Light scattering detection device and light scattering detection method

Abstract

Provided are a light scattering detection device and a light scattering detection method, which can well maintain molecular weight calculation accuracy and particle size calculation accuracy, for example, regardless of the arrangement angle of the detector. The light scattering detection device comprises: a sample analysis chamber; a light source, which irradiates coherent light to the sample analysis chamber; a plurality of detectors, which receive scattered light scattered from the sample analysis chamber to the surrounding at different scattering angles; and a plurality of apertures, which limit the scattered light, wherein the sample analysis chamber has a sample channel sealed with a liquid sample, the light source is arranged so that the coherent light enters from one end side of the sample channel and passes through the sample channel, the plurality of detectors are arranged on the same circumference centered on the central axis of the sample analysis chamber extending in the vertical direction (Z-axis direction), and the opening width of each aperture is the largest when the arrangement angle is 90°, and becomes smaller as the arrangement angle is away from 90°.

Description

Technical Field

[0001] The invention relates to a light scattering detection device and a light scattering detection method. Background Art

[0002] As a method for separating microparticles such as proteins dispersed in a liquid sample, size exclusion chromatography (SEC) and gel filtration chromatography (GPC) are known. In recent years, as a chromatographic detection device, in addition to using an ultraviolet (UV) absorbance detection device and a differential refractive index detection device, a multi-angle light scattering (MALS) detection device is also used. The multi-angle light scattering detection device has the characteristic of being able to calculate the molecular weight and particle size of the measurement sample.

[0003] As a multi-angle light scattering detection device, a detection device is known, which includes: an analysis chamber, which has a through hole, which is formed to penetrate in the radial direction and is used to fill a liquid sample; a light source, which irradiates a light beam toward the through hole; and a plurality of detectors, which are arranged at intervals along the outer circumference of the analysis chamber and receive scattered light scattered from the analysis chamber (liquid sample) (for example, refer to Patent Document 1).

[0004] Prior art literature

[0005] Patent Literature

[0006] Patent Document 1: Japanese Patent Application Laid-Open No. 61-120947 Summary of the invention

[0007] Problem that the invention aims to solve

[0008] There is also a detection device having a structure in which a plurality of apertures are added to the multi-angle light scattering detection device described in Patent Document 1 to limit the scattered light incident on the detector (see Fig.10 ). Fig.10 The conventional multi-angle light scattering detection device 1000 shown in the figure comprises: an analysis chamber 1002, which has a through hole 1001, which is formed to penetrate in the radial direction (X-axis direction) and is used to fill the liquid sample Q; a light source 1003, which irradiates the light beam BM toward the through hole 1001; a focusing lens 1007, which is arranged between the analysis chamber 1002 and the light source 1003; a plurality of detectors 1004, which are arranged at intervals along the outer periphery of the analysis chamber 1002 to receive scattered light scattered from the analysis chamber 1002 (liquid sample Q); and a plurality of apertures 1006, which are arranged between the analysis chamber 1002 and each detector 1004, and use the width of the opening 1005 to limit the scattered light incident on the detector 1004.

[0009] In addition, Fig.10 , as the detector 1004 and the aperture 1006, the detector 1004A, the first aperture 1006A-1, and the second aperture 1006A-2 located at the configuration angle θ1, and the detector 1004B, the first aperture 1006B-1, and the second aperture 1006B-2 located at the configuration angle θ2 larger than the configuration angle θ1 are representatively depicted. Furthermore, the widths of the openings 1005 of the first aperture 1006A-1, the second aperture 1006A-2, the first aperture 1006B-1, and the second aperture 1006B-2 are all the same.

[0010] like Fig.11 As shown in FIG. 1 , in the multi-angle light scattering detection device 1000, the light receiving area of ​​the detector 1004A configured at the angle θ1 overlaps with the scattered light generating area more than the light receiving area of ​​the detector 1004B configured at the angle θ2 overlaps with the scattered light generating area. Therefore, it can be said that the multi-angle light scattering detection device 1000 has the following tendency: as the configuration angle moves away from 90 degrees, the scattered light generating area of ​​the scattered light received by the detector 1004 becomes larger. Therefore, as Fig.12 The graph of is as follows: even if each detector is arranged at an equal distance from the center of the analysis chamber, as long as the arrangement angle is different, the scattered light generation area of ​​the scattered light received by each detector will be deviated. This deviation becomes an error in calculating, for example, molecular weight and particle size, and therefore, it is difficult to perform correct calculations.

[0011] An object of the present invention is to provide a light scattering detection device and a light scattering detection method that can maintain good molecular weight calculation accuracy and particle size calculation accuracy, for example, regardless of the arrangement angle of the detector.

[0012] Solutions for solving problems

[0013] A first aspect of the present invention is related to a light scattering detection device, which is used to detect microparticles in a liquid sample, and the light scattering detection device comprises: a transparent sample analysis chamber, which is used to hold the liquid sample; a light source, which irradiates coherent light to the sample analysis chamber; a plurality of detectors, which receive scattered light scattered from the sample analysis chamber to the surrounding at different scattering angles; and a plurality of apertures, which are arranged between the sample analysis chamber and each of the detectors and limit the scattered light incident on the detector by using the opening width, wherein the sample analysis chamber has a sample channel, which is formed to pass through the sample analysis chamber in a straight line, and the sample The channel is sealed with the liquid sample, the light source is configured to allow the coherent light to be incident from one end side of the sample channel and pass through the sample channel, the multiple detectors are configured on the same circumference centered on the central axis of the sample analysis chamber extending in the vertical direction, the multiple detectors include a first detector and a second detector, and a reference position is taken as a position at an angle of 90° relative to the incident direction of the coherent light incident on the sample analysis chamber, the first detector is configured at a position closer to the reference position, and the second detector is configured at a position farther from the reference position, and the opening width of the diaphragm of the first detector is larger than the opening width of the diaphragm of the second detector.

[0014] A second aspect of the present invention is related to a light scattering detection method for detecting microparticles in a liquid sample, the light scattering detection method comprising the following steps: sealing the liquid sample in a sample channel, the sample channel being formed to linearly penetrate a transparent sample analysis chamber for holding the liquid sample; irradiating coherent light from a light source from one end side of the sample channel so that the coherent light passes through the sample channel; and a scattered light receiving step, in which a plurality of detectors are used to receive scattered light scattered from the sample analysis chamber to the surroundings at different scattering angles, the plurality of detectors being arranged at a center extending in a vertical direction from the sample analysis chamber. The scattered light receiving process includes the following process: limiting the scattered light incident on each of the detectors by using the opening width of a plurality of apertures arranged between the sample analysis chamber and each of the detectors, the plurality of detectors including a first detector and a second detector, taking a position at an angle of 90° relative to the incident direction of the coherent light incident on the sample analysis chamber as a reference position, the first detector being arranged at a position closer to the reference position, the second detector being arranged at a position farther from the reference position, and the opening width of the aperture of the first detector being larger than the opening width of the aperture of the second detector.

[0015] Effects of the Invention

[0016] According to the present invention, the size of the overlapped range between the light receiving area of ​​each detector and the scattered light generating area can be made consistent, i.e., the same, regardless of the configuration angle. As a result, the light intensity at each detector is substantially the same, i.e., within the range of the allowable error, and therefore, for example, the molecular weight calculation accuracy and the particle size calculation accuracy can be well maintained regardless of the configuration angle of the detector. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 It is a plan view showing a first embodiment of the light scattering detection device of the present invention.

[0018] Figure 2 It shows that the Figure 1 FIG. 5 is a graph showing relative values ​​of the intensity of scattered light received by each detector at each arrangement angle in the case of the light scattering detection device shown.

[0019] Figure 3 1 is a flowchart showing the process sequence of the light scattering detection method of the present invention.

[0020] Figure 4 It is a plan view showing a second embodiment of the light scattering detection device of the present invention.

[0021] Figure 5 This indicates that the moving mechanism is used when it is stopped. Figure 2 FIG. 5 is a graph showing light intensity at various configuration angles in the case of the light scattering detection device shown.

[0022] Figure 6 It shows that the moving mechanism is used in the state of motion. Figure 2 FIG. 5 is a graph showing relative values ​​of the intensity of scattered light received by each detector at each arrangement angle in the case of the light scattering detection device shown.

[0023] Figure 7 It is a plan view showing a third embodiment of the light scattering detection device of the present invention.

[0024] Figure 8 It is a plan view showing a fourth embodiment of the light scattering detection device of the present invention.

[0025] Fig. 9 It shows that the Figure 8 FIG. 5 is a graph showing relative values ​​of the intensity of scattered light received by each detector at each arrangement angle in the case of the light scattering detection device shown.

[0026] Fig.10 It is a plan view showing the structure of a conventional light scattering detection device.

[0027] Fig.11 It is used to illustrate Fig.10FIG. 5 is a diagram showing the difference in scattered light generation areas of scattered light received by each detector in the light scattering detection device shown.

[0028] Fig.12 It shows that the Fig.10 FIG. 5 is a graph showing relative values ​​of the intensity of scattered light received by each detector at each arrangement angle in the case of the light scattering detection device shown.

[0029] Description of Reference Numerals

[0030] 1: light scattering detection device; 2: sample analysis chamber; 21: cylindrical portion; 211: outer peripheral surface; 22: sample channel; 221: one end; 222: the other end; 3: light source; 4, 4A, 4B: detector; 5: aperture; 5A-1, 5B-1: first aperture; 5A-2, 5B-2: second aperture; 51: opening; 6: focusing lens; 61: convex surface; 62: plane; 7A, 7B: moving unit; 71: moving mechanism; 72: control unit; 73: storage unit; 74: base; 8A, 8B: rotating unit; 81: rotating mechanism; 82: control unit; 83: storage unit; 84: light adjustment member; 1000: multi-angle light scattering detection device; 1001: through hole; 1002: analysis chamber; 1003: light source; 1004, 1004A, 1004B: detector; 1005: opening; 1006: aperture; 1006A-1, 1006B-1: first aperture; 1006A-2, 1006B-2: second aperture; 1007: focusing lens; BM: light beam; L1: coherent light; L2: scattered light; O 21 : Central axis; O 84 : rotation axis; Q: liquid sample; R: radius; W 51 : opening width; θ, θ1, θ2: configuration angles. DETAILED DESCRIPTION

[0031] Hereinafter, a light scattering detection device and a light scattering detection method according to the present invention will be described in detail based on preferred embodiments shown in the drawings.

[0032] <First Embodiment>

[0033] Figure 1 It is a plan view showing a first embodiment of the light scattering detection device of the present invention. Figure 2 It shows that the Figure 1 FIG. 5 is a graph showing relative values ​​of the intensity of scattered light received by each detector at each arrangement angle in the case of the light scattering detection device shown. Figure 3: is a flowchart showing the process sequence of the light scattering detection method of the present invention. In addition, hereinafter, for the convenience of explanation, one direction in the horizontal direction is referred to as the "X-axis direction", the direction in the horizontal direction perpendicular to the X-axis direction is referred to as the "Y-axis direction", and the vertical direction, that is, the direction perpendicular to the X-axis direction and the Y-axis direction is referred to as the "Z-axis direction". In addition, sometimes the arrow side of each axis direction is referred to as the "positive side", and the side opposite to the arrow is referred to as the "negative side". In addition, Figure 2 , Figure 6 , Fig. 9 , Fig.12 The graphs shown are all results obtained by setting the light intensity obtained by receiving scattered light generated at the center of the analysis chamber (x=0) as 1 to calculate the analysis chamber position dependence (x dependence) of the relative value of the intensity of the received scattered light.

[0034] Figure 1 The light scattering detection device 1 of the present invention shown is a multi-angle light scattering (MALS) detection device for detecting the molecular weight and the radius of rotation (size) of microparticles such as proteins dispersed in a liquid sample Q. The light scattering detection device 1 includes: a transparent sample analysis chamber 2 for holding the liquid sample Q; a light source 3 for irradiating the sample analysis chamber 2 with coherent light L1; a focusing lens 6 for focusing the coherent light L1 incident on the sample analysis chamber 2; a plurality of detectors 4 for receiving scattered light L2 scattered from the sample analysis chamber 2 (liquid sample Q); and a plurality of apertures 5 for limiting the scattered light L2 incident on the detector 4.

[0035] In addition, the light scattering detection method of the present invention is a method of detecting the molecular weight and the rotation radius of microparticles such as proteins dispersed in the liquid sample Q using the light scattering detection device 1. Figure 3 As shown, the light scattering detection method includes a liquid sample sealing step (first step), a coherent light irradiation step (second step), and a scattered light receiving step (third step), and these steps are performed sequentially.

[0036] like Figure 1 As shown, the sample analysis chamber 2 is cylindrical and has a central axis O 21 The cylindrical portion 21 is arranged parallel to the Z-axis direction. The cylindrical portion 21 has a sample channel 22, which is formed to pass through the cylindrical portion 21 (sample analysis chamber 2) in a straight line along the X-axis direction, and the sample channel 22 is sealed with a liquid sample Q. In addition, the sample channel 22 is preferably parallel to the central axis O 21 cross.

[0037] The columnar portion 21 is made of a transparent material. The material of the cylindrical portion 21 is not particularly limited, and an example thereof is colorless and transparent quartz glass.

[0038] In the liquid sample sealing step, the liquid sample Q is sealed in the sample channel 22. This sealing operation may be performed automatically by a sealing device or manually by an operator, for example.

[0039] The light source 3 irradiates the sample analysis chamber 2 with coherent light L1. "Coherent light" refers to light in which the phase relationship between light waves at any two points in the light beam does not change in time and remains fixed, and after the light beam is split by any method, it shows complete interference even when a large optical path difference is applied and the light beam is put together again. As the light source 3, for example, a laser light source for irradiating visible light laser can be used. Completely coherent light L1 does not exist in nature, and laser oscillating in a single mode is light close to a coherent state.

[0040] The light source 3 is arranged on the negative side in the X-axis direction relative to the sample analysis chamber 2, and faces the one end 221 side of the sample channel 22. Thus, the coherent light L1 emitted from the light source 3 enters from the one end 221 side of the sample channel 22. Then, the coherent light L1 passes through the sample channel 22 and is emitted from the other end 222 side of the sample channel 22.

[0041] In the coherent light irradiation step, the sample analysis chamber 2 is irradiated with coherent light L1 using the light source 3 . Thus, the sample channel 22 is irradiated with coherent light L1 from the light source 3 from the one end 221 side thereof so that the coherent light L1 passes through the sample channel 22 .

[0042] A condenser lens 6 is disposed between the light source 3 and the sample analysis chamber 2. The condenser lens 6 is a plano-convex lens, and the incident side of the coherent light L1 is a convex surface 61 and the emitting side is a flat surface 62.

[0043] In addition, the light scattering detection device 1 may be provided with a condensing optical system composed of a plurality of compound lenses and condensing lenses instead of the condensing lens 6. This condensing optical system can also condense the coherent light L1 incident on the sample analysis chamber 2 in the same manner as the condensing lens 6.

[0044] A plurality of detectors 4 are arranged around the sample analysis chamber 2 so as to be spaced apart from the outer peripheral surface 211 of the cylindrical portion 21. As described above, the coherent light L1 passes through the sample channel 22. Moreover, during the passage, the coherent light L1 is scattered from the sample analysis chamber 2 to the surroundings at different scattering angles due to the liquid sample Q, thereby becoming scattered light L2. Each detector 4 is capable of receiving the scattered light L2. In addition, in the light scattering detection device 1, the outer peripheral surface 211 of the cylindrical portion 21 plays the role of a lens, and the light receiving surface of each detector 4 is located at its focal position. Therefore, the plurality of detectors 4 are in the following state: around the central axis O extending in the vertical direction of the sample analysis chamber 2 21 The light receiving surface is arranged on the same circumference with φ as the center, that is, on the circumference of the radius R.

[0045] In addition, Figure 1 In the figure, when the position at an angle of 90° relative to the incident direction of the coherent light L1 incident on the sample analysis chamber 2 is set as the reference position, as the detector 4, a detector (first detector) 4A arranged at a position farther from the reference position, that is, at a configuration angle θ1, and a detector (second detector) 4B arranged at a position closer to the reference position, that is, at a configuration angle θ2 larger than the configuration angle θ1 are representatively depicted.

[0046] In addition, although a photodiode is used as the detector 4 in the present embodiment, the present invention is not limited to this, and for example, an array detector such as a two-dimensional CMOS may be used.

[0047] In the scattered light receiving step, the scattered light L2 can be received by the plurality of detectors 4. The plurality of detectors 4 are arranged on the XY plane with the central axis O of the sample analysis chamber 2 being the same as that of the sample analysis chamber 2. 21 on the same circle with the center being the same circle.

[0048] Between the sample analysis chamber 2 and each detector 4, a plurality of apertures 5 are arranged at intervals along the optical axis direction of the scattered light L2. Each aperture 5 has an opening 51 formed to penetrate along the optical axis direction of the scattered light L2. At least the sides of the opening 51 along the vertical direction are straight lines, preferably in the shape of a rectangle that is long in the vertical direction (Z-axis direction). Moreover, each aperture 5 can utilize the opening width W of the opening 51. 51 The scattered light L2 incident on the detector 4 corresponding to the aperture 5 is limited.

[0049] Furthermore, the scattered light receiving step includes a scattered light limiting step (refer to Figure 3 ), in the scattered light limiting step, the opening width W of the plurality of apertures 5 disposed between the sample analysis chamber 2 and each detector 4 is used. 51 , to limit the scattered light L2 incident on each detector 4.

[0050] In addition, Figure 1 In the figure, as multiple apertures 5, a first aperture plate 5A-1 located on the sample analysis chamber 2 side and a second aperture plate 5A-2 located on the detector 4 side, which are arranged between the sample analysis chamber 2 and the detector 4A, and a first aperture plate 5B-1 located on the sample analysis chamber 2 side and a second aperture plate 5B-2 located on the detector 4 side, which are arranged between the sample analysis chamber 2 and the detector 4B.

[0051] The opening width W of the first aperture plate 5A-1 and the second aperture plate 5A-2 at the arrangement angle θ1 51The aperture widths W of the first aperture plate 5B-1 and the second aperture plate 5B-2 at the configuration angle θ2 are the same as each other and are used to limit the scattered light L2 incident on the detector 4A in a stepwise manner. 51 They are also the same as each other and are used to limit the scattered light L2 incident on the detector 4B in a step-by-step manner.

[0052] In the light scattering detection device 1, the opening width W of each aperture 5 is 51 The opening width W of each aperture 5 is different depending on the arrangement angle θ. 51 It is the largest when the configuration angle θ is 90°, and becomes smaller as the configuration angle θ moves away from 90°. Figure 1 As shown, the opening width W at the configuration angle θ1 is 51 The opening width W at the configuration angle θ2 51 In addition, in the present embodiment, when the configuration angle θ is set to 90°, the opening width W 51 Set to W 51(MAX) In the case of 51 The opening width W 51(MAX) Multiply the value from the central axis O of the sample analysis chamber 2 21 The value obtained by the distance to each detector 4, that is, the radius R and the sine value of the arrangement angle θ of each detector 4 (=W 51(MAX) ×R×sinθ). In addition, as long as the object of the present invention can be achieved, a value obtained by adding a slight correction to this value is also included in the scope of the present invention.

[0053] As described above, the following result is obtained: regardless of the arrangement angle θ, the opening width W 51 If all are the same, even if each detector 4 is arranged at a distance O from the central axis of the sample analysis chamber 2 21 Even at positions of equal distance, deviations occur in the scattered light generation regions of the scattered light received by each detector 4 (see Fig.10 , Fig.12 ). Moreover, this deviation becomes an error in calculating the molecular weight and particle size, for example, and therefore, it is difficult to perform accurate calculations.

[0054] On the other hand, in the light scattering detection device 1, the opening width W is set as described above. 51 In this case, the size of the overlapped range between the light receiving area of ​​each detector 4 and the scattered light generating area can be made uniform, that is, the same, regardless of the configuration angle θ. Therefore, as the detection result of each detector 4, Figure 2 The curve diagram of Figure 2 As shown in the graph of 21In the case of positions at equal distances, the light intensity at each detector 4 is substantially the same, that is, within the range of the allowable error. Thus, in the light scattering detection device 1, regardless of the configuration angle θ of the detector 4, for example, the molecular weight and particle size can be accurately calculated, that is, the molecular weight calculation accuracy and particle size calculation accuracy can be well maintained. In addition, Figure 2 The graph of is the result when the refractive index of the solvent in the liquid sample Q is the same as the refractive index of the sample analysis chamber 2 (columnar portion 21 ) (eg, a refractive index of 1.46).

[0055] <Second Embodiment>

[0056] Figure 4 It is a plan view showing a second embodiment of the light scattering detection device of the present invention. Figure 5 This indicates that the moving mechanism is used when it is stopped. Figure 2 FIG. 5 is a graph showing relative values ​​of the intensity of scattered light received by each detector at each arrangement angle in the case of the light scattering detection device shown. Figure 6 It shows that the moving mechanism is used in the state of motion. Figure 2 FIG. 5 is a graph showing relative values ​​of the intensity of scattered light received by each detector at each arrangement angle in the case of the light scattering detection device shown.

[0057] Hereinafter, a second embodiment of the light scattering detection device and the light scattering detection method of the present invention will be described with reference to these drawings, but the description will focus on the differences from the above-described embodiment, and description of the same matters will be omitted.

[0058] This embodiment is the same as the first embodiment except that it includes a moving mechanism for moving the first aperture plate.

[0059] like Figure 4 As shown in FIG. 1 , the light scattering detection device 1 of the present embodiment includes a moving unit 7A for moving the first aperture plate 5A-1 and a moving unit 7B for moving the first aperture plate 5B-1. The moving unit 7A and the moving unit 7B have the same structure except that the aperture 5 to be moved is different, so the moving unit 7A will be described as a representative example below. In addition, in the light scattering detection device 1, the moving unit 7B can also be omitted depending on the size of the configuration angle θ2.

[0060] The moving unit 7A includes a moving mechanism 71 that moves the first aperture plate 5A- 1 relative to the second aperture plate 5A- 2 , a control unit 72 that controls the operation of the moving mechanism 71 , and a storage unit 73 that stores the refractive index information of the solvent in the liquid sample Q.

[0061] The moving mechanism 71 is connected to the first aperture plate 5A-1 and is composed of, for example, a motor, a ball screw, a linear guide, etc. The moving mechanism 71 can move the first aperture plate 5A-1 in parallel with the second aperture plate 5A-2 and in the horizontal direction, that is, in the direction of the tangent of the tangent at the intersection of the outer peripheral surface 211 of the sample analysis chamber 2 on the outer peripheral surface 211 and the optical axis of the scattered light L2 to the detector 4A.

[0062] The storage unit 73 stores refractive index information of the solvent in each liquid sample Q.

[0063] The control unit 72 extracts the refractive index information of the solvent in the liquid sample Q to be analyzed from the storage unit 73. Then, the control unit 72 operates the moving mechanism 71 based on the extracted refractive index information of the solvent to control the movement amount (movement distance) of the first aperture plate 5A-1.

[0064] Furthermore, in the scattered light limiting step, based on the refractive index information of the solvent in the liquid sample Q, the first aperture plate 5A- 1 can be moved in parallel with the second aperture plate 5A- 2 in the horizontal direction.

[0065] In addition, when the refractive index of the solvent in the liquid sample Q is different from the refractive index of the sample analysis chamber 2 (cylindrical portion 21) (for example, the refractive index of the solvent is 1.333, and the refractive index of the sample analysis chamber 2 is 1.46), if the moving units 7A and 7B are kept stopped, the following is obtained: Figure 5 The results are as shown in the graph. Figure 5 As can be seen from the graph, the light intensity at each detector 4 has the following tendency: the smaller the configuration angle θ (for example, when the configuration angle θ is 28 degrees), the more it deviates from the light intensity when the configuration angle θ is large.

[0066] Therefore, even when the refractive index of the solvent in the liquid sample Q is different from the refractive index of the sample analysis chamber 2 (cylindrical portion 21), the light scattering detection device 1 can move the first aperture plate 5A-1 located at the configuration angle θ1 where the configuration angle θ is small relative to the second aperture plate 5A-2 as described above to prevent the deviation of the light intensity caused by the size of the configuration angle θ. Figure 6 The results are as shown in the graph. Figure 6 As can be seen from the graph of , no matter how the configuration angle θ is, the graphs of light intensity corresponding to each configuration angle θ are substantially overlapped with each other, preventing the above-mentioned deviation from occurring. Thus, regardless of the configuration position of the detector 4, for example, molecular weight and particle size can be accurately calculated.

[0067] <Third Embodiment>

[0068] Figure 7It is a plan view showing a third embodiment of the light scattering detection device of the present invention.

[0069] Hereinafter, a third embodiment of the light scattering detection device and the light scattering detection method of the present invention will be described with reference to the drawing, but the description will be mainly focused on the differences from the above-mentioned embodiments, and the description of the same matters will be omitted.

[0070] This embodiment is the same as the second embodiment except that the moving object of the moving unit is different.

[0071] like Figure 7 As shown, in this embodiment, the moving unit 7A is configured to move the second aperture plate 5A-2 and the detector 4A, and the moving unit 7B is configured to move the second aperture plate 5B-2 and the detector 4B. In this embodiment, the moving unit 7A and the moving unit 7B are also the same in structure except that the aperture 5 as the moving object is different, so the moving unit 7A is described as a representative example below.

[0072] The moving mechanism 71 of the moving unit 7A is connected to a base 74 on which the second aperture plate 5A-2 and the detector 4A are placed, and can move the second aperture plate 5A-2 and the detector 4 together in the horizontal direction parallel to the first aperture plate 5A-1.

[0073] The control unit 72 operates the moving mechanism 71 based on the refractive index information of the solvent extracted from the storage unit 73 to control the movement amount (movement distance) of the second aperture plate 5A- 2 and the detector 4A.

[0074] Furthermore, in the scattered light limiting step, based on the refractive index information of the solvent in the liquid sample Q, the second aperture plate 5A- 2 and the detector 4A can be moved in parallel with the first aperture plate 5A- 1 in the horizontal direction.

[0075] According to the above configuration, regardless of the size of the arrangement angle θ, it is possible to prevent the above-mentioned deviation between the light intensities detected by the detectors 4. Therefore, regardless of the arrangement position of the detectors 4, for example, the molecular weight and particle size can be accurately calculated.

[0076] <Fourth Embodiment>

[0077] Figure 8 It is a plan view showing a fourth embodiment of the light scattering detection device of the present invention. Fig. 9 It shows that the Figure 8 FIG. 5 is a graph showing relative values ​​of the intensity of scattered light received by each detector at each arrangement angle in the case of the light scattering detection device shown.

[0078] Hereinafter, a fourth embodiment of the light scattering detection device and the light scattering detection method of the present invention will be described with reference to these drawings, but the description will focus on the differences from the above-mentioned embodiments, and description of the same matters will be omitted.

[0079] This embodiment is the same as the second embodiment except that a rotation unit is provided instead of a moving unit.

[0080] like Figure 8 As shown in FIG. 1 , the light scattering detection device 1 of the present embodiment includes a rotating unit 8A and a rotating unit 8B. The rotating unit 8A and the rotating unit 8B have the same structure except for the different configuration positions, so the rotating unit 8A will be described as a representative example below. In addition, in the light scattering detection device 1, the rotating unit 8B can also be omitted depending on the size of the configuration angle θ2.

[0081] The rotation unit 8A has: a light adjustment component 84, which is arranged between the first aperture plate 5A-1 and the second aperture plate 5A-2; a rotation mechanism 81, which rotates the light adjustment component 84; a control unit 82, which controls the action of the rotation mechanism 81; and a storage unit 83, which stores the refractive index information of the solvent in the liquid sample Q.

[0082] The rotation mechanism 81 is connected to the light adjustment member 84 and is composed of, for example, a motor, a reducer, etc. The rotation mechanism 81 can make the light adjustment member 84 rotate around a rotation axis O parallel to the Z-axis direction. 84 , that is, rotate in the horizontal direction.

[0083] The light adjustment member 84 rotates around the rotation axis O 84 By rotating, the position of the scattered light L2 (light) from the first aperture plate 5A-1 to the second aperture plate 5A-2 can be adjusted. In addition, the light adjustment member 84 is composed of parallel plate glass. Thus, the light adjustment member 84 can be set to a simple structure, so, for example, the manufacturing cost of the light adjustment member 84 can be suppressed.

[0084] The storage unit 83 stores refractive index information of the solvent in each liquid sample Q.

[0085] The control unit 82 extracts the refractive index information of the solvent in the liquid sample Q to be analyzed from the storage unit 83. Then, the control unit 82 operates the rotation mechanism 81 based on the extracted refractive index information of the solvent to control the rotation amount (rotation angle) of the light adjusting member 84.

[0086] Furthermore, in the scattered light limiting step, the light beam adjusting member can be rotated in the horizontal direction based on the refractive index information of the solvent in the liquid sample Q.

[0087] In the light scattering detection device 1 having the above structure, the following is obtained: Fig. 9 The results are as shown in the graph. Fig. 9 As can be seen from the curve graph, no matter how large the configuration angle θ is, the curve graphs of light intensity corresponding to each configuration angle θ are substantially overlapped with each other, thus preventing the above-mentioned deviation from occurring. Thus, regardless of the configuration position of the detector 4, for example, molecular weight and particle size can be accurately calculated.

[0088] The light scattering detection device and the light scattering detection method of the present invention have been described above with respect to the illustrated embodiments, but the present invention is not limited thereto. In addition, each part constituting the light scattering detection device can be replaced with any structure that can perform the same function. In addition, any structure can be added. In addition, the light scattering detection device and the light scattering detection method of the present invention can also be obtained by combining any two or more structures (features) in the above-mentioned embodiments.

[0089] [Way]

[0090] It will be understood by those skilled in the art that the above-mentioned multiple exemplary embodiments are specific examples of the following aspects.

[0091] (Item 1) A light scattering detection device according to one embodiment is used to detect microparticles in a liquid sample, and includes:

[0092] A transparent sample analysis chamber, which is used to hold the liquid sample;

[0093] a light source for irradiating coherent light to the sample analysis chamber;

[0094] a plurality of detectors for receiving scattered light scattered from the sample analysis chamber to surroundings at different scattering angles; and

[0095] a plurality of apertures, the plurality of apertures being arranged between the sample analysis chamber and each of the detectors and limiting the scattered light incident on the detector by using an opening width;

[0096] The sample analysis chamber has a sample channel, the sample channel is formed to linearly penetrate the sample analysis chamber, and the liquid sample is sealed in the sample channel.

[0097] The light source is configured to allow the coherent light to enter from one end side of the sample channel and pass through the sample channel.

[0098] The plurality of detectors are arranged on the same circumference centered on a central axis of the sample analysis chamber extending in a vertical direction.

[0099] The opening width of each of the apertures is maximum when the arrangement angle with respect to the incident direction of the coherent light into the sample analysis chamber is 90°, and becomes smaller as the arrangement angle is farther from 90°.

[0100] According to the light scattering detection device described in the first item, the size of the overlapped range between the light receiving area of ​​each detector and the scattered light generating area can be made uniform, i.e., the same, regardless of the arrangement angle. As a result, the light intensity at each detector is substantially the same, i.e., within the range of the allowable error, and therefore, for example, the molecular weight accuracy and the particle size calculation accuracy can be well maintained regardless of the arrangement angle of the detector.

[0101] (Item 2) In the light scattering detection device described in Item 1,

[0102] The opening width of each of the apertures is a value obtained by multiplying the distance from the central axis of the sample analysis chamber to each detector and the sine value of the arrangement angle of each detector.

[0103] According to the light scattering detection device described in the second aspect, the sizes of the overlapping ranges of the light receiving regions of the respective detectors and the scattered light generating regions can be made more accurately consistent, that is, the same, regardless of the arrangement angle.

[0104] (Item 3) In the light scattering detection device described in Item 1 or Item 2,

[0105] Each of the apertures includes a first aperture plate disposed on the sample analysis chamber side and a second aperture plate disposed on the detector side.

[0106] According to the light scattering detection device described in the third aspect, the scattered light incident on the detector can be limited neither too much nor too little.

[0107] (Item 4) In the light scattering detection device described in Item 3,

[0108] The device further includes a moving mechanism that moves the first aperture plate in a horizontal direction in parallel with the second aperture plate.

[0109] According to the light scattering detection device described in the fourth item, the position of the first aperture plate can be adjusted.

[0110] (Item 5) In the light scattering detection device described in Item 4,

[0111] The moving mechanism moves the first aperture plate based on refractive index information of a solvent in the liquid sample.

[0112] According to the light scattering detection device described in item 5, when the refractive index of the solvent and the refractive index of the sample analysis chamber are different, for example, the position of the first aperture plate can be adjusted so that the light intensity of the light received by each detector is consistent.

[0113] (Item 6) In the light scattering detection device described in Item 3,

[0114] The device further includes a moving mechanism that moves the second aperture plate and the detector in a horizontal direction in parallel with the first aperture plate.

[0115] According to the light scattering detection device described in the sixth item, the positions of the second aperture plate and the detector can be adjusted together.

[0116] (Item 7) In the light scattering detection device described in Item 6,

[0117] The moving mechanism moves the second aperture plate and the detector based on refractive index information of a solvent in the liquid sample.

[0118] According to the light scattering detection device described in item 7, when the refractive index of the solvent is different from the refractive index of the sample analysis chamber, for example, the positions of the second aperture plate and the detector can be adjusted so that the light intensity received by each detector is consistent.

[0119] (Item 8) In the light scattering detection device described in Item 3,

[0120] Each of the apertures has:

[0121] a light adjustment member disposed between the first aperture plate and the second aperture plate and configured to adjust a position of light traveling from the first aperture plate to the second aperture plate; and

[0122] The rotating mechanism rotates the light adjusting member in a horizontal direction.

[0123] According to the light scattering detection device described in the eighth aspect, it is possible to perform fine adjustment of the position of the light beam from the first aperture plate to the second aperture plate.

[0124] (Item 9) In the light scattering detection device described in Item 8,

[0125] The rotating mechanism rotates the light beam adjusting member based on refractive index information of a solvent in the liquid sample.

[0126] According to the light scattering detection device described in item 9, for example, when the refractive index of the solvent is different from the refractive index of the sample analysis chamber, the position of the light from the first aperture plate to the second aperture plate can be adjusted to make the light intensity received by each detector consistent.

[0127] (Item 10) In the light scattering detection device described in Item 9,

[0128] The light adjustment member is composed of parallel plate glass.

[0129] According to the light scattering detection device described in the tenth item, the light beam adjusting member can be made into a simple structure, so that, for example, the manufacturing cost of the light beam adjusting member can be suppressed.

[0130] (Item 11) A light scattering detection method according to one embodiment is used to detect microparticles in a liquid sample, the light scattering detection method comprising the following steps:

[0131] The liquid sample is sealed in a sample channel, wherein the sample channel is formed to linearly penetrate a transparent sample analysis chamber for holding the liquid sample;

[0132] irradiating coherent light from a light source from one end side of the sample channel to allow the coherent light to pass through the sample channel; and

[0133] a scattered light receiving step, receiving scattered light scattered from the sample analysis chamber to the surroundings at different scattering angles by using a plurality of detectors, wherein the plurality of detectors are arranged on the same circumference centered on a central axis extending in a vertical direction of the sample analysis chamber,

[0134] The scattered light receiving step includes a scattered light limiting step, in which the scattered light incident on each of the detectors is limited by using the opening width of a plurality of apertures arranged between the sample analysis chamber and each of the detectors.

[0135] The opening width of each of the apertures is maximum when the arrangement angle with respect to the incident direction of the coherent light into the sample analysis chamber is 90°, and becomes smaller as the arrangement angle is farther from 90°.

[0136] According to the light scattering detection method described in the eleventh item, the size of the overlapped range between the light receiving area of ​​each detector and the scattered light generating area can be made uniform, i.e., the same, regardless of the arrangement angle. As a result, the light intensity at each detector is substantially the same, i.e., within the range of the allowable error, and therefore, for example, the molecular weight accuracy and particle size calculation accuracy can be well maintained regardless of the arrangement angle of the detector.

[0137] (Item 12) In the light scattering detection method described in Item 11,

[0138] The opening width of each of the apertures is a value obtained by multiplying the distance from the central axis of the sample analysis chamber to each detector and the sine value of the arrangement angle of each detector.

[0139] According to the light scattering detection method described in the twelfth aspect, the sizes of the overlapping ranges of the light receiving regions of the detectors and the scattered light generating regions can be made more accurately consistent, that is, the same, regardless of the arrangement angle.

[0140] (Item 13) In the light scattering detection method described in Item 11 or Item 12,

[0141] Each of the apertures includes a first aperture plate disposed on the sample analysis chamber side and a second aperture plate disposed on the detector side.

[0142] According to the light scattering detection method described in the thirteenth item, the scattered light incident on the detector can be limited neither too much nor too little.

[0143] (Item 14) In the light scattering detection method described in Item 13,

[0144] In the scattered light limiting step, the first aperture plate is moved in a horizontal direction in parallel with the second aperture plate based on the refractive index information of the solvent in the liquid sample.

[0145] According to the light scattering detection method described in item 14, for example, when the refractive index of the solvent and the refractive index of the sample analysis chamber are different, the position of the first aperture plate can be adjusted so that the light intensity of the light received by each detector is consistent.

[0146] (Item 15) In the light scattering detection method described in Item 13,

[0147] In the scattered light limiting step, the second aperture plate and the detector are moved in a horizontal direction parallel to the first aperture plate based on the refractive index information of the solvent in the liquid sample.

[0148] According to the light scattering detection method described in item 15, for example, when the refractive index of the solvent is different from the refractive index of the sample analysis chamber, the positions of the second aperture plate and the detector can be adjusted so that the light intensity received by each detector is consistent.

[0149] (Item 16) In the light scattering detection method described in Item 13,

[0150] Each of the apertures has a light adjustment member, which is disposed between the first aperture plate and the second aperture plate and is used to adjust the position of light from the first aperture plate to the second aperture plate.

[0151] In the scattered light limiting step, the light beam adjusting member is rotated in the horizontal direction based on the refractive index information of the solvent in the liquid sample.

[0152] According to the light scattering detection method described in Item 16, for example, when the refractive index of the solvent is different from the refractive index of the sample analysis chamber, the position of the light from the first aperture plate to the second aperture plate can be adjusted so that the light intensity received by each detector is consistent.

[0153] (Item 17) In the light scattering detection method described in Item 16,

[0154] The light adjustment member is composed of parallel plate glass.

[0155] According to the light scattering detection method described in the seventeenth aspect, the light beam adjusting member can be made into a simple structure, so that, for example, the manufacturing cost of the light beam adjusting member can be suppressed.

Claims

1. A light scattering detection device for detecting microparticles in a liquid sample, the light scattering detection device comprising: A transparent sample analysis chamber, which is used to hold the liquid sample; a light source for irradiating coherent light to the sample analysis chamber; a plurality of detectors for receiving scattered light scattered from the sample analysis chamber to surroundings at different scattering angles; and a plurality of apertures, the plurality of apertures being arranged between the sample analysis chamber and each of the detectors and limiting the scattered light incident on the detector by using an opening width; in, The sample analysis chamber has a sample channel formed to linearly penetrate the sample analysis chamber, and the liquid sample is sealed in the sample channel. The light source is configured to allow the coherent light to enter from one end side of the sample channel and pass through the sample channel. The plurality of detectors are arranged on the same circumference centered on a central axis of the sample analysis chamber extending in a vertical direction. The plurality of detectors include a first detector and a second detector, wherein a position at which an angle of 90° relative to an incident direction of the coherent light incident on the sample analysis chamber is used as a reference position, the first detector is arranged at a position closer to the reference position, and the second detector is arranged at a position farther from the reference position. The aperture width of the first detector is larger than the aperture width of the second detector. The opening width of each aperture is a value obtained by multiplying the distance from the central axis of the sample analysis chamber to each detector and the sine value of the angle of the detector corresponding to the aperture with respect to the incident direction of the coherent light into the sample analysis chamber.

2. The light scattering detection device according to claim 1, characterized in that: Each of the apertures includes a first aperture plate disposed on the sample analysis chamber side and a second aperture plate disposed on the detector side.

3. The light scattering detection device according to claim 2, characterized in that: The device further includes a moving mechanism that moves the first aperture plate in a horizontal direction in parallel with the second aperture plate.

4. The light scattering detection device according to claim 3, characterized in that: The moving mechanism moves the first aperture plate based on refractive index information of a solvent in the liquid sample.

5. The light scattering detection device according to claim 2, characterized in that: The device further includes a moving mechanism that moves the second aperture plate and the detector in a horizontal direction in parallel with the first aperture plate.

6. The light scattering detection device according to claim 5, characterized in that: The moving mechanism moves the second aperture plate and the detector based on refractive index information of a solvent in the liquid sample.

7. The light scattering detection device according to claim 2, characterized in that: Each of the apertures has: a light adjustment member disposed between the first aperture plate and the second aperture plate and configured to adjust a position of light traveling from the first aperture plate to the second aperture plate; and The rotating mechanism rotates the light adjusting member in a horizontal direction.

8. The light scattering detection device according to claim 7, characterized in that: The rotating mechanism rotates the light beam adjusting member based on refractive index information of a solvent in the liquid sample.

9. The light scattering detection device according to claim 7 or 8, characterized in that: The light adjustment member is composed of parallel plate glass.

10. A light scattering detection method for detecting microparticles in a liquid sample, the light scattering detection method comprising the following steps: The liquid sample is sealed in a sample channel, wherein the sample channel is formed to linearly penetrate a transparent sample analysis chamber for holding the liquid sample; irradiating coherent light from a light source from one end side of the sample channel so that the coherent light passes through the sample channel; as well as a scattered light receiving step, receiving scattered light scattered from the sample analysis chamber to the surroundings at different scattering angles by using a plurality of detectors, wherein the plurality of detectors are arranged on the same circumference centered on a central axis extending in a vertical direction of the sample analysis chamber, The scattered light receiving step includes a scattered light limiting step, in which the scattered light incident on each of the detectors is limited by using the opening width of a plurality of apertures arranged between the sample analysis chamber and each of the detectors. The plurality of detectors include a first detector and a second detector, wherein a position at which an angle of 90° relative to an incident direction of the coherent light incident on the sample analysis chamber is used as a reference position, the first detector is arranged at a position closer to the reference position, and the second detector is arranged at a position farther from the reference position. The aperture width of the first detector is larger than the aperture width of the second detector. The opening width of each aperture is a value obtained by multiplying the distance from the central axis of the sample analysis chamber to each detector and the sine value of the angle of the detector corresponding to the aperture with respect to the incident direction of the coherent light into the sample analysis chamber.

11. The light scattering detection method according to claim 10, characterized in that: Each of the apertures includes a first aperture plate disposed on the sample analysis chamber side and a second aperture plate disposed on the detector side.

12. The light scattering detection method according to claim 11, characterized in that: In the scattered light limiting step, the first aperture plate is moved in a horizontal direction in parallel with the second aperture plate based on the refractive index information of the solvent in the liquid sample.

13. The light scattering detection method according to claim 11, characterized in that: In the scattered light limiting step, the second aperture plate and the detector are moved in a horizontal direction parallel to the first aperture plate based on the refractive index information of the solvent in the liquid sample.

14. The light scattering detection method according to claim 11, characterized in that: Each of the apertures has a light adjustment member, which is disposed between the first aperture plate and the second aperture plate and is used to adjust the position of light from the first aperture plate to the second aperture plate. In the scattered light limiting step, the light beam adjusting member is rotated in the horizontal direction based on the refractive index information of the solvent in the liquid sample.

15. The light scattering detection method according to claim 14, characterized in that: The light adjustment member is composed of parallel plate glass.

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