JP2022171444A - Desktop sample pretreatment device - Google Patents

Abstract

To provide a desktop sample pretreatment device that can automatically perform dispensing operations in succession from a plurality of small containers and microplates, and to facilitate cleaning and operation of the device and operate the device at low running cost.SOLUTION: A desktop sample pretreatment device can be easily removably attached to small plate groups that can hold a plurality of small containers and a micropipette chip group at a predetermined position. These small plate groups are assigned with IDs associated with information on nozzle control. A control unit reading the IDs and the information associated therewith continuously performs dispensing and mixing operations by using a nozzle head group having needle nozzles and micropipette chip nozzles moving on an X or Y axis and a Z axis.SELECTED DRAWING: Figure 1

Description

In the present invention, liquids such as specimens and reagents contained in small containers are sequentially sucked and discharged by a plurality of dispensing nozzles into desired microplate holes (wells) and small containers such as microtubes. The present invention relates to a desk-top size sample pretreatment device that can be installed even in a safety cabinet, which automatically prepares a sample liquid, and a pipetting operation method using the same.

In some cases, it is required to dispense and mix reagents and sample liquids contained in small containers such as microplate holes and microtubes for sample pretreatment. For example, when analyzing with a PCR device for identifying the target bacterial or viral nucleic acids contained in a minute amount of cell fragments collected from a patient, the sample solution from the container containing the sample, the reaction reagent from the reaction reagent container group into the test container and use the mixed sample.

In the pretreatment operation, for example, 6.5 μL, 6.5 μL, and 2.0 μL of three types of test reagents are dispensed from a plurality of containers, and mixed reagent solutions and 5 μL of specimen solutions are dispensed. In this way, the amount of specimens and reagents to be dispensed is very small, on the order of microliters. Additionally, if the specimen is infectious, pretreatment operations should be performed in a secure area.

When the number of pretreatment samples is small, a series of operations may be performed manually. In that case, a skilled technique is required for the pretreatment work because the amount to be dispensed is small. Therefore, pretreatment sample preparation must be performed by a skilled person.

On the other hand, there is an automatic pretreatment device as a method that does not involve a skilled person. The automatic pretreatment apparatus consists of a container group containing a plurality of containers containing a plurality of specimens and various reagents, a plurality of nozzle heads for dispensing from the containers, a dispensing pump for aspirating and discharging with the nozzles, and a nozzle. to a predetermined position in the horizontal or vertical axis and the vertical axis, and a control unit for controlling the pump and the moving mechanism. In such a pretreatment device, a micropipette tip is used to dispense samples that require high carryover accuracy, and is used to dispense reagents that do not require as high carryover accuracy as samples but require high dispensing accuracy. used a needle nozzle. In such a device, the micropipette tip head and the needle nozzle head operate independently, so to prevent interference between the operations of each nozzle head, secure an installation area that matches the operation range of each nozzle head. When handling infectious specimens, it was necessary to secure a large dedicated safety compartment and prepare a large safety cabinet.

In order to reduce the installation area of the device while ensuring dispensing accuracy, it has multiple nozzle heads that can be equipped with multiple micropipette tips with different dispensing accuracy according to the dispensing volume. By using a mechanism that simultaneously moves the horizontal or vertical axis and the vertical axis, it is possible to use different nozzles depending on the dispensing accuracy. In such a pretreatment device, a plurality of chips are consumed for each pipetting operation for samples requiring extremely high carryover accuracy and for reagents requiring less carryover accuracy than the samples. Since a large amount of discarded tips is generated during the pipetting process, used micropipette tips are discarded in the disposal section. There was a possibility of Moreover, when the sample is an infectious solution, the inside of the device may be contaminated by the scattered chips. Moreover, since a large amount of tips are used, there is a potential risk that the micropipette tips may come off. In addition, since infectious tips are mixed with used tips, all discarded tips must be treated as infectious waste, which increases the cost of disposal as infectious waste in addition to the cost of consumable micropipette tips. There was a problem.

In small-lot pretreatment work, the types of reagents and containers may differ depending on the test content and test equipment. The above-mentioned pretreatment device automatically repeats the dispensing operation using the same sample container group or reagent container group, or can quickly dispense a large amount of samples and their reagents. It is necessary to individually change the setting of the operating position of the nozzle head group for each container each time the type of reagent or specimen or the container group changes. This operation can only be performed by a skilled person, thus placing a heavy burden on the user and posing a problem that prompt changes cannot be made.

JP 2000-105248 WO2018-038019 JP 2008-39471

The present invention has been made to solve the above problems, and the first purpose thereof is to reduce the installation area of the device and reduce the amount of consumable micropipette tips used while achieving high carryover. To provide a desktop sample pretreatment device capable of maintaining accuracy.

A second object is to provide a tabletop sample pretreatment apparatus having a structure that allows easy cleaning of the main body of the apparatus so that it can be put into and taken out of a general-purpose safety cabinet.

A third object is to provide a desktop sample pretreatment apparatus that can easily change a series of dispensing operations when a reagent or sample container group to be dispensed is changed.

A first invention comprises a nozzle head group in which a plurality of nozzles are arranged, a suction and discharge mechanism for sucking and discharging a liquid through the nozzle group, a container group and a micropipette chip group containing the liquid, and the nozzles. a moving mechanism capable of relatively moving vertically or vertically and vertically between a group of heads; A mechanism is provided so that the micropipette tip can be attached and detached relative to the head, the remaining nozzle head group among the plurality of nozzle heads has fixed needle nozzles, and the fixed needle nozzles are washed. The sample pretreatment apparatus has a mechanism for controlling the movement of the nozzle group, suction/discharge, and washing.

Here, the container group may be a cartridge plate containing a plurality of small containers such as microtubes in a row or matrix, or a microplate arranged in a predetermined matrix, such as a plate with 16 or 256 holes. be.

Here, the micropipette tip group is in a state in which the tip group is arranged in a row or in a matrix at regular intervals and inserted into the plate.

Here, the nozzle head group includes at least one nozzle head to which a micropipette chip can be attached and at least one nozzle head to which a fixed needle nozzle is attached. It is ready to move.

A nozzle head equipped with a needle nozzle has, for example, a needle with a needle length of 15 mm, an inner diameter of 1 mm, and an outer diameter of 1.6 mm. movement allows fluid to be aspirated and ejected through the needle. The needle nozzle attached to the needle head can be used repeatedly by cleaning the inside and outside of the needle nozzle each time the object to be dispensed changes. In the cleaning process, for example, the needle nozzle is moved to the nozzle cleaning area, the needle nozzle is inserted into a container filled with the cleaning liquid, the surface of the needle is immersed in the cleaning liquid, and the inner wall of the needle is also washed away with the cleaning liquid. Make an operation to aspirate the remaining liquid.

As a mechanism that makes it possible to attach and detach the tip relative to the micropipette tip nozzle head, the tip is inserted into the head by lowering the tip nozzle head above the tip of the plate, and then raised. This allows the tip to be attached to the nozzle head. As a mechanism for detaching the tip, the tip attached to the nozzle head is laterally moved to the clamp that holds the tip, and the nozzle head and the tip are separated by raising the nozzle head at the position where the tip is clamped. To separate. The separated chips are automatically stored in a chip collection container under the clamp, and preferably the collection container has an indication when the stored chips are full.

In the nozzle head with the needle nozzle attached, the tip of the needle nozzle is at a position higher than the tip of the tip nozzle head with the micropipette tip attached, and at a position lower than the tip of the nozzle head with the micropipette tip removed. When aspirating and discharging with the micropipette tip attached, the needle nozzle is attached at a position that does not contact the container protrusion, but when the micropipette tip is removed, the micropipette nozzle does not touch the container. It is preferable that the needle nozzle is arranged so as not to come into contact with the projection, and that the needle nozzle can be sucked and discharged. Further, the nozzles of the nozzle head group may have a structure in which each nozzle has an independent vertical axial movement mechanism.

In the present invention, high carryover accuracy is required, for example, a sample is dispensed with a nozzle equipped with a micropipette tip, and an accurate amount of dispensing is required although the accuracy of carryover is relatively low. In the dispensing, a needle nozzle that has high dispensing accuracy and can be used repeatedly is used.

Even if independent pumps corresponding to the number of nozzle heads are used for the dispensing and washing, a switching valve is attached between the nozzle head group and a single pump, and a single pump can be used by switching the valve. may be used to dispense each nozzle head.

It is desirable that the floor space of the device be within a width of 600 mm, a depth of 500 mm, and a height of 600 mm, so that it can be stored in a general safety cabinet. In order to avoid contamination from the outside air and carryover, the entire sample pretreatment device is not housed in a safety cabinet, but in a sealed structure, and housed in a dedicated structure where only air filtered by a HEPA filter or the like flows in and out. can be

A second invention is a holding mechanism in which a group of containers for samples and reagents and a group of micropipette chips are housed in a small plate classified in advance by use, and the plate group can be easily attached and detached at a predetermined position. It is a desktop sample pretreatment device with For example, as shown in FIG. 3, a group of containers and a group of micropipette chips are housed in functional plates such as a reagent set, a sample set, and a micropipette chip set. When cleaning the desktop sample pretreatment device, each plate group is separated from the main body, and after cleaning the device main body and the plate group separately, the plate group is attached to the main body again.

The plate group may be a microplate instead of a plate on which a container group is arranged.

In the third invention, the plate group is classified by function such as a reagent set, a specimen set, a microplate set, and a micropipette chip set, and a unique ID is assigned to each plate, and the ID is associated with the plate. Information such as the type, quantity, and position of the containers and chips received is stored in advance in the control device. On the screen of the control device, the ID can be displayed in the menu to be selected from the plate group. This is a sample pretreatment apparatus that can determine the pipetting operation of a plate group to be used by selecting information such as the reagent container group to be used and the type and quantity of micropipette tips using an ID. The ID of the plate group is not determined by the operator, but a storage mechanism such as a barcode or an IC tag that stores the ID information of the plate group is attached to the plate group, and the information of the storage mechanism is sent to the control device. , the dispensing operation information of the plate group may be automatically read.

According to the present invention, a micropipette tip is used to dispense only into a liquid that requires high carryover accuracy, such as a sample, and although dispensing accuracy is required, relatively high carryover accuracy is not required. A reusable needle nozzle can be used for dispensing liquids such as reagents. As a result, the consumption of micropipette tips can be reduced by 50% or more while maintaining the required carryover accuracy and dispensing accuracy. This not only reduces the running cost of the sample pretreatment operation, but also prevents contamination inside the device and, for example, when dispensing an infectious solution, it reduces the amount of infectious waste that requires high management and disposal costs. can be reduced. In addition, since the number of chips used is reduced, the occurrence of potential chip dropouts can be reduced.

In addition, by using a mechanism that moves the needle nozzle head and the micropipette tip nozzle head simultaneously in the horizontal or vertical axis and a group of dedicated compact trays, the size of the device can be reduced. It eliminates the need to prepare equipment and equipment, and it is possible to reduce equipment costs and save space. In addition, cleaning the device is essential when moving the device from the safety cabinet to another location, and the ability to easily separate the device main body from the small plate significantly reduces the time and effort required for cleaning the device. In addition, when the device is moved in and out of the safety cabinet, it is possible to expect an improvement in the operating rate of the sample pretreatment device due to the ease of cleaning.

Even if the type of reagent or specimen container changes, the operating conditions of the sample pretreatment device can be easily changed by selecting the plate group ID. no longer need someone. Therefore, the sample pretreatment work can be entrusted to an unskilled worker, so that the efficiency of the pretreatment worker can be improved and the multiskilled worker can be achieved.

FIG. 4 is a perspective view showing the main part of the sample pretreatment according to the mode of practice of the present invention; FIG. 2 is an enlarged perspective view showing a nozzle portion of the sample pretreatment shown in FIG. 1; FIG. 2 is a diagram showing a layout of a container group for sample pretreatment shown in FIG. 1; FIG. 4 is a diagram showing operation items on an operation screen;

BEST MODE FOR CARRYING OUT THE INVENTION Embodiments of the present invention will be described below based on examples shown in the accompanying drawings.
A micropipette tip head 28 is moved within the apparatus via the lateral and longitudinal axis drives 4 . When attaching a micropipette tip, the micropipette tip head stage 1 lowers the micropipette tip head 28 to attach the tip to the head. Similarly, when aspirating and discharging with the micropipette tip nozzle, the micropipette tip head 28 is moved horizontally or vertically and vertically to a predetermined position, and the micropipette tip attached to the micropipette tip head 28 is used to perform microscopic injection. A pipette syringe pump 9 is used to suck and discharge the solution.

The needle nozzle 27 is attached to the side of the micropipette tip head 28 and moves inside the device via the lateral and vertical axis drive units 4 like the micropipette tip head 28 . Similarly to the micropipette tip nozzle, the needle nozzle 27 also moves to a predetermined position when aspirating and discharging into a sample container (micropube or microplate), and the needle nozzle 27 is used by the needle nozzle syringe pump 8 to extract the solution. Aspirate and dispense.

Using a needle nozzle 27, a predetermined volume of reagent is dispensed from an arbitrary reagent container 30 into a hole of a microplate 22 into which a specimen is placed. After dispensing, the needle nozzle 27 is washed in the needle nozzle washing tank 15, and dispensing of the liquid of another reagent container 30 is started. A necessary reagent is dispensed by a series of operations of the needle nozzle 27 .

Samples requiring high carryover accuracy are pipetted from any container of any sample set to any hole of the microplate 22 by the micropipette tip nozzle. When the dispensing is completed, the micropipette tip is removed by the tip holding mechanism 17 and stored in the tip collection container 18 .

The reagent container 30 and the sample container 34 are inserted into the plate as the reagent set 19 and the sample set 21, and the micropipette tip 32 is inserted as the micropipette tip set 20 into the plate as shown in FIG. Each of the plate group and the microplate 22 has an ID for identification, and the plate group is installed by a holder so as to be fixed and detachable at a predetermined position.

In the dispensing operation, the target plate ID is selected from the plate selection menu 37 shown in the plate selection area 38, and the sample dispensing hole selection menu 39 is used to dispense each sample into the hole of the destination microplate. is set in the sample dispensing order 42 , and the dispensing amount of each reagent and sample is input in the reagent dispensing amount input menu 40 and the sample dispensing amount input menu 41 . When the start button is pressed, the total aspirated amount of each reagent is calculated according to the number of specimens selected in advance, and the needle nozzles 27 aspirate the reagents and discharge them individually into the holes of the microplate 22 in the order indicated. After that, the specimen is aspirated by the micropipette nozzle and dispensed into the indicated holes of the microplate 22 .

1 Micropipette tip head stage 2 Needle nozzle head stage 3 Vertical axis drive 4 Horizontal and vertical axis drive 5 Needle nozzle liquid feed tube 6 Micropipette liquid feed tube 7 Shielding cover 8 Needle nozzle syringe pump 9 For micropipette Syringe pump 10 Syringe pump unit 11 Control unit 12 Cleaning solution pump 13 Cleaning solution 14 Cleaning solution transport tube 15 Needle nozzle cleaning tank 16 Cleaning solution drain 17 Tip holding mechanism 18 Tip collection container 19 Reagent set 20 Micropipette tip set 21 Specimen set 22 Microplate 23 Container set base 24 Body base 25 Communication cable 26 PC
27 needle nozzle 28 micropipette tip head 29 needle nozzle head 30 reagent container 31 reagent container group 32 micropipette tip 33 micropipette tip group 34 sample container 35 sample container group 36 nozzle washing tank 37 plate selection menu 38 plate selection area 39 specimen amount Injection hole selection menu 40 Reagent dispensing amount input menu 41 Sample dispensing amount input menu 42 Sample dispensing order

Claims (4)

A sample pretreatment device equipped with a multi-nozzle head having a mechanism for sucking and discharging a solution in a small container such as a microtube or a microplate, and a moving mechanism for freely moving in the horizontal or vertical axis direction, The desktop sample pretreatment device, wherein the plurality of nozzle heads move simultaneously, and the group of nozzle heads includes a nozzle head having a mechanism for attaching and detaching a micropipette tip, and at least one of which is equipped with a needle nozzle. . 2. The sample pretreatment apparatus according to claim 1, wherein a group of target vessels to be discharged and aspirated by this sample pretreatment apparatus is attached to a small plate on which individual vessels can be placed for each function, and a micropipette A desktop sample pretreatment apparatus, wherein a group of chips is attached to a dedicated small plate, and the small plate group has a position determined for each function. In the sample pretreatment apparatus according to claim 2, the container group of the small plate group is assigned in advance an ID associated with information necessary for dispensing operation such as container position information and container type information. A desktop sample pretreatment device characterized by automatically setting dispensing operations for a selected group of small plates by selecting an assigned ID from a screen of a control device. A small, high-speed readable medium that can store the name of the small plate group, information necessary for dispensing control such as container position and type, a small plate group attached with this medium, and a mechanism for reading these media. 3. The tabletop sample pretreatment device according to claim 2, wherein setting of the small container group and the micropipette tip group is automatically performed by having the device.

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