JP2009128125A - Liquid analyzer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To detect efficiently scattered light or fluorescence by a small-sized device while coping with the reduction of the quantity of a sample solution. <P>SOLUTION: In consideration of each refractive index of a transparent container storing the sample solution and air, scattered light or fluorescence emitted within a Brewster angle from the container side having a large refractive index to the atmospheric air side having a small refractive index is condensed on the front side and on the rear side by each different rotating elliptic mirror, and is allowed to enter a detector at a smaller angle than a Brewster angle. <P>COPYRIGHT: (C)2009,JPO&amp;INPIT

Description

  The present invention relates to an analyzer that detects the amount of a component contained in a sample, and relates to a liquid analyzer that can stably analyze a very small amount of sample.

  As an analyzer for detecting the amount of components contained in a sample, the sample solution is irradiated with white light from a halogen lamp, etc., and the light transmitted through the sample solution is dispersed by a diffraction grating to extract the necessary wavelength component. A spectroscopic analyzer that measures the amount of a target component by determining the absorbance and a fluorometer that measures the amount of the target component by determining the amount of fluorescence that is excited by irradiation light are widely used.

  When measuring scattered light or fluorescence with these analyzers, first, when measuring scattered light with a spectroscopic analyzer, the sample solution is irradiated with white light and the light transmitted through the sample solution is dispersed with a diffraction grating or the like. The amount of light attenuated by scattering was calculated by measuring the amount of light at a specific wavelength, and the amount of scattered light was derived. Next, when measuring fluorescence with a fluorescence spectrophotometer, white light from a halogen lamp or the like is split by a diffraction grating, light of a specific wavelength is irradiated as excitation light, and the fluorescence excited thereby is emitted. Detection was performed from an angle different from the irradiation direction of the excitation light, and in order to improve the analysis accuracy of the fluorescence wavelength, the fluorescence was measured with a diffraction grating. In the case of fluorescence measurement, there is a method in which only the fluorescence is detected by switching the irradiation timing of the irradiation light and the detection timing of the fluorescence with a chopper in consideration of the time for emitting the fluorescence.

  In these analyzers, a sample solution is often dispensed into a plastic or glass container, and the amount of components is measured by irradiating the sample solution with light. In addition, the sample solution to be used is many tens to hundreds of microliters, and an optical system that collects and irradiates light such as a halogen lamp with a lens is used. Each of the sample solution and the diffraction grating and the detector from the diffraction grating has a length of about 100 mm.

  However, in recent years, in order to reduce reagent costs and reduce the burden on the environment, there has been a demand for a minute amount of sample solution used for analysis, and the smaller the amount, the more difficult it is to squeeze light into the liquid. There is a problem that it is difficult to efficiently detect scattered light and fluorescence emitted from a minute region. Regarding the detection of scattered light, the method of calculating the amount attenuated by the scattered light from the amount of transmitted light can be detected if the light can be narrowed down to the sample solution, but a halogen lamp or diffraction grating as described above is used. The optical system had a large size, and it was difficult to cope with the downsizing of the device required as the sample solution became smaller.

  Therefore, as a method of downsizing the optical system and detecting scattered light and fluorescence efficiently, first, downsizing of the light source can be considered. Light emitting diodes have been raised as compact and high-luminance light sources, and measurement methods using light emitting diodes as light sources have also been reported in Patent Document 1, Patent Document 2, Patent Document 3, and the like. However, these are used to detect transmitted light, and unlike transmitted light, scattered light emitted at a specific scattering angle in the same direction as the irradiation direction of irradiation light and in a direction opposite to the irradiation direction of irradiation light, or irradiation It is not an optical system suitable for detecting fluorescence emitted in almost all directions from a region irradiated with light.

  Next, lenses and mirrors are conceivable as methods for condensing light. Unlike transmitted light, scattered light has a specific scattering angle in the same direction as the irradiation direction of irradiation light and in the direction opposite to the irradiation direction of irradiation light. It is emitted at. In addition, fluorescence is emitted almost in all directions from the region irradiated with the irradiation light. Therefore, in order to efficiently collect and detect scattered light and fluorescence emitted from a very small amount of sample solution, it is necessary to use a concave mirror that simultaneously reflects and collects light, and collects and detects it on one detector. It is considered suitable. There are reports in Patent Literature 4, Patent Literature 5 and the like as methods for condensing light using a concave mirror. However, in an analyzer that detects the amount of components contained in a sample, the sample contained in a transparent glass or plastic container is irradiated with excitation light to separate scattered light, fluorescence, and excitation light. It is necessary, and it is necessary to capture only scattered light and fluorescence that diverges, avoiding portions such as excitation light, transmitted light, and members that absorb transmitted light. The path of light when light travels from the transparent glass or plastic container side to the atmosphere or from the air to the transparent glass or plastic container side is obtained from Snell's Law or Fresnel's formula. It is done.

  On the other hand, in the examples of Patent Document 4 and Patent Document 5, since light that diverges from one point, including its light source, is condensed to another point without avoiding other objects, excitation light is applied to the sample. There are many problems to apply to an analyzer that irradiates and detects the amount of components contained in a sample.

Japanese Patent Application No. 11-340964 Japanese Patent Application 2001-146695 Japanese Patent Application No. 2004-151177 JP 2006-108521 A JP 2006-126013 A

  In the above-mentioned conventional technology, it is difficult to cope with the minute amount of the sample solution and the miniaturization of the apparatus or the optical system, and it is not suitable for measurement of scattered light or fluorescence only by detecting transmitted light. Because it is not structured to irradiate a sample contained in a transparent glass or plastic container with excitation light and to detect scattered light, fluorescence and excitation light separately, only scattered light and fluorescence can be detected efficiently. There was a problem that it was not suitable to do.

  In order to solve the above problems, the present invention takes into account the refractive index of a transparent glass or plastic container containing the sample solution and the air, and the scattering emitted from the glass or plastic container side within the Brewster angle. Light emitted from the front side, which is the irradiation direction side of the excitation light, and the rear side, which is opposite to the irradiation direction of the excitation light, are collected by separate spheroid mirrors, and the light is emitted to the detector. A configuration in which the incident light is smaller than the star angle is adopted.

  According to the present invention, when the detection optical system is downsized in accordance with the miniaturization of the sample solution or downsizing of the apparatus, the fluorescence or scattered light emitted from the sample can be efficiently collected and detected. It is possible to perform a stable analysis on the samples.

  Embodiments of the present invention will be described below with reference to the drawings.

  A liquid analysis method in this embodiment will be described. First, a case where scattered light is used will be described. Examples of a method for measuring the amount of a specific component contained in a sample solution by irradiating the sample solution with light and measuring the amount of light scattered by scattering particles in the sample solution include an immunoturbidimetric method using latex agglutination. In this method, latex is agglomerated by reaction of latex with antibody, which is a reagent component in the sample solution, and antigen in the specimen consisting of blood components, etc., and the light irradiated to the sample solution depends on the degree of latex aggregation. The amount of antigen in the sample is measured by utilizing the change in the degree of scattering of the sample.

  Based on this principle, the liquid analysis method by measuring scattered light according to this example irradiates the sample solution in the container with light, except for the amount of the irradiated light refracted and transmitted through the sample solution and the container. Forward scattered light scattered in the same direction as the direction of light irradiation is collected on a detector with a reflecting mirror, and the amount of antigen and the like are measured.

  Next, a case where fluorescence is used will be described. There is a fluorescence measurement method for measuring fluorescence emitted by irradiating a sample solution with excitation light and exciting a phosphor inside the sample. In some cases, the amount of phosphor is simply measured, but the antibody to which the phosphor is attached reacts with the antigen in the sample consisting of blood components, etc., and after removing the unreacted components, the excitation light is irradiated and the amount of fluorescence Is used to measure the amount of antigen in the specimen.

  Based on this principle, the liquid analysis method by fluorescence measurement according to the present embodiment irradiates the sample solution placed in the container with excitation light, except for the amount of the irradiated excitation light refracted and transmitted through the sample solution and the container. The fluorescence emitted in the same direction as the direction of light irradiation is collected on the detector by a reflecting mirror, and the amount of antigen and the like are measured.

  The optical system of this embodiment has a structure that can handle both the case of using scattered light and the case of using fluorescence, and selects the reagent to be used and the wavelength of light to be irradiated according to the purpose. Just do it.

  FIG. 1 shows a measuring unit of a liquid analyzer according to the present invention. A sample solution 1 consisting of a specimen and a reagent is placed in a transparent container 2. In FIG. 1, the sample solution 1 and the container are shown in cross section, but are shown without hatching to avoid complicating the drawing and hindering understanding. A lens 5 for narrowing the light 4 emitted from the light source 3 and the light source 3 to the sample solution 1 is arranged on the left side of the container 2 in the figure. On the right side of the container 2, the transmitted light absorber 6 and the light 4, which are narrowed down to the sample solution 1 by the lens 5 and absorb and capture the light transmitted through the sample solution 1, are scattered by the components contained in the sample solution 1. The first reflecting mirror 8 that reflects and collects the scattered light 7 emitted forward, which is the right direction of the solution 1, that is, the same direction as the irradiation direction of the light 4, or the fluorescence 7 ′ that is excited by the light 4 and emitted forward. A detector 9 for detecting the reflected and condensed light and converting it into an electrical signal is disposed. These are arranged on the optical axis 10 and are held by holding members, but the holding members are omitted from the drawings in order to avoid making the drawing complicated and obstructing the understanding.

  Next, the path of light and the configuration of the apparatus at the time of liquid analysis according to this embodiment will be described. At the time of liquid analysis, first, the sample solution 1 is put in the container 2 and set on the optical axis as shown in FIG. The container 2 is made of a material such as glass or plastic that transmits light 4 with low loss. In this state, light 4 is emitted from the light source 3 by energizing the light source 3 or the like. The light 4 is narrowed down by the lens 5 and irradiated to the sample solution 1.

  At that time, in the case of the scattered light measurement, the degree of aggregation of the latex contained in the reagent in the sample solution differs depending on the amount of antibody contained in the sample in the sample solution, and the scattered light 7 is scattered according to the degree of aggregation. The liquid component can be analyzed by condensing the scattered light 7 by the first reflecting mirror 8 and measuring the amount of light by the detector 9. The scattered light 7 scatters a part of the light 4, and the unscattered part of the light 4 passes through the sample solution 1 and is absorbed by the transmitted light absorber 6. In this case, the scattering direction is scattered light emitted to the right side of the sample solution 1, that is, in the same direction as the irradiation direction of the light 4.

  In the case of fluorescence measurement, when the amount of phosphor is simply measured, fluorescence corresponding to the amount of phosphor in the sample solution is emitted, and the amount of fluorescence is measured. Further, when measuring the amount of antigen in the sample, etc., fluorescence is emitted according to the amount of antibody phosphor bound in response to the antigen in the sample, so that the fluorescence is collected by the first reflector 8. The liquid component can be analyzed by measuring the amount of light with the light detector 9. In this case, the light 4 is used as excitation light, and the portion not used for excitation passes through the sample solution 1 and is absorbed by the transmitted light absorber 6.

  The transmitted light absorber 6 has a cylindrical cup shape with the optical axis as the central axis. The left side of FIG. 1 has an opening on the left side. 4 is absorbed and converted into heat. Further, by replacing the transmitted light absorber 6 with a transmitted light detector and comparing the amount of transmitted light with the amount of scattered light or fluorescence, the amount of light lost to transmitted light, scattered light or other than fluorescence is calculated. Can be made, and a more institutional analysis can be made possible.

  As described above, the present embodiment has a structure that can handle both the case where scattered light is used and the case where fluorescence is used. Will be described.

  Scattered light 7 emitted from the sample solution 1 is emitted from the container 2 into the atmosphere. In this embodiment, the light is emitted into the atmosphere. However, in order to easily maintain the sample solution 1 or the like at a constant temperature, the entire measurement unit or a part thereof may be in constant temperature water or the like. In this embodiment, for the sake of simplicity, the refractive index of the atmosphere is assumed to be 1.0, the refractive index of the liquid sample solution 1, the container 2, and the light receiving portion of the detector 9 is assumed to be about 1.5. This is also shown in the case of representing the refraction of light.

  When the scattered light 7 is emitted from the container 2 into the atmosphere, it is divided into reflected light and transmitted light at the interface between the container 2 and the atmosphere, and only the scattered light that is refracted and becomes transmitted light is emitted into the atmosphere. The ratio of reflected light and transmitted light will be described with reference to FIGS.

  FIG. 2 shows a simplified state in which scattered light 7 travels from the container 2 side to the atmosphere side. In FIG. 2, the incident angle when the scattered light 7 is incident on the boundary surface between the container 2 and the atmosphere as φ is φ, and the reflection of the p-polarized light and the s-polarized component when φ is varied from 0 ° to 90 °. FIG. 3 shows the reflectance and transmittance of light and transmitted light. FIG. 3 shows a result obtained by adding the value of the refractive index shown in FIG. 2 to an expression obtained from Snell's law and Fresnel's expression, and is a general result.

  FIG. 4 shows a simplified state in which the scattered light 7 travels from the atmosphere side to the container 2 side. In FIG. 4, the incident angle when the scattered light 7 is incident on the boundary surface between the atmosphere and the container 2 is φ, and the reflection of the p-polarized light and the s-polarized light component when φ is varied from 0 ° to 90 °. FIG. 5 shows the reflectance and transmittance of light and transmitted light. FIG. 5 is a general result obtained by adding the refractive index value shown in FIG. 4 to an expression obtained from Snell's law and Fresnel's expression as in FIG.

In both FIG. 3 and FIG. 5, when the incident angle exceeds a certain value, the transmittance rapidly decreases and the reflectance increases. In particular, the p-polarized component has an incident angle at which the reflectance is 0% and the transmittance is 100%. When the incident angle is larger than that angle, the transmittance is rapidly decreased and the reflectance is increased. The angle is called the Brewster angle φ _b and can be expressed by the following equation (1). n ₁ is the refractive index on the atmosphere side, and n ₂ is the refractive index on the container side.

  In other words, collecting scattered light incident at an angle smaller than the Brewster angle is an efficient detection method. Incidentally, when the refractive index of each part in the present embodiment defined above is substituted into the above equation (1), the Brewster angle when the scattered light travels from the container 2 side to the atmosphere side is as shown in the following equation (2). The Brewster angle when the scattered light travels from the atmosphere side to the detector 9 side is about 56.3 ° as shown in the following equation (3).

  Therefore, in the present invention, when the scattered light 7 travels from the container 2 side to the atmosphere side, or travels from the atmosphere side to the detector 9 side, a region that transmits through the boundary surface at an incident angle smaller than the Brewster angle is efficiently obtained. An optical system suitable for condensing was used, and the first reflecting mirror defined as follows was adopted.

  Details of the first reflecting mirror 8 will be described with reference to FIGS. 1, 6, 7, and 8. 6 is a partially enlarged view from the vicinity of the emission center of the scattered light 7 in FIG. 1 to the vicinity of the center of the detector 9, FIG. 7 is an enlarged view of the vicinity of the emission center of the scattered light 7 in FIG. 1 shows a partially enlarged view of the vicinity of the detector 9 in FIG. 6, 7, and 8, in order to make it easy to understand the intersection position of the light beams, the hatching representing the light is omitted.

  As shown in FIG. 6, the first reflecting mirror 8 is composed of a part of a spheroid with the optical axis 10 as the center of rotation. The shape determines the two focal points 12 and 13 and the passing point 14 which define the ellipse 11 as follows. That is, as shown in the partial enlarged views of FIGS. 6, 7, and 8, the light 16 that is refracted at the Brewster angle from the emission center 15 of the scattered light 7 is refracted and transmitted at the boundary between the container 2 and the atmosphere. Before the light beam which is incident on the detector 9 at the Brewster angle and is refracted and reaches the detection surface center 17 is refracted on the surface of the detector 9. The light beam 18 is extended in the traveling direction, and the position intersecting the optical axis 10 is defined as the focal point 13. The reason why the focal point 12 is not provided at the emission center 15 of the scattered light 7 and the focal point 13 is not provided at the detection surface center 17 is that the refraction of the container 2 and the atmosphere and the refraction of the atmosphere and the surface of the detector 9 are taken into consideration. . Here, the reason why it is necessary to enter the detector 9 at an angle smaller than the Brewster angle is that the light receiving surface of a semiconductor detector such as a photodiode is covered with a transparent insulating film such as silicon oxide. This is because reflection, refraction, and transmission need to be taken into consideration, and the same applies to the case where it is protected with a transparent resin or glass. Moreover, even if it is not a semiconductor detector but a photomultiplier tube, the light incident port of a light-receiving part is glass.

  Next, the passing point 14 is defined as light out of light that can be radiated from the emission center 15 of the scattered light 7 and refracted at the boundary between the container 2 and the atmosphere and transmitted without hitting the transmitted light absorber 6. The light beam 19 having the smallest angle with the axis 10, or the light beam 19 ′ having the largest angle with the optical axis 10 out of the light 4 transmitted through the sample solution 1 and the container 2, and the light beam 18. An ellipse 11 is defined as a position extending in the direction opposite to the axis 10 and intersecting, and the reflecting mirror 8 is composed of a part of a spheroid in which the ellipse 11 is rotated about the optical axis 10.

  Further, a part of the spheroid means a position where the light beam 16 extends in the traveling direction and intersects the ellipse 11 as a start position 20, and a position where the light beam 18 extends in the direction opposite to the traveling direction and intersects the ellipse 11. An end portion 21 is an intermediate portion obtained by cutting the spheroid in a plane perpendicular to the optical axis 10 into three at a start position 20 and an end position 21. If the length of the reflecting mirror 8 in the direction of the optical axis is at least within this range, except for the area of the light 4 out of the scattered light 7 emitted forward, the incident angle is smaller than the Brewster angle from the container 2 to the atmosphere side. It is possible to condense all the exiting regions, and to enter the detector 9 at an incident angle smaller than the Brewster angle, so that the collected scattered light 7 can be detected efficiently. Since the reflecting mirror 8 defined above is located between the focal point 12 and the focal point 13, both the scattered light 7 incident on the reflecting mirror 8 and the scattered light 7 reflected by the reflecting mirror 8 on the detector 9 are both in the optical axis direction. The direction with reference to is not reversed. That is, the detection surface of the detector 9 only needs to face the emission center 15 of the scattered light 7, and no detection surface for detecting light from the opposite direction is necessary, and only one detector is required. effective.

  In order to explain the efficiency of the reflector according to the above definition, the positions of the focal point 12 and the focal point 13 are the same and are made of a larger spheroid, and the focal point 12 and the focal point 13 are also the same and the rotation is smaller. The case of an ellipsoid will be described with reference to FIGS.

  First, the case where the positions of the focal point 12 and the focal point 13 are the same and are made of a larger spheroid will be described with reference to FIG. 9 is the same as that of the first reflecting mirror 8 in the positions of the focal points 12 and 13 in FIG. 6, and the reflecting mirror is a first reflecting mirror made of a spheroid based on an ellipse 11 ′ larger than the first reflecting mirror 8. The path of light when replaced with 8 'is shown. A light beam 16 refracted and transmitted through the boundary between the container 2 and the atmosphere from the container 2 at the Brewster angle is reflected on the right side as compared with the case of the reflecting mirror 8 in FIG. Is possible. However, as the first reflecting mirror 8 ′ increases, the scattered light 7 emitted from the container 2 cannot enter the detector 9 from the inside near the optical axis 10 and cannot be detected. The reason for this is that light emitted from the emission center 15 of the scattered light 7 is refracted at the boundary between the container 2 and the atmosphere at the intersection 22 where the extended line of the light ray 18 incident on the detector 9 at the Brewster angle intersects the ellipse 11 ′. Of the light 4 that passes through the sample solution 1 and the container 2 and is emitted through the sample solution 1 and the container 2. , The light 24 (the area shown by hatching in FIG. 9) reflected from the intersection 23 where the light beam 19 ′ having the maximum angle with the optical axis 10 intersects the ellipse 11 ′ and directed to the detector 9 is detected by the detector 9 This is because most of the light is incident on the light receiving surface of the detector 9 and cannot enter the detector 9. Therefore, it can be said that the first reflecting mirror 8 defined by the ellipse 11 is the largest reflecting mirror that can efficiently collect the scattered light 7 when the positions of the focal point 12 and the focal point 13 are fixed.

  Next, the case where the positions of the focal point 12 and the focal point 13 are the same and are made of smaller spheroids will be described with reference to FIG. 10 is the same as the first reflecting mirror 8 in the positions of the focal points 12 and 13 in FIG. 6, and the reflecting mirror is a first reflecting mirror made of a spheroid based on an ellipse 11 ″ smaller than the first reflecting mirror 8. The path of light when replaced with 8 ″ is shown. A light beam 16 refracted and transmitted through the boundary between the container 2 and the atmosphere from the container 2 at the Brewster angle is reflected by a portion closer to the container 2 on the left side than the case of the reflecting mirror 8 in FIG. , Can enter the detector 9. Also, the light beam 19 or the light beam 19 ′ can be incident on the detector 9 at an angle smaller than the Brewster angle after being reflected by the first reflecting mirror 8 ″. The ellipse 11 ″ to be defined can be made smaller if the positions of the focal point 12 and the focal point 13 are the same. The limit of making the ellipse 11 ”is refracted at the incident surface of the detector 9 and incident on the center of the detector 9. Spheroid consisting of an ellipse 11 ″ that can be incident on the transmitted light absorber 6 without being incident on the transmitted light absorber 6 and defined as a passing point 14 at the intersection 26 of the light beam 25 and the light beam 16 having the smallest angle with the optical axis 10. This is the first reflecting mirror 8 ″ using a body. However, when the transmitted light absorber 6 is sufficiently small, the passing point 14 is a position where the light emitted from the container 2 at the Brewster angle intersects the surface of the container 2. Therefore, the first defined by the ellipse 11 described above is used. The reflector 8 is the largest reflector that can efficiently collect the scattered light 7 when the positions of the focal point 12 and the focal point 13 are fixed, and the first reflecting mirror 8 ″ defined by the ellipse 11 ″ described above is It can be said that it is the smallest reflector.

  In the present embodiment, the first reflecting mirror 8 is used in consideration of the simultaneous use of the second reflecting mirror described in the second and later embodiments. Further, since the actual light source is not a point light source and has a diameter of about 1 mm, it includes a light beam deviated from the light beam position described so far, but the size of the detection surface of the detector 9 is larger than the size of the light source. Since it is sufficiently large, the light beam shifted from the light beam position is ignored.

  The liquid analysis method in this example is almost the same as in Example 1. The difference from Example 1 is that in Example 1, the sample solution placed in the container is irradiated with light, and the same direction as the direction of light irradiation. In this embodiment, scattered light scattered in the direction side and fluorescence emitted in the same direction as the direction of light irradiation are collected on the detector by a reflecting mirror and the amount of antigen is measured. Then, the scattered light scattered in the direction opposite to the direction of light irradiation or the fluorescence emitted in the direction opposite to the direction of light irradiation is condensed on the detector with a reflector to measure the amount of antigen, etc. It is in. For this reason, in this embodiment, the part relating to the principle of liquid analysis and the optical law will be omitted. In addition, since the present embodiment also has a structure that can handle both the case where scattered light is used and the case where fluorescence is used, the following description will be made simply as scattered light including fluorescence.

  FIG. 11 shows a measuring unit of the liquid analyzer according to the present invention. A sample solution 1 consisting of a specimen and a reagent is placed in a transparent container 2. In FIG. 11, the sample solution 1 and the container are shown in cross section, but are shown without hatching in order to avoid complicating the drawing and hindering understanding. A lens 5 for narrowing the light 4 emitted from the light source 3 and the light source 3 to the sample solution 1 is arranged on the left side of the container 2 in the figure. On the right side of the container 2, the transmitted light absorber 6 and the light 4, which are narrowed down to the sample solution 1 by the lens 5 and absorb and capture the light transmitted through the sample solution 1, are scattered by the components contained in the sample solution 1. The second reflecting mirror 28 that reflects and collects the scattered light 27 that is emitted to the left side of the solution 1, that is, the direction opposite to the irradiation direction of the light 4, and the scattered light 27 that is collected by the second reflecting mirror 28 are detected. A detector 9 for converting into an electric signal is arranged. These are arranged on the optical axis 10 and are held by holding members, but the holding members are omitted from the drawings in order to avoid making the drawing complicated and obstructing the understanding.

  Next, the path of light and the configuration of the apparatus at the time of liquid analysis according to this embodiment will be described. At the time of liquid analysis, first, the sample solution 1 is put in the container 2 and set on the optical axis as shown in FIG. The container 2 is made of a material such as glass or plastic that transmits light 4 with low loss. In this state, light 4 is emitted from the light source 3 by energizing the light source 3 or the like. The light 4 is narrowed down by the lens 5 and irradiated to the sample solution 1. As a result, the scattered light 27 emitted from the sample solution 1 is collected by the second reflecting mirror 28, and the amount of light is measured by the detector 9, whereby the liquid component can be analyzed in the same manner as in the first embodiment.

  Details of the second reflecting mirror 28 will be described with reference to FIGS. FIG. 12 is an enlarged view from the vicinity of the emission center of the scattered light 27 in FIG. 11 to the second reflecting mirror 28. In FIG. 11 and FIG. 12, the second reflecting mirror 28 is composed of a part of a spheroid with the optical axis 10 as the center of rotation. As for the shape, two focal points 30, a focal point 31, and a passing point 32 that define an ellipse 29 are determined as follows. That is, in FIG. 11 and FIG. 12, the light 33 radiated backward from the emission center 15 of the scattered light 27 at the Brewster angle is refracted and transmitted at the boundary between the container 2 and the atmosphere, and extends in the direction opposite to the traveling direction. , The position intersecting the optical axis 10 is the focal point 30, and the focal point 31 is the same position as in the first embodiment, and the light beam incident on the detector 9 at the Brewster angle and refracted and reaches the detection surface center 17 is the surface of the detector 9. The light beam 18 before being refracted by is extended in the direction of travel and is set to a position intersecting the optical axis 10.

  Next, the definition of the passing point 32 can be made without being kicked by the inner diameter of the first reflecting mirror 8 among the rays that enter the detector 9 and refract and reach the detection surface center 17. An ellipse 29 is defined as a position where the light ray 34 in which the maximum light ray has traveled in the direction of the second reflecting mirror 28 and the light ray 33 intersect. The second reflecting mirror 28 has the ellipse 29 centered on the optical axis 10. It consists of a part of a spheroid rotated in the direction of

  Further, the part of the spheroid is a range in which the radius of the outer diameter around the optical axis of the second reflecting mirror is larger than the distance from the optical axis to the passing point 32. Further, the angle with the optical axis through which the light passing through the irradiation light 4 centered on the optical axis 10 or the light incident on the emission center 15 of the scattered light 27 can be incident without being kicked by the transmitted light absorber is minimized. An opening having a radius from the optical axis at the intersection of the light beam and the ellipse 29 is provided.

  Since the reflecting mirror 28 according to the above definition is located on the left side of the positions of the focal point 30 and the focal point 31 in FIG. 11, the reflected light 27 incident on the reflecting mirror 28 is reflected only once, and the orientation is based on the optical axis direction. Invert. That is, the detection surface of the detector 9 only needs to be directed toward the emission center 15 of the scattered light 27, and a detector for detecting the scattered light 27 emitted backward is not necessary. It has the effect of being finished.

  In order to explain the efficiency of the reflector according to the above definition, the positions of the focal point 30 and the focal point 31 are the same and are made of larger spheroids, and the focal point 30 and the focal point 31 are also at the same position and smaller in rotation. The case of an ellipsoid will be described with reference to FIGS.

  First, the case where the positions of the focal points 30 and 31 are the same and are formed of a larger spheroid will be described with reference to FIG. In FIG. 13, the positions of the focal points 30 and 31 are the same as those of the second reflecting mirror 28, and the reflecting mirror is changed to a second reflecting mirror 28 ′ composed of a spheroid based on an ellipse 29 ′ larger than the second reflecting mirror 28. The path of light when replaced is shown. A light beam 33 refracted and transmitted through the Brewster angle from the container 2 rearward at the boundary between the container 2 and the atmosphere is reflected on the outer peripheral side compared to the case of the reflecting mirror 28 in FIG. As the reflecting mirror 28 'increases, the area kicked by the first reflecting mirror 8 before entering the detector 9 increases. A hatched portion along the light beam 33 and the light beam 34 in FIG. 13 indicates the region. Therefore, the second reflecting mirror 28 defined by the ellipse 29 can be said to be the largest reflecting mirror that can efficiently collect the scattered light 27 emitted backward when the positions of the focal point 30 and the focal point 31 are fixed. .

  Next, the case where the positions of the focal point 30 and the focal point 31 are the same and are made of smaller spheroids will be described with reference to FIG. In FIG. 14, the positions of the focal points 30 and 31 are the same as those of the second reflecting mirror 28, and the reflecting mirror is changed to a second reflecting mirror 28 ″ composed of a spheroid based on an ellipse 29 ″ smaller than the second reflecting mirror 28. The path of light when replaced is shown. Of the light emitted backward from the container 2, the light that enters the irradiation light 4 centered on the optical axis 10 or the light incident on the emission center 15 of the scattered light 27 is not kicked by the transmitted light absorber. Compared with the case where the light 35 hitting the aperture whose radius from the optical axis is the intersection of the light beam having the smallest angle with the optical axis and the ellipse 29 hits the second reflecting mirror 28 defined by the ellipse 29, 14, the second reflecting mirror 28 ″ hits on the right side and the optical axis side. Therefore, as the ellipse 29 ″ becomes smaller, it is kicked by the transmitted light absorber 6 and does not reach the detector 9. The area will increase. In FIG. 14, a hatched portion along the light beam 36 that goes to the detector without being kicked by the transmitted light absorber 6 indicates the region.

  However, when the transmitted light absorber 6 is sufficiently small, this is not limited to this, but the incident is incident on the detector 9 as the ellipse 29 ″ becomes smaller because the focal point 30 is output at the Brewster angle. Therefore, when the position of the focal point 30 and the focal point 31 is fixed by the second reflecting mirror 28 defined by the ellipse 29, the scattered light 27 is emitted backward. Can be said to be the smallest reflecting mirror that can efficiently collect the light, but depending on the size of the transmitted light absorber, it can be reduced in the direction of the ellipse 29 ″. In the present embodiment, the second reflecting mirror 28 defined by the ellipse 29 is used.

  FIG. 15 shows a measuring unit of the liquid analyzer according to the present invention. In this embodiment, the scattered light or fluorescence emitted forward shown in the first embodiment and the scattered light or fluorescence emitted backward shown in the second embodiment are simultaneously detected. Since the analysis method, the light detection method, and the like are the same as those in the first and second embodiments, the description of the contents is omitted.

  16, FIG. 17, FIG. 18 and FIG. 19 show the measuring unit and the like of the liquid analyzer according to the present invention. FIG. 16 shows a measuring unit of the liquid analyzer according to the present embodiment, and shows only two of a plurality of measuring units arranged side by side. FIG. 17 is a plan view showing a part of the shutter used in this embodiment. FIG. 18 shows only one of the measurement units shown in FIG. 16 together with scattered light and fluorescence indicated by hatching. FIG. 19 shows a part of the measurement unit of the liquid analyzer according to this embodiment in a three-dimensional view excluding the first reflecting mirror 8, the second reflecting mirror 28, and the transmitted light absorber 6. Since the present embodiment also has a structure that can handle both the case of using scattered light and the case of using fluorescence, the following description will be made simply as scattered light including fluorescence.

  The sample solution 1 in this example is placed in a container composed of a first transparent substrate 37 and a second transparent substrate 38 arranged in parallel with a certain gap as shown in FIGS. It is held in the corresponding device 39. The device 39 corresponds to the container 2 shown in Example 1, Example 2 and Example 3, and the sample solution 1 is applied to the plurality of transport electrodes 40 provided in the device 39 inside the device 39. By sequentially switching the voltage, it is conveyed by electrostatic force, and is sequentially measured by a plurality of measuring units. In FIGS. 16 and 18, the optical axis 10 is arranged in the vertical direction. However, these figures may be plan views as viewed from above, and the optical axis may be horizontal.

  The liquid analysis method in this embodiment is the same as that in Embodiment 3, and both scattered light emitted forward and scattered light emitted backward are detected. The difference from the third embodiment is that both the scattered light emitted forward and the scattered light emitted backward are detected by the shutter 41 shown in FIGS. 16, 17 and 18, and the scattering emitted forward or backward is performed. There is a switch between detecting either one of the lights or hiding both scattered lights. This is an effective means for obtaining more detailed detection data especially when detecting scattered light. The reason for this is that when the immunoturbidimetric method using latex agglutination described in detail in Example 1 is used, depending on the size of latex particles used and the degree of aggregation between latex particles, This is because the angle changes.

  In this embodiment, when detecting both the scattered light emitted forward and the scattered light emitted backward, when detecting either the scattered light emitted forward or backward, or hiding both scattered lights A method for switching between cases will be described. In this case, the scattered light emitted forward is the scattered light 7 collected by the first reflecting mirror 8 as shown in FIG. 18, and the scattered light emitted backward is caused by the second reflecting mirror 28. This is the condensed scattered light 27.

  FIG. 17 shows a part of the shutter 41 viewed from the plane direction. The shutter 41 has three optical openings 42 for each measurement position. The hatched portions in FIG. 17 block light, and the other white portions pass light. The optical opening 42 includes an opening 43 that transmits both scattered light emitted forward and scattered light emitted backward, an opening 44 that transmits only scattered light emitted forward, and only scattered light emitted backward. Three places of the opening 45 which permeate | transmit are one set. However, it goes without saying that if there are unnecessary openings in the three places, they can be excluded. At the center of the opening 44 is a shutter 46 that blocks only scattered light emitted backward, and the opening 44 is a ring-shaped opening. The shutter is manufactured by pasting or vapor-depositing a light blocking material on a material such as glass or plastic that transmits light, or by creating an opening by etching or the like in a light blocking material such as metal. Just make it. When the opening is formed by etching or the like, the portion that becomes the shutter 46 may be connected to the inner diameter of the opening 44 by a thin rib-shaped bridge. When both the scattered light emitted forward and the scattered light emitted backward are detected by the shutter 41, when either the scattered light emitted forward or backward is detected, or when both scattered lights are hidden Is switched, the shutter is moved in the direction of the arrow shown in FIGS. 16 and 18 so that the center of the desired opening is positioned on the optical axis 10. The arrangement of the openings of the opening 42 need not be in accordance with the present embodiment, and may be arbitrary. Incidentally, if the arrangement in the openings 42 changes by 90 degrees, the direction of the arrows shown in FIGS. 16 and 18 becomes a direction perpendicular to the paper surface, and there may be changes other than 90 degrees. Further, the position of the shutter 41 in the present embodiment is between the first reflecting mirror 8 and the detector 9, but if the size and arrangement of the openings are determined according to the arrangement location, the shutter 41 is arranged at another location. It is possible.

  The present invention relates to an analyzer that detects the amount of a component contained in a liquid sample, and is capable of detecting fluorescent light and scattered light emitted from a sample in a small size and efficiently condensing and detecting a small amount of sample. Enables stable analysis. Therefore, it can contribute not only to the miniaturization of the sample solution but also to miniaturization of the apparatus.

The figure which shows the measurement part of a liquid analyzer. Schematic of the state in which scattered light or fluorescence travels from the container side to the atmosphere side. The graph showing the reflectance and transmittance of light traveling from the container side to the atmosphere side. Schematic of a state in which scattered light or fluorescence travels from the atmosphere side to the container side. The graph showing the reflectance and transmittance | permeability of the light which progresses from the atmosphere side to the container side. The elements on larger scale of FIG. The elements on larger scale of FIG. The elements on larger scale of FIG. The figure which shows the example which enlarged the reflective mirror of FIG. The figure which shows the example which made the reflective mirror of FIG. 6 small. The figure which shows the measurement part of a liquid analyzer. The elements on larger scale of FIG. The figure which shows the example which enlarged the reflective mirror of FIG. The figure which shows the example which made the reflective mirror of FIG. 11 small. The figure which shows the measurement part of a liquid analyzer. The figure which shows the measurement part of a liquid analyzer. The top view of a shutter. The elements on larger scale of FIG. The three-dimensional figure of a part of measuring part of a liquid analyzer.

Explanation of symbols

  DESCRIPTION OF SYMBOLS 1 ... Sample solution, 2 ... Container, 3 ... Light source, 4 ... Light, 5 ... Lens, 6 ... Transmitted light absorber, 7 ... Scattered light, 7 '... Fluorescence, 8 ... 1st reflective mirror, 8' ... 1st 1 reflecting mirror, 8 "... 1st reflecting mirror, 9 ... detector, 10 ... optical axis, 11 ... ellipse, 11 '... ellipse, 11" ... ellipse, 12 ... focus, 13 ... focus, 14 ... pass point , 15 ... emission center, 16 ... light beam, 17 ... detection surface center, 18 ... light beam, 19 ... light beam, 19 '... light beam, 20 ... start position, 21 ... end position, 22 ... intersection point, 23 ... intersection point, 24 ... light , 25 ... light rays, 26 ... intersections, 27 ... scattered light, 27 '... fluorescence, 28 ... second reflecting mirror, 28' ... second reflecting mirror, 28 "... second reflecting mirror, 29 ... ellipse, 29 '... ellipse, 29 "... ellipse, 30 ... focus, 31 ... focus, 32 ... pass point, 33 ... light, 34 ... light, 35 ... light, 36 ... light, 37 ... first transparent substrate 38 ... second transparent substrate, 39 ... device, 40 ... carrier electrode, 41 ... shutter, 42 ... opening, 43 ... opening, 44 ... opening, 45 ... opening, 46 ... shutter.

Claims (14)

A sample holder having at least two wall surfaces for holding the sample solution;
An irradiation optical system for irradiating the sample solution in the sample holder with light;
A transmitted light capturing device for capturing light transmitted through the sample solution in the sample holder;
A first reflecting mirror for condensing forward scattered light scattered on the light irradiation direction side by the sample solution in the sample holder;
A second reflecting mirror for condensing backscattered light scattered on the opposite side of the light irradiation direction by the sample solution in the sample holder;
A liquid analyzer comprising: a first reflecting mirror; and a detector that detects scattered light collected by the second reflecting mirror.   2. The liquid analyzer according to claim 1, wherein the first reflecting mirror and the second reflecting mirror are spheroid mirrors made of a part of a spheroid.   3. The liquid analyzer according to claim 2, wherein the scattered light collected by the second reflecting mirror is collected on a detector through an opening of the first reflecting mirror.   4. The liquid analyzer according to claim 3, wherein the forward scattered light is reflected once by the first reflecting mirror and collected by the detector, and the backward scattered light is reflected once by the second reflecting mirror. Then, the liquid analyzer is focused on the detector.   The liquid analyzer according to claim 3, wherein the first reflecting mirror does not invert a component in an optical axis direction of a traveling direction of the forward scattered light by reflection.   4. The liquid analyzer according to claim 3, wherein focal positions of the first reflecting mirror and the second reflecting mirror on the detector side are the same. The liquid analyzer according to claim 6,
The first focal position of the first reflecting mirror is an optical axis when the optical path of the light beam emitted from the emission center of the forward scattered light at the Brewster angle on the wall surface of the sample holder is extended in the direction opposite to the traveling direction. Matches the point where
The first focal position of the second reflecting mirror is an optical axis when the optical path of the light beam emitted from the emission center of the backscattered light on the wall surface of the sample holder at a Brewster angle is extended in the direction opposite to the traveling direction. Matches the point where
The second focal positions of the first reflecting mirror and the second reflecting mirror are incident on the detector at a Brewster angle and travel toward the detector at the detection surface center of the detector. A liquid analyzer characterized by being coincident with a point intersecting the optical axis when the optical path is extended in the traveling direction as it is. 8. The liquid analyzer according to claim 7, wherein an ellipse defining a reflecting surface of the first reflecting mirror is
Out of the light beams that can reach the detection surface center of the detector without hitting the transmitted light capture device, the light path of the light beam having the smallest angle with the optical axis, and the Brewster angle emitted from the emission center of the forward scattered light A first ellipse having a point of intersection with the reflected ray as a passing point;
Out of the forward scattered light that travels out from the emission center of the forward scattered light and travels without hitting the transmitted light capturing device, a light beam having a minimum angle with the optical axis is incident on the detector at a Brewster angle. A liquid analyzer characterized by being in the range of a second ellipse having an intersection with an optical path of a light beam reaching the center of the detection surface as a passing point.   9. The liquid analyzer according to claim 8, wherein the length of the reflecting surface of the first reflecting mirror in the optical axis direction is at least a Brewster angle from the emission center of the forward scattered light to the wall surface of the sample holder. The length of the forward scattered light that can be reflected from the light beam emitted from the step up to the light beam having the smallest angle with the optical axis out of the forward scattered light that travels from the emission center of the forward scattered light and does not hit the transmitted light capturing device. A liquid analyzer characterized by having a range of   8. The liquid analyzer according to claim 7, wherein an ellipse defining a reflection surface of the second reflecting mirror is a light beam emitted from the emission center of the backscattered light at a wall surface of the sample holder at a Brewster angle, and An ellipse having, as a passing point, an intersection with a light path of a light beam having a maximum angle with the optical axis among light beams that can enter the detection surface center of the detector at an angle smaller than the Brewster angle without being kicked by the first reflecting mirror. A liquid analyzer characterized by that.   11. The liquid analyzer according to claim 10, wherein an outer diameter centered on the optical axis of the second reflecting mirror is larger than a distance from the optical axis to the passing point, and passes irradiation light centered on the optical axis. An ellipse that defines a light ray having a minimum angle with an optical axis at which the light incident on the aperture or the second focal position can be incident without being kicked by the transmitted light capturing device and the reflecting surface of the second reflecting mirror A liquid analyzer having an opening having a radius from the optical axis as an intersection with the liquid crystal.   2. The liquid analyzer according to claim 1, wherein the sample solution is irradiated with excitation light, and the fluorescence emitted from the sample solution to the light irradiation direction side and the fluorescence emitted to the opposite side of the light irradiation direction are condensed. A liquid analyzer.   2. The liquid analyzer according to claim 1, wherein the transmitted light capturing device also serves as a transmitted light detector for detecting a transmitted light amount.   8. The liquid analyzer according to claim 7, wherein a first shutter that blocks forward scattered light collected by the first reflecting mirror and a back scattered light collected by the second reflecting mirror are blocked. 2 shutters, or one of the first shutter and the second shutter, and by switching the shutter, both the forward scattered light and the back scattered light, or the forward scattered light or A liquid analyzer that detects only one of the backscattered light.

Images