JP2007101318A - Analyzer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an analyzer capable of mixing/stirring surely a magnetic carrier and a liquid sample, in a micro reaction space. <P>SOLUTION: This analyzer X is provided with two magnetic field generation means 10a, 10b, and a microchip A provided with an analytical part 1 for introducing the magnetic carrier 40 and a measured substance to analyze the measured substance, the two magnetic field generation means 10a, 10b are opposedly arranged under the condition where the same polarity sides thereof are directed respectively toward microchip A with the microchip A therebetween, and the analyzer is further provided with driving mechanisms 20a, 20b for approaching and separating respectively the two magnetic field generation means 10a, 10b to/from the the microchip A, and a control means 30 for controlling an operation of the driving mechanisms 20a, 20b. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an analyzer including a microchip provided with an analysis unit for analyzing a substance to be measured when a liquid sample containing the substance to be measured flows down.

  Conventionally, a technique using a magnetic carrier (magnetic carrier) is known in the case of analyzing a liquid sample containing a biological substance such as nucleic acid, protein, or fungus body as a substance to be measured.

For example, Patent Document 1 discloses a method of isolating a target biological material from a liquid sample containing the biological material using a magnetic carrier.
In this document, a magnetic carrier containing ferromagnetic iron oxide particles is used, the magnetic carrier and a biological substance are combined to form a complex, and the complex is separated from the liquid sample by an external magnetic field and separated. The biological material is isolated by eluting the biological material from the complex.

  In order to separate the complex from the liquid sample, a magnet is used as an external magnetic field. The magnet is brought close to the side wall of the reaction tube containing the liquid sample containing the formed complex, and the complex is collected on the side wall to be separated from the solution. As a magnet used as an external magnetic field, a magnet having a magnetic flux density of 2000 gauss to 3000 gauss is used.

  And in the said literature, if it is a magnetic support | carrier which has a coercive force and saturation magnetization in a specific range in a magnetic characteristic, and has an average particle size in a specific range, it will depend on a binding property and a magnetic field with a biological substance. It is described that the collection property, the dispersibility in an aqueous solution and the elution property of the biological material are compatible, and the biological material isolation performance is excellent. Such a magnetic carrier improves the isolation / purification efficiency of the biological material.

  Patent Document 2 discloses the use of a magnetic carrier having a magnetic material as a cell culture carrier. Cells can adhere to the surface of the magnetic carrier. Then, the magnetic carrier is accommodated in the culture solution in the culture vessel, and the magnetic carrier is moved by a magnetic field generator such as an electromagnet or a permanent magnet. Thereby, the culture solution can be stirred uniformly and gently, and the cells can be efficiently attached to the surface of the magnetic carrier and proliferated.

JP 2004-65132 A JP 2004-313007 A

  In Patent Document 1, a biological material is isolated using a reaction tube as a reaction vessel, and the reaction tube is overturned and stirred when mixing the solution. Moreover, in patent document 2, cell culture is performed using a culture container. That is, when a reaction tube or culture vessel is used, the reaction volume is generally several hundred microliters to several milliliters.

  In the technique of using a magnetic carrier when analyzing a liquid sample, if the reaction volume is several tens of microliters or less, there are restrictions on how to apply a magnetic field by a magnetic force generator and the recovery rate of the magnetic carrier. There are problems such as low. Therefore, it is difficult for the biological material analysis technique using a magnetic carrier to cope with a low-volume reaction system having a reaction volume of several tens of microliters or less.

  Further, for example, an immune reaction is performed in a reaction system having a reaction volume of about several tens of microliters in a well such as a microtiter plate. In general, reagents used for an immune reaction are expensive, and therefore it is required to reduce the reaction volume without lowering the detection sensitivity. If the reaction volume performed in the well is reduced, it becomes difficult to obtain a desired detection sensitivity due to evaporation of the reaction solution, and problems such as difficulty in handling due to a reduction in volume occur. There is a limit to reducing the capacity in open systems such as

  On the other hand, in recent years, microchips (biochips) manufactured by applying microfabrication technology (MEMS) such as semiconductors have been used in fields such as biochemistry and medicine. A microchip is, for example, a microanalysis device in which a structure such as a sample injection hole, a sample discharge hole, a minute capillary fluid channel, or a reaction chamber as a reaction region connected to this channel is formed. Indicates. This microchip is mainly used for analyzing biological materials.

  In the above-described microchip, a fluid channel and a reaction chamber are usually finely processed on a base material having a thickness of about 1 mm. Therefore, the thickness dimension of the reaction chamber is about several hundred micrometers, and the volume of the reaction chamber itself can be set to several tens of microliters or less.

  However, while the reaction volume in such a minute space can reduce the reaction volume, it is difficult for the user to agitate the liquid sample from the outside to the reaction chamber because it is a substantially closed space. It is. Furthermore, since the reaction chamber is a minute space, liquid convection hardly occurs even if it is mixed by inversion. Therefore, it is difficult to reliably mix and agitate the reagent and the liquid sample in the reaction chamber, and it becomes difficult to cause the reaction to proceed uniformly, making it difficult to obtain a desired detection sensitivity.

  Therefore, when a liquid sample containing a biological material is analyzed using a magnetic carrier in a minute space such as in a reaction chamber of a microchip, it is difficult to mix and agitate the magnetic carrier and the liquid sample. is there.

  Accordingly, an object of the present invention is to provide an analyzer capable of reliably mixing and stirring a magnetic carrier and a liquid sample in a fine reaction space.

  In order to achieve the above object, a first characteristic configuration of an analyzer according to the present invention is provided with two magnetic field generating means, and an analyzer for analyzing the substance to be measured by introducing a magnetic carrier and the substance to be measured. The two magnetic field generating means are arranged opposite to each other with the same polarity side facing the microchip and sandwiching the microchip, and further, the two magnetic field generating means are arranged in the microchip. And a control mechanism for controlling the operation of the drive mechanism.

  In this configuration, since the two magnetic field generating means are arranged opposite to each other with the microchip sandwiched therebetween, when the two magnetic field generating means are simultaneously close to the analysis part of the microchip, the magnetic field lines of the respective magnetic field generating means are The magnetic carrier extends from the opposite direction to the microchip. Since these magnetic field generating means are arranged with the same polarity side facing the microchip, the analysis unit is located in a region where the magnetic fields generated by the two magnetic field generating means are repelled. In this way, the magnetic carrier can be moved by applying a repulsive force that causes the magnetic field of the magnetic field generating means to deflect from the substantially vertical direction to the horizontal direction with respect to the microchip.

  For this purpose, in this configuration, each magnetic field generating means is moved close to and away from the microchip by the drive mechanism. Thereby, the state which exerts a magnetic force with respect to the magnetic carrier which exists in an analysis part, and the state which does not exert a magnetic force are switched.

For example, when one magnetic field generating means is moved from the separated state to the close state with respect to the microchip, the magnetic carrier existing in the analysis unit is magnetized. The magnetized magnetic carriers are aligned with polarity. Since the two magnetic field generating means are arranged opposite to each other with the same polarity side facing the microchip, the polarity state of this magnetic carrier can be maintained by bringing either one of the magnetic field generating means close to each other.
In this state, another magnetic field generating means different from the magnetic field generating means for maintaining the polarity state of the magnetic carrier is brought close to the magnetic carrier. At this time, since the magnetized magnetic carrier and the adjacent magnetic field generation means are in the same polarity, the magnetized magnetic carrier repels the adjacent magnetic field generation means while maintaining the polarity state, and magnetic The carrier moves in the horizontal direction to the range covered by the magnetic field lines of the adjacent magnetic field generating means.

  Then, by controlling the operation of the drive mechanism by the control means, the strength of the magnetic force acting on the magnetic carrier can be controlled, so that the magnetic carrier can be moved to a desired state.

  By moving the magnetic carrier in the analyzer in this way, the magnetic carrier in the analyzer and the liquid sample can be reliably mixed and stirred. That is, it is possible to increase the contact opportunities between various biological substances bound to the surface of the magnetic carrier and the substance to be measured contained in the liquid sample, and it is possible to make the reaction progress uniformly and increase the reaction speed. .

  The second characteristic configuration of the analyzer according to the present invention is that the two magnetic field generating units have different magnetic forces, and the control unit causes the first magnetic field generating unit of the two magnetic field generating units to be close to the analyzing unit. And aggregating the magnetic carrier, separating the first magnetic field generating means from the analyzing unit, and further, of the two magnetic field generating means, a second magnetic field generating means having a weaker magnetic force than the first magnetic field generating means. After being brought close to the agglomerated magnetic carrier, in this state, the first magnetic field generating means is brought close to the agglomerated magnetic carrier to diffuse the magnetic carrier.

  First, in a state where the magnetic carrier and the sample to be measured are introduced into the analysis unit, they coexist in the analysis unit, but reliable mixing is not performed. For example, in the initial state, the two magnetic field generating means are respectively separated from the microchip. In this state, the two magnetic field generating means do not exert a magnetic force on the magnetic carrier existing in the analysis unit.

Then, the first magnetic field generating means is brought close to the analysis unit. At this time, the magnetic force from the first magnetic field generating means reaches the magnetic carrier, and the magnetic carrier is attracted to the first magnetic field generating means and aggregates.
Here, the first magnetic field generating means is separated from the analysis unit, and control is performed so that the magnetic force of the first magnetic field generating means does not reach the aggregated magnetic carrier.

  Next, the second magnetic field generating means is brought close to the aggregated magnetic carrier. Since the two magnetic field generating means are arranged opposite to each other with the same polarity side facing the microchip, the agglomerated magnetic carrier exerts the same magnetic force as that when the first magnetic field generating means is brought close to the analyzer. It is. At this time, the magnetic carrier maintains an aggregated state.

  In this state, the first magnetic field generating means is brought close to the agglomerated magnetic carrier. At this time, the second magnetic field generating means tries to maintain the polarity of the magnetic carrier. However, since the magnetic force of the first magnetic field generating means is stronger than the magnetic force of the second magnetic field generating means, the repulsive force between the magnetic carrier and the first magnetic field generating means is greater than the force with which the second magnetic field generating means attracts the magnetic carrier. . For this reason, the agglomerated magnetic carrier is strongly pressed against the second magnetic field generating means, and accordingly, the magnetic carrier is also spread in the in-plane direction of the microchip.

  Therefore, according to the second feature configuration, the magnetic carrier existing in the analysis unit can be aggregated and diffused. By repeating the series of operation control described above, the magnetic carrier can be repeatedly agglomerated and diffused, so that the mixing and stirring of the magnetic carrier and the liquid sample are further ensured.

  A third characteristic configuration of the analyzer according to the present invention is that the area along the microchip in the two magnetic field generating means is configured such that the second magnetic field generating means is larger than the first magnetic field generating means.

  When the first magnetic field generating means approaches the aggregated magnetic carrier, the magnetic carrier and the first magnetic field generating means repel each other and the magnetic carrier moves. In particular, when the magnetic carrier deviates from the surface of the second magnetic field generating means along the microchip, the direction of the polarity generated in the magnetic carrier changes along the direction of the magnetic field lines of the second magnetic field generating means. As a result, the repulsive force between the first magnetic field generating means having a strong magnetic force and the magnetic carrier is also reduced, and the effect of diffusing the magnetic carrier is reduced. In some cases, the magnetic carrier may be adsorbed on the first magnetic field generating means side.

  However, if the second magnetic field generating means is configured to be larger than the first magnetic field generating means in the area along the microchip as in the present configuration, the surface on which the second magnetic field generating means can maintain the polarity state of the magnetic carrier. Can be set large. Therefore, since the repulsive force between the magnetic carrier and the first magnetic field generating means can secure at least the surface along the microchip of the second magnetic field generating means, the extent to which the agglomerated magnetic carrier diffuses can be increased. .

  A fourth characteristic configuration of the analyzer according to the present invention is that it includes a slide mechanism that moves at least one of the magnetic field generating means along the microchip.

  According to the fourth characteristic configuration, the magnetic carrier can be guided to a desired position by moving the magnetic field generating means along the microchip in the vicinity of the position where the magnetic carrier is present. it can.

  In addition, when the two magnetic field generating means are arranged to face each other and at least one of the magnetic field generating means is slightly slid along the microchip by the slide mechanism, the state of the magnetic field can be changed. The movement can be performed efficiently.

  A fifth characteristic configuration of the analyzer according to the present invention is that the drive mechanism includes positioning means for setting a distance of the magnetic field generating means with respect to the microchip.

  When the positioning means is provided as in the fifth characteristic configuration, the position of the magnetic field generating means can be adjusted so that the strength of the lines of magnetic force exerted on the magnetic carrier existing in the microchip is optimized.

  The sixth characteristic configuration of the analyzer according to the present invention comprises two magnetic field generating means, and a microchip provided with an analysis unit for introducing the magnetic carrier and the substance to be measured to analyze the substance to be measured, The two magnetic field generating means are disposed opposite each other with the same polarity side facing the microchip and sandwiching the microchip, and the magnetic field generating means is composed of an electromagnet, and controls the current flowing through the electromagnet. Thus, magnetic field control means for separately controlling the magnetic field strengths of the two magnetic field generating means is provided.

  In this configuration, the two magnetic field generating means are electromagnets, and these electromagnets are opposed to each other with the microchip sandwiched therebetween, so that when the two electromagnets simultaneously exert a magnetic force on the analysis part of the microchip, The field lines of the electromagnet extend from the opposite direction to the microchip to the magnetic carrier. Since these electromagnets are arranged with the same polarity side facing the microchip, the analysis unit is located in a region where the magnetic fields generated by the two electromagnets repel. Thus, the magnetic carrier can be moved by applying a repulsive force that causes the magnetic field of the electromagnet to be deflected from the substantially vertical direction to the horizontal direction with respect to the microchip.

  For this purpose, in this configuration, each electromagnet is individually controlled to be turned on and off by magnetic field control means for controlling the current flowing in the electromagnet and controlling the strength of the magnetic fields of the two electromagnets. Thereby, the state which exerts a magnetic force with respect to the magnetic carrier which exists in an analysis part, and the state which does not exert a magnetic force are switched.

For example, when one electromagnet is turned on, the magnetic carrier present in the analysis unit is magnetized. The magnetized magnetic carriers are aligned with polarity. Since the two electromagnets are opposed to each other with the same polarity facing the microchip, the polarity state of the magnetic carrier can be maintained by turning on one of the electromagnets.
In this state, another electromagnet different from the electromagnet that maintains the polarity of the magnetic carrier is turned on. At this time, since the magnetized magnetic carrier and the electromagnet operated on are in the same polarity, the magnetized magnetic carrier repels the electromagnet operated on while maintaining the state of polarity, and the magnetic carrier is It moves in the horizontal direction to the range covered by the magnetic field lines of the electromagnet turned on.

  And by controlling the magnetic field control means, it is possible to control the strength of the magnetic force acting on the magnetic carrier, so that the magnetic carrier can be moved to a desired state.

  By moving the magnetic carrier in the analyzer in this way, the magnetic carrier in the analyzer and the liquid sample can be reliably mixed and stirred. That is, it is possible to increase the contact opportunities between various biological substances bound to the surface of the magnetic carrier and the substance to be measured contained in the liquid sample, and it is possible to make the reaction progress uniformly and increase the reaction speed. .

Embodiments of the present invention will be described below with reference to the drawings.
The present invention is an analyzer including a microchip provided with an analysis unit for analyzing a substance to be measured when a liquid sample containing the substance to be measured flows down. 1 and 2 show an analyzer X of the present embodiment.

The analyzer X of the present embodiment includes two magnetic field generating units 10a and 10b, a microchip A provided with an analysis unit 1 that introduces a magnetic carrier 40 and a substance to be measured and analyzes the substance to be measured. The two magnetic field generating means 10a and 10b are opposed to each other with the same polarity side facing the microchip A and sandwiching the microchip A therebetween.
And the drive mechanism 20 which makes two magnetic field generation | occurrence | production means 10a, 10b approach and separate separately with respect to the microchip A, and the control means 30 which controls operation | movement of the drive mechanism 20 are provided.

(Magnetic field generation means)
In the present embodiment, two magnetic field generating means 10a and 10b having different magnetic forces are provided.
The magnetic field generation means 10 is a device that generates a magnetic field for moving the magnetic carrier 40 introduced from the outside, for example, in the analysis unit 1, and a permanent magnet, an electromagnet, or the like can be applied.

In this embodiment, the case where a permanent magnet is applied as the magnetic field generation means 10 is illustrated. As the permanent magnet, a neodymium magnet, a samarium cobalt magnet, a ferrite magnet, an alnico magnet or the like can be applied. These permanent magnets can be applied in various shapes such as a round shape, a square shape, and a ring shape. Permanent magnets are small and lightweight, are not easily affected by the environment of use such as temperature, humidity, vibration, and impact, and have excellent characteristics such as no need to supply energy for generating a magnetic field from the outside. .
Since neodymium magnets and samarium cobalt magnets are powerful rare earth magnets, it is easy to improve the performance and miniaturization of the magnetism generating means. Neodymium magnets are more powerful permanent magnets than samarium cobalt magnets and can produce a strong magnetic field.

The two magnetic field generating means 10a and 10b are opposed to each other with the same polarity side facing the microchip A and sandwiching the microchip A therebetween.
That is, the magnetic field generating means 10a and 10b are opposed to each other, for example, with the N pole side facing each other, and the microchip A is disposed between the magnetic field generating means 10a and 10b.

  Since the magnetic forces of the two magnetic field generating means 10 are different, the magnetic field generating means have different magnetic flux densities. Further, since the two magnetic field generating means 10a and 10b are opposed to each other with the microchip A sandwiched therebetween, when the two magnetic field generating means 10a and 10b are close to the analyzing unit 1 of the microchip A at the same time, respectively. Magnetic field lines of the magnetic field generating means 10a and 10b extend from the opposite direction to the microchip A to the magnetic carrier 40. And since these magnetic field generation means 10 are arrange | positioned toward the microchip A with the same polarity side, the analysis part 1 will be located in the area | region where the magnetic field by two magnetic field generation means 10a, 10b repels ( FIG. 3). In this way, the magnetic carrier 40 can be moved by applying a repulsive force to the magnetic carrier 40 so that the magnetic field of the magnetic field generating means 10a, 10b is deflected from the substantially vertical direction to the horizontal direction with respect to the microchip A. it can.

  In this embodiment, the areas along the microchip A of the two magnetic field generating means 10a and 10b are different from each other. For example, the second magnetic field generating unit 10b is configured larger than the first magnetic field generating unit 10a. A permanent magnet (first magnetic field generating means) 10a having a strong magnetic force is designed to have a diameter of about 3 mm, and a permanent magnet (second magnetic field generating means) 10b having a low magnetic force is set to have a diameter of about 6 mm. The magnetic force of the first magnetic field generating means 10a is about 350 millitesla, and the magnetic force of the second magnetic field generating means 10b is about 100 millitesla. However, the magnitude | size and magnetic force of each magnetic field generation | occurrence | production means 10a, 10b are not restricted to these.

  Hereinafter, a neodymium magnet and a samarium cobalt magnet are applied as the two magnetic field generating means 10a and 10b. The first magnetic field generating means is a neodymium magnet having a strong magnetic force, and the second magnetic field generating means is a samarium cobalt magnet having a weak magnetic force compared to the first magnetic field generating means.

(Microchip)
The microchip A is manufactured by applying a microfabrication technology (MEMS) such as a semiconductor, and is used for measuring a measurement substance contained in a liquid sample in fields such as biochemistry and medicine. As shown in FIG. 2, the microchip A includes, for example, an injection hole 2 for injecting a liquid sample or the like into a member, minute capillary fluid flow paths 3a to 3b, a liquid sample flowing down the fluid flow path 3, or the like. The structure of the discharge part 4 which discharges | emits, and the analysis part 1 (reaction chamber) as an analysis area | region connected with the fluid flow path 3 is formed.

Glass, quartz, silicon resin, or the like can be used as a constituent material of the microchip A, but is not limited thereto. Of the silicon resins, it is preferable to use a PDMS substrate a1 mainly composed of polydimethylsiloxane (hereinafter abbreviated as PDMS) from the viewpoint of easy molding and optical characteristics.
The PDMS substrate a1 is excellent in adhesion with a glass plate or the like. Therefore, the fluid flow path 3, the analysis unit 1, and the like can be formed by bringing a flat glass plate into contact with the finely processed PDMS substrate a <b> 1. Hereinafter, in the present embodiment, a microchip A in which a flat glass plate a2 is brought into contact with a finely processed PDMS substrate a1 will be exemplified.

  In the analysis unit 1, the liquid sample introduced from the injection hole 2 is mixed with the carrier-containing solution containing the magnetic carrier 40, and the substance to be measured contained in the liquid sample is analyzed using an optical technique or the like.

The inner surface of the analysis unit 1 (that is, the surface of the PDMS base material a1 and the glass plate a2 that forms the analysis unit 1) can be coated. Examples of the coating material include BSA and gas-impermeable polymers such as polyparaxylene resins. By applying the coating material to the inner surface, it is possible to prevent the substance to be measured and the magnetic carrier 40 in the liquid sample from being adsorbed on the inner surface in advance. Thereby, since noise at the time of detection can be reduced, accurate analysis can be performed.
The coating may be applied not only to the inner surface of the analysis unit 1 but also to the entire surfaces of the PDMS substrate a1 and the glass plate a2. In this case, for example, the substance to be measured and the magnetic carrier 40 can be prevented from being adsorbed on the inner surface of the fluid channel 3 in advance.

  In addition, the microchip A can be provided with chambers 5 to 6 in which a reaction buffer, a washing buffer, and the like are previously stored. At this time, each of the chambers 5 to 6 is connected to the fluid flow path 3a via the flow paths 7 to 8, respectively.

The dimensions of the microchip A can be set as appropriate. For example, in the case of a rectangular shape, the long dimension is about 3 to 8 cm and the short dimension is about 2 to 5 cm. If it is circular, the diameter is about 5 to 10 cm. The thickness dimensions of the PDMS substrate a1 and the glass plate a2 can be about 0.5 to 1 mm.
When the thickness dimension of the PDMS substrate a1 and the glass plate a2 is about 1 mm, the analysis unit 1 is finely processed to have a height of about 0.1 to 0.5 mm, for example. For example, the planar size of the analysis unit 1 is 5 × 18 mm, but is not limited thereto. At this time, the volume of the analysis unit 1 is a fine space of several tens of microliters or less.

  The fluid flow path 3 can be set, for example, to a width of about 0.4 mm and a height of about 0.1 mm. However, the size is not limited to this as long as the magnetic carrier 40 has a size that allows the fluid carrier 3 to flow down smoothly.

(Liquid sample)
The liquid sample refers to a liquid sample containing or possibly containing the substance to be measured. As long as the sample satisfies this requirement, it may be derived from any source. For example, it can be obtained from cells, cultures, tissues, body fluids, urine, serum, environmental samples, biopsy samples, and the like.

(Substance to be measured)
The substance to be measured can be any substance to be measured such as microorganisms or viruses and fragments thereof, DNA or RNA fragments, chemical substances, polymers such as proteins, and the like. For example, pathogenic bacteria such as E. coli contained in a sample collected from a river or the like, harmful chemical substances such as DDT contained in a sample collected from soil, etc. are exemplified.

(Magnetic carrier)
The magnetic carrier 40 is contained in a carrier-containing solution. The magnetic carrier 40 may be made of any material as long as it has magnetic responsiveness. For example, metal particles such as iron, cobalt, and nickel, oxides such as iron oxide and chromium dioxide, and composites of these oxides can be used. In particular, granular materials (ferromagnetic oxide particles) obtained by oxidizing metal particles containing iron oxide as the main component are excellent in stability when dispersed in various chemicals and are sensitive to magnetic fields. Therefore, it is widely used as a magnetic carrier.

  As the ferromagnetic iron oxide particles, for example, ferrite particles such as magnetite particles, maghemite particles, and manganese zinc ferrite particles are suitable.

  The shape of the magnetic carrier 40 can take various shapes such as a granular shape, a spherical shape, an elliptical shape, a rectangular shape, and a plate shape. The average particle size of the magnetic carrier 40 is preferably about 5 to 10 μm. Thus, the magnetic carrier 40 can be set to a desired surface area by variously changing the shape and particle size of the magnetic carrier 40.

The magnetic carrier 40 is magnetized to some extent when the magnetic force of the magnetic field generating means 10 reaches. Therefore, when the magnetic carrier 40 is agglomerated and diffused by the magnetic field generating means 10, it is necessary to set a holding force that does not increase the cohesive force between the magnetized magnetic carriers 40.
Further, the greater the saturation magnetization, the more the response to the magnetic force of the magnetic field generating means 10 is improved, which affects the efficiency when the magnetic carrier 40 is aggregated and diffused.

  The surface of the magnetic carrier 40 can be coated with various inorganic compounds and organic compounds as long as the magnetic response is not affected. For example, a silica coating or a silane coating can be formed on the iron oxide particles. When nucleic acid is bound to the surface of the magnetic carrier 40 as a biological material, it is effective to deposit silica on the surface of the magnetic carrier 40.

In addition, when proteins such as antibodies and enzymes are bound as biological substances to the surface of the magnetic carrier 40, these proteins can be directly bound to the magnetic carrier 40. Further, if various organic compounds are bonded to the particle surface of the magnetic carrier 40, the protein can be bonded more efficiently. Examples of organic compounds that can be used include albumin, carbodiimide, glutaraldehyde, streptavidin, biotin, and a silane coupling agent having a functional group.
Thereby, the magnetic support | carrier 40 can exhibit the function as a support | carrier for biological material coupling | bonding effectively.

If an immunological technique is used to detect the substance to be measured, an antibody against the substance to be measured (antigen) is supported on the magnetic carrier 40. Then, by means of a so-called “sandwich method”, the substance to be measured is sandwiched between the labeled antibody and the antibody immobilized on the surface of the magnetic carrier, so that an immunospecific complex is formed in the analysis unit 1 and the substance to be measured is measured. The substance can be captured.
Thus, the presence of the substance to be measured can be detected or quantitatively measured by labeling the antibody or the like that forms the immunospecific complex. The labeling may be performed on either the substance to be measured or the antibody.
In this way, the substance to be measured in the liquid sample can be detected with high sensitivity.

  Although the case where an immunological technique is used for detecting a substance to be measured has been described, the present invention is not limited to such a form, and a substance that can recognize a substance to be measured, that is, a substance to be measured is selectively detected. It is possible to support a binding substance having molecular affinity to be obtained on a magnetic carrier. Examples of the binding substance having the ability to discriminate the molecule to be measured include not only the specific binding ability between antigen and antibody, but also the complementary binding ability between nucleic acids and the biological response ability between ligand and receptor. Illustrated.

Detection of immune-specific complex formed by antigen-antibody reaction is performed by, for example, labeling an antibody with a fluorescent (luminescent) substance and directly detecting the fluorescence (luminescent) intensity, or by binding an enzyme to the antibody and chemically An optical change is detected by carrying out an enzymatic reaction using a luminescent substrate.
For example, when the fluorescence intensity of a fluorescent substance is measured with a fluorescence measurement device, the measured labeling amount is compared with the labeling intensity when a known amount of “measuring substance” is measured, thereby determining the amount of the measuring substance in the liquid sample. Can be determined.

(Drive mechanism)
The drive mechanism 20 is configured so that the two magnetic field generating means 10a and 10b are closely spaced from the microchip A. For example, the first magnetic field generating means 10a is brought close to the microchip A by the driving mechanism 20a provided with the arm means 22 for holding the first magnetic field generating means 10a and the support means 23 for supporting the arm means 22. Can be separated. On the other hand, the second magnetic field generating means 10b can be moved close to and away from the microchip A by the drive mechanism 20b housed in the mounting table 24 on which the microchip A is mounted. The drive mechanism 20 may have any configuration as long as each magnetic field generating unit 10 can be moved close to and away from the microchip A.

  That is, by moving the magnetic field generating means 10a and 10b separately to and from the microchip A by the driving mechanisms 20a and 20b, for example, the state and magnetic force exerted on the magnetic carrier 40 existing in the analysis unit 1 Can be switched to a state that does not affect. For example, the distance between the first magnetic field generating means 10a and the microchip A may be set to about 0.5 to 1 mm, or the magnetic field generating means 10a and the microchip A may be brought into contact with each other.

  Further, the magnetic field generating means 10a and 10b can be separated by the drive mechanisms 20a to 20b until the magnetic carrier 40 is not affected. For example, the distance between the first magnetic field generating means 10a and the microchip A can be set to be separated by about 4 to 6 mm or more.

The drive mechanism 20 includes positioning means 21 for setting the distance of the magnetic field generating means 10 with respect to the microchip A. The positioning means 21 can be applied in any form as long as the distance between the microchip A and the magnetic field generation means 10 is maintained. For example, the proximity position of the first magnetic field generation means 10a can be set by positioning means 21 such as a stopper provided in the drive mechanism 20a.
When the positioning means 21 is provided in this way, the position of the magnetic field generating means 10 can be adjusted so that the strength of the magnetic lines of force exerted on the magnetic carrier 40 existing in the microchip A is optimized.

(Control means)
The control unit 30 controls the operation of the drive mechanism 20.
The carrier-containing solution is a suspension solution of the magnetic carrier 40 containing a large number of minute magnetic carriers 40. Then, by controlling the operation of the drive mechanism 20 by the control unit 30, the magnetic carrier 40 can be controlled, so that the magnetic carrier 40 can be moved to a desired state.

  By moving the magnetic carrier 40 in the analyzer 1 in this way, the magnetic carrier 40 in the analyzer 1 and the liquid sample can be reliably mixed and stirred. That is, the contact opportunity between the magnetic carrier 40 and the substance to be measured contained in the liquid sample can be increased.

  For example, the control unit 30 drives the magnetic field generation units 10a and 10b to be close to the microchip A for a desired time, and then separates the magnetic field generation units 10a and 10b from the microchip A for a desired time. It can be configured by a microcomputer or the like provided with software capable of controlling the operation of the mechanism 20.

(Operation control of magnetic field generation means)
The control unit 30 controls the operation of the drive mechanisms 20a to 20b so that the magnetic carrier 40 is aggregated and diffused, for example. This operation control will be described below.

  First, a carrier-containing solution containing the magnetic carrier 40 and a liquid sample containing the sample to be measured are respectively introduced from the liquid injection hole 2, and the fluid flow path 3 a is caused to flow down and stored in the analysis unit 1. At this time, the carrier-containing solution and the liquid sample coexist in the analysis unit 1 but are not reliably mixed.

  In the analysis apparatus X of the present invention, the two magnetic field generating means 10a and 10b are arranged to face each other with the same polarity side (for example, the N pole side) facing the microchip A and the microchip A sandwiched therebetween. In FIG. 1, the first magnetic field generating means 10a having a strong and small magnetic force (diameter 3 mm) is placed on the upper side of the microchip A, and the second magnetic field generating means 10b having a weak magnetic force (diameter 6 mm) is placed on the lower side of the microchip A. It is arranged.

  In the initial state, the two magnetic field generating means 10a and 10b are separated from the microchip A (FIG. 4A). At this time, the two magnetic field generating means 10 a and 10 b do not exert a magnetic force on the magnetic carrier 40 existing in the analysis unit 1.

  Of the two magnetic field generating means 10a and 10b, the first magnetic field generating means 10a having a strong magnetic force is brought close to the analysis unit 1. At this time, the magnetic force from the first magnetic field generating unit 10a is attracted to the magnetic carrier 40 and the magnetic carrier 40 and aggregates (FIGS. 4B and 5A).

  Here, the 1st magnetic field generation means 10a is separated from the analysis part 1 (FIG.4 (c)). At this time, the magnetic force of the first magnetic field generating means 10a does not reach the aggregated magnetic carrier 40.

  Next, the second magnetic field generating means 10b having a weak magnetic force out of the two magnetic field generating means 10a and 10b is brought close to the aggregated magnetic carrier 40 (FIG. 4D). Since the two magnetic field generating means 10a and 10b are arranged opposite to each other with the same polarity side facing the microchip A, when the first magnetic field generating means 10a is brought close to the analyzing unit 1 in the aggregated magnetic carrier 40 A magnetic force having the same polarity as that in FIG. At this time, the magnetic carrier 40 maintains an aggregated state.

  In this state, the first magnetic field generating means 10a is brought close to the agglomerated magnetic carrier 40 (FIG. 4E). At this time, the second magnetic field generating means 10b tries to maintain the polarity state of the magnetic carrier 40. However, since the magnetic force of the first magnetic field generating means 10a is stronger than the magnetic force of the second magnetic field generating means 10b, the repulsive force between the magnetic carrier 40 and the first magnetic field generating means 10a is the second magnetic field generating means 10b. It becomes larger than the force to attract. For this reason, the agglomerated magnetic carrier 40 is strongly pressed against the second magnetic field generating means 10b, and accordingly, the magnetic carrier 40 is also spread in the in-plane direction of the microchip A. (FIG. 4 (e), FIG. 5 (b)).

  In this way, the magnetic carrier 40 present in the analysis unit 1 can be aggregated and diffused. The series of operation control of the magnetic field generation means 10 can be performed in about 30 seconds, for example, but is not limited thereto. By repeating the above-described series of operation control, the aggregation and diffusion movement of the magnetic carrier 40 can be repeated, so that the mixing and stirring of the magnetic carrier and the liquid sample are further ensured.

  This aggregation / diffusion movement of the magnetic carrier 40 is considered to be caused because the magnetic carrier 40 is magnetized to some extent when the first magnetic field generating means 10a having a strong magnetic force is brought close to the analysis unit 1.

That is, for example, when the magnetic carrier 40 is magnetized when the first magnetic field generating means 10a with the N pole facing the microchip A is brought close to the analysis unit 1, the magnetized magnetic carrier 40 causes the S pole to be the first magnetic field. Aggregates on the generation means 10a side with the N pole directed toward the second magnetic field generation means 10b (FIG. 4B).
Then, the second magnetic field generating means 10b with the north pole facing the microchip A is brought close to the agglomerated magnetic carrier 40 (FIG. 4D). At this time, the magnetized magnetic carrier 40 maintains an agglomerated state, but the polarity toward the second magnetic field generating means 10b is reversed from the N pole to the S pole due to the influence of the second magnetic field generating means 10b. That is, the magnetic carrier 40 is in a state in which the N pole is directed toward the first magnetic field generating means 10a.
In this state, the first magnetic field generating means 10a with the N pole facing the microchip A is brought close to the agglomerated magnetic carrier 40 (FIG. 4E). At this time, since the magnetic carrier 40 is in a state where the N pole is directed to the first magnetic field generating means 10a, the first magnetic field generating means 10a and the magnetic carrier 40 repel each other, and the magnetic carrier 40 generates the first magnetic field. It spreads in the horizontal direction to the range covered by the magnetic field lines of the means 10a.

  Normally, convection hardly occurs in a liquid stored in a fine space such as the analysis unit 1 of the microchip A. For this reason, it is difficult to reliably mix the two liquids in the analysis unit 1, and it is difficult to cause the reaction to proceed uniformly.

However, in the analyzer X of the present invention, since the magnetic carrier 40 can be aggregated and diffused in a fine space (analyzer 1) provided in the microchip A, the carrier containing the magnetic carrier 40 is contained. Mixing and stirring of the solution and the liquid sample can be performed reliably.
As a result, various biological substances (for example, antibodies) bound to the surface of the magnetic carrier 40 can be efficiently brought into contact with a substance to be measured (for example, an antigen against the antibody) contained in the liquid sample. It is possible to advance the reaction uniformly and increase the reaction rate.

[Another embodiment 1]
In the embodiment described above, it is possible to provide a slide mechanism (not shown) that moves at least one of the permanent magnets along the microchip A.

  For example, the slide mechanism is configured so that one of the magnetic field generating means 10 can be slid from the injection hole 2 to the analysis unit 1 along the fluid flow path 3a. Then, the magnetic carrier 40 injected into the injection hole 2 is attracted by moving the magnetic field generation means 10 from the injection hole 2 along the fluid flow path 3 to the analysis unit 1 in a state where the magnetic field generation means 10 is in proximity to the microchip A. The analysis unit 1 can be led.

  In addition, when the two magnetic field generating means 10a and 10b are arranged to face each other and at least one of the permanent magnets is slightly slid along the microchip A by the slide mechanism, the state of the magnetic field can be changed, Aggregation and diffusion movement of the magnetic carrier 40 can be performed efficiently.

[Another embodiment 2]
In the above-described embodiment, the case where a permanent magnet is used as the magnetic field generating means 10 has been described, but the case where an electromagnet is used will be described below.
The electromagnet includes a metal core and a copper wire wound around the core. By controlling the current flowing through the electromagnet, the strength of the magnetic force generated from the electromagnet and the presence or absence of the magnetic force can be changed. The current is controlled by magnetic field control means. The magnetic field control means is configured to be able to control the intensity of the current flowing through the electromagnet or the on / off of the current.

In this way, the two electromagnets are configured to have different magnetic forces by the magnetic field control means, or to turn on and off the two electromagnets separately by the magnetic field control means.
That is, after the first electromagnet of two electromagnets is turned on to aggregate the magnetic carrier, the first electromagnet is turned off, and the second electromagnet whose magnetic force is weaker than the first electromagnet of the two electromagnets In this state, the first electromagnet is turned on to set the magnetic field control means to diffuse the magnetic carrier.

  Thereby, similarly to the above-described embodiment, the aggregation and diffusion of the magnetic carrier existing in the analysis unit can be repeatedly performed.

[Another embodiment 3]
In the embodiment described above, the case where the microchip A provided with the analysis unit 1 which is a minute space is applied as the reaction chamber is illustrated. However, the present invention is not limited to a microchip provided with a reaction chamber, and can be applied to any container that contains a magnetic carrier and a substance to be measured in a fine closed space.
Furthermore, even in a reaction vessel provided with a reaction chamber that is not a fine closed space, if it is necessary to contain a magnetic carrier and a substance to be measured and mix and agitate them, apply the reaction vessel. Can do.

[Another embodiment 4]
In the above-described embodiment, the analysis apparatus X to which one microchip A is applied is illustrated, but the present invention is not limited to this, and a plurality of microchips can be applied.
At this time, a plurality of microchips A are placed, for example, in a rotatable disc-like plate shape, and one microchip A is transported to a position where the magnetic field generating means 10 is provided. Then, after the magnetic carrier 40 is appropriately aggregated and diffused by the drive mechanism 20 and the control unit 30, another microchip is transported to a position where the magnetic field generating unit 10 is provided.

  In this way, by transferring a plurality of microchips A sequentially to the position where the magnetic field generating means 10 is provided, a large number of liquid samples can be analyzed efficiently and the analyzer X can be easily automated.

  Further, when the analysis apparatus X is automated, for example, a PCR reaction apparatus, another apparatus such as a detection apparatus that can detect the label intensity, and the analysis apparatus X are combined, and a control unit that can control these apparatuses in association is provided. Thus, it is possible to construct an analysis system useful for the experimenter. That is, it is possible to reduce the chance of contact with reagents, samples, and the like that are harmful to the experimenter, and more efficiently analyze the samples.

  When using a microchip or the like provided with an analysis unit for analyzing a substance to be measured using a magnetic carrier, the analyzer of the present invention ensures that the magnetic carrier and the substance to be measured are in a small space. It can be used for mixing and stirring.

Schematic diagram of the analyzer of the present invention Schematic diagram of microchip Diagram showing the concept when two magnetic field generating means exert magnetic force on the magnetic carrier Schematic showing a series of operation control of magnetic field generation means Diagram showing the moving state of the magnetic carrier

Explanation of symbols

A Microchip X Analyzer 1 Analyzer 10a, 10b Magnetic field generator 20a-20b Drive mechanism 21 Positioner 30 Controller 40 Magnetic carrier

Claims (6)

Two magnetic field generating means, and a microchip provided with an analysis unit for introducing the magnetic carrier and the substance to be measured and analyzing the substance to be measured, each of the two magnetic field generating means having the same polarity side of the microchip. It is arranged facing the chip with the microchip sandwiched therebetween, and
An analysis apparatus comprising: a driving mechanism that moves the two magnetic field generating units close to and away from the microchip; and a control unit that controls the operation of the driving mechanism. The two magnetic field generating means have different magnetic forces,
The control means includes
After aggregating the magnetic carrier by bringing the first magnetic field generating means close to the analyzing unit out of the two magnetic field generating units, the first magnetic field generating unit is separated from the analyzing unit, and
Of the two magnetic field generating means, a second magnetic field generating means having a lower magnetic force than the first magnetic field generating means is brought close to the aggregated magnetic carrier, and in this state, the first magnetic field generating means is moved to the aggregated magnetic carrier. The analyzer according to claim 1, wherein the analyzer is set so as to diffuse the magnetic carrier in the vicinity.   The analyzer according to claim 2, wherein an area along the microchip in the two magnetic field generation units is configured such that the second magnetic field generation unit is larger than the first magnetic field generation unit.   The analyzer according to any one of claims 1 to 3, further comprising a slide mechanism that moves at least one of the magnetic field generating units along the microchip.   The analyzer according to any one of claims 1 to 4, wherein the driving mechanism includes positioning means for setting a distance of the magnetic field generating means with respect to the microchip. Two magnetic field generating means, and a microchip provided with an analysis unit for introducing the magnetic carrier and the substance to be measured and analyzing the substance to be measured, each of the two magnetic field generating means having the same polarity side of the microchip. It is arranged facing the chip with the microchip sandwiched therebetween, and
An analysis apparatus, wherein the magnetic field generating means comprises an electromagnet, and magnetic field control means for controlling the current flowing through the electromagnet to control the strength of the magnetic fields of the two magnetic field generating means separately.