Sample pretreatment device

Abstract

Provided is a sample pretreatment device that enables efficient dispensing of a sample solution when multiple sample solutions are used. One embodiment of the present invention includes: a container storage section that can store multiple containers including a first well plate and a second well plate, the first well plate being formed by forming wells in an N row by M column matrix, and the second well plate being formed with N elongated wells having a length corresponding to the M wells in one row of the first well plate, or being formed with M elongated wells having a length corresponding to the N wells in one column of the first well plate; a dispensing section including a worktable on which the containers can be placed, a pipette section including M aligned pipette tips corresponding to the M wells in one row of the first well plate or N aligned pipette tips corresponding to the N wells in one column of the first well plate, and a pump section that aspirates and ejects a liquid in the container placed on the worktable via the pipette section; a conveyance section that conveys the containers between the container storage section and the worktable; and a control section that controls the operation of the dispensing section and the conveyance section.

Description

TECHNICAL FIELD

[0001] The present application relates to a sample pretreatment device that performs pretreatment of a sample as an object of analysis and measurement, and more particularly, to a sample pretreatment device that performs pretreatment of a plurality of liquid samples using a reagent or a reagent solution or the like. BACKGROUND

[0002] One of the peptides produced in the human body, Amyloid β (hereinafter sometimes referred to as "Aβ"), is considered to be closely related to the onset of Alzheimer's disease, which is the main cause of dementia. It is said that if Aβ accumulates in the entire brain, healthy nerve cells change or fall off to promote brain atrophy, and it is important to correctly determine the accumulation state of Aβ in the brain in performing early diagnosis of Alzheimer's disease.

[0003] As one of the methods of determining the presence or absence of accumulation of Aβ in the brain, in recent years, IP-MS method obtained by combining ImmunoPrecipitation (IP) method and Matrix Assisted Laser Desorption / Ionization Mass Spectrometry (hereinafter sometimes referred to as "MALDI-MS") has been proposed and attracted attention. For example, in Non-Patent Literature 1, 2 and the like, it is reported that a composite biomarker obtained by combining Aβ-related peptide APP669-711 / Aβ1-42 ratio and Aβ1-40 / Aβ1-42 ratio is promising as a blood biomarker of amyloid accumulation in the brain (in addition, "APP" is Amyloid precursor protein, which is a precursor protein of Aβ).

[0004] In order to implement detection of Aβ using IP-MS method on a large scale, it is necessary to process a plurality of specimens (plasma samples) simultaneously and rapidly using IP method, and therefore, a sample pretreatment device having a dispensing mechanism for automatically dispensing a specimen or a plurality of reagent solutions is indispensable. In addition, in order to improve reproducibility when the same operation is repeatedly performed regardless of the skill and proficiency of the operator, it is also important to implement processing related to IP method using an automatic sample pretreatment device.

[0005] In the past, as an automatic dispensing device for such processing, "Bravo" manufactured by Agilent, "NIMBUS" manufactured by Thermo Fisher Scientific, "epMotion" manufactured by Eppendorf, "Microlab" manufactured by Hamilton, "Biomak" manufactured by Beckman Coulter, and the like are known. In such a conventional automatic dispensing device, it is possible to perform operations such as dispensing from each well of a widely used 96-well plate to each well of another 96-well plate, dispensing from a reservoir to each well of a 96-well plate, or dispensing from a microtube or a conical tube to each well of a 96-well plate.

[0006] Prior Art Documents

[0007] Patent Documents

[0008] Non-Patent Document 1: Kaneko N et al., "Novel plasma biomarker surrogating cerebral amyloid deposition", Proc. Jpn. Acad., Ser. B, Phys. Biol. Sci., 2014, Vol. 90, No. 9, pp. 353-364

[0009] Non-Patent Document 2: Nakamura A et al., "High performance plasma amyloid-β biomarkers for Alzheimer's disease", Nature, 2018, Vol. 554, No. 7691, pp. 249-254 SUMMARY

[0010] PROBLEMS TO BE SOLVED BY THE INVENTION

[0011] As disclosed in non-patent literature 1, 2, etc., the IP method in the above-mentioned IP-MS method sometimes carries out two-stage affinity purification, and its process is complicated and requires multiple test solutions. Moreover, with respect to several test solutions therein, if they are not transferred to each hole of a 96-well plate (or 384-well plate) in advance, they cannot be processed simultaneously. If the operator performs the operation of dispensing multiple test solutions to multiple 96-well plates respectively by manual operation, it is not only very troublesome, but also risks such as error in the injection or mixing of the test solution are generated. If an automatic dispensing device as described above is used, although it is possible to reliably dispense test solutions from a microtube or conical tube containing each test solution to each hole on a 96-well plate, dispensing requires a long time. If the type of test solution increases, the dispensing time is further extended, and the efficiency of pretreatment is reduced.

[0012] The present invention is completed to solve the above-mentioned problems. Its main purpose is to provide a sample pretreatment device that can efficiently dispense multiple test solutions into each well on different well plates, thereby implementing efficient pretreatment by shortening the time required for the dispensing operation.

[0013] Solutions for solving problems

[0014] One embodiment of the sample preprocessing device according to the present invention, which has been made to solve the above-mentioned problems, comprises:

[0015] A container storage unit capable of storing a plurality of containers, the plurality of containers comprising a first orifice plate and a second orifice plate, the first orifice plate being an orifice plate in which holes are formed in a matrix of N rows by M columns, and the second orifice plate being formed with N elongated holes having a length equivalent to the M holes in a row of the first orifice plate, or being formed with M elongated holes having a length equivalent to the N holes in a column of the first orifice plate, wherein both N and M are integers greater than or equal to 2;

[0016] The dispensing operation unit includes an operation table, a pipette unit, and a pump unit. The operation table is capable of placing the container. The pipette unit includes M rows of pipette heads corresponding to the M wells in a row of the first well plate or N rows of pipette heads corresponding to the N wells in a column of the first well plate. The pump unit sucks and ejects liquid from the container placed on the operation table through the pipette unit.

[0017] a conveying unit that conveys the container between the container storage unit and the workbench; and

[0018] The control unit controls the actions of the dispensing operation unit and the conveying unit so as to perform the following processes: a conveying process of conveying the second orifice plate prepared in the container storage unit and containing the L elongated holes of the test solution for pretreatment and the L empty first orifice plates from the container storage unit to the workbench; a test solution dispensing process of using the M-row pipette head or the N-row pipette head on the workbench to perform the following actions on the L prepared test solutions, respectively, which is to suck one of the L test solutions contained in each elongated hole of the second orifice plate containing the test solution and dispense the one test solution into one of the L empty first orifice plates. The invention relates to a method for preparing a molten salt solution and a molten salt solution, comprising: a step of injecting a sample into each hole of the first orifice plate dispensed with the first test solution on the workbench; a step of injecting a sample into each hole of the first orifice plate dispensed with the first test solution on the workbench; and a step of injecting a mixed solution, wherein after the test solution dispensing step, when the first orifice plate dispensed with the second test solution used in the second stage of pretreatment is returned from the workbench to the container storage part, the first orifice plate containing the second test solution is transported from the container storage part to the workbench, and then, on the workbench, the mixed solution containing the first test solution and the sample is injected from each hole of the first orifice plate into each hole of the first orifice plate containing the second test solution, wherein L is an integer greater than 2 and less than N or M.

[0019] For example, when a standard 96-well plate is used as the first well plate, N=8, M=12 (or vice versa), and when a 384-well plate is used as the first well plate, N=16, M=24 (or vice versa).

[0020] Effects of the Invention

[0021] In one embodiment of the sample pretreatment device of the present invention, an M-piece or N-piece pipette head can be used to simultaneously dispense test solutions contained in each of the multiple elongated wells of a second well plate into M or N wells of a first well plate. This process can be repeated multiple times to dispense a single test solution into (N×M) wells of a first well plate. This process can then be repeated for each of L test solutions, allowing different test solutions to be contained in each of the L first well plates.

[0022] As described above, according to one embodiment of the sample pretreatment device of the present invention, even when a large number of test solutions are used in pretreatment, each test solution can be efficiently dispensed into each well of a different well plate, thereby reducing the time required for the test solution dispensing operation.

[0023] Furthermore, if the number of test solutions is large, the number of orifice plates to which the test solutions are dispensed also increases. Orifice plates dispensed with test solutions other than the most recently used test solution or the test solution to be used at the start of pretreatment are temporarily returned to the container storage unit. Whenever a test solution is needed for pretreatment, the orifice plates dispensed with the required test solution are transported from the container storage unit to the dispensing unit. This allows complex pretreatment using multiple test solutions to be performed on a relatively small workbench. This reduces the size of the sample pretreatment apparatus, particularly its installation area.

[0024] Furthermore, according to one embodiment of the sample pretreatment device according to the present invention, a highly versatile sample pretreatment device can be provided that can flexibly cope with various pretreatments having different numbers of work stages or different amounts of test solutions used. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1A This is a front view schematically showing the appearance of a sample preprocessing device according to one embodiment of the present invention.

[0026] Figure 1B This is a top view schematically showing the appearance of the sample pretreatment device according to this embodiment.

[0027] Figure 2 This is a block diagram of the control system of the sample preprocessing device according to this embodiment.

[0028] Figure 3 This is a flowchart showing an example of the process of pretreatment (IP method treatment) and sample preparation for Aβ measurement using the IP-MS method.

[0029] Figure 4 It is an explanatory diagram of the dispensing state of the test solution when the IP method is performed using the sample pretreatment device of this embodiment.

[0030] Figure 5 This is a schematic plan view of a 96-well plate used in the sample pretreatment device of this embodiment.

[0031] Figure 6 This is a schematic plan view of an 8-well plate (8-channel reservoir plate) used in the sample pretreatment device of this embodiment.

[0032] Figure 7 This is a schematic plan view of a 12-well plate (12-channel reservoir plate) used in the sample pretreatment device of this embodiment. DETAILED DESCRIPTION

[0033] Before describing the sample pretreatment device according to the present invention, an example of the IP method processing procedure when Aβ is measured by the IP-MS method will be described with reference to the drawings.

[0034] [IP Law Process]

[0035] Figure 3 This is a flowchart showing an example of the processing procedure of the IP method for Aβ measurement. This procedure itself is disclosed in Non-Patent Documents 1 and 2, etc.

[0036] The sample (specimen) is plasma from the blood collected from the subject. Seven test solutions are used for pretreatment: solution A, solution B, solution C, solution D, solution E, solution F, and solution G. In addition, two types of microbeads, microbeads A and microbeads B, are used as microbeads for immobilizing the specified antibodies used for affinity purification. Since the test solution is not the subject of the invention, a detailed description of the test solution is omitted here. The specific types of the test solution can be those disclosed in non-patent documents 1 and 2, etc.

[0037] First, the sample is mixed with liquid A (step S1), and the mixture of the sample and liquid A is introduced into a container (well) containing a microbeads for mixing (step S2). By incubating the container containing the microbeads for an appropriate time (for example, one hour) in this state, the reaction between the target component in the sample (strictly speaking, the compound containing the target component) and the antibody immobilized on the microbeads is promoted (step S3). As a result, the target component in the sample is carried by the microbeads. Afterwards, the microbeads are left and the supernatant in the container, that is, the mixture of the sample and liquid A, is removed (step S4).

[0038] Next, the a-beads, still carrying the target component, are washed with solutions B and C (step S5). This washing removes most of the impurities that are simply attached to the beads. Afterward, solution D, the eluent, is introduced into the container containing the washed a-beads and mixed, eluting the target component carried by the a-beads into solution D (step S6). This completes the first stage of affinity purification.

[0039] Next, liquid D, which is an eluent, containing the target component is collected from the container and mixed with liquid E (step S7). Then, the mixture of the eluent and liquid E is introduced into a container containing b microbeads for mixing (step S8). Similarly to step S3, by incubating the container containing b microbeads for an appropriate time (for example, one hour) in this state, the reaction between the target component in the sample (strictly speaking, a compound containing the target component) and the antibody immobilized on the b microbeads is promoted (step S9). Thus, the target component in the eluent is carried by the b microbeads. Afterwards, the b microbeads are left in the container and the supernatant, that is, the mixture of the eluent and liquid E is removed (step S10).

[0040] Next, the b-beads, still carrying the target component, are washed using Liquid F, Liquid C, and Liquid G, respectively, in that order (Step S11). The washed b-beads are then mixed with Liquid H, eluting the target component carried by the b-beads into Liquid H (Step S12). Furthermore, volatile acetonitrile is added to Liquid H immediately before the elution process. This completes the second stage of affinity purification, resulting in an eluate in which the target component is dissolved in Liquid H. This eluate is then added dropwise to a MALDI sample plate, and after adding a matrix, it is dried and solidified to prepare a MALDI sample (Step S13).

[0041] As described above, in the Aβ measurement using the IP-MS method, extracting the Aβ-related substance to be analyzed from plasma requires very complicated operations. In a sample pretreatment device according to one embodiment of the present invention, this series of operations can be performed automatically without human intervention.

[0042] [Structure of the Sample Pretreatment Device of the Present Embodiment]

[0043] Figure 1A and Figure 1B This is a schematic external view of the sample pretreatment device of this embodiment. Figure 1A This is the main view. Figure 1B It is roughly a top view. Figure 2 This is a block diagram of the control system of the sample preprocessing device according to this embodiment. Figure 1A and Figure 1B This is a schematic diagram that only records the components necessary for explanation. Figure 1A and Figure 1B As shown in , three mutually orthogonal axes, X, Y, and Z, are defined in space, and the installation surface of the device is a surface parallel to the XY plane.

[0044] like Figure 1A and Figure 1B As shown, the base plate 1 of the sample pretreatment device of this embodiment includes a container storage unit 2, a dispensing operation unit 3, and a container transport unit 4 for transporting containers such as well plates between the container storage unit 2 and the dispensing operation unit 3. Figure 2 As shown, the sample preprocessing device of this embodiment includes a control unit 5 and an operation unit 6 as a user interface.

[0045] The container storage unit 2 is a type of rack capable of accommodating a plurality of various containers such as well plates, reservoirs, or tube racks having sizes compliant with SBS (Society for Biomolecular Screening) standards.

[0046] The dispensing unit 3 comprises a workbench 30 capable of holding multiple containers, such as well plates; a suction and discharge unit 31, mounted on the workbench 30, for sucking liquid from or discharging liquid into the wells of the well plates; and a suction and discharge mechanism moving unit 32, which moves the suction and discharge unit 31 within the XY plane and along the Z axis, or height. The suction and discharge unit 31 comprises a pipette unit 312, attached to the tip of which is a suction head 311 that contacts the liquid to be sucked or discharged; and a pump unit 313, which uses the pipette unit 312 to suck or discharge the liquid. Here, a 12-piece stack of 12 integrated suction heads is used as the suction head 311 to simultaneously dispense from an 8-well plate, described later. The workbench 30 is equipped with multiple plates, including a magnetic plate and a cooling plate, described later, to hold the well plates.

[0047] The container transport unit 4 includes an arm mechanism 41 for gripping a container such as a well plate, and an arm moving unit 42 for moving the arm mechanism within a predetermined range in the three-axis directions of X, Y, and Z.

[0048] The control unit 5 drives the arm moving unit 42 , the suction and discharge unit 31 (pump unit 313 ), and the suction and discharge mechanism moving unit 32 in accordance with a pre-stored operation program 51 , thereby executing a series of pre-processing including a dispensing operation described later.

[0049] Furthermore, at least a part of the functions of the control unit 5 can be realized by using a personal computer as a hardware resource and having the computer execute dedicated software pre-installed on the computer.

[0050] [Container used in pretreatment]

[0051] Here, several types of containers are used to perform the series of processes described above using the sample preprocessing apparatus of this embodiment. Figures 5-7 is a top view showing a typical orifice plate. Figure 5 This is a top view of a 96-well plate. Figure 6 This is a top view of an 8-well plate. Figure 7 This is a top view of a 12-well plate.

[0052] Figure 5The 96-well plate 7 shown is a well plate compliant with the SBS standard. It has 96 wells 71 of equal diameter and depth formed in a matrix of 8 rows by 12 columns on a flat synthetic resin member. Examples of the 96-well plate 7 include "Eppendorf twin.tec (registered trademark) PCR Plates" manufactured by Eppendorf or "Axygen (registered trademark) 96-well PCR Plates" manufactured by Corning. These 96-well plates 7 come in two types: deep well and PCR well. The deep well has deeper wells and a larger capacity than the PCR well.

[0053] Figure 6 The 8-well plate 8 shown is also a well plate that complies with the SBS standard. The 8-well plate 8 has eight slot-shaped elongated holes 81 formed in a flat plate-shaped member of the same size as the 96-well plate 7. The elongated holes 81 have a length that covers the entire 12 wells 71 arranged in a row in the 96-well plate 7. As the 8-well plate 8, for example, the "Axygen 8-Channel Reagent Reservoir High Configuration ( Multiple Well Reagent Reservoir with 8-Channel Trough, HighProfile)" etc.

[0054] Likewise, Figure 7 The 12-well plate 9 shown is also a well plate that complies with the SBS standard. The 12-well plate 9 has 12 slot-shaped elongated holes 91 formed in a flat plate-shaped member of the same size as the 96-well plate 7. The elongated holes 91 have a length that covers the entire 8 wells 71 arranged in a row in the 96-well plate 7. As the 12-well plate 9, for example, the "Axygen 12-channel Reagent Reservoir High Configuration ( Multiple Well Reagent Reservoir with 12-ChannelTrough, High Profile)" etc.

[0055] In the treatment described later, in addition to the orifice plate, a large-capacity reservoir having dimensions conforming to the SBS standard is used.

[0056] [Preprocessing Operation in the Sample Preprocessing Device of the Present Embodiment]

[0057] Next, the sample pretreatment device of this embodiment is used to perform Figure 3 The operations and device actions during IP processing are described below.

[0058] As a preparation step, the operator injects different types of test solutions required for pretreatment into each well 81 of an 8-well plate 8 (or a 12-well plate 9). When using an 8-well plate 8, a maximum of 8 test solutions can be used, and when using a 12-well plate 9, a maximum of 12 test solutions can be used. Figure 4 As shown, five test solutions, namely, solution A, a microbeads (suspension), solution E, b microbeads (suspension), and solution H, are respectively contained in five of the eight elongated wells 81 of the 8-well plate 8. Furthermore, the remaining test solutions, namely, solution B, solution C, solution D, solution F, and solution G, are respectively contained in the reservoirs.

[0059] The operator places an 8-well plate (hereinafter referred to as a "test solution plate") 8 containing the five test solutions, a reservoir containing the five test solutions, five empty 96-well plates 7 (the same number as the number of test solutions), and a rack containing 96 specimens (analytes) in designated locations in the container storage unit 2. Two of the five empty 96-well plates are deep well type, and three are PCR well type. This completes preparations for automatic pretreatment, and the operator then uses the operating unit 6 to perform the prescribed operations to start the process.

[0060] Upon receiving the instruction to start processing, the control unit 5 drives the arm moving unit 42 to move the test plate and five empty 96-well plates 7 from the container storage unit 2 to predetermined positions on the workbench 30 of the dispensing operation unit 3. If there is space on the workbench 30 for placing five empty 96-well plates 7, or if five empty 96-well plates 7 can be stacked, all five empty 96-well plates 7 may be transferred to the workbench 30 before dispensing the test solution, which will be described later. On the other hand, if there is no such space on the workbench 30, the following operation may be repeated: one empty 96-well plate 7 is transferred to the workbench 30, and when dispensing of the test solution is completed, the 96-well plate 7 that has been dispensed with the test solution is returned to the container storage unit 2, and the remaining empty 96-well plates 7 are transferred from the container storage unit 2 to the workbench 30.

[0061] When the test plate and an empty 96-well plate 7 are placed at predetermined positions on the workbench 30, the control unit 5 drives the suction and discharge mechanism moving unit 32 to move the suction and discharge unit 31 to above the test plate. The suction and discharge unit 31 then descends, immersing the suction heads (12-well rows of suction heads) 311 in the same test solution within a single elongated well of the test plate. The pump unit 313 of the suction and discharge unit 31 then operates, sucking and retaining the test solution into each of the 12-well rows of suction heads. The control unit 5 then raises the suction and discharge unit 31 via the suction and discharge mechanism moving unit 32, this time moving it to above an empty 96-well plate 7. The suction and discharge unit 31 then descends, dispensing the retained test solution into the twelve wells 71 arranged in a row in the empty 96-well plate 7. Thus, one test solution is dispensed simultaneously into the twelve wells 71 in one 96-well plate 7. By repeating this operation eight times, the same test solution is dispensed into all the wells 71 in one 96-well plate 7.

[0062] Then, the other four test solutions prepared in the test solution plate are dispensed into the holes 71 of the other empty 96-well plates 7 in the same manner. Figure 4 As shown, different types of test solutions, namely solution A, microbeads a, solution E, microbeads b and solution H, are dispensed into the prepared five empty 96-well plates.

[0063] After that, the control unit 5 drives the arm moving unit 42 to move the test plate after the test solution dispensing is completed and the five 96-well plates 7 containing the test solution from the workbench 30 to the container storage unit 2. However, if necessary (for example, when the pretreatment is continued without a break), the test solution dispensed in the first step of the pretreatment ( Figure 3 The 96-well plate 7 containing the test solution (here, solution A) used in step S1) is still placed on the workbench 30. Alternatively, as described above, the following operation may be repeated: empty 96-well plates 7 are sequentially transferred from the container storage unit 2 to the workbench 30 one by one to dispense the test solution, and when the dispensing is completed, the 96-well plates 7 are temporarily returned to the container storage unit 2.

[0064] After the dispensing of the test solution is completed, the control unit 5 drives the arm moving unit 42 to dispense the test solution in the first stage ( Figure 3 The 96-well plate 7 containing the test solution, i.e., solution A, used in step S1) and the tube rack containing the sample (analyte) are moved from the container storage unit 2 to the workbench 30. Of course, if the 96-well plate 7 containing solution A is already on the workbench 30, only the tube rack may be moved.

[0065] Afterwards, the control unit 5 drives the suction and ejection mechanism moving unit 32 and the suction and ejection unit 31 to suck a specified amount of sample from each tube in the tube rack through the suction and ejection unit 31, and inject the sample into each well of the 96-well plate 7 containing liquid A. At this time, as needed, the cycle of suctioning and ejecting the liquid in the well (a mixture of liquid A and sample) is performed more than once, thereby promoting the mixing of liquid A and the sample. Then, by repeating the same action, different samples are mixed with the liquid A contained in all the wells of the 96-well plate 7. As a result, the solution in each well of the 96-well plate 7 containing only liquid A becomes a mixture of liquid A and the sample. The tube rack from which the sample has been sucked is returned to the container storage unit 2 by the container transport unit 4.

[0066] Furthermore, as described above, an 8-row or 12-row pipette head is used as the pipette head 311 when initially dispensing the test solution. However, a pipette head other than the 8-row or 12-row pipette head may be used as the pipette head 311 for each treatment after mixing the sample with liquid A. For example, a 96-row pipette head may be used that can simultaneously aspirate and eject liquid from all wells.

[0067] Next, the control unit 5 drives the arm moving unit 42 to move the cell containing the affinity purification ( Figure 3 The 96-well plate 7 containing the test solution, i.e., a microbead, used in steps S2 to S6 in the step (step S2 to S6) is moved from the container storage unit 2 to the magnetic plate on the workbench 30. In the case where the 96-well plate 7 containing a microbeads is already on the magnetic plate (in the case where the 96-well plate 7 containing a microbeads is not returned to the container storage unit 2 after dispensing), this movement is not required. The magnetic plate is a loading platform equipped with a magnet, which has the function of attracting the microbeads in the hole by magnetic force. Therefore, the microbeads in each hole of the 96-well plate 7 placed on the magnetic plate (not only a microbeads, but also b microbeads) are attached to the inner bottom or side of the hole.

[0068] The control unit 5 drives the suction and ejection mechanism mobile unit 32 and the suction and ejection unit 31 to inject the mixed solution formed by mixing each sample with liquid A not long ago into each hole of the 96-well plate 7 containing a microbeads. At this time, it is also possible to repeat the cycle of suction and ejection of the liquid in the hole as needed to promote mixing. In addition, when the test solution used here is a microbead suspension containing a microbeads, it is also possible to first suction and remove the supernatant of the suspension. At this time, the microbeads are attached to the bottom of the hole by magnetic force, so that the microbeads can be left and only the supernatant can be sucked.

[0069] As described above, after the mixture of liquid A and the sample is injected into the 96-well plate 7 in which a microbeads are respectively contained in each hole, the 96-well plate 7 is incubated for a predetermined time (e.g., one hour) on a cooling plate provided on the workbench 30 to promote the antigen-antibody reaction. During the incubation period, the suction and ejection action of the liquid in the hole by the suction and ejection portion 31 is continuously and continuously implemented for a predetermined time, or the suction and ejection action of the liquid in the hole by the suction and ejection portion 31 is intermittently implemented after a certain period of time, so that the microbeads are maintained in a suspended state in each hole of the 96-well plate 7. In addition, it is also possible to set a space for incubation different from the workbench 30 instead of incubating on the workbench 30. The space can also be provided in the container storage portion 2. Thus, while the sample in a certain 96-well plate 7 is incubated, other samples can be processed in steps S1 and S2.

[0070] After incubation of one 96-well plate 7 is completed, the 96-well plate 7 is returned to the workbench 30, and the processes corresponding to steps S4 to S12 are sequentially performed on the workbench 30 using Solution B, Solution C, Solution D, Solution E, Solution F, Solution G, Solution H, and the b-beads. In any operation, the following steps are performed: the well plate or reservoir containing the test solution required for the operation is transferred from the container storage unit 2 to the workbench 30, and when the operation is completed, the well plate or reservoir is returned to the container storage unit 2.

[0071] At the time point when the processing corresponding to step S12 is completed, a 96-well plate 7 is placed on the workbench 30, with each well containing b microbeads and H solution as an eluent. In the H solution in the well, the Aβ-related peptides derived from the sample carried by the b microbeads are in an eluted state. At this time, the eluent in the well is the sample solution after the pretreatment based on the IP method. Next, the control unit 5 drives the suction and discharge mechanism moving unit 32 and the suction and discharge unit 31 to aspirate a specified amount of each solution in each well of the 96-well plate 7 after the above-mentioned treatment, and respectively add each solution dropwise to the prepared MALDI sample plate. After that, the pre-prepared matrix is ​​added to each point on the sample plate, and the matrix is ​​air-dried to prepare the sample. In this way, by integrating the function of the spotting device into the sample pretreatment device of this embodiment, it is possible to continuously perform the processing until the MALDI sample is prepared. Alternatively, the sample can be prepared by first dropping a matrix onto a MALDI sample plate and then dropping the treated solution in each well of the 96-well plate 7 thereon.

[0072] Alternatively, the 96-well plate 7 after the above-mentioned treatment may be transported to a spotting device that is separately provided from the sample pretreatment device of the present embodiment, for example, by other processing mechanisms, and each solution in the well may be dripped onto a sample plate for MALDI in the spotting device, and a matrix may be added, followed by air drying, thereby preparing the sample.

[0073] As described above, in the sample pretreatment device of the present embodiment, a series of processes of the IP method for Aβ measurement can be automatically performed.

[0074] As described above, in the sample pretreatment device of this embodiment, when dispensing multiple test solutions into the wells of a well plate, an 8-piece pipette tip strip or a 12-piece pipette tip strip is used. Therefore, compared to the conventional method of dispensing test solutions into the wells of a 96-well plate from multiple microtubes or conical tubes, the time required for test solution dispensing can be reduced to 1 / 8 when using a 12-well plate and an 8-piece pipette tip strip. Furthermore, the time required for test solution dispensing can be reduced to 1 / 12 when using an 8-well plate and a 12-piece pipette tip strip. The greater the number of test solutions to be dispensed, the greater the reduction in dispensing time.

[0075] Furthermore, in the sample pretreatment device of this embodiment, pretreatment is performed using the following process: well plates or reservoirs each containing a plurality of test solutions are stored in the container storage unit 2, only the containers required for the operation are transported to the workbench 30, and when the operation is completed, the required containers are returned to the container storage unit 2. Therefore, even for complex pretreatment using multiple test solutions, the workspace for dispensing or injecting the liquids can be small, thereby reducing the size of the pretreatment device and enabling installation of the device even in a small space.

[0076] Furthermore, the above-described embodiment and variations are merely examples of the present invention. Any appropriate variations, modifications, and additions within the scope of the present invention are also encompassed by the claims of this application. For example, while the sample pretreatment device of the above-described embodiment utilizes a standard 96-well plate, a 384-well plate, which has four times the number of wells, can also be configured to utilize a 384-well plate. Furthermore, by changing the operating program, various types and processes of pretreatment can be accommodated.

[0077] [Various methods]

[0078] It should be understood by those skilled in the art that the above exemplary embodiments are specific examples of the following aspects.

[0079] (Item 1) One embodiment of a sample preprocessing device according to the present invention comprises:

[0080] A container storage unit capable of storing a plurality of containers, the plurality of containers comprising a first orifice plate and a second orifice plate, the first orifice plate being an orifice plate in which holes are formed in a matrix of N rows by M columns, and the second orifice plate being formed with N elongated holes having a length equivalent to the M holes in a row of the first orifice plate, or being formed with M elongated holes having a length equivalent to the N holes in a column of the first orifice plate, wherein both N and M are integers greater than or equal to 2;

[0081] The dispensing operation unit includes an operation table, a pipette unit, and a pump unit. The operation table is capable of placing the container. The pipette unit includes M rows of pipette heads corresponding to the M wells in a row of the first well plate or N rows of pipette heads corresponding to the N wells in a column of the first well plate. The pump unit sucks and ejects liquid from the container placed on the operation table through the pipette unit.

[0082] a conveying unit that conveys the container between the container storage unit and the workbench; and

[0083] The control unit controls the actions of the dispensing operation unit and the conveying unit so as to perform the following processes: a conveying process of conveying the second orifice plate prepared in the container storage unit and containing the L elongated holes of the test solution for pretreatment and the L empty first orifice plates from the container storage unit to the workbench; a test solution dispensing process of using the M-row pipette head or the N-row pipette head on the workbench to perform the following actions on the L prepared test solutions, respectively, which is to suck one of the L test solutions contained in each elongated hole of the second orifice plate containing the test solution and dispense the one test solution into one of the L empty first orifice plates. The invention relates to a method for preparing a molten salt solution and a molten salt solution, comprising: a step of injecting a sample into each hole of the first orifice plate dispensed with the first test solution on the workbench; a step of injecting a sample into each hole of the first orifice plate dispensed with the first test solution on the workbench; and a step of injecting a mixed solution, wherein after the test solution dispensing step, when the first orifice plate dispensed with the second test solution used in the second stage of pretreatment is returned from the workbench to the container storage part, the first orifice plate containing the second test solution is transported from the container storage part to the workbench, and then, on the workbench, the mixed solution containing the first test solution and the sample is injected from each hole of the first orifice plate into each hole of the first orifice plate containing the second test solution, wherein L is an integer greater than 2 and less than N or M.

[0084] The sample pretreatment device according to the first aspect can efficiently dispense each of the test solutions into the respective wells of the different well plates, even when the number of test solutions used in the pretreatment is large, and can shorten the time required for the dispensing operation. In addition, the sample pretreatment device according to the first aspect can provide a sample pretreatment device that is highly versatile and can flexibly cope with various pretreatments that differ in the number of operation stages or the number of test solutions used.

[0085] (Second aspect) In the sample pretreatment device according to the first aspect, the control section can be configured to control the operation of the dispensing operation section and the conveyance section so that, after the test solution dispensing process, a returning process of returning the first well plate in which the test solution other than the first test solution used in the first stage of the pretreatment is dispensed to the container storage section from the operation table is performed, and in the mixed solution injection process, the first well plate containing the second test solution is conveyed from the container storage section to the operation table.

[0086] The sample pretreatment device according to the second aspect can perform a complex pretreatment using a plurality of test solutions on an operation table in a relatively narrow space. Thus, the size of the sample pretreatment device, particularly the installation area, can be suppressed.

[0087] (Third aspect) In the sample pretreatment device according to the first or second aspect, the control section can be configured to control the operation of the dispensing operation section and the conveyance section so that, after the dispensing of the test solution from the second well plate containing the test solution to the empty first well plate is completed, the second well plate is returned from the operation table to the container storage section, and the container containing the test sample is conveyed from the container storage section to the operation table to perform the dispensing of the test sample.

[0088] The sample pretreatment device according to the third aspect returns the used second well plate to the container storage section, so that the operator can immediately recover the second well plate for the next operation as needed. In addition, the container containing the test sample can be placed together with other containers in the container storage section, so that the operation is simplified.

[0089] (Fourth aspect) In the sample pretreatment device according to any one of the first to third aspects, the control section can be configured to perform a mixing promotion process by performing a suction and discharge operation of liquid with respect to the well of the first well plate containing the first test solution when the test sample is dispensed into the well.

[0090] (Fifth aspect) In the sample pretreatment device according to any one of the first to fourth aspects, the control section can be configured to perform a mixing promotion process by performing a suction and discharge operation of liquid with respect to the well of the first well plate containing the second test solution when the mixed solution is injected into the well.

[0091] The sample pretreatment device according to item 4 or 5 can fully mix the liquid contained in the well with the liquid added to the well in a short period of time, for example, promoting a reaction between components in the two liquids. This can shorten the time required for pretreatment or improve reaction efficiency, thereby achieving optimal pretreatment.

[0092] (Item 6) In the sample pretreatment device according to any one of Items 1 to 5, it can be set that the sample pretreatment device further includes a sample preparation unit, which drops a plurality of solutions that have been pretreated in multiple stages after the second stage on the workbench onto a sample plate for matrix-assisted laser desorption ionization, and mixes the solutions with a specified matrix on the sample plate to prepare samples for matrix-assisted laser desorption ionization (MALDI).

[0093] In the sample preparation section, the pre-treated solution can be added dropwise onto the sample plate and then the matrix can be added dropwise onto a point on the solution to mix the two. Alternatively, the matrix can be added dropwise onto the sample plate and then the pre-treated solution can be added dropwise onto the sample plate before or after the matrix is ​​dried and solidified to mix the two.

[0094] According to the sample pretreatment device described in Item 6, operations up to MALDI sample preparation can be performed by a single device, and the efficiency of the MALDI mass spectrometry operation can be further improved.

[0095] (Item 7) In the sample preprocessing apparatus according to any one of Items 1 to 6, N can be 8 and M can be 12, or N can be 16 and M can be 24.

[0096] According to the sample pretreatment device described in Item 7, pretreatment corresponding to a container such as a well plate conforming to widely used standard specifications can be performed.

[0097] Description of Reference Numerals

[0098] 1: Substrate; 2: Container storage unit; 3: Dispensing operation unit; 30: Workbench; 31: Suction and ejection unit; 311: Suction head; 312: Pipette unit; 313: Pump unit; 32: Suction and ejection mechanism moving unit; 4: Step S; 41: Arm mechanism; 42: Arm moving unit; 5: Control unit; 6: Operation unit; 7: 96-well plate; 71: Hole; 8: 8-well plate; 9: 12-well plate; 81, 91: Elongated holes.

Claims

1. A sample pretreatment device comprising: A container storage unit capable of storing a plurality of containers, wherein the plurality of containers includes a first orifice plate and a second orifice plate, wherein the first orifice plate is a orifice plate formed with holes in a matrix of N rows × M columns, and the second orifice plate is formed with N elongated holes having a length equivalent to the M holes in a row of the first orifice plate, or is formed with M elongated holes having a length equivalent to the N holes in a column of the first orifice plate, wherein: N and M are both integers greater than 2; The dispensing operation unit includes an operation table, a pipette unit, and a pump unit. The operation table is capable of placing the container. The pipette unit includes M rows of pipette heads corresponding to the M wells in a row of the first well plate or N rows of pipette heads corresponding to the N wells in a column of the first well plate. The pump unit sucks and ejects liquid from the container placed on the operation table through the pipette unit. a conveying unit configured to convey the container between the container storage unit and the workbench; as well as The control unit controls the actions of the dispensing operation unit and the conveying unit so as to perform the following processes: a conveying process of conveying the second orifice plate prepared in the container storage unit and containing the L elongated holes of the pretreatment test solution and the L empty first orifice plates from the container storage unit to the workbench; a test solution dispensing process of using the M-row pipette tip or the N-row pipette tip on the workbench to perform the following actions on the L prepared test solutions, respectively, which is an action of sucking one of the L test solutions contained in the elongated holes of the second orifice plate containing the test solution and dispensing the one test solution into each hole of one of the L empty first orifice plates; a sample injection process of injecting the sample into each hole of the first orifice plate containing the first test solution on the workbench; and a mixed solution injection process, wherein after the test solution dispensing process, when the first orifice plate dispensed with the second test solution used in the second stage of pretreatment is returned from the workbench to the container storage part, the first orifice plate containing the second test solution is transported from the container storage part to the workbench, and then, on the workbench, the mixed solution of the first test solution and the sample is respectively injected from each hole of the first orifice plate into each hole of the first orifice plate containing the second test solution, wherein L is an integer greater than 2 and less than N or M.

2. The sample pretreatment device according to claim 1, characterized in that: The control unit controls the actions of the dispensing operation unit and the conveying unit, so that after the test solution dispensing process, a return process is performed to return the first orifice plate dispensed with a test solution other than the first test solution used in the first stage of pretreatment from the workbench to the container storage unit, and in the mixed solution injection process, the first orifice plate containing the second test solution is conveyed from the container storage unit to the workbench.

3. The sample pretreatment device according to claim 1, characterized in that: The control unit controls the actions of the dispensing unit and the conveying unit so that after the dispensing of the test solution from the second orifice plate filled with the test solution to the empty first orifice plate is completed, the second orifice plate is returned from the workbench to the container storage unit, and the container containing the sample is conveyed from the container storage unit to the workbench for sample dispensing.

4. The sample pretreatment device according to claim 1, characterized in that: The control unit performs mixing promotion processing by performing liquid suction and discharge operations on each well of the first well plate containing the first test solution when dispensing the sample into the well.

5. The sample pretreatment device according to claim 1, characterized in that: The control unit performs mixing promotion processing by sucking and ejecting liquid from each well of the first well plate containing the second test solution when the mixed solution is injected into the well.

6. The sample pretreatment device according to claim 1, characterized in that: The sample pretreatment device also includes a sample preparation unit, which drips multiple solutions that have completed pretreatment in multiple stages after the second stage on the workbench onto a sample plate for matrix-assisted laser desorption ionization, and mixes the solutions with a specified matrix on the sample plate to prepare samples for matrix-assisted laser desorption ionization.

7. The sample pretreatment device according to claim 1, characterized in that: N is 8 and M is 12, or N is 16 and M is 24.

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