阅读说明：本技术 磁性体颗粒操作用装置 (Magnetic particle manipulation device ) 是由 村松晃 四方正光 山野彩花 于 2019-06-14 设计创作，主要内容包括：提供一种磁性体颗粒操作用装置,即使在无法从外部视觉识别磁体的位置的状态下也能够判别正在执行的处理工序的种类。磁体以与保持在容器保持部中的容器的外侧接近的方式配置,通过沿容器进行相对移动,从而利用磁力使该容器内的磁性体颗粒移动。壳体(100a)以无法从外部视觉识别沿容器进行相对移动的磁体的位置的状态在内部容纳磁体和容器保持部。显示部(140)以能够从外部视觉识别的方式设置于壳体(100a),具有能够发光的发光区域(140a～140c)。处理工序执行部通过使磁体进行相对移动来依次执行多个处理工序。发光控制部在依次执行多个处理工序时,根据磁体的位置使发光区域(140a～140c)发光。(Provided is a magnetic particle manipulation device which can determine the type of a processing step being executed even in a state where the position of a magnet cannot be visually recognized from the outside. The magnet is disposed so as to be close to the outside of the container held by the container holding portion, and moves the magnetic particles in the container by magnetic force by moving the magnet relative to the container. The housing (100a) accommodates the magnet and the container holding portion therein in a state where the position of the magnet relatively moving along the container cannot be visually recognized from the outside. The display unit (140) is provided on the housing (100a) so as to be visually recognizable from the outside, and has light emitting regions (140 a-140 c) that can emit light. The processing step execution unit sequentially executes a plurality of processing steps by relatively moving the magnet. When a plurality of processing steps are sequentially executed, the light emission control unit causes the light emission regions (140 a-140 c) to emit light according to the position of the magnet.)

1. A magnetic particle manipulation device for moving magnetic particles in a tubular device in which a gel-like medium layer and a liquid layer are alternately stacked in a container and the magnetic particles are packed, in a state in which a target substance is fixed to the magnetic particles, the device comprising:

a container holding portion for holding the container;

a magnet disposed so as to be close to the outside of the container held by the container holding portion, and configured to move the magnetic particles in the container by a magnetic force by moving the magnet relative to the container;

a drive unit that relatively moves the magnet along the container;

a housing in which the magnet and the container holding portion are accommodated in a state in which the position of the magnet relatively moving along the container cannot be visually recognized from the outside;

a display unit provided in the housing so as to be visually recognizable from the outside, the display unit having a light emitting region that can emit light;

a storage unit which holds details of a processing step of relatively moving the magnet in advance;

a processing step execution unit that sequentially executes a plurality of processing steps by relatively moving the magnet; and

and a light emission control unit that causes the light emission region to emit light in accordance with a position of the magnet when the plurality of processing steps are sequentially executed.

2. The device for manipulating magnetic particles according to claim 1,

the object is a tubular device having the following features: the liquid layer includes a layer made of a cleaning liquid for removing impurities other than the target substance.

3. The device for manipulating magnetic particles according to claim 1,

the object is a tubular device having the following features: the liquid layer includes a layer composed of a reagent that acts on the target substance.

4. The magnetic particle manipulation device according to any one of claims 1 to 3,

the display portion has a plurality of the light emitting regions capable of independently emitting light.

5. The magnetic particle manipulation apparatus according to claim 4,

the display unit is configured by arranging a plurality of the light emitting regions in parallel.

6. The magnetic particle manipulation apparatus according to claim 4,

when the plurality of processing steps are sequentially executed, the light emission control unit sequentially causes the light emitting regions corresponding to the respective processing steps to emit light.

7. The device for manipulating magnetic particles according to claim 6,

further comprising an operation section operated to omit a part of the plurality of process steps sequentially executed by the process step execution section and execute the next process step,

when the next process step is executed by the operation of the operation unit, the light emission control unit causes the light emission region corresponding to the process step to emit light.

8. The device for manipulating magnetic particles according to claim 6,

when a specific process step, which is a process step predetermined not to be stopped halfway, among the plurality of process steps sequentially executed by the process step execution unit is executed, the light emission control unit causes the light emitting region corresponding to the specific process step to emit light in a different manner from the light emitting regions corresponding to the other process steps.

Technical Field

The present invention relates to a magnetic particle manipulation device for moving magnetic particles in a tubular device in which a gel-like medium layer and a liquid layer are alternately stacked in a container and magnetic particles are filled, in a state in which a target substance is fixed to the magnetic particles.

Background

In medical examinations, food safety and hygiene control, monitoring for environmental protection, and the like, it is required to extract a target substance from a sample containing various foreign substances and subject the substance to detection or reaction. For example, in medical examination, it is necessary to detect, identify, and quantify nucleic acids, proteins, sugars, lipids, bacteria, viruses, radioactive substances, and the like contained in blood, serum, cells, urine, feces, and the like obtained by separation from animals and plants. In these inspections, the target substance may be separated and purified in order to eliminate adverse effects such as background due to foreign substances.

In order to separate and purify a target substance in a sample, a method of using magnetic particles obtained by imparting a chemical affinity and/or a molecular recognition function with respect to the target substance to the surface of a magnetic material having a particle diameter of about 0.5 μm to tens of μm has been developed and put into practical use. In this method, the following steps are repeatedly performed: after the target substance is fixed to the surface of the magnetic particles, the magnetic particles are separated/recovered from the liquid phase by magnetic field operation, and the recovered magnetic particles are dispersed in a liquid phase such as a cleaning liquid as needed, thereby separating/recovering the magnetic particles from the liquid phase. Then, the magnetic particles are dispersed in the eluent, thereby releasing the target substance immobilized on the magnetic particles in the eluent, and the target substance in the eluent is recovered. By using magnetic particles, recovery of target substances can be performed using a magnet, and thus has a feature advantageous to automation of chemical extraction/purification.

Magnetic particles capable of selectively immobilizing a target substance have been marketed as part of separation/purification kits. The reagent kit contains a plurality of reagents in separate containers, and a user uses a pipette or the like to dispense the reagents when using the reagent kit. A device for automating such pipette operation and magnetic field operation is also commercially available (patent document 1). On the other hand, the following method is proposed: instead of the pipette operation, a tubular device in which liquid layers such as a dissolving/fixing liquid, a cleaning liquid, and an eluent, and a gel-like medium layer are alternately stacked in a tubular container such as a capillary tube is used, and magnetic particles are moved in the longitudinal direction of the container in the tubular device to separate and purify a target substance (patent document 2).

In the structure in which the magnetic particles are moved in the tubular container as described above, the magnetic field is changed by moving a magnet serving as a magnetic field applying unit provided outside the container in the longitudinal direction of the container. Following the change in the magnetic field, the magnetic particles also move in the longitudinal direction of the container, and the magnetic particles sequentially move in the liquid layer and the gel-like medium layer which are alternately stacked. In this way, while the magnetic particles move through the plurality of liquid layers, different processing steps are performed in the respective liquid layers, and a plurality of processing steps corresponding to the positions of the magnets are performed.

Patent document 1: international publication pamphlet of WO97/44671

Patent document 2: international publication pamphlet of WO2012/086243

Disclosure of Invention

Problems to be solved by the invention

When the magnet is moved in the longitudinal direction of the container, if the distance between the magnetic particles in the container and the magnet is too long, the magnetic particles cannot be moved well in the longitudinal direction of the container. Since the container is elongated and generally warped, the warpage of the container is corrected in the present situation in order to keep the distance between the magnetic particles and the magnet in the container at a constant short distance. Specifically, the flat contact surface is pressed against the container from the side opposite to the magnet, and the container is corrected to extend linearly.

However, in this case, the container is in a state in which the container is not visually recognized from the outside due to the contact surface of the container being pressed, and therefore the position of the magnet moving along the container is also not visually recognized from the outside. The position of the magnet along the longitudinal direction of the container corresponds to the position of the layer (liquid layer or gel-like medium layer) in which the magnetic particles are present in the container, and therefore there are the following problems: in a state where the position of the magnet cannot be visually recognized from the outside, the type of the process step being executed cannot be determined.

The present invention has been made in view of the above circumstances, and an object thereof is to provide a device for manipulating magnetic particles, which can discriminate the type of a process being performed even in a state where the position of a magnet cannot be visually recognized from the outside.

Means for solving the problems

(1) The device for manipulating magnetic particles according to the present invention is used in a tubular device in which a gel-like dielectric layer and a liquid layer are alternately stacked in a container and magnetic particles are filled, and the magnetic particles are moved in a state in which a target substance is fixed to the magnetic particles, and the device for manipulating magnetic particles includes a container holding unit, a magnet, a driving unit, a casing, a display unit, a storage unit, a processing step execution unit, and a light emission control unit. The container holding portion is for holding the container. The magnet is disposed so as to be close to the outside of the container held by the container holding portion, and moves the magnetic particles in the container by magnetic force by moving the magnet relative to the container. The drive section relatively moves the magnet along the container. The inside of the housing accommodates the magnet and the container holding portion in a state where the position of the magnet relatively moving along the container cannot be visually recognized from the outside. The display unit is provided in the housing so as to be visually recognizable from the outside, and has a light emitting region that can emit light. The storage unit holds details of a processing step of relatively moving the magnet. The processing step execution unit sequentially executes a plurality of processing steps by relatively moving the magnet. When the plurality of processing steps are sequentially executed, the light emission control unit causes the light emission region to emit light in accordance with the position of the magnet.

According to such a configuration, when the plurality of processing steps are sequentially executed, the light emitting region that emits light in accordance with the position of the magnet can be visually recognized from the outside. Therefore, even in a state where the position of the magnet cannot be visually recognized from the outside, the type of the processing step being executed can be determined.

(2) The liquid layer may include a layer formed of a cleaning liquid for removing impurities other than the target substance.

With such a configuration, it is possible to determine whether or not the process step (cleaning step) for removing impurities with the cleaning liquid is being performed even in a state where the position of the magnet cannot be visually recognized from the outside. Therefore, it is possible to accurately determine the time when the cleaning step is being performed or the time when the cleaning step is not being performed, and take out the container in the middle of any one of the processing steps.

(3) Alternatively, the liquid layer may include a layer formed of a reagent that acts on the target substance.

With such a configuration, it is possible to determine whether or not a processing step (reagent step) for causing a reagent to act on a target substance is being performed even in a state where the position of the magnet cannot be visually recognized from the outside. Therefore, it is possible to accurately determine the time when the reagent step is being performed or the time when the reagent step is not being performed, and take out the container in the middle of any one of the processing steps.

The reagent process may include: a restriction enzyme step of allowing a restriction enzyme for fragmenting nucleic acid to act on a target substance as a reagent; and a reaction enzyme step of reacting the fragmented nucleic acids with a reaction enzyme serving as a reagent to act on the target substance. In this case, the time when the restriction enzyme step is performed and the time when the reaction enzyme step is performed can be accurately determined. Therefore, the container can be taken out after the restriction enzyme step and before the reaction enzyme step, for example.

(4) The display unit may have a plurality of light emitting regions that can independently emit light.

According to this configuration, when a plurality of processing steps are sequentially executed, the plurality of light-emitting regions can be independently caused to emit light according to the position of the magnet, and thus the type of the processing step being executed can be easily determined.

(5) The display unit may be configured by arranging a plurality of the light emitting regions in parallel.

According to such a configuration, by independently emitting light from the plurality of light emitting regions arranged side by side, a process of sequentially performing the plurality of processing steps can be displayed easily.

(6) When the plurality of processing steps are sequentially executed, the light emission control unit may sequentially cause the light emitting region corresponding to each processing step to emit light.

With this configuration, the light-emitting regions corresponding to the respective processing steps are sequentially emitted, whereby the process in which the plurality of processing steps are sequentially performed can be easily displayed.

(7) The magnetic particle manipulation device may further include a manipulation unit that is manipulated to omit a part of the plurality of processing steps sequentially performed by the processing step execution unit and to execute a subsequent processing step. In this case, when the next process step is executed by the operation of the operation unit, the light emission control unit may cause the light emission region corresponding to the process step to emit light.

According to such a configuration, by operating the operation unit, it is possible to omit a part of the plurality of process steps sequentially executed and execute the next process step. Since the light emitting region corresponding to the subsequent process step is caused to emit light when the subsequent process step is performed, the type of the process step being performed can be accurately determined even when a part of the process steps is omitted.

For example, when the reagent step includes the restriction enzyme step and the reaction enzyme step as described above, it may be desirable to perform the reaction with the reaction enzyme without fragmenting the nucleic acid by omitting the restriction enzyme step and performing the reaction enzyme step. In this case, the restriction enzyme step can be omitted and the reaction enzyme step can be performed by operating the operation unit. Even in this case, by visually recognizing the light emission of the light emitting region, it is possible to accurately judge whether or not the restriction enzyme step is omitted and the reaction enzyme step is performed.

(8) In the light emitting device according to the present invention, the light emitting control unit may cause the light emitting region corresponding to the specific process step to emit light in a different manner from the light emitting regions corresponding to the other process steps, when the specific process step is executed among the plurality of process steps sequentially executed by the process step execution unit, the specific process step being a process step which is determined in advance not to be stopped halfway.

With this configuration, it is possible to easily determine whether the currently executed process is a process that should not be stopped halfway or another process according to the light emission pattern of the light emitting region. Therefore, it is possible to prevent the container from being taken out by erroneously stopping the processing step during execution of the processing step that should not be stopped halfway.

ADVANTAGEOUS EFFECTS OF INVENTION

According to the present invention, even in a state where the position of the magnet cannot be visually recognized from the outside, when a plurality of processing steps are sequentially executed, the type of the processing step being executed can be determined by visually recognizing the light emitting region that emits light in accordance with the position of the magnet.

Drawings

Fig. 1 is a front view showing a structural example of a tubular apparatus.

Fig. 2 is a cross-sectional view a-a of the tubular apparatus of fig. 1.

Fig. 3 is a front view showing a configuration example of a magnetic particle manipulation device according to an embodiment of the present invention.

Fig. 4 is a B-B sectional view of the magnetic particle manipulation device of fig. 3.

Fig. 5 is a schematic view for explaining an aspect when the magnetic particles are handled.

Fig. 6 is a schematic front view for explaining the appearance of the magnetic particle manipulation device.

Fig. 7 is a block diagram showing an example of an electrical configuration of the magnetic particle manipulation device.

Description of the reference numerals

1: a tubular device; 11: a liquid layer; 12: a gel-like medium layer; 13: magnetic particles; 20: a container; 100: a device for handling magnetic particles; 100 a: a housing; 101: a main body; 102: a container pressing portion; 110: a container holding section; 130: a magnet; 140: a display unit; 140a to 140 c: a light emitting region; 150: an operation section; 150 a: a start key; 150 b: a skip key; 200: a control unit; 201: a processing procedure execution part; 202: a light emission control unit; 300: a storage section.

Detailed Description

1. Tubular device

Fig. 1 is a front view showing a structural example of a tubular apparatus 1. Fig. 2 is a cross-sectional view a-a of the tubular device 1 of fig. 1. The tubular device 1 is used for extracting/purifying a target substance from a liquid sample, and includes a tubular container 20 extending in a straight line.

A plurality of liquid layers 11 and a plurality of gel-like medium layers 12 are formed in the container 20. Specifically, the liquid layer 11 is formed in the lowermost part of the container 20, and the gel medium layers 12 and the liquid layers 11 are alternately stacked upward in the longitudinal direction. In this example, 4 liquid layers 11 and 3 gel medium layers 12 are alternately formed in the longitudinal direction, but the present invention is not limited thereto, and the number of the liquid layers 11 and the gel medium layers 12 can be set arbitrarily.

The liquid layer 11 at the uppermost portion of the container 20 is a solution filled with a plurality of magnetic particles 13. The dissolving solution is, for example, water or a surfactant, and a liquid sample containing the target substance is mixed with the dissolving solution. The dissolving solution may be mixed with the liquid sample and then injected into the container 20. The lowermost liquid layer 11 of the container 20 is an eluent for eluting the target substance in the liquid sample. The liquid layer 11 of 1 or more (2 in this example) in the middle portion of the container 20 is a cleaning liquid for removing impurities (foreign substances) other than the target substance contained in the liquid sample. These liquid layers 11 are separated from each other by a gel-like medium layer 12. In a state where the target substance contained in the liquid sample is immobilized on the magnetic particles 13, an operation (particle operation) of moving the magnetic particles 13 from the uppermost portion to the lowermost portion of the container 20 by changing the magnetic field is performed, and during this time, the target substance is eluted in the eluent at the lowermost portion after being washed with the washing liquid.

The magnetic particles 13 are particles capable of specifically immobilizing a target substance such as a nucleic acid or an antigen on the surface or inside thereof. The magnetic particles 13 are dispersed in the liquid layer 11 at the uppermost portion of the container 20, whereby the target substance contained in the liquid layer 11 is selectively fixed to the magnetic particles 13.

The method of immobilizing the target substance to the magnetic particles 13 is not particularly limited, and various known immobilization mechanisms such as physical adsorption and chemical adsorption can be applied. For example, the target substance is fixed to the surface or the interior of the magnetic particles 13 by various intermolecular forces such as van der waals force, hydrogen bond, hydrophobic interaction, ionic interaction, and pi-pi stacking.

The particle diameter of the magnetic particles 13 is preferably 1mm or less, more preferably 0.1 to 500 μm, and still more preferably 3 to 5 μm. The shape of the magnetic particles 13 is desirably spherical with a uniform particle diameter, but may have an irregular shape or a certain degree of particle diameter distribution as long as the particle manipulation is possible. The structural component of the magnetic particles 13 may be a single substance or may be composed of a plurality of components.

The magnetic particles 13 may be composed of only a magnetic material, but it is preferable to use magnetic particles coated on the surface of the magnetic material to specifically immobilize the target substance. Examples of the magnetic material include iron, cobalt, nickel, and compounds, oxides, and alloys thereof. Specifically, magnetite (Fe) may be mentioned₃O₄) Hematite (Fe)₂O₃Or alpha Fe₂O₃) Maghemite (gamma Fe)₂O₃) Titanomagnetite (xFe)₂TiO₄·(1-x)Fe₃O₄) Ilmenohemite (xFeTiO), ilmenite₃·(1-x)Fe₂O₃) Pyrrhotite (Fe)_1-xS(x＝0～0.13)‥Fe₇S₈(x-0.13)), greigite (Fe)₃S₄) Goethite (alpha FeOOH), chromium oxide (CrO)₂) Permalloy, alnico, stainless steel, samarium, neodymium, and barium magnets.

Examples of the target substance selectively immobilized on the magnetic particles 13 include a substance derived from a living body such as a nucleic acid, a protein, a sugar, a lipid, an antibody, a receptor, an antigen, and a ligand, and a cell itself. When the target substance is a substance derived from a living body, the target substance can be immobilized in the magnetic particles 13 or on the particle surfaces by molecular recognition or the like. For example, when the target substance is a nucleic acid, magnetic particles having a silica coating on the surface thereof, or the like, are preferably used as the magnetic particles 13. When the target substance is an antibody (e.g., a labeled antibody), a receptor, an antigen, a ligand, or the like, the target substance can be selectively immobilized on the particle surface by using an amino group, a carboxyl group, an epoxy group, avidin, biotin, digoxin, protein a, protein G, or the like on the surface of the magnetic particle 13. As the magnetic particles 13 capable of selectively immobilizing a specific target substance, for example, commercially available magnetic beads attached to Dynabeads (registered trademark) sold by Thermo Fisher Scientific, or the like can also be used.

When the target substance is a nucleic acid, the washing solution may be any one that can release impurities such as components (for example, proteins, sugars, and the like) other than the nucleic acid contained in the liquid sample, reagents used in a process such as nucleic acid extraction, and the like into the washing solution so as to maintain the state in which the nucleic acid is immobilized on the surface of the magnetic particle 13. Examples of the cleaning liquid include a high-salt aqueous solution such as sodium chloride, potassium chloride, and ammonium sulfate, and an alcohol aqueous solution such as ethanol and isopropyl alcohol.

As an eluent for eluting nucleic acids (nucleic acid eluent), water or a buffer containing a salt at a low concentration can be used. Specifically, Tris buffer, phosphate buffer, distilled water, etc. can be used, and Tris buffer of 5mM to 20mM adjusted to pH7 to 9 is usually used. By dispersing the magnetic particles 13 having the nucleic acid immobilized thereon in the eluting solution, the nucleic acid can be eluted freely in the nucleic acid eluting solution. The recovered nucleic acid can be subjected to an operation such as concentration and drying, and then subjected to analysis, reaction, or the like, as necessary.

The gel-like medium layer 12 is in a gel or paste form prior to granulation. The gel medium layer 12 is insoluble or poorly soluble in the adjacent liquid layer 11, and is preferably made of a chemically inert substance. Here, the term "insoluble or poorly soluble in a liquid" means that the solubility in a liquid at 25 ℃ is substantially 100ppm or less. The chemically inert substance is a substance that does not chemically affect the liquid layer 11, the magnetic particles 13, and the substance fixed to the magnetic particles 13 in contact with the liquid layer 11 or in an operation of the magnetic particles 13 (i.e., an operation of moving the magnetic particles 13 in the gel-like medium layer 12).

The material, composition, and the like of the gel medium layer 12 are not particularly limited, and may be a physical gel or a chemical gel. For example, as described in WO2012/086243, a liquid material that is not water-soluble or water-insoluble is heated, a gelling agent is added to the heated liquid material to completely dissolve the gelling agent, and then the liquid material is cooled to a temperature not higher than the sol-gel transition temperature to form a physical gel.

The filling of the liquid layer 11 and the gel-like medium layer 12 into the container 20 can be carried out by an appropriate method. When a tubular container 20 is used as in the present embodiment, it is preferable that the opening at one end (for example, the lower end) of the container 20 is sealed before filling, and the liquid layer 11 and the gel medium layer 12 are filled in this order from the opening at the other end (for example, the upper end).

The capacities of the liquid layer 11 and the gel medium layer 12 filled in the container 20 can be set as appropriate according to the amount of the magnetic particles 13 to be handled, the kind of the handling, and the like. In the case where a plurality of liquid layers 11 and gel medium layers 12 are provided in the container 20 as in the present embodiment, the volumes of the respective layers may be the same or different. The thickness of each layer can also be set appropriately. The thickness of each layer is preferably, for example, about 2mm to 20mm in consideration of workability and the like.

The uppermost portion of the container 20 is a bulge 21 having an inner diameter and an outer diameter larger than those of other portions. The upper surface of the bulging portion 21 serves as an opening, and the opening can be sealed with a cap 30 that is detachable from the bulging portion 21. The liquid layer 11 at the uppermost portion of the container 20 is formed by injecting a liquid sample into the bulging portion 21 with the cap 30 removed.

The portion of the container 20 located below the bulging portion 21 is a linear portion 22, and the cross-sectional shape of the linear portion 22 perpendicular to the longitudinal direction is a fixed shape as shown in fig. 2. The bulging portion 21 and the linear portion 22 are connected by a tapered portion 23, and the tapered portion 23 is tapered from the bulging portion 21 side toward the linear portion 22 side. An opening is formed at the lower end of the linear portion 22 (the bottom surface of the container 20), and the opening is sealed by a film member 40. The target substance eluted from the eluent can be sucked out into a pipette by inserting the pipette into the liquid layer 11, i.e., the eluent, at the lowermost portion of the container 20 so as to penetrate the membrane member 40. The film member 40 is formed of, for example, aluminum or the like, but is not limited thereto.

The material of the container 20 is not particularly limited as long as it can move the magnetic particles 13 in the container 20 and can hold the liquid layer 11 and the gel-like medium layer 12. In order to move the magnetic particles 13 in the container 20 by an operation of changing a magnetic field (magnetic field operation) from outside the container 20, a magnetically permeable material such as plastic is preferable, and examples thereof include polyolefins such as polypropylene and polyethylene, fluorine-based resins such as tetrafluoroethylene, and resin materials such as polyvinyl chloride, polystyrene, polycarbonate, and cyclic polyolefin. As the material of the container 20, ceramics, glass, silicone, nonmagnetic metal, and the like can be used in addition to the above-described materials. In order to improve the water repellency of the inner wall surface of the container 20, coating with a fluorine-based resin, silicone, or the like may be performed.

As shown in fig. 2, the cross-sectional shape (cross-sectional shape perpendicular to the longitudinal direction) of the linear portion 22 of the container 20 located below the expanded portion 21 is asymmetrical with respect to the center C, as the shape of the container 20. Specifically, the outer peripheral surface of the straight portion 22 on the front side is a flat surface 221, and the outer peripheral surface on the rear side opposite to the center C is a convex curved surface 222. The shape of the container 20 is not limited to the above shape, and may be, for example, a shape (e.g., a circle) in which the cross-sectional shape of the linear portion 22 is symmetrical with respect to the center C.

2. Magnetic particle manipulation device

Fig. 3 is a front view showing a configuration example of the magnetic particle manipulation device 100 according to the embodiment of the present invention. Fig. 4 is a B-B sectional view of the magnetic particle manipulation device 100 of fig. 3. The magnetic particle manipulation apparatus 100 (hereinafter referred to as "apparatus 100") is used in a state in which the tubular device 1 shown in fig. 1 and 2 is fixed, and is used for performing a particle manipulation on a target substance contained in a liquid sample in the container 20 of the tubular device 1.

The outer shape of the device 100 is defined by a housing 100 a. The housing 100a includes: a main body 101 formed with a container holding portion 110 that holds the tubular device 1; and a container pressing portion 102 for pressing and fixing the container 20 of the tubular device 1 held by the container holding portion 110. In this example, the container pressing portion 102 is formed of a door that is pivotally attached to the main body 101 by a hinge (not shown). However, the container pressing portion 102 is not limited to a structure that can pivot with respect to the main body 101, and may be a structure that can slide with respect to the main body 101, a structure that can be attached to and detached from the main body 101, or the like, as long as it can fix the tubular device 1 held by the container holding portion 110.

The container holding portion 110 is formed of a recess formed in the front surface 120 of the main body 101. The container holding portion 110 is formed as follows: the first accommodating portion 111 accommodating the bulging portion 21 of the container 20 of the tubular apparatus 1 and the second accommodating portion 112 accommodating the linear portion 22 extend continuously in the up-down direction D1. Further, the width of the container holding portion 110 in the lateral direction D2, which is a direction perpendicular to the direction in which the linear portion 22 extends (the vertical direction D1) and parallel to the front surface 120 of the main body 101, corresponds to the width of the tubular device 1.

Specifically, width W1 in lateral direction D2 of first container 111 is slightly larger than the width of bulge 21 of container 20. On the other hand, width W2 in lateral direction D2 of second container 112 is slightly larger than the width of linear portion 22 of container 20 and smaller than the width of bulging portion 21. The first housing portion 111 and the second housing portion 112 are connected by a narrowed portion 113 inclined at an angle corresponding to the tapered portion 23 of the container 20. Thus, in a state where the container 20 is accommodated in the container holding portion 110, the tapered portion 23 of the container 20 is caught by the narrowed portion 113 of the container holding portion 110 and held in a suspended state.

As shown in fig. 4, the container 20 is accommodated in the container holding portion 110 such that the flat surface 221 extends in the lateral direction D2 and the convex curved surface 222 is located on the back surface side of the flat surface 221. A step 114 is formed on the inner surface of the second housing portion 112 of the container holding portion 110 so as to protrude inward from both sides in the lateral direction D2. The width W3 in the lateral direction D2 of the first container part 111 at this stepped part 114 is smaller than the width W2 on the front surface 120 side and smaller than the width in the lateral direction D2 of the linear part 22 of the container 20.

Therefore, the linear portion 22 of the container 20 accommodated from the front surface 120 side into the container holding portion 110 is in a state where the convex curved surface 222 side thereof abuts against the stepped portion 114. At this time, the flat surface 221 of the container 20 projects forward from the container holding portion 110 than the front surface 120 of the main body 101. By closing the door constituting the container pressing portion 102 in this state, as shown in fig. 4, the abutment surface 121 facing the front surface 120 of the main body 101 can be brought into abutment with the flat surface 221 of the container 20 and pressed toward the back surface side. This allows the linear portion 22 of the container 20 to be sandwiched between the abutment surface 121 and the stepped portion 114, thereby firmly fixing the linear portion 22.

An opening is provided on the back side of the container holding portion 110, and a magnet 130 is disposed so as to face the container holding portion 110. The magnet 130 approaches the container 20 held by the container holding portion 110 from the outside (back surface side). The magnet 130 is formed of a permanent magnet and is held so as to be slidable in the vertical direction D1.

The magnet 130 attracts the magnetic particles 13 in the container 20 by magnetic force. Thereby, as shown in fig. 4, the magnetic particles 13 are gathered on the convex curved surface 222 side. By moving the magnet 130 in the vertical direction D1 in a state where the magnetic particles 13 are attracted to the magnet 130 side in this manner, the magnetic particles 13 in the container 20 can be moved in the vertical direction D1 by the magnetic force.

In this manner, the magnet 130 constitutes a magnetic field applying portion that moves the magnetic particles 13 in the container 20 by changing the magnetic field. The magnet 130 can be slid by a driving unit such as a motor. In the example of fig. 4, the opposing surface 131 of the magnet 130 opposing the container 20 is formed of a concave curved surface. The opposing surface 131 is a concave curved surface having a radius of curvature corresponding to the convex curved surface 222 of the container 20. The opposing surface 131 is not limited to the concave curved surface, and may be a flat surface, for example.

The shape, size, and material of the magnet 130 are not particularly limited as long as the magnetic particles 13 can be handled. As the magnet 130, an electromagnet can be used in addition to a permanent magnet. In addition, the magnet 130 may be provided in plurality. The magnet 130 may be configured to be movable relative to the container 20 to change the magnetic field, and is not limited to the configuration in which the magnet 130 is moved as in the present embodiment, and may be configured to move the container 20.

3. Manipulation of magnetic particles

Fig. 5 is a schematic diagram for explaining an aspect when the magnetic particles 13 are handled. In fig. 5, the shape of the tubular device 1 is shown simplified for ease of understanding of the description. In a of fig. 5, a plurality of magnetic particles 13 are contained in the liquid layer 11 at the uppermost portion of the container 20. By thus dispersing the magnetic particles 13 in the liquid layer 11, the target substance contained in the liquid layer 11 is selectively fixed to the magnetic particles 13.

Then, as shown in B of fig. 5, when the magnet 130 as the magnetic force source is brought close to the outer peripheral surface of the container 20, the magnetic particles 13 to which the target substance is fixed are gathered on the magnet 130 side (convex curved surface 222 side) in the container 20 by the action of the magnetic field. Then, as shown in C of fig. 5, when the magnet 130 is moved in the longitudinal direction (vertical direction) of the container 20 along the outer peripheral surface of the container 20, the magnetic particles 13 also move in the longitudinal direction of the container 20 following the change in the magnetic field, and sequentially move in the liquid layer 11 and the gel medium layer 12 which are alternately stacked.

Most of the liquid physically adhering to the periphery of the magnetic particles 13 in the form of droplets is released from the surfaces of the magnetic particles 13 when the magnetic particles 13 enter the gel medium layer 12. The gel medium layer 12 is perforated by the entry and movement of the magnetic particles 13 into the gel medium layer 12, but the pores of the gel medium layer 12 are immediately closed by the self-repairing action based on the restoring force of the gel. Therefore, the liquid hardly flows into the gel medium layer 12 through the through holes formed by the magnetic particles 13.

By dispersing the magnetic particles 13 in the liquid layer 11, the magnetic particles 13 are brought into contact with the liquid in the liquid layer 11, whereby operations such as fixation of the target substance to the magnetic particles 13, a cleaning operation for removing impurities (foreign substances) adhering to the surfaces of the magnetic particles 13, a reaction of the target substance fixed to the magnetic particles 13, and elution of the target substance fixed to the magnetic particles 13 into the liquid are performed.

4. Appearance of device for manipulating magnetic particles

Fig. 6 is a schematic front view for explaining the appearance of the magnetic particle manipulation device 100. As shown in fig. 6, in a state where the container 20 of the tubular device 1 is fixed in the main body 101 by pressing the container pressing portion 102 with the door as the container pressing portion 102 closed, the front of the container 20 is covered with the container pressing portion 102. In the state of fig. 6, the magnet 130 and the container holding portion 110 are accommodated in the case 100a (the main body 101 and the container pressing portion 102), and the periphery of the container 20 is completely covered with the case 100a, so that the magnet 130 relatively moving along the container 20 cannot be visually recognized from the outside.

When the magnetic particles 13 are sequentially moved in the plurality of liquid layers 11 by moving the magnet 130, different processing steps are performed in the respective liquid layers 11. In the present embodiment, a dissolving step of stirring a solution containing a target substance mixed therein to dissolve the target substance is performed in a state where the magnet 130 faces the uppermost liquid layer 11 of the container 20. The elution step for eluting the target substance into the eluent is performed in a state where the magnet 130 faces the liquid layer 11 at the lowermost portion of the container 20. The cleaning step for removing impurities other than the target substance is performed in a state where the magnet faces 1 or more (2 in this example) liquid layers 11 in the middle portion of the container 20. In this manner, in the present embodiment, a plurality of processing steps (dissolving step, washing step, and eluting step) corresponding to the position of the magnet 130 are sequentially performed.

A display unit 140 having a plurality of light-emitting regions 140a to 140c is provided on the front surface of the casing 100 a. Each of the light emitting regions 140a to 140c includes, for example, an LED (light emitting diode), and can independently emit light. Since the display unit 140 is provided on the front surface of the casing 100a, the user can visually recognize the light emitting regions 140a to 140c of the display unit 140 from the outside of the casing 100 a. However, the display unit 140 may be visually recognized from the outside of the casing 100a, and the display unit 140 may be provided on other outer surfaces (side surfaces, upper surfaces, and the like) of the casing 100a, without being limited to the front surface of the casing 100 a. Not only the display based on the LEDs arranged side by side as described above, but also the display may be performed by a device such as a liquid crystal display, and they may be a progress bar display. In this case, the progress bar may be displayed in association with the processing time, not limited to the processing step. In this case, the length of the progress bar may be associated with the processing time. The plurality of light-emitting regions 140a to 140c may not be associated with each process step. The number of the light-emitting regions may be 1, and the light-emitting regions may be made to emit light in a color corresponding to a process step being performed, for example.

An operation unit 150, such as a start key 150a and a skip key 150b, which can be operated by a user, is provided on the front surface of the casing 100a in addition to the display unit 140. As with the display unit 140, the operation unit 150 is not limited to the configuration provided on the front surface of the casing 100a, and may be provided on other outer surfaces (side surfaces, upper surfaces, etc.) of the casing 100 a. The user operates the start key 150a with the door constituting the container pressing portion 102 closed, thereby starting the pellet operation of the magnetic pellets 13 and sequentially performing a plurality of processing steps. Further, the user can execute the next process step while omitting a part of the process steps by operating the skip key 150b in the middle of sequentially executing a plurality of process steps.

5. Electrical structure of device for operating magnetic particles

Fig. 7 is a block diagram showing an example of an electrical configuration of the magnetic particle manipulation device 100. The magnetic particle manipulation device 100 includes a drive unit 160, an open/close sensor 170, a control unit 200, a storage unit 300, and the like, in addition to the display unit 140 and the manipulation unit 150.

The driving unit 160 is a mechanism for moving the magnet 130 in the vertical direction D1 along the container 20, and includes, for example, a motor. The open/close sensor 170 detects whether or not the door constituting the container pressing portion 102 is closed. The control unit 200 includes, for example, a CPU (central processing unit), and functions as a processing step execution unit 201, a light emission control unit 202, and the like by running a program on the CPU. The storage unit 300 is composed of, for example, a RAM (random access memory), a ROM (read only memory), a hard disk, or the like, and holds details of a processing step (for example, processing contents, processing time, or the like) for moving the magnet 130.

The process step execution unit 201 controls the drive unit 160 to move the magnet 130 from the upper side to the lower side, thereby executing a plurality of process steps corresponding to the positions of the magnet 130. The process execution unit 201 starts control of the drive unit 160 in response to the start key 150a of the operation unit 150 being operated. When the skip key 150b is operated during the sequential execution of a plurality of processing steps, the processing step execution unit 201 controls the drive unit 160 to move the magnet 130, thereby executing the processing step subsequent to the processing step being executed at that time.

When the door constituting the container holding portion 102 is opened while the plurality of processing steps are sequentially executed, the processing step execution portion 201 stops the control of the driving portion 160 based on the detection signal from the open/close sensor 170, and interrupts the processing step being executed. Then, when the door is closed, the process step execution portion 201 restarts the control of the driving portion 160 based on the detection signal from the open/close sensor 170.

The light emission control unit 202 controls the operation of the display unit 140 to cause the plurality of light emission regions 140a to 140c to emit light independently. In the present embodiment, the light-emitting region 140a corresponds to the dissolution step, the light-emitting region 140b corresponds to the cleaning step, and the light-emitting region 140c corresponds to the elution step. That is, the light emitting regions 140a to 140c corresponding to the respective processing steps executed in sequence are arranged in the same order as the execution order of the respective processing steps (see fig. 6). When a plurality of processing steps are sequentially executed, the light emission control unit 202 sequentially causes the light emission regions 140a to 140c corresponding to the respective processing steps to emit light.

Accordingly, when a plurality of processing steps corresponding to the positions of the magnets 130 are sequentially performed, the light emitting regions 140a to 140c sequentially emitting light corresponding to the respective processing steps can be visually recognized from the outside. Therefore, even in a state where the position of the magnet 130 cannot be visually recognized from the outside, the type of the processing step being executed can be determined.

Specifically, when the light-emitting regions 140a to 140c corresponding to the currently performed process are turned on and the next process is performed, the light-emitting regions 140a to 140c corresponding to the process are turned on, and the light-emitting regions 140a to 140c corresponding to the already performed process are maintained in the turned-on state. Therefore, the following states are achieved: only the light-emitting region 140a is turned on in the dissolving step, the light-emitting regions 140a and 140b are turned on in the cleaning step, and the light-emitting regions 140a to 140c are turned on in the elution step.

However, only the light-emitting regions 140a to 140c corresponding to the currently performed process may be turned on, and the light-emitting regions 140a to 140c corresponding to the already performed process may be turned off. The light emitting regions 140a to 140c are not limited to lighting, and may emit light in other patterns such as blinking.

In particular, in the present embodiment, it is possible to determine whether or not the process step (cleaning step) for removing impurities with the cleaning liquid is being performed even in a state where the position of the magnet 130 cannot be visually recognized from the outside. Therefore, it is possible to accurately determine the time when the cleaning process is performed or the time when the cleaning process is not performed, and take out the container 20 in the middle of any one of the processing processes.

The light emission control unit 202 may cause the plurality of light emission regions 140a to 140c to emit light in different patterns. For example, when a specific process step, which is a process step predetermined not to be stopped halfway, among a plurality of process steps sequentially executed is executed, the light-emitting regions 140a to 140c corresponding to the specific process step may emit light in a different manner from the light-emitting regions 140a to 140c corresponding to other process steps. This makes it possible to easily determine whether the processing step being executed is a processing step that should not be stopped halfway or is other processing steps based on the light emission pattern of the light-emitting regions 140a to 140 c. Therefore, it is possible to prevent the container 20 from being taken out by erroneously stopping the processing step during execution of the processing step that should not be stopped halfway.

The processing step that should not be stopped in the middle is arbitrary, and may be, for example, a step of reciprocating the magnet 130 at a high speed in the vertical direction D1. In addition, regarding the different patterns, for example, a light-emitting pattern that is more noticeable to the user than the light-emitting regions 140a to 140c corresponding to other processing steps, such as lighting in different colors or blinking at short intervals, is preferable.

In the present embodiment, the light emitting regions 140a to 140c are arranged in the horizontal direction, but the present invention is not limited thereto, and the light emitting regions 140a to 140c may be arranged in other directions such as the vertical direction. In addition, in the case where the cleaning process is performed in a plurality of liquid layers 11, light emitting regions may be provided independently of each other in association with the cleaning process in each liquid layer 11.

When skip key 150b is operated during the sequential execution of a plurality of processing steps to execute the next processing step, light emission control unit 202 causes light emission regions 140a to 140c corresponding to the processing step to emit light. That is, the light emitting regions 140a to 140c which the light emission control unit 202 emits light always correspond to the currently executed process step even when the skip key 150b is operated.

As described above, in the present embodiment, by operating the operation unit 150 (skip key 150b), a part of a plurality of process steps sequentially executed can be omitted and the next process step can be executed. Since the light-emitting regions 140a to 140c corresponding to the subsequent process steps are caused to emit light when the process steps are performed, the type of the process step being performed can be accurately determined even when a part of the process steps is omitted.

6. Modification example

In the above embodiment, the case where a part of the plurality of liquid layers 11 is a layer formed of a cleaning liquid for removing impurities other than the target substance has been described. However, the liquid constituting the liquid layer 11 is arbitrary, and a part of the plurality of liquid layers 11 may be a layer made of, for example, a reagent that acts on a target substance. Examples of the reagent include, but are not limited to, enzymes such as restriction enzymes for fragmenting nucleic acids and reaction enzymes for reacting fragmented nucleic acids, and other reagents may be used.

In this case, even in a state where the position of the magnet 130 cannot be visually recognized from the outside, it is possible to determine whether or not the processing step (reagent step) for causing the reagent to act on the target substance is being performed. Therefore, the time when the reagent step is performed or the time when the reagent step is not performed can be accurately determined, and the container 20 can be taken out in the middle of a certain processing step.

The reagent process may include: a restriction enzyme step of allowing a restriction enzyme to act as a reagent on a target substance; and a reaction enzyme step of allowing a reaction enzyme to act on the target substance as a reagent. In this case, the time at which the restriction enzyme step is performed and the time at which the reaction enzyme step is performed can be accurately determined by sequentially emitting light from the light-emitting regions corresponding to the restriction enzyme step and the reaction enzyme step. Therefore, the container 20 can be taken out after the restriction enzyme step and before the reaction enzyme step, for example.

When the reagent step includes the restriction enzyme step and the reaction enzyme step as described above, the user may want to perform the reaction with the reaction enzyme without fragmenting the nucleic acid by omitting the restriction enzyme step and performing the reaction enzyme step. In this case, the restriction enzyme step can be omitted and the reaction enzyme step can be performed by operating the operation unit 150 (skip key 150 b). Even in this case, by visually recognizing the light emission of the light emitting region, it is possible to accurately judge whether or not the restriction enzyme step is omitted and the reaction enzyme step is performed.