CN112469985A - Light scattering detection device - Google Patents
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- 238000003384 imaging method Methods 0.000 claims abstract description 54
- 239000007788 liquid Substances 0.000 claims abstract description 17
- 230000001427 coherent effect Effects 0.000 claims abstract description 11
- 239000010419 fine particle Substances 0.000 claims description 5
- 230000001678 irradiating effect Effects 0.000 claims description 3
- 239000007787 solid Substances 0.000 abstract description 15
- 239000011521 glass Substances 0.000 description 8
- 238000000569 multi-angle light scattering Methods 0.000 description 7
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- 238000005259 measurement Methods 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 239000002245 particle Substances 0.000 description 4
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- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
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- 239000011859 microparticle Substances 0.000 description 2
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- 238000001542 size-exclusion chromatography Methods 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
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- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
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Abstract
The invention provides a light scattering detection device, which can set the solid angle of the vertical direction of the entrance side slit of an imaging lens to be larger without setting the inclination angle alpha of the incident light to be larger, and can measure with high S/N ratio and high detection precision. A light scattering detection device (1) is provided with: a transparent sample cell (10) for holding a liquid sample; a light source (20) that irradiates the sample cell (10) with coherent light; an imaging optical system (50) for collecting light scattered from the sample cell (10) at different scattering angles toward the surroundings; a slit plate (40) for limiting a scattering angle range incident to the imaging optical system (50); and a detector (70) that receives the light collected from the imaging optical system (50), wherein the center axis (41S) of the slit (41) is arranged so as to be vertically off-center from the center axis (50S) of the imaging optical system (50).
Description
Technical Field
The present invention relates to a light scattering detection device used in a microparticle detection device for measuring a molecular weight, a radius of rotation (size), and the like of microparticles dispersed in a liquid sample.
Background
Size Exclusion Chromatography (SEC) is known as a method for separating fine particles such as proteins dispersed in a liquid sample. In recent years, as a chromatography detection apparatus, a multi-angle light scattering (MALS) detection apparatus is used in addition to an Ultraviolet (UV) absorbance detection apparatus and a differential refractive index detection apparatus. The MALS detection apparatus has an advantage of calculating the molecular weight and the particle size of a measurement sample (see patent documents 1 and 2).
Fig. 7 shows a coordinate system of the scattered light radiation direction in the case where the scattered light generation light source is disposed at the origin. As shown in fig. 7, light enters the positive X-direction on the XY plane, the scattering angle from the traveling direction of light on the XY plane is defined as θ, and the angle from the XY plane is defined as Φ.
Next, fig. 8 shows a plan view of a basic configuration example of the MALS detection apparatus, and fig. 9 shows a side view. In fig. 8 and 9, 110 denotes a sample cell, 111 denotes a liquid sample, 120 denotes a light source, 121 denotes a condenser lens, 140 denotes a slit plate, 150 denotes an imaging lens, 160 denotes an aperture plate, and 170 denotes a detector. As shown in fig. 8 and 9, the liquid sample 111 is passed through the cylindrical sample cell 110, and light is irradiated from the light source 120 so as to pass through the sample cell 110 and the center of the flow channel. As the light source, a visible laser is generally used. The angle θ from the light traveling direction is defined as a scattering angle on a horizontal plane (on the XY plane), and a plurality of detectors 170 are arranged on a horizontal plane (on the XY plane) passing through the sample cell 110 and the flow path center so as to detect different scattering angles. Fig. 8 shows an example in which 2 detectors 170 are arranged at an arrangement angle of θ 1 and θ 2.
In the case of the cylindrical sample cell 110, there are the following technical problems: reflected light at the interface between the glass and the air and the interface between the glass and the flow path enters the detector 170 as stray light, deteriorating the detection accuracy of the detector 170. As a solution to this problem, the inventors have found a method of reducing the stray light by inclining the incident light to the sample cell 110 (at an angle α), as shown in fig. 9 (see non-patent document 1).
Next, fig. 10 and 11 show the relationship between the scattered light intensity and the scattering angle. That is, fig. 10 and 11 are results of calculating the scattering pattern of particles having a refractive index of 1.59 in a solvent having an incident wavelength of 660nm and a refractive index of 1.33 according to the theoretical formula of Mie scattering. The particle diameters were 1nm, 100nm, and 500 nm. Fig. 10 is a scattering angle pattern in the horizontal direction (theta direction), and fig. 11 is a scattering angle pattern in the vertical direction (phi direction). When the wavelength of the sample is sufficiently smaller than the wavelength of the incident light, scattered light in the θ direction occurs isotropically, and the intensity of the scattered light does not depend on the scattering angle. As the particle size of the sample increases, scattering of scattered light in the forward direction (direction in which θ is small) becomes strong. The scattered light intensity tends to decrease as phi increases in the phi direction.
Further, for the MALS detection apparatus, an apparatus capable of measuring a sample of low concentration and having a high S/N ratio is desired. For this reason, an optical system for efficiently allowing scattered light generated from a sample to enter a detector is required. In other words, the solid angle of the scattered light entering each detector needs to be made large. When the solid angle of the detector is increased, the angular resolution of each detector is deteriorated if the solid angle in the horizontal direction (on the optical axis plane) is increased, and therefore, it is desirable to increase the solid angle in the vertical direction. However, increasing the detector size to increase the solid angle is not preferable because it increases the dark current.
In order to increase the solid angle without increasing the size of the detector, a method of condensing scattered light with a lens or the like is employed (non-patent document 1). Specifically, the arrangement is such that scattered light generated in the cell channel is imaged on the detector via the imaging lens. Therefore, in order to reduce the solid angle in the horizontal direction and increase the solid angle in the vertical direction, a slit having a narrow opening width in the horizontal direction and a wide opening length in the vertical direction is provided on the incident side of the imaging lens. This enables efficient detection of scattered light without degrading the angular resolution.
Documents of the prior art
Patent document
Patent document 1: japanese laid-open patent publication No. H07-72068
Patent document 2: japanese laid-open patent publication No. 2015-111163
Non-patent document 1: analysis of Absolute molecular weight of protein and Complex formation by light scattering method, tail elegant article, Biotechnology 89 Vol
Disclosure of Invention
Technical problem to be solved by the invention
In order to increase the amount of light entering the detector, it is necessary to set the opening length in the vertical direction of the slit disposed on the incident side of the imaging lens to be large. However, if the opening length of the slit in the vertical direction is set to be large, reflected light at the interface between the glass and the air of the sample cell and the interface between the glass and the flow path enters the detector as stray light. Since the intensity of the reflected light is large compared to the scattered light, the dynamic range of the detector is reduced, and the detection accuracy of the detector is deteriorated due to the intensity variation of the stray light. On the other hand, if the inclination angle α of the incident light is set to be large in order to remove the stray light, as shown in fig. 11, the scattering angle Φ in the vertical direction becomes large, the intensity of the scattered light entering the slit decreases, and the S/N ratio decreases.
Therefore, an object of the present invention is to provide a light scattering detection device capable of setting a solid angle in a vertical direction of an entrance-side slit of an imaging lens to be large without setting an inclination angle α of incident light to be large, and capable of performing measurement with a high S/N ratio and high detection accuracy.
Solution for solving the above technical problem
A light scattering detection device according to an aspect of the present invention is a light scattering detection device for detecting fine particles in a liquid sample, including: a transparent sample cell for holding a liquid sample; a light source for irradiating the sample cell with coherent light; an imaging optical system for condensing light scattered from the sample cell at different scattering angles toward the surroundings; a slit plate for limiting a scattering angle range incident to the imaging lens; a detector for receiving the collected light from the imaging optical system; the central axis of the slit is arranged to be eccentric in one vertical direction from the central axis of the imaging optical system.
In the configuration of the light scattering detection device, the slit is preferably vertically elongated, and at least one side along the vertical direction is preferably linear.
Further, it is preferable that the slit is arranged to be eccentric from a center axis of the imaging optical system to an upper side in a vertical direction.
Further, it is preferable that the slit has a < b where a lower length of the slit in a vertical direction from a central axis of the imaging optical system is a and an upper length thereof is b.
Preferably, a plurality of detection optical systems extending from the sample cell to the detectors are arranged around the sample cell at equal intervals from a cell center axis, and a length ratio between a lower length a and an upper length b of the slit is adjusted under a condition that a is equal to or less than b according to an arrangement angle of each detector with respect to the sample cell.
In addition, it is preferable that the light source is arranged such that an optical axis of coherent light entering the sample cell from the light source is inclined at a predetermined angle from a plane including the sample cell and the detector.
Effects of the invention
According to the present invention, it is possible to provide a light scattering detection device capable of setting a solid angle in the vertical direction of an entrance-side slit of an imaging lens to be large without setting the inclination angle α of incident light to be large, and capable of performing measurement with a high S/N ratio and high detection accuracy.
Drawings
Fig. 1 is a side view of an embodiment of the light scattering detection device of the present invention.
Fig. 2 is a partially enlarged side view of the light scattering detection device of the present embodiment.
Fig. 3 is a plan view of the light scattering detection device of the present embodiment.
Fig. 4 is a side view for explaining the configuration and the component size in the ray tracing simulation.
Fig. 5 is a plan view for explaining a ray tracing simulation result in the case where the detector is disposed at an angle θ of 15 degrees.
Fig. 6 is a side view for explaining a ray tracing simulation result in the case where the detector is arranged at θ 15 degrees.
Fig. 7 is a coordinate system of the scattered light radiation direction in the case where the scattered light generation light source is disposed at the origin.
Fig. 8 is a plan view of a basic configuration example of the MALS detection apparatus.
Fig. 9 is a side view of a basic configuration example of the MALS detection apparatus.
Fig. 10 is an explanatory diagram of the relationship between the scattered light intensity and the scattering angle (scattering angle pattern in the horizontal direction).
Fig. 11 is an explanatory diagram of the relationship between the scattered light intensity and the scattering angle (scattering angle pattern in the vertical direction).
Detailed Description
Hereinafter, an embodiment of the light scattering detection device according to the present invention will be described with reference to the drawings. In addition, components denoted by the same reference numerals in the drawings have the same or similar configurations.
[ constitution of light scattering detection device ]
First, the configuration of an embodiment of the light scattering detection device of the present invention will be described with reference to fig. 1 to 3. Fig. 1 is a side view of an embodiment of the light scattering detection device of the present invention. Fig. 2 is a partially enlarged side view of the light scattering detection device of the present embodiment. As shown in fig. 1 and 2, the light scattering detection device 1 of the present embodiment is a device for detecting the molecular weight and the radius of rotation (size) of fine particles such as proteins dispersed in a liquid sample. The light scattering detection device 1 includes a sample cell 10, a light source 20, a slit plate 40, an imaging optical system 50, an aperture plate 60, and a detector 70. Hereinafter, each of the components will be described.
The sample cell 10 is a transparent cylindrical cell for holding a liquid sample in an internal flow path. The sample cell 10 is formed of, for example, colorless transparent quartz glass.
The light source 20 irradiates the sample cell 10 with coherent light. "coherent light" refers to light that: the phase relationship of the light waves at 2 arbitrary points in the light beam is kept constant without changing with time, and complete coherence is shown even when a large optical path difference is applied and the light waves are overlapped again after being divided by an arbitrary method. As the light source 20, for example, a laser light source for irradiating visible light laser light is used. There is no completely coherent light in nature, and laser light oscillated in a single mode is light in a nearly coherent state.
A condensing optical system 21 is disposed on an optical path L1 of incident light from the light source 20 to the sample cell 10. As the condensing optical system 21, for example, a single condensing lens is used. The condenser lens is a plano-convex lens, and the incident side of the light from the light source 20 is formed as a convex surface and the exit side is formed as a flat surface. In the present embodiment, a single condenser lens is used as the condenser optical system 21, but the condenser optical system 21 may be configured by combining a plurality of compound lenses and condenser lenses.
The light source 20 and the condensing optical system 21 are arranged such that the optical axis of coherent light entering the sample cell 10 from the light source 20 is inclined at a predetermined angle (inclination angle α) from a plane (XY plane) including the sample cell 10 and the detector 50. Specifically, the light source 20 and the light collecting optical system 21 are arranged so that the incident light enters the sample cell 10 from obliquely above. By inclining the incident light with respect to the sample cell 10 (angle α), stray light caused by reflected light at the interface between the glass and the air and the interface between the glass and the flow path of the sample cell 10 (hereinafter, collectively referred to as "cell interface") can be reduced. The laser light emitted from the light source 20 passes through the condensing optical system 21 and is then condensed near the central axis of the sample cell 10.
The detection optical system 30 is disposed on an optical path L2 of light emitted from the sample cell 10. The detection optical system 30 of the present embodiment is composed of a slit plate 40, an imaging optical system 50, an aperture plate 60, and a detector 70.
The imaging optical system 50 condenses light scattered from the sample cell 10 at different scattering angles toward the surroundings. As the imaging optical system 50, for example, a single imaging lens is used. The imaging lens is a plano-convex lens, and the incident side of the scattered light from the sample cell 10 is formed as a plane and the exit side is formed as a convex surface. In the present embodiment, a single imaging lens is used as the imaging optical system 50, but the imaging optical system 50 may be configured by combining a plurality of compound lenses and imaging lenses.
The slit plate 40 is disposed between the sample cell 10 and the imaging optical system 50 on the optical path L2 of the light emitted from the sample cell 10. The slit plate 40 limits the range of the scattering angle incident to the imaging optical system 50. That is, the slit 41 opened in the slit plate 40 is vertically long and has a straight line shape along at least the vertical side in order to restrict the scattering angle in the horizontal direction and to absorb a large amount of the light flux in the vertical direction. Specifically, the slit 41 has a rectangular shape or a long hole shape elongated in the vertical direction.
The center axis 41S of the slit 41 is arranged to be eccentric in the vertical direction from the center axis 50S of the imaging optical system 50. Since the light emitted from the light source 20 enters the sample cell 10 from an obliquely upper side, the reflected light RL generated at the interface between the air and the glass and the interface between the glass and the liquid (hereinafter referred to as "cell interface") in the sample cell 10 tends to be shifted downward as stray light. Therefore, in order to limit the reflected light (stray light) RL, the central axis 41S of the slit 41 of the present embodiment is arranged to be vertically eccentric upward from the central axis 50S of the imaging optical system 50.
Specifically, as shown in fig. 2, when the lower length of the slit 41 in the vertical direction from the central axis 50S of the imaging optical system 50 is a and the upper length is b, the relationship between the lower length a and the upper length b of the slit 41 is set to a < b. The incident light may be incident upward from below, and in this case, the relationship between the lower length a and the upper length b of the slit 41 is set to a > b.
The aperture plate 60 is disposed on the optical path L2 of the light emitted from the sample cell 10, closer to the imaging optical system than the detector 70. The aperture plate 60 has a function of restricting stray light, and the aperture 61 is opened in front of the light receiving surface of the detector 70.
The detector 70 receives the collected light from the imaging optical system 50. That is, the light receiving surface of the detector 70 is located at the focal point of the imaging optical system 50. The detector 70 of the present embodiment may be a Photodiode (PD) or an array detector such as a two-dimensional CMOS.
Fig. 3 is a plan view of an embodiment of the light scattering detection device of the present embodiment. As shown in fig. 3, a plurality of detection optical systems 30 extending from the sample cell 10 to the detector 70 are arranged around the sample cell 10 at equal intervals d from the cell center axis S. An angle θ from the traveling direction of light is defined as a scattering angle on a horizontal plane (on the XY plane). A plurality of detectors 70 are disposed on a horizontal plane (XY plane) passing through the sample cell 10 and the cell center axis S so as to be able to detect different scattering angles. In the embodiment of fig. 3, 2 sets of detection optical systems 30 and 31 are arranged at arrangement angles θ 1 and θ 2.
When a plurality of detectors 70 are provided at equal intervals d around the sample cell 10, the length ratio between the lower length a and the upper length b of the slit 41 is adjusted under the condition that a is not more than b according to the arrangement angle of each detector 70 with respect to the sample cell 10. That is, in fig. 3, the influence of the reflected light at the cell interface by the detection optical system 31 arranged at the scattering angle θ 2 is smaller than that by the detection optical system 30 arranged at the scattering angle θ 1. Since the influence of the reflected light at the cell interface by the detection optical system 31 having the scattering angle θ 2 of 90 degrees is small, the solid angle in the vertical direction can be optimized by setting a large value close to b.
[ Effect of light scattering detection device ]
Next, the operation of the light scattering detection device of the present embodiment will be described with reference to fig. 1 to 6.
As shown in fig. 1 and 2, a liquid sample 11 is passed through a channel of a cylindrical sample cell 10. When the liquid sample 11 is completely passed through, a visible laser beam as coherent light is irradiated from the light source 20 through the condensing optical system 21. The visible laser light advances along the optical path L1, and the laser light is incident on the liquid sample 11 in the flow path of the sample cell 10. When laser light is incident on the liquid sample 11, the light is irradiated on the fine particles contained in the liquid sample 11 and scattered at a predetermined scattering angle. The scattered light emitted from the sample cell 10 passes through the slit 41 of the slit plate 40, passes through the imaging optical system 50 and the aperture plate 60, and is received on the light receiving surface of the detector 70.
When the scattered light is emitted from the sample cell 10, reflected light RL is generated as stray light at the cell interface of the sample cell 10. Since the light emitted from the light source 20 is incident from the obliquely upper side of the sample cell 10 at the inclination angle α, the reflected light (stray light) generated at the cell interface of the sample cell 10 tends to be shifted downward. Since the slit 41 of the present embodiment is disposed so as to be vertically eccentric upward from the central axis 50S of the imaging optical system 50, the reflected light (stray light) RL that is offset downward can be restricted by the plate portion of the slit plate 40.
In this manner, by disposing the slit 31 of the slit plate 30 so as to be vertically off-center from the central axis 40S of the imaging optical system 40, scattered light beneficial to analysis can be positively incident on the imaging optical system 40. The imaging optical system 40 images on the light receiving surface of the detector 60, and an aperture plate 50 is disposed in front of the light receiving surface. This allows the aperture plate 50 to further limit stray light, and allows scattered light necessary for analysis to be received on the light-receiving surface of the detector 70.
Specifically, when the lower length of the slit 41 in the vertical direction from the central axis 50S of the imaging optical system 50 is a and the upper length is b, the relationship between the lower length a and the upper length b of the slit 41 is set to a < b. That is, by setting the lower length a of the slit 41 to be small, the reflected light RL at the cell interface can be blocked by the plate portion of the slit plate 40. The vertical solid angle can be increased by setting the upper length b to be large with the effective diameter of the imaging optical system 50 as an upper limit.
As shown in fig. 3, a plurality of detection optical systems 30 extending from the sample cell 10 to the detector 70 are arranged around the sample cell 10 at equal intervals d from the cell center axis S. In the case where the light scattering detection apparatus 1 includes a plurality of detection optical systems 30 at equal intervals d around the sample cell 10, the length ratio between the lower length a and the upper length b of the slit 41 is adjusted under the condition that a is equal to or less than b according to the arrangement angle of each detector 70 with respect to the sample cell 10. That is, in the detection optical system 31 disposed at the scattering angle θ 2 of 90 degrees, which is less affected by the reflected light RL at the cell interface, by setting a to be large and approaching b, the solid angle in the vertical direction can be optimized.
[ ray tracing simulation ]
In order to confirm the operation and effect of the above embodiment, ray tracing simulation was performed. Fig. 4 is a side view for explaining the configuration and the component size in the ray tracing simulation. Fig. 5 is a plan view and fig. 6 is a side view for explaining the result of ray tracing simulation in the case where the detector (PD) is arranged at θ ═ 15 degrees.
As shown in FIG. 4, the sample cell 10 is, for example, a transparent cylindrical cell having an inner diameter of 1.6mm and an outer diameter of 8.0 mm. An imaging lens (plano-convex lens) is disposed at a position distant from the center axis S47mm of the sample cell 10. The imaging lens is formed, for example, with a convex diameter of phi 12.7mm and a focal length of 38 mm. The aperture plate 60 and the PD of 2.4mm are disposed at a position apart from the center axis S140mm of the sample cell 10. The slit 41 of the slit plate 40 is formed as an opening having a width in the horizontal direction (XY direction) of 3mm and a length in the vertical direction (Z direction) of 8.5mm, for example. Regarding the vertical direction opening of the slit 41, for example, a lower length a from an XY plane including the central axis 50S of the imaging optical system 50 is set to 2.5mm in the-Z direction, and an upper length b is set to 6.0mm in the + Z direction.
As shown in fig. 5 and 6, by setting the relationship between the lower length a and the upper length b to a < b, the reflected light RL generated at the cell interface of the sample cell 10 is blocked by the plate portion of the slit plate 41 and does not reach the detector 70, so that high-precision measurement can be performed.
Further, stray light caused by reflected light RL at the cell interface depends on the arrangement angle θ of the detector 70. In the case where θ is small, since the influence of the reflected light RL at the cell interface is large, it is necessary to shorten a, and in the case where θ is close to 90 degrees, the reflected light RL at the cell interface is hardly generated. The slits 41 are optimized under the condition that a is not more than b according to the arrangement angle of the detector 70, whereby reduction of stray light and improvement of the S/N ratio can be achieved.
As described above, according to the light scattering detection device 1 of the present embodiment, it is possible to provide a light scattering detection device capable of setting a solid angle in the vertical direction of the slit 31 of the slit plate 30 disposed on the incident side of the imaging optical system 40 to be large without setting the inclination angle α of the incident light to be large, and capable of performing measurement with a high S/N ratio and high detection accuracy.
The above-described embodiments are intended to facilitate understanding of the present invention, and are not intended to limit the present invention. The elements and their arrangement, materials, conditions, shapes, dimensions, and the like included in the embodiments are not limited to the examples, and can be appropriately modified. Further, the configurations shown in the different embodiments may be partially substituted or combined with each other.
Description of the reference numerals
1 light scattering detection device
10 sample cell
20 light source
30 detection optical system
40 slit plate
41 slit
Center axis of 41S slit
50 imaging optical system
Center axis of 51S imaging optical system
70, a detector.
Claims (6)
1. A light scattering detection device for detecting fine particles in a liquid sample, comprising:
a transparent sample cell for holding a liquid sample;
a light source for irradiating the sample cell with coherent light;
an imaging optical system that condenses light scattered from the sample cell at different scattering angles toward the surroundings;
a slit plate for limiting a scattering angle range incident to the imaging optical system;
a detector for receiving the collected light from the imaging optical system,
the central axis of the slit is configured to be eccentric in the vertical direction from the central axis of the imaging optical system.
2. The light scatter detection device of claim 1,
the slit is vertically long, and at least one side in the vertical direction is linear.
3. The light scatter detection device of claim 1 or claim 2,
the slit is configured to be eccentric from a center axis of the imaging optical system to an upper side in a vertical direction.
4. The light scatter detection device of claim 3,
when a lower length of the slit in a vertical direction from a central axis of the imaging optical system is defined as a and an upper length thereof is defined as b, a < b is given.
5. The light scatter detection device of claim 4,
a plurality of detection optical systems extending from the sample cell to the detector are arranged around the sample cell at equal intervals from the cell center axis,
the length ratio between the lower length a and the upper length b of the slit is adjusted under the condition that a is not more than b according to the arrangement angle of each detector relative to the sample cell.
6. The light scattering detection device as claimed in any of claims 1 to 5,
the light source is arranged such that an optical axis of coherent light incident on the sample cell from the light source is inclined at a predetermined angle from a plane including the sample cell and the detector.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| PCT/JP2018/028124 WO2020021682A1 (en) | 2018-07-26 | 2018-07-26 | Light scattering detection device |
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| CN112469985A true CN112469985A (en) | 2021-03-09 |
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| WO (1) | WO2020021682A1 (en) |
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Also Published As
| Publication number | Publication date |
|---|---|
| JP7078115B2 (en) | 2022-05-31 |
| JPWO2020021682A1 (en) | 2021-04-30 |
| WO2020021682A1 (en) | 2020-01-30 |
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