Analytical device and analytical method

Abstract

To provide an analysis device capable of determining whether or not a particulate substance to be analyzed is an expected particulate substance.SOLUTION: An analysis device 10 includes a magnetic field generation unit 20, an observation unit 30, a processing unit 42 and a storage unit 41. The magnetic field generation unit 20 generates a magnetic field to subject a particulate substance p to be analyzed to magnetic migration. The observation unit 30 observes the substance p to be analyzed. The processing unit 42 measures a magnetic migration speed of the substance p to be analyzed from the observation result by the observation unit 30 to measure volume susceptibility of the substance p to be analyzed on the basis of the magnetic migration speed and a particle size of the substance p to be analyzed. The storage unit 41 stores reference data 43 indicating volume susceptibility of a reference particulate substance. The processing unit 42 compares the volume susceptibility of the substance p to be analyzed with the reference data 43 to analyze the substance p to be analyzed.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to an analytical device and an analytical method. [Background technology]

[0002] The present inventors have previously proposed a method for quantitatively evaluating the affinity of particles for a medium using the volume magnetic susceptibility of particles (Patent Document 1).The present inventors have also previously proposed a method for analyzing the amount of functional groups on particles using the volume magnetic susceptibility of particles (Patent Document 2). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. 2016 / 208723 [Patent Document 2] International Publication No. 2016 / 208724 Summary of the Invention [Problem to be solved by the invention]

[0004] The present inventors have conducted extensive research into methods for analyzing particulate matter and have completed the present invention. The present invention aims to provide an analytical device and analytical method that can determine whether particulate matter to be analyzed is the intended particulate matter. [Means for solving the problem]

[0005] The analysis device of the present invention includes a magnetic field generation unit, an observation unit, a processing unit, and a storage unit. The magnetic field generation unit generates a magnetic field to is Particle size less than 1 μm extracellular vesicles The mixture is subjected to magnetic migration. The reference data includes the volume magnetic susceptibility and particle size of multiple types of extracellular vesicles.The observation unit observes the subject to be analyzed. The processing unit measures the magnetophoretic velocity of the subject to be analyzed from the observation result of the observation unit, and measures the volume magnetic susceptibility of the subject to be analyzed based on the magnetophoretic velocity of the subject to be analyzed and a particle diameter calculated based on the Brownian motion of the subject to be analyzed. The memory unit stores reference data indicating the volume magnetic susceptibility of a reference particulate matter. The processing unit: The data acquired by the processing unit By comparing the volume magnetic susceptibility with the reference data, Identifying the type of extracellular vesicles to be analyzed .

[0014] In one embodiment, The reference data includes information on extracellular vesicles derived from cancer cells, The processing unit determines whether the analysis target is an extracellular vesicle derived from a cancer cell.

[0020] In one embodiment, the storage unit further stores cell-identification data, which is data indicating the relationship between the volume magnetic susceptibility of at least one type of extracellular vesicle and the cell that releases the extracellular vesicle. When the processing unit determines that the analyte corresponds to one of the at least one type of extracellular vesicles, it refers to the cell identification data and determines the cell that released the analyte.

[0024] The analytical method of the present invention comprises: The subject of analysis Particle size less than 1 μm extracellular vesicles a first measuring step of measuring the magnetophoretic velocity of the analyte from the observation results, a second measuring step of measuring the volume magnetic susceptibility of the analyte based on the magnetophoretic velocity of the analyte and the particle diameter calculated based on the Brownian motion of the analyte, and an analyzing step of analyzing the analyte by comparing the volume magnetic susceptibility of the analyte with reference data. Includes volume magnetic susceptibility and particle size of multiple types of extracellular vesicles. In the analysis step, the type of extracellular vesicles to be analyzed is determined by comparing the volume magnetic susceptibility obtained in the second measurement step with the reference data. Identify.

[0033] In one embodiment, the reference data includes information on extracellular vesicles derived from cancer cells, In the analysis step, it is determined whether the analysis target is an extracellular vesicle derived from a cancer cell.

[0041] In one embodiment, The reference data includes cell-identifying data, which is data showing the relationship between the volume magnetic susceptibility of at least one type of extracellular vesicle and the cell that releases the extracellular vesicle;In the analysis step, when it is determined that the analyte corresponds to one of the at least one type of extracellular vesicles, by referring to data showing the relationship between the volume magnetic susceptibility of the at least one type of extracellular vesicles and the cells that release the extracellular vesicles, extracellular vesicles The cells that released the antibody are determined. [Effects of the Invention]

[0042] According to the analysis device and analysis method of the present invention, it is possible to determine whether or not the particulate matter to be analyzed is the intended particulate matter. [Brief explanation of the drawings]

[0043] [Figure 1] 1 is a schematic diagram of an analysis device according to a first embodiment of the present invention. [Figure 2] 1(a) and 1(b) are diagrams showing the movement of particulate matter according to the first embodiment of the present invention. [Figure 3] 1 is a diagram showing the configuration of an analysis device according to a first embodiment of the present invention. [Figure 4] FIG. 3 is a diagram showing an example of the measurement results of the volume magnetic susceptibility of the particulate matter according to the first embodiment of the present invention. [Figure 5] 1 is a flowchart showing an analysis method according to a first embodiment of the present invention. [Figure 6] FIG. 10 is a diagram showing the configuration of a modified example of the analyzer according to the first embodiment of the present invention. [Figure 7] FIG. 10 is a diagram showing an example of measurement results of volume magnetic susceptibility of a plurality of types of reference particulate matter according to the second embodiment of the present invention. [Figure 8] FIG. 10 is a schematic diagram of an analysis device according to a third embodiment of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0044] Hereinafter, embodiments of the present invention will be described with reference to the drawings. However, the present invention is not limited to the following embodiments. Note that duplicated explanations may be omitted as appropriate. In addition, the same or equivalent parts in the drawings will be designated by the same reference numerals, and explanations will not be repeated.

[0045] [Embodiment 1] First, an analysis device 10 according to the present embodiment will be described with reference to Fig. 1. Fig. 1 is a schematic diagram of the analysis device 10 according to the present embodiment. The analysis device 10 analyzes particulate matter p. More specifically, the analysis device 10 analyzes the state of the surface film of the particulate matter p.

[0046] In this embodiment, the particulate matter p is a micelle. The surface membrane of a micelle has a hydrophilic portion and a lipophilic (hydrophobic) portion. More specifically, the surface membrane of a micelle has a structure in which the hydrophilic groups face outward and the lipophilic groups face inward. For example, a micelle has a structure in which an oily component is covered with a surfactant (surface membrane).

[0047] As shown in FIG. 1, the analysis device 10 includes a magnetic field generation unit 20, an observation unit 30, and a calculation unit 40. A cell 21 is disposed near the magnetic field generation unit 20. The magnetic field generation unit 20 generates a magnetic field to magnetophoretically migrate particulate matter p in the cell 21. The observation unit 30 observes the particulate matter p in the cell 21. The calculation unit 40 measures the particle diameter and magnetophoretic velocity of the particulate matter p from the results of observation by the observation unit 30. The calculation unit 40 also measures the volume magnetic susceptibility of the particulate matter p based on the particle diameter and magnetophoretic velocity of the particulate matter p. The calculation unit 40 then analyzes the state of the surface of the particulate matter p based on the volume magnetic susceptibility of the particulate matter p. The analysis device 10 will be described in more detail below.

[0048] The magnetic field generating unit 20 generates a magnetic field gradient (a gradient of magnetic flux density) to apply a magnetic force to the particulate matter p in the cell 21. As a result, the particulate matter p undergoes magnetophoresis. In this embodiment, the magnetic field generating unit 20 includes a pair of permanent magnets that generate a magnetic field gradient. The two permanent magnets that make up the pair of permanent magnets are arranged with a gap of a fixed distance, for example, not less than 100 μm and not more than 500 μm. The cell 21 is arranged in the gap between the two permanent magnets.

[0049] In this embodiment, the cell 21 is a capillary tube. A capillary tube is an example of a tubular member. The material of the cell 21 is not particularly limited as long as it is a material that can transmit visible light or laser light L. For example, the cell 21 can be made of glass or plastic.

[0050] The particulate matter p is introduced into the cell 21 together with the medium m by, for example, a microsyringe, a micropump, or an autosampler. Alternatively, the particulate matter p can be introduced into the cell 21 together with the medium m based on the siphon principle. Alternatively, droplets containing the particulate matter p may be introduced into the cell 21 (capillary tube) by capillary action. When the droplets containing the particulate matter p are dropped onto one end of the capillary tube, the droplets flow through the capillary tube by capillary action.

[0051] Particulate matter p is present in medium m. One particulate matter p may be present in medium m, or multiple particulate matters p may be present in medium m. When multiple particulate matters p are present in medium m, the multiple particulate matters p may be dispersed in medium m or unevenly distributed in medium m. Medium m may be a liquid or a gas. For example, medium m is an aqueous electrolyte solution, a culture medium, an organic solvent, or air. The analysis device 10 measures the volume magnetic susceptibility of each of the multiple particulate matters p present in medium m.

[0052] The volume magnetic susceptibility of the particulate matter p varies depending on the state of the surface film of the particulate matter p. Specifically, the volume magnetic susceptibility of the particulate matter p varies depending on the amount of the surface film. Alternatively, the volume magnetic susceptibility of the particulate matter p varies depending on whether the surface film has disappeared or not. Therefore, the analysis device 10 can analyze whether the surface film of the particulate matter p is in the intended state based on the volume magnetic susceptibility of the particulate matter p. Alternatively, the analysis device 10 can determine whether the surface film of the particulate matter p has disappeared or not based on the volume magnetic susceptibility of the particulate matter p.

[0053] The observation unit 30 observes the particulate matter p in the cell 21 and generates a signal indicating the observation result. The calculation unit 40 measures the particle diameter and magnetophoretic velocity of the particulate matter p based on the signal generated by the observation unit 30. The calculation unit 40 includes a memory unit 41 and a processing unit 42.

[0054] The memory unit 41 stores programs, setting information, etc. The memory unit 41 may be configured, for example, by a storage device and a semiconductor memory. The storage device is, for example, a hard disk drive (HDD). The memory unit 41 may have, as semiconductor memory, for example, a random access memory (RAM) and a read only memory (ROM). The processing unit 42 executes programs stored in the memory unit 41 to perform various processes such as numerical calculations, information processing, and device control. The processing unit 42 is configured, for example, by a processor such as a central processing unit (CPU). A general-purpose computer such as a personal computer is used as the calculation unit 40.

[0055] The processing unit 42 measures the change over time in the position of the particulate matter p in the cell 21 from the observation results of the observation unit 30. For example, the processing unit 42 measures the position of the particulate matter p in the cell 21 at predetermined time intervals. In other words, the processing unit 42 measures the position of the particulate matter p at different times. The processing unit 42 measures the magnetophoretic velocity of the particulate matter p from the change over time in the position of the particulate matter p.

[0056] The processing unit 42 also measures the particle diameter of the particulate matter p from the signal generated by the observation unit 30. The processing unit 42 further measures the volume magnetic susceptibility of the particulate matter p based on the particle diameter and magnetophoretic velocity of the particulate matter p.

[0057] For example, the processing unit 42 calculates the volume magnetic susceptibility of the particulate matter p based on the following formula (1). v={2(χs-χm)r 2 / 9ημ o}B(dB / dx)···(1)

[0058] In equation (1), v is the magnetophoretic velocity of particulate matter p, χs is the volume magnetic susceptibility of particulate matter p, χm is the volume magnetic susceptibility of medium m, r is the radius of particulate matter p, η is the viscosity of medium m, and μ o is the magnetic permeability of a vacuum, B is the magnetic flux density, and dB / dx is the magnetic field gradient (gradient of the magnetic flux density). Equation (1) is derived from the fact that the difference in the magnetic force acting on the particulate matter p and the medium m in the axial direction (x direction) of the cell 21 (capillary tube) is approximately equal to the viscous drag force.

[0059] The magnetophoretic velocity v and the volume magnetic susceptibility χs of the particulate matter p are measured values ​​by the processing unit 42. The radius r of the particulate matter p is calculated, for example, by the processing unit 42 from the measured particle diameter of the particulate matter p. The volume magnetic susceptibility χm of the medium m, the viscosity η of the medium m, and the magnetic permeability μ of a vacuum are o , the magnetic flux density B, and the magnetic field gradient dB / dx are stored in advance in the storage unit 41. For example, the user can operate an input device such as a keyboard, a mouse, or a touch display to input the volume magnetic susceptibility χm of the medium m, the viscosity η of the medium m, the magnetic permeability μ of a vacuum, o , the magnetic flux density B, and the magnetic field gradient dB / dx can be stored in the storage unit 41. The volume magnetic susceptibility χm of the medium m, the viscosity η of the medium m, the magnetic permeability μ of a vacuum o are, for example, literature values. The magnetic flux density B and the magnetic field gradient dB / dx are, for example, measured values.

[0060] The memory unit 41 stores reference data 43 indicating the volume magnetic susceptibility of a reference particulate matter. In this embodiment, the reference data 43 indicates the volume magnetic susceptibility of the particulate matter p when the surface film is in a desired state. The processing unit 42 analyzes the state of the surface film of the particulate matter p (analysis target) by comparing the volume magnetic susceptibility of the particulate matter p (analysis target) with the reference data 43. More specifically, the processing unit 42 analyzes whether the surface film of the particulate matter p (analysis target) is in a desired state. Alternatively, the processing unit 42 analyzes whether the surface film of the particulate matter p (analysis target) has disappeared.

[0061] The reference data 43 is created by measuring the volume magnetic susceptibility of the desired particulate matter p. The reference data 43 of this embodiment is created by measuring the volume magnetic susceptibility of the particulate matter p when the state of the surface film is in a desired state.

[0062] Next, the movement of particulate matter p will be described with reference to Figures 2(a) and 2(b). Figures 2(a) and 2(b) are diagrams showing the movement of particulate matter p. In detail, Figures 2(a) and 2(b) show the relationship between the volume magnetic susceptibility of particulate matter p and medium m and the movement direction of particulate matter p. As shown in Figures 2(a) and 2(b), the magnetic field generation unit 20 includes a permanent magnet 20a with a north magnetic pole and a permanent magnet 20b with a south magnetic pole. The two permanent magnets 20a and 20b face each other with a cell 21 between them.

[0063] As shown in Figure 2(a), when the volume magnetic susceptibility of the particulate matter p is smaller than that of the medium m, the particulate matter p moves in a direction away from the magnetic field (magnetic field generating unit 20). On the other hand, as shown in Figure 2(b), when the volume magnetic susceptibility of the particulate matter p is larger than that of the medium m, the particulate matter p moves in a direction toward the magnetic field (magnetic field generating unit 20).

[0064] 2(a) and 2(b), the movement of the particulate matter p is determined according to the volume magnetic susceptibility of the particulate matter p and the medium m. The particulate matter p is subjected to a force in the vicinity of the ends of the permanent magnets 20a and 20b. For example, the particulate matter p is subjected to the force within a range of about ±200 μm from the vicinity of the ends of the permanent magnets 20a and 20b.

[0065] Next, the analysis device 10 will be further described with reference to Fig. 3. Fig. 3 is a diagram showing the configuration of the analysis device 10. As shown in Fig. 3, the analysis device 10 further includes a light source 50. In addition, the observation unit 30 includes a magnification unit 32 and an imaging unit 34.

[0066] The light source 50 emits relatively high-intensity light containing a visible light component. The light source 50 irradiates the cell 21 with the light. As a result, the particulate matter p is irradiated with the light. The wavelength spectrum of the light emitted from the light source 50 may be relatively broad. For example, a halogen lamp is suitably used as the light source 50.

[0067] The particulate matter p introduced into the cell 21 is magnified at an appropriate magnification by the magnifying unit 32 and imaged by the imaging unit 34. The position of the particulate matter p can be identified from the imaging result of the imaging unit 34 (the image captured by the imaging unit 34). For example, the magnifying unit 32 includes an objective lens, and the imaging unit 34 includes a charge coupled device (CCD). Alternatively, each pixel of the imaging unit 34 may be composed of a photodiode or a photomultiplier tube. The imaging unit 34 images the particulate matter p, for example, at predetermined time intervals. Note that the imaging unit 34 may image light emitted from the light source 50 and transmitted through the cell 21, or may image light emitted from the light source 50 and scattered by the particulate matter p.

[0068] The calculation unit 40 (processing unit 42) measures the change in the position of the particulate matter p over time from the imaging results of the imaging unit 34, and measures the magnetophoretic velocity of the particulate matter p from the change in the position of the particulate matter p over time.

[0069] Furthermore, the calculation unit 40 (processing unit 42) measures the particle diameter of the particulate matter p from the imaging results of the particulate matter p. For example, the calculation unit 40 (processing unit 42) executes the following process. That is, first, the image captured by the imaging unit 34 is converted to monochrome and its brightness is quantified. Next, the differential value of the brightness value is compared with a threshold value to set the boundary of the particulate matter p. Next, the area of ​​the particulate matter p is detected from the set boundary, and the particle diameter is calculated from the radius of a circle corresponding to that area. Alternatively, the center of the particulate matter p is defined, multiple straight lines passing through the center of the particulate matter p are drawn, and the average distance between two points where each straight line intersects with the boundary of the particulate matter p is calculated.

[0070] Next, the measurement results of the volume magnetic susceptibility of particulate matter p will be described with reference to Fig. 4. Fig. 4 is a diagram showing an example of the measurement results of the volume magnetic susceptibility of particulate matter p. In detail, Fig. 4 shows the measurement results of the volume magnetic susceptibility of a first particulate matter p1 and the measurement results of the volume magnetic susceptibility of a second particulate matter p2.

[0071] The first particulate matter p1 is particulate matter p whose surface film state is in a desired state. In other words, the first particulate matter p1 is the reference particulate matter. On the other hand, the second particulate matter p2 is particulate matter p whose surface film state has changed from the desired state. Alternatively, the second particulate matter p2 is particulate matter p whose surface film has disappeared.

[0072] In Fig. 4, the horizontal axis represents volume magnetic susceptibility and the vertical axis represents the proportion of particle number. Also in Fig. 4, graph 61 (solid line) represents the measurement results of the volume magnetic susceptibility of the first particulate matter p1, and graph 62 (dashed line) represents the measurement results of the volume magnetic susceptibility of the second particulate matter p2.

[0073] Graph 61 was created by measuring the volume magnetic susceptibility of each of the plurality of first particulate matter p1 introduced into cell 21. Similarly, graph 62 was created by measuring the volume magnetic susceptibility of each of the plurality of second particulate matter p2 introduced into cell 21. Specifically, graph 61 was created by measuring the volume magnetic susceptibility of each of the first particulate matter p1 and calculating the proportion of the number of first particulate matter p1 for each measured volume magnetic susceptibility. Similarly, graph 62 was created by measuring the volume magnetic susceptibility of each of the second particulate matter p2 and calculating the proportion of the number of second particulate matter p2 for each measured volume magnetic susceptibility.

[0074] Specifically, the first particulate matter p1 is fresh cream containing a dispersant. Fresh cream containing a dispersant has a micelle structure in the intended state. Specifically, the dispersant forms a surface film. The second particulate matter p2 was produced by leaving fresh cream containing a dispersant in a room temperature environment for 30 minutes. If fresh cream containing a dispersant is left in a room temperature environment for a certain period of time or longer, the dispersant will no longer be able to maintain the surface film, and as a result, the state of the surface film will change from the intended state. Specifically, the amount of the surface film will decrease. Alternatively, the surface film will disappear.

[0075] As shown in FIG. 4, the first particulate matter p1 and the second particulate matter p2 have different volume magnetic susceptibilities. The difference in volume magnetic susceptibility is due to a change in the state of the surface film from the intended state. Specifically, this is due to a change in the amount of the surface film. Alternatively, this is due to the disappearance of the surface film. Therefore, by comparing the volume magnetic susceptibility of the second particulate matter p2 (analysis target) with the volume magnetic susceptibility (reference data 43) of the first particulate matter p1 (reference particulate matter), the state of the surface film of the second particulate matter p2 (analysis target) can be analyzed. Alternatively, it can be analyzed whether the surface film has disappeared.

[0076] The above has described the analysis device 10 of this embodiment. According to this embodiment, it is possible to analyze the state of the surface film of particulate matter p. Furthermore, according to this embodiment, it is possible to analyze and evaluate food materials having a micelle structure.

[0077] The particulate matter p may be any particle having a surface membrane and is not limited to micelles. For example, the particulate matter p may be a vesicle. More specifically, the particulate matter p may be, for example, a liposome or an extracellular vesicle (EV). The surface membrane of a vesicle is made of lipids. More specifically, the surface membrane of a vesicle has a lipid bilayer. A liposome is a vesicle whose surface membrane is made of phospholipids. Liposomes are used as encapsulating materials in DDS (drug delivery systems). An extracellular vesicle is a vesicle released from a cell. An extracellular vesicle contains a portion of the intracellular components of the cell that released the extracellular vesicle.

[0078] In this embodiment, the state of the surface film of the particulate matter p is analyzed, but the internal state of the particulate matter p may also be analyzed. Specifically, it may be analyzed whether the internal state of the particulate matter p is in a desired state. In this case, the reference data 43 indicates the volume magnetic susceptibility of the particulate matter p when the internal state is in the desired state. In other words, the reference particulate matter is particulate matter p whose internal state is in the desired state.

[0079] Alternatively, it may be analyzed whether the particulate matter p is a desired particulate matter, in which case the reference data 43 indicates the volume magnetic susceptibility of the desired particulate matter.

[0080] For example, encapsulation materials (capsule-shaped products) used in DDSs encapsulate specific components (drugs or physiologically active substances), but due to manufacturing variations, the specific components may not be encapsulated. The volume magnetic susceptibility of encapsulation materials encapsulating specific components differs from that of encapsulation materials not encapsulating specific components. Therefore, by creating reference data 43 using the volume magnetic susceptibility of an encapsulation material encapsulating a specific component and comparing the volume magnetic susceptibility of the encapsulation material to be analyzed with the reference data 43, it is possible to analyze whether the specific component is encapsulated in the encapsulation material (whether the internal state of the particulate matter p is as expected). In other words, it is possible to analyze whether the encapsulation material is of good quality. The reference data 43 may also be created using the volume magnetic susceptibility of an encapsulation material not encapsulating a specific component.

[0081] For example, extracellular vesicles derived from cancer cells may contain tumor antigens. Alternatively, the types of proteins constituting the extracellular vesicles may differ between extracellular vesicles derived from cancer cells and those derived from non-cancerous cells. Alternatively, the amount of a specific protein constituting the extracellular vesicles may differ between extracellular vesicles derived from cancer cells and those derived from non-cancerous cells. As a result, extracellular vesicles derived from cancer cells have a volume magnetic susceptibility different from that of extracellular vesicles derived from non-cancerous cells. Therefore, by creating reference data 43 using the volume magnetic susceptibility of extracellular vesicles derived from cancer cells and comparing the volume magnetic susceptibility of the extracellular vesicles to be analyzed with the reference data 43, it is possible to analyze whether the extracellular vesicles are extracellular vesicles derived from cancer cells (whether the particulate matter p is the intended particulate matter). In other words, it is possible to diagnose whether a patient has cancer. The reference data 43 may also be created using the volume magnetic susceptibility of extracellular vesicles derived from non-cancerous cells.

[0082] Next, the analysis method of this embodiment will be described with reference to Fig. 5. Fig. 5 is a flowchart showing the analysis method of this embodiment. The analysis method of this embodiment can be performed using the analysis device 10 described with reference to Figs. 1 to 4.

[0083] As shown in FIG. 5, first, magnetophoretic particulate matter p (analysis target) is observed (step S1). Next, the magnetophoretic velocity and particle diameter of the particulate matter p are measured from the observation results (step S2). Next, the volume magnetic susceptibility of the particulate matter p is measured based on the measured magnetophoretic velocity and particle diameter (step S3). Next, the measured volume magnetic susceptibility is compared with reference data 43 to analyze the particulate matter p (step S4). More specifically, the state of the surface film of the particulate matter p is analyzed. Alternatively, the internal state of the particulate matter p is analyzed. Alternatively, it is analyzed whether the particulate matter p is the desired particulate matter.

[0084] When measuring the volume magnetic susceptibility of the particulate matter p, the magnetic field generating unit 20 magnetophores the particulate matter p in the cell 21, and the observing unit 30 observes the particulate matter p during magnetophoresis. Then, the processing unit 42 measures the volume magnetic susceptibility of the particulate matter p from the results of observation by the observing unit 30.

[0085] When analyzing the particulate matter p, the processing unit 42 compares the volume magnetic susceptibility of the particulate matter p with the reference data 43 stored in the memory unit 41. As already explained, the reference data 43 indicates the volume magnetic susceptibility of the particulate matter p when the surface film is in a desired state. Alternatively, the reference data 43 indicates the volume magnetic susceptibility of the particulate matter p when the internal state is in a desired state. Alternatively, the reference data 43 indicates the desired volume magnetic susceptibility of the particulate matter p.

[0086] The first embodiment of the present invention has been described above with reference to FIGS. 1 to 5. According to this embodiment, it is possible to analyze particulate matter p that is an analysis target. Specifically, it is possible to analyze whether the state of the surface film of the analysis target is a desired state. It is also possible to analyze whether the inside of the analysis target is a desired state. It is also possible to analyze whether the analysis target is a desired particulate matter.

[0087] When measuring particle diameters of less than 1 μm, laser light L is irradiated onto the particulate matter p, and the Brownian motion of the particulate matter p is analyzed. For example, vesicles such as liposomes and extracellular vesicles have particle diameters of less than 1 μm.

[0088] FIG. 6 is a diagram showing the configuration of a modified example of the analysis device 10 of this embodiment. As shown in FIG. 6, the analysis device 10 may further include a laser device 60. The laser device 60 emits laser light L. The laser device 60 irradiates the cell 21 with the laser light L. As a result, the particulate matter p is irradiated with the laser light L. The observation unit 30 observes the particulate matter p using the laser light L (scattered light) scattered by the particulate matter p in the cell 21. Specifically, the image capture unit 34 captures an image of the laser light L scattered by the particulate matter p via the magnification unit 32.

[0089] The calculation unit 40 (processing unit 42) measures the particle diameter from the imaging results of the imaging unit 34. Specifically, a diffusion coefficient is calculated from the variance of the change (displacement) in the position of the particulate matter p in the direction (y direction) perpendicular to the axial direction (x direction) of the cell 21 (capillary tube), and the particle diameter of the particulate matter p is measured from this diffusion coefficient. More specifically, the particulate matter p is affected by the magnetic field gradient in the axial direction (x direction) of the cell 21 (capillary tube), but is hardly affected by the magnetic field gradient in the direction (y direction) perpendicular to the axial direction of the cell 21. Therefore, the diffusion coefficient D can be calculated from the variance of the displacement of the position of the particulate matter p in the y direction. Specifically, the diffusion coefficient D can be calculated by dividing the square of the movement distance in the y direction of the particulate matter p undergoing Brownian motion by twice the time.

[0090] The calculation unit 40 (processing unit 42) measures the particle diameter of the particulate matter p from the diffusion coefficient D based on the following equation (2): In equation (2), d is the particle diameter of the particulate matter p, k is the Boltzmann constant, T is the absolute temperature, and η is the viscosity of the medium m. d=kT / (3πηD) (2)

[0091] In addition, the analysis device 10 shown in Figure 6 stops emitting laser light L from the laser device 60 when emitting light from the light source 50, and stops emitting light from the light source 50 when emitting laser light L from the laser device 60.

[0092] [Embodiment 2] Next, a second embodiment of the present invention will be described with reference to Figures 1 to 3, 5, and 7. However, differences from the first embodiment will be described, and a description of the same aspects as in the first embodiment will be omitted. The second embodiment differs from the first embodiment in that the type of analysis target is analyzed.

[0093] First, the analysis device 10 of this embodiment will be described with reference to FIG. 1. In this embodiment, the reference data 43 indicates the volume magnetic susceptibility of multiple types of reference particulate matter. The processing unit 42 analyzes the type of particulate matter p to be analyzed by comparing the volume magnetic susceptibility of the particulate matter p to be analyzed with the reference data 43. More specifically, the processing unit 42 determines whether the analysis target corresponds to one of the multiple types of reference particulate matter. If the analysis target corresponds to one of the multiple types of reference particulate matter, the processing unit 42 identifies the type of the analysis target from the type of the corresponding reference particulate matter. In this embodiment, the processing unit 42 further analyzes the particle diameter of the analysis target.

[0094] More specifically, the reference data 43 of this embodiment indicates the volume magnetic susceptibility and particle diameter of each of multiple types of reference particulate matter. The processing unit 42 analyzes the type of particulate matter p to be analyzed by comparing the volume magnetic susceptibility and particle diameter of the particulate matter p to be analyzed with the reference data 43. Furthermore, when the particulate matter p to be analyzed corresponds to one of the multiple types of reference particulate matter, the processing unit 42 identifies the particle diameter of the particulate matter p to be analyzed. For example, the processing unit 42 acquires a measured value of the particle diameter of the particulate matter p to be analyzed as the particle diameter of the particulate matter p to be analyzed.

[0095] Fig. 7 is a diagram showing an example of the measurement results of the volume magnetic susceptibility of a plurality of types of reference particulate matter. In other words, Fig. 7 shows an example of the reference data 43 of this embodiment. In detail, Fig. 7 shows the measurement results of the volume magnetic susceptibility of each of the first to third reference particulate matter.

[0096] In Fig. 7, the horizontal axis represents particle diameter and the vertical axis represents volume magnetic susceptibility. Also in Fig. 7, graph 71 represents the measurement results of the volume magnetic susceptibility of the first reference particulate matter pr1, graph 72 represents the measurement results of the volume magnetic susceptibility of the second reference particulate matter pr2, and graph 73 represents the measurement results of the volume magnetic susceptibility of the third reference particulate matter pr3.

[0097] Graph 71 was created by measuring the volume magnetic susceptibility and particle diameter of each of the plurality of first reference particulate matter pr1 introduced into cell 21. Specifically, graph 71 was created by measuring the volume magnetic susceptibility and particle diameter of each of the first reference particulate matter pr1 and calculating the average value of the volume magnetic susceptibility and the average value of the particle diameter in increments of 0.5 μm in particle diameter. Graphs 72 and 73 were also created in the same way as graph 71.

[0098] In detail, graph 71 shows the measurement results of the volume magnetic susceptibility and particle size of commercially available coffee drink A, graph 72 shows the measurement results of the volume magnetic susceptibility and particle size of commercially available coffee drink B, and graph 73 shows the measurement results of the volume magnetic susceptibility and particle size of commercially available coffee drink C. Commercially available coffee drinks A to C were all products that contained milk components.

[0099] As shown in FIG. 7, commercially available coffee drinks A to C had different relationships between volume magnetic susceptibility and particle size. Therefore, by comparing the volume magnetic susceptibility and particle size of the analysis target with reference data 43, it can be determined whether the analysis target corresponds to one of commercially available coffee drinks A to C. Furthermore, if the analysis target corresponds to one of commercially available coffee drinks A to C, the type of analysis target can be identified as one of commercially available coffee drinks A to C. Furthermore, after identifying the type of analysis target, the particle size of the analysis target can be analyzed. More specifically, the particle size distribution of the analysis target can be identified.

[0100] Next, the analysis method of this embodiment will be described with reference to Fig. 5. As shown in Fig. 5, first, magnetophoretic particulate matter p (analysis target) is observed (step S1). Next, the magnetophoretic velocity and particle diameter of the particulate matter p are measured from the observation results (step S2). Next, the volume magnetic susceptibility of the particulate matter p is measured based on the measured magnetophoretic velocity and particle diameter (step S3). Next, the measured volume magnetic susceptibility and particle diameter are compared with reference data 43 to analyze the type of particulate matter p to be analyzed and the particle diameter of the analysis target (step S4).

[0101] The second embodiment of the present invention has been described above with reference to Figures 1 to 3, 5, and 7. According to this embodiment, the type and particle size of particulate matter p (analysis target) can be analyzed.

[0102] Note that one or more types of analytes may be introduced into cell 21. According to the present embodiment, it is possible to introduce multiple types of analytes into cell 21 and analyze the type and particle diameter of each analyte.

[0103] Furthermore, in this embodiment, the reference data 43 indicates the volume magnetic susceptibility and particle diameter of a plurality of types of reference particulate matter, but the reference data 43 may indicate the volume magnetic susceptibility and particle diameter of one type of reference particulate matter.

[0104] In addition, in this embodiment, the processing unit 42 compares the volume magnetic susceptibility and particle diameter of the analysis target with the reference data 43, but the processing unit 42 may compare the volume magnetization of the analysis target with the reference data 43.

[0105] Furthermore, in this embodiment, the reference data 43 indicates the volume magnetic susceptibility and particle diameter of the reference particulate matter, but the reference data 43 may indicate only the volume magnetic susceptibility of the volume magnetic susceptibility and particle diameter of the reference particulate matter.

[0106] In the present embodiment, the processing unit 42 acquires the measured value of the particle diameter of the analysis target as the particle diameter of the analysis target, but the particle diameter of the analysis target is not limited to the measured value of the particle diameter of the analysis target. For example, the processing unit 42 may acquire the particle diameter of the analysis target from the reference data 43.

[0107] Furthermore, in this embodiment, the processing unit 42 analyzes the particle diameter of the analysis target, but the process of analyzing the particle diameter of the analysis target may be omitted.

[0108] [Embodiment 3] Next, a third embodiment of the present invention will be described with reference to Figures 5 and 8. However, differences from the first and second embodiments will be described, and a description of the same aspects as the first and second embodiments will be omitted. The third embodiment differs from the first and second embodiments in that it determines cells that release extracellular vesicles.

[0109] 8 is a schematic diagram of the analysis device 10 of this embodiment. In this embodiment, the reference data 43 indicates the volume magnetic susceptibility and particle diameter of multiple types of extracellular vesicles as the volume magnetic susceptibility and particle diameter of multiple types of reference particulate matter.

[0110] As in the second embodiment, the processing unit 42 determines whether the analysis target corresponds to one of a plurality of types of reference particulate matter (extracellular vesicles) by comparing the volume magnetic susceptibility and particle diameter of the analysis target particulate matter p with the reference data 43. In this embodiment, the analysis target particulate matter p is extracellular vesicles.

[0111] When the analyte corresponds to one of multiple types of reference particulate matter (extracellular vesicles), the processing unit 42 identifies the type of the analyte (extracellular vesicles) from the type of the corresponding reference particulate matter (extracellular vesicles), as in embodiment 2. In this embodiment, the processing unit 42 determines the type of cell that released the analyte from the identified type of analyte (extracellular vesicles).

[0112] Specifically, as shown in FIG. 8, the memory unit 41 further stores cell identification data 44. The cell identification data 44 indicates the relationship between the type of extracellular vesicles (reference particulate matter) and the type of cell that releases the extracellular vesicles. When the processing unit 42 identifies the type of analysis target (extracellular vesicles), it refers to the cell identification data 44 to determine the type of cell that released the analysis target. For example, the reference data 43 may indicate the volume magnetic susceptibility of extracellular vesicles derived from plankton. In this case, the cell identification data 44 indicates the relationship between the type of extracellular vesicles and the type of plankton that releases the extracellular vesicles. Alternatively, the reference data 43 may indicate the volume magnetic susceptibility of extracellular vesicles derived from lactic acid bacteria. In this case, the cell identification data 44 indicates the relationship between the type of extracellular vesicles and the type of lactic acid bacteria that release the extracellular vesicles.

[0113] Next, the analysis method of this embodiment will be described with reference to Fig. 5. As shown in Fig. 5, first, the particulate matter p (extracellular vesicles) to be analyzed undergoing magnetophoresis are observed (step S1). Next, the magnetophoretic velocity and particle diameter of the particulate matter p are measured from the observation results (step S2). Next, the volume magnetic susceptibility of the particulate matter p is measured based on the measured magnetophoretic velocity and particle diameter (step S3). Next, the measured volume magnetic susceptibility and particle diameter are compared with reference to reference data 43 to analyze the type of the particulate matter p (extracellular vesicles) and analyze the particle diameter of the particulate matter p (step S4). In this embodiment, the type of cell that released the particulate matter p is determined from the type of the particulate matter p (extracellular vesicles) identified with reference to cell identification data 44 (step S4).

[0114] The third embodiment of the present invention has been described above with reference to Figures 5 and 8. According to this embodiment, the type and particle size of the analysis target (extracellular vesicles) can be analyzed.

[0115] Note that one or more types of analytes (extracellular vesicles) may be introduced into the cell 21. According to this embodiment, multiple types of analytes can be introduced into the cell 21, and the type and particle size of each analyte (extracellular vesicle) can be analyzed.

[0116] In addition, in this embodiment, the reference data 43 indicates the volume magnetic susceptibility and particle diameter of multiple types of reference particulate matter (extracellular vesicles), but the reference data 43 may also indicate the volume magnetic susceptibility and particle diameter of one type of reference particulate matter (extracellular vesicles).

[0117] In addition, in this embodiment, the processing unit 42 compared the volume magnetic susceptibility and particle diameter of the object to be analyzed (extracellular vesicles) with the reference data 43, but the processing unit 42 may also compare the volume magnetization of the object to be analyzed with the reference data 43.

[0118] In addition, in this embodiment, the reference data 43 indicates the volume magnetic susceptibility and particle diameter of the reference particulate matter (extracellular vesicles), but the reference data 43 may indicate only the volume magnetic susceptibility of the volume magnetic susceptibility and particle diameter of the reference particulate matter.

[0119] In this embodiment, the processing unit 42 acquires the measured particle diameter of the analyte (extracellular vesicles) as the particle diameter of the analyte, but the particle diameter of the analyte is not limited to the measured particle diameter of the analyte. For example, the processing unit 42 may acquire the particle diameter of the analyte (extracellular vesicles) from the reference data 43.

[0120] Furthermore, in this embodiment, the processing unit 42 analyzes the particle diameter of the analysis target (extracellular vesicles), but the process of analyzing the particle diameter of the analysis target may be omitted.

[0121] The embodiments of the present invention have been described above with reference to the drawings. According to the embodiments of the present invention, it is possible to determine whether or not particulate matter to be analyzed is the intended particulate matter. Note that the present invention is not limited to the above embodiments, and can be embodied in various aspects without departing from the spirit and scope of the present invention.

[0122] For example, in the embodiment of the present invention, the magnetic field generating unit 20 includes a pair of permanent magnets 20a, 20b. Alternatively, the magnetic field generating unit 20 may include a pair of magnetic pole pieces to generate a magnetic field gradient. Alternatively, the magnetic field generating unit 20 may include an electromagnet, a magnetic circuit, or a superconducting magnet to generate a magnetic field gradient. When the magnetic field generating unit 20 includes a pair of magnetic pole pieces, the two magnetic pole pieces constituting the pair are spaced apart by a gap of, for example, 100 μm or more and 500 μm or less. The cell 21 is disposed in the gap between the two magnetic pole pieces. The magnetic pole pieces may be, for example, magnetized iron pieces. The iron pieces may be magnetized by, for example, a permanent magnet, an electromagnet, a magnetic circuit, or a superconducting magnet.

[0123] Furthermore, although the cell 21 is a capillary tube in the embodiment of the present invention, the cell 21 may be a glass cell or a plastic cell. The glass cell and the plastic cell have a recess for holding the medium m containing the particulate matter p. Alternatively, the glass cell and the plastic cell have a channel through which the medium m containing the particulate matter p flows. When the cell 21 is a glass cell or a plastic cell having a microchannel, when a droplet containing the particulate matter p is dropped onto one end of the microchannel, the droplet flows through the microchannel by capillary action.

[0124] Furthermore, in the embodiment described with reference to FIG. 6, the analysis device 10 is equipped with the light source 50 and the laser device 60, but the analysis device 10 may be equipped with only the laser device 60 out of the light source 50 and the laser device 60.

[0125] When using the laser device 60, the particle diameter of the particulate matter p may be measured, for example, by dynamic light scattering or static light scattering. When irradiating the particulate matter p with laser light L, the capillary tube is preferably a square capillary having a square cross section perpendicular to its axial direction. Using a square capillary makes it easy to mirror-finish the side surface of the cell 21 that is irradiated with the laser light L.

[0126] Furthermore, in the embodiment of the present invention, the calculation unit 40 (processing unit 42) measures the particle diameter of the particulate matter p, but the image captured by the imaging unit 34 may be displayed on a display, and the analyst may measure the particle diameter of the particulate matter p from the image displayed on the display. Alternatively, the image captured by the imaging unit 34 may be printed, and the analyst may measure the particle diameter of the particulate matter p from the printed image. In this case, the analyst operates the input device to store data indicating the particle diameter in the memory unit 41.

[0127] In the embodiment of the present invention, the particle diameters of the reference particulate matter and the analysis target are measured, but a literature value may be used for at least one of the particle diameters of the reference particulate matter and the analysis target. In this case, the analyst operates the input device to store data indicating the particle diameter (literature value) in the memory unit 41.

[0128] Furthermore, in the embodiment of the present invention, the calculation unit 40 (processing unit 42) measured the volume magnetic susceptibility of the reference particulate matter, but the volume magnetic susceptibility of the reference particulate matter may also be measured using a SQUID element, a magnetic balance, or the like. In this case, the analyst operates the input device to store data indicating the volume magnetic susceptibility of the reference particulate matter in the memory unit 41. Alternatively, the volume magnetic susceptibility of the reference particulate matter may be a literature value. In this case, the analyst operates the input device to store data indicating the volume magnetic susceptibility (literature value) of the reference particulate matter in the memory unit 41.

[0129] In addition, in the embodiment of the present invention, the imaging unit 34 measures the magnetophoretic velocity of the particulate matter p by imaging the particulate matter p at predetermined time intervals, but the magnetophoretic velocity of the particulate matter p may also be measured using a laser device 60, for example, based on the laser Doppler method. [Industrial Applicability]

[0130] The present invention is useful in the fields of pharmaceuticals, environmental chemistry, food, cosmetics, regenerative medicine, and the like. [Explanation of symbols]

[0131] 10 Analyzer 20 Magnetic field generation section 21 cells 30 Observation section 40 Arithmetic section 41 Storage section 42 Processing section 43 Reference Data m medium p particulate matter

Claims

1. A magnetic field generating unit that generates a magnetic field and magnetically migrates extracellular vesicles having a particle diameter of less than 1 μm that are the analysis target; an observation unit for observing the analysis target; a processing unit that measures the magnetophoretic velocity of the analyte from the observation result of the observation unit, and measures the volume magnetic susceptibility of the analyte based on the magnetophoretic velocity of the analyte and a particle diameter calculated based on the Brownian motion of the analyte; a storage unit that stores reference data indicating the volume magnetic susceptibility of reference particulate matter; Equipped with The reference data includes volume magnetic susceptibility and particle diameter of multiple types of extracellular vesicles, The processing unit identifies the type of extracellular vesicles to be analyzed by comparing the volume magnetic susceptibility acquired by the processing unit with the reference data.

2. The reference data includes information on extracellular vesicles derived from cancer cells, The analyzer according to claim 1 , wherein the processing unit determines whether the analysis target is an extracellular vesicle derived from a cancer cell.

3. The memory unit further stores cell identification data, which is data indicating a relationship between the volume magnetic susceptibility of at least one type of extracellular vesicle and a cell that releases the extracellular vesicle; The analysis device according to claim 1 or 2, wherein when the processing unit determines that the analyte corresponds to one of the at least one type of extracellular vesicles, the processing unit refers to the cell identification data to determine the cell that released the analyte.

4. A step of observing extracellular vesicles with a particle diameter of less than 1 μm that are the subject of analysis undergoing magnetic migration; a first measurement step of measuring the magnetophoretic velocity of the analyte from the observation results; a second measuring step of measuring the volume magnetic susceptibility of the analyte based on the magnetophoretic velocity of the analyte and the particle diameter calculated based on the Brownian motion of the analyte; an analyzing step of analyzing the analyte by comparing the volume magnetic susceptibility of the analyte with reference data; It encompasses The reference data includes volume magnetic susceptibility and particle diameter of multiple types of extracellular vesicles, In the analysis step, the type of extracellular vesicles to be analyzed is identified by comparing the volume magnetic susceptibility acquired in the second measurement step with the reference data. Analysis method.

5. The reference data includes information on extracellular vesicles derived from cancer cells, The analysis method according to claim 4, wherein in the analysis step, it is determined whether the analysis target is an extracellular vesicle derived from a cancer cell.

6. The reference data includes cell-identification data, which is data indicating a relationship between the volume magnetic susceptibility of at least one type of extracellular vesicle and a cell that releases the extracellular vesicle; 6. The analytical method according to claim 4 or 5, wherein, when it is determined in the analysis step that the analyte corresponds to one of the at least one type of extracellular vesicles, the cell that released the extracellular vesicles is determined by referring to data showing the relationship between the volume magnetic susceptibility of the at least one type of extracellular vesicles and the cell that released the extracellular vesicles.

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