tetraacetic acid, sodium dodecyl sulfate.

In one embodiment, the micropipette further comprises a second seal (V) closing off a third port on the adapter.

In one embodiment, a plurality of the micropipettes may be combined together, preferably for use with a gun or multichannel pipette.

According to a second aspect of the present invention, there is provided a method of dispensing a liquid using the micropipette of the present invention, comprising the steps of:

step S1: removing a sealing part on the pipetting device and connecting external sub-packaging equipment through a joint;

step S2: adding a volume of liquid to the micro-storage tube by using external packaging equipment, wherein the liquid is water-insoluble liquid or water-containing liquid;

step S3: next, a volume of liquid is added to the micro-reservoir, wherein when the liquid in step S2 is a water-immiscible liquid, the liquid in step S3 is a water-containing liquid; when the liquid in step S2 is a liquid containing water, the liquid in step S3 is a liquid that is not water-miscible;

step S4: repeating steps S2 and S3 such that the water-immiscible liquid and the water-containing liquid are alternately arranged in the micro-storage tube;

step S5: sealing the micro-storage tube with a sealing portion;

step S6: and storing the micropipette according to the requirement.

According to the dispensing method provided by the invention, various reagents can be contained in a liquid containing water, and can be stored for a long period of time because they are separated by a liquid (e.g., mineral oil, silicone oil, alkane, or liquid paraffin) which is not compatible with water, and do not contact each other, and therefore do not react with each other.

In one embodiment, the external dispensing device may be a micropump, an automated filling machine, or the like.

In preferred embodiments, the micropump may include a microinjection pump, a gear pump, a peristaltic pump, a piston pump, and the like.

In one embodiment, the micropipette after being sub-packaged by the above method is stored at room temperature, 2-8℃or frozen, preferably at-15 to-25℃or less than-70℃and more preferably at-20℃or-80 ℃.

According to a third aspect of the present invention, there is provided a method of micropipetting using the micropipette apparatus of the present invention, comprising the steps of:

step S7: if the micropipette is stored frozen, thawing the liquid in step S6, otherwise, directly performing the following step S8;

step S8: opening the sealing part;

step S9: connecting a pipettor of an automated workstation with a joint of the micropipette;

step S10: moving the micropipette to a desired position;

step S11: the liquid in the micro-storage tube is completely discharged to a required position through a liquid dispenser of an automatic workstation, so that water-containing liquids which are not in direct contact with each other originally are fused with each other to form mixed liquid, and the automatic configuration process of a reaction system is realized;

step S12: and separating the pipettor of the automatic workstation from the micropipette.

In one embodiment, in the using method, step S9 may be performed first, and then step S8 may be performed.

The invention provides the micro-pipetting device and the using method thereof, which obtain excellent technical effects:

1. according to the invention, accurate split charging of the trace liquid is finished through an external split charging device (such as a trace pump (including a trace injection pump, a gear pump, a peristaltic pump, a piston pump and the like), a trace liquid dispenser, an automatic filling machine and the like) with extremely high precision, and when the automatic split charging device is used, only the pre-charged liquid in the trace liquid charging device is required to be completely discharged through the liquid charging device, so that the requirement on the precision of the liquid charging device is reduced, the automatic configuration and use of trace reagents are realized, and the problem of insufficient liquid charging precision of an automatic workstation is avoided.

2. According to the invention, through the mode of alternately pre-packaging the required multiple reagents according to the specified volume, all the reagents are discharged from the micro-storage tube by using the pipette, and each reagent is not required to be sucked out from an independent reagent tube, so that the waste caused by dead volume is reduced.

3. According to the invention, the required multiple reagents are stored in the same micropipette in an alternating and pre-split manner according to the specified volume, and all the reagents are discharged from the micropipette when required, so that the various reagents are mutually fused, the automatic configuration and use of the micropipette are realized, the plate positions are saved, and more operation flows can be realized on an automatic workstation.

Drawings

In the drawings of the present application, I-micro-reservoir, II-connector, III-first seal, IV-pre-filled liquid, V-second seal, VI-pipette; 1-a first interface, 2-a second interface and 3-a 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.

Fig. 1 is an exemplary configuration of a micropipette apparatus suitable for use in an automated workstation according to the present invention.

Fig. 2 illustrates an exemplary manner of connection of the micropipette to the pipette.

Fig. 3 is a diagram illustrating a combination of a plurality of exemplary micropipettes.

Fig. 4 is a diagram showing the constitution of a pre-filled liquid in an exemplary micropipette.

Fig. 5 is a schematic illustration of an exemplary micropipette dispensing and use method. A) An initial configuration of an exemplary micropipette; b) Opening the sealing part; c) The pipettor of the automatic workstation is connected with the joint of the micro pipetting device; d) All the liquid in the micro-storage tube is discharged, and the phase solution bodies which are not in direct contact with each other originally are mutually mixed to form mixed liquid, so that the automatic configuration process of a reaction system is realized; e) And separating the pipettor of the automatic workstation from the micropipette.

Detailed Description

For a better understanding of the present invention, the following examples are further illustrated, but are not limited to the following examples. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to fall within the scope of the invention. The experimental procedures described in the following examples, unless otherwise specified, were all conventional; the reagents and materials, unless otherwise specified, are commercially available.

Example 1. Automated configuration, micropipette and method of use and basic use thereof suitable for automated workstations

As shown in fig. 1, an exemplary micropipette device suitable for automated configuration of an automated workstation comprises at least the following components: i-micro-reservoir, II-joint, III-first seal. Wherein the I-micro-storage tube is connected with the II-joint through a 2-second interface. III-first seal closes the I-micro-reservoir via the 1-first port.

Further, the micropipette may also include a V-second seal. The V-second seal closes the II-joint via the 3-third interface.

IV-pre-filled fluids may be included in the micropipette. IV-prefilled liquid is stored in an I-micro-storage tube. The IV-pre-filled liquid is packaged in the I-micro-storage tube by the III-first seal and, optionally, the V-second seal, avoiding leakage of the IV-pre-filled liquid.

In some cases, the III-first seal, V-second seal are separate components; in other cases, the III-first seal and the V-second seal may not be integrated as a single piece at the 1-first port of the I-micro-storage tube and the 3-third port of the II-joint, and the III-first seal and the V-second seal may be opened by external force.

In the case where no V-second seal encloses the 3-third port and no III-first seal encloses the 1-first port, the II-connector may be connected to a VI-pipette (as shown in fig. 2) so that the micropipette may act as a pipette tip, thereby achieving transfer, aspiration, drainage, mixing, pipetting, etc. of liquid through the pipette.

In some cases, as shown in FIG. 3, multiple micropipettes can be combined together for use, further, for use with a gun or multichannel pipette.

Example 2 implementation of liquid Pre-dispensing and storage

The precision of the conventional microinjection pump can reach pL or nL, so that the dispensing of trace liquid can be realized by means of external dispensing equipment such as the microinjection pump, thereby solving the problem that the conventional automatic workstation is difficult to accurately configure trace (less than or equal to 1 mu L) of liquid.

In this embodiment, the micro pipetting device provided by the invention is used for realizing pre-filling of liquid, and comprises the following steps:

step S1: the interface is connected with external split charging equipment, and liquid, including micro liquid split charging, is realized under the help of the external split charging equipment.

By the method, liquid (containing the required reagent) is packaged into the micropipette through external packaging equipment, and the liquid in the I-micro-storage tube is completely discharged during use, so that the reagent is not required to be sucked from the reagent tank independently, and the generation of dead volume is reduced.

Further, in some cases, pre-packaging of multiple liquids may be achieved, including the steps of:

step S2: firstly sub-packaging 4-liquid A1 with the help of external sub-packaging equipment;

step S3: sub-packaging 5-liquid B1;

step S4: the steps are repeated in a circulating way until the required various liquids are equally packaged (IV-pre-packaged liquid after being packaged is shown in figure 4);

the amount and sequence of dispensed liquid can be precisely controlled by external dispensing equipment. Wherein the liquid a and the liquid B are mutually immiscible, the liquids A1, A2, A3 are mutually compatible, and the liquids B1, B2, B3 are mutually compatible in the composition shown in the example of fig. 4. B1, B2 and B3 represent different liquids or the same liquid, and it should be noted that 1, 2 and 3 are only examples, and any liquid can be used in the actual process according to the need; a represents a liquid which is not compatible with B.

In some cases, liquid a is a water-immiscible liquid such as mineral oil, silicone oil, alkane, liquid paraffin, and the like; liquid B is a water-containing liquid that contains the various components required for the experiment (e.g., the required reagents). Different liquids B (e.g., liquids B1, B2, B3) are separated by liquid a, avoiding mixing by direct contact between liquids B1, B2, B3. In other cases, liquid B is a water-immiscible liquid such as mineral oil, silicone oil, alkane, liquid paraffin, and the like; liquid a is a water-containing liquid that contains the various components required for the experiment (e.g., the required reagents). Different liquids a (such as liquids A1, A2, A3) are separated by liquid B, avoiding mixing by direct contact between liquids A1, A2, A3.

Step S5: closing the micropipette with a III-first seal, and optionally, a V-second seal;

by this method, all the components (A1, A2, A3, etc. or B1, B2, B3, etc.) required for one reaction mixture can be stored in the same micropipette, so that the components can be stably stored for a long time, and the storage time is shortened due to the fact that the components are mixed together in advance.

Step S6: the micropipette is stored under appropriate conditions.

Depending on the storage requirements of the various components required for the experiment (e.g., the reagents required), micropipettes are stored in: room temperature, 2-8deg.C or freezing (e.g., -15-25deg.C or +.70C, e.g., -20deg.C or-80deg.C).

Example 3 implementation of liquid Automation configuration

The embodiment provides an exemplary manner (as shown in fig. 5) for implementing the automatic configuration of the liquid by the micropipette device, which comprises the following steps:

step S7: thawing the liquid in step S6; (if the micropipette is not stored frozen, thawing is not necessary and step S8 is performed directly) (FIG. 5A);

step S8: opening the III-first seal, and optionally, the V-second seal (fig. 5B);

step S9: the VI-pipette of the automated workstation is connected to the II-junction of the micropipette (fig. 5C);

step S10: moving the micro-pipetting device to a required position under the drive of the VI-pipetting device;

step S11: all of the liquids (e.g., 4-liquid A1, 5-liquid B1, 6-liquid A2, 7-liquid B2, 8-liquid A3, 9-liquid B3) in the I-micro-storage tube are discharged to a desired position, and the compatible liquids (e.g., A1, A2, A3 and B1, B2, B3) which are not in direct contact with each other are mutually fused into a mixed liquid (e.g., 10-mixed liquid A, 11-mixed liquid B), so that an automatic configuration process of the reaction system is realized (FIG. 5D);

further, 11-mixed liquid B may be sucked for a stock-building reaction. Since the 11-mixed liquid B is formed by the 5-liquid B1, 7-liquid B2, and 9-liquid B3 being fused with each other into a mixed liquid, in the case where the single-step dead volume of the pipette is fixed, the generation of the dead volume can be reduced. For example, assuming a single-step dead volume of 10. Mu.L of the pipette, each of the 5-liquid B1, 7-liquid B2 and 9-liquid B3 would leave a dead volume of 10. Mu.L if the 11-mixed liquid B were separately disposed, resulting in a total dead volume of 30. Mu.L. By using the micropipette of the present invention, 5-liquid B1, 7-liquid B2 and 9-liquid B3 are dispensed together in advance in the micropipette of the present invention, and all of them are discharged into one reagent tank at the time of use, the dead volume left by each of the liquids separately disposed can be reduced, so that the disposable arrangement generates only a dead volume of, for example, 10. Mu.L.

In FIG. 5, 11-mixed liquid B is a reaction system formed after automatic configuration. This is only a schematic diagram of a case, and in other cases, the mixed liquid a may be a reaction system formed after the automatic arrangement.

Step S12: the VI-pipettor of the automated workstation was separated from the micropipette (fig. 5E).

Completion of the liquid automation configuration is achieved by the use method as described above.

The present invention will be described in further detail with reference to the drawings and examples, in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.

Claims (21)

1. A micropipette for an automated workstation, the device comprising a joint, a micropipette and a first seal, wherein the first seal is connected to a first port on the micropipette to enclose the micropipette, the joint is connected to a second port on the micropipette, and the joint is for connection to a pipette of an automated workstation through a third port.

2. The micropipette of claim 1 wherein said micropipette is a micropipette containing a pre-dispensed liquid, preferably said pre-dispensed liquid is contained in said micropipette.

3. The micropipette of claim 1 wherein the micropipette inner diameter is in the range of 0.2mm to 5mm, preferably in the range of 0.5mm to 2.5 mm;

wherein the tube length of the micro-storage tube is in the range of 1cm to 20cm, preferably in the range of 2cm to 10 cm.

4. The micropipette of claim 1 wherein said adapter is connected to a pipette such that the micropipette acts as a pipette tip.

5. The micropipette of claim 2 wherein the liquid is one liquid or a mixture comprising multiple liquids.

6. The micropipette of claim 5 wherein said mixture comprises a water-immiscible liquid and a water-containing liquid; preferably, in the micro-storage tube, the water-immiscible liquid and the water-containing liquid are alternately arranged in the micro-storage tube; preferably, in the micro-reservoir tube, the different water-containing liquids are separated by a water-immiscible liquid, the different water-containing liquids not being in direct contact with each other.

7. The micropipette of claim 6 wherein said water-immiscible liquid has a density different from a density of said water-containing liquid, wherein said water-immiscible liquid and said water-containing liquid delaminate due to a density difference after a pre-filled liquid exits said micropipette.

8. The micropipette of claim 6 wherein said water-containing fluids merge with one another to form a mixed fluid after a pre-filled fluid exits said micropipette.

9. The micropipette of claim 6 wherein said water-immiscible liquid is selected from the group consisting of: mineral oil, alkane, silicone oil and liquid paraffin.

10. The micropipette of claim 6 wherein the water-containing liquid is a water-based liquid and contains a desired reagent, preferably one water-containing liquid contains at least one desired reagent; preferably, the water-containing liquid also contains other components.

11. The micropipette of claim 10 wherein said aqueous liquid further comprises other components including one or more of acids, bases, salts, buffer substances, enzymes, polymers, nucleotides, nucleic acids, proteins, organics.

12. The micropipette of claim 11 wherein said high molecular polymer is one or more of polyethylene glycol, polyethylenimine, dextran.

13. The micropipette of claim 11 wherein the organic substance is one or more of glycerol, dithiothreitol, tris, ethylenediamine tetraacetic acid, sodium dodecyl sulfate.

14. The micropipette of any one of claims 1-13 wherein said micropipette further comprises a second seal that encloses a third port on said joint.

15. The micropipette of any one of claims 1-13 wherein a plurality of said micropipettes are combined together for use, preferably for use with a lance or multichannel pipette.

16. A method of dispensing a liquid using the micropipette device of any one of claims 1-15, comprising the steps of:

step S1: removing the sealing part on the micropipette, and connecting external split charging equipment through a joint;

step S2: adding a volume of liquid to the micro-storage tube by using external packaging equipment, wherein the liquid is water-insoluble liquid or water-containing liquid;

step S3: next, a volume of liquid is added to the micro-reservoir, wherein when the liquid in step S2 is a water-immiscible liquid, the liquid in step S3 is a water-containing liquid; when the liquid in step S2 is a liquid containing water, the liquid in step S3 is a liquid that is not water-miscible;

step S4: repeating steps S2 and S3 such that the water-immiscible liquid and the water-containing liquid are alternately arranged in the micro-storage tube;

step S5: sealing the micro-storage tube with a sealing portion;

step S6: storing the micropipette.

17. The method of claim 16, wherein the external dispensing device is selected from the group consisting of a micropump, and an automated filling machine.

18. The method of claim 17, wherein the micropump is selected from the group consisting of a microinjection 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 micropipette is stored at room temperature, 2-8 ℃ or frozen, preferably at-15 to-25 ℃ or-70 ℃, more preferably at-20 ℃ or-80 ℃.

20. A method of micropipetting using the micropipette of any one of claims 1-15 wherein said micropipette contains a pre-packaged liquid, said method comprising the steps of:

step S7: if the micropipette is stored frozen, thawing the liquid in the micropipette, otherwise, directly performing the following step S8;

step S8: opening the sealing part;

step S9: connecting a pipettor of an automated workstation with a joint of the micropipette;

step S10: moving the micropipette to a desired position;

step S11: the liquid in the micro-storage tube is completely discharged to a required position through a liquid dispenser of an automatic workstation, so that water-containing liquids which are not in direct contact with each other originally are fused with each other to form mixed liquid, and the automatic configuration process of a reaction system is realized;

step S12: and separating the pipettor of the automatic workstation from the micropipette.

21. The method according to claim 20, wherein in the steps of the method, step S9 may be performed before step S8.