JPH0478138B2 - - Google Patents

Description

Detailed Description of the Invention [Industrial Application Field] The present invention is directed to the production of physical or chemical reactions between fine particles or reactive molecules, such as physical adsorption or immune reactions based on antigen-antibody reactions. The present invention relates to a method and apparatus for measuring using light scattered by particles.

[Conventional technology]

Generally, a reaction liquid containing fine particles is irradiated with a light beam such as a laser beam, and the reaction between the fine particles is measured by measuring the scattered light of the radiation by the fine particles aggregated by the reaction between the fine particles contained in the reaction liquid. is being measured.

Taking immunoassay as an example, an immunoassay that utilizes selective immune reactions has been proposed as a method for measuring trace components in the body such as immune substances, hormones, medicines, and immunomodulation. Two methods are well known: labeled immunoassay, which uses enzymes or radioisotopes as labeling substances, and unlabeled immunoassay, which directly measures antigen-antibody complexes. ,
"Immunochemistry", Vol.12, No.4 (1975), No.
On pages 349 to 351, particles carrying antibodies or antigens on their surfaces are reacted with antigens or antibodies, and the average diffusion constant, which is an index of Brownian motion, which decreases in proportion to the size of aggregated particles, is calculated in response to irradiated laser light. A method for quantitatively analyzing antigens or antibodies by determining from changes in the spectral width of scattered light is disclosed.

In addition to these, there are other non-labeled immunoassay methods such as immunoelectrophoresis, immunodiffusion, and precipitation, and these immunoassays are described in the "Recommendations for Clinical Testing Methods" (authored by Izumihara Kanai, written by Izumihara Kanai). (edited by Masamitsu, Kanehara Publishing)
``Clinical Examination'' Vol. 22, No. 5 (1978), No. 471~
It is explained in detail on page 487.

In addition, examples of labeled immunoassay include radioimmunoassay (RIA), enzyme immunoassay (EIA), and fluorescence immunoassay (FIA).

[Problem that the invention seeks to solve]

However, with the reaction measurement method described above, when measuring scattered light, it is difficult to prevent ambient stray light from being mixed in as noise, and it is difficult to eliminate the influence of scattered light due to impurity particles in the reaction solution. I couldn't do it.

Regarding immunoassays, although labeled immunoassay methods are highly sensitive, they have various limitations such as handling of isotopes and waste disposal, and also require a long time for measurement and the labeling reagents are expensive. There are disadvantages such as high inspection cost. Furthermore, although conventional non-labeled immunoassays are simple analytical methods, they are insufficient as precise measurement methods in terms of sensitivity, quantitative performance, and reproducibility. Furthermore, although the analytical method for determining the average diffusion constant has the advantage of not using a labeling reagent, it uses a spectrometer to detect the broadening of the spectrum of incident light due to the Doppler effect caused by the Brownian motion of particles. There are drawbacks that the device is large and expensive, and errors occur when mechanically driving the spectrometer, resulting in poor accuracy and reproducibility. Furthermore, this method only calculates the average diffusion constant from the spectral width of light, and has the disadvantage that the amount of information is small.

The present invention was made to solve the above-mentioned problems, and involves irradiating a reaction liquid or a reaction gas containing fine particles or reaction molecules with a light beam such as a laser beam, thereby causing the fine particles contained in the reaction liquid to react with each other. By measuring the scattered light of the radiation by microparticles aggregated with each other or microparticles generated by a reaction between reactant molecules, it is possible to detect physical or chemical reactions between the microparticles or reactant molecules, such as physical adsorption or antigen - When measuring immune reactions based on antibody reactions, etc., it prevents ambient stray light from entering as noise, and
By removing the influence of scattered light caused by impurity particles, it is possible to perform sequential measurements efficiently with high accuracy and reproducibility without using an expensive and large spectrometer, and the measurement time is shortened. The object of the present invention is to provide a reaction measurement method and apparatus that can automate reaction measurement and improve reaction detection sensitivity.

[Means for solving problems]

To this end, the reaction measurement method and apparatus using synchronous detection of orthogonal polarization components of the present invention apply an electric field whose size or direction changes over time to a reaction liquid or reaction gas containing fine particles or reaction molecules.
Radiation having a linearly polarized wave is projected, and scattered light of the radiation by fine particles aggregated by a reaction between fine particles or fine particles generated by a reaction between reaction molecules is directed to the polarization plane of the radiation. measuring the reaction by homodyne or heterodyne detection through a polarizer having orthogonal polarization planes, and synchronously detecting the detection output with a signal synchronized with the change in the electric field; or a cell containing a reaction liquid or a reaction gas containing reaction molecules, a means for applying an electric field whose magnitude or direction changes over time to the reaction liquid or reaction gas in the cell, and linearly polarized light. a light source device that makes the radiation incident on the cell; and a light source device that makes the scattered light of the radiation by fine particles aggregated by the mutual reaction of the fine particles or fine particles generated by the mutual reaction of the reactive molecules with the linearly polarized light. A photodetection device that receives light in a homodyne or heterodyne manner through a polarizer having a polarization plane orthogonal to the polarization plane of the photodetection device, and synchronous detection that synchronously detects the output from the photodetection device with a signal synchronized with the change in the electric field. The present invention is characterized in that it includes a data processing device and a data processing device to which the output of the synchronous detection device is input.

[Effect]

The reaction measurement method and apparatus using synchronous detection of orthogonally polarized components of the present invention provides a reaction solution containing fine particles or reaction molecules to which an electric field whose size or direction changes over time has a linearly polarized wave. Microparticles that aggregate due to reactions between microparticles that are exposed to radiation,
Alternatively, the scattered light of the radiation by fine particles generated by a mutual reaction between reactant molecules is detected via a polarizer having a polarization plane perpendicular to the polarization plane of the radiation, and this detection output is used as the By performing synchronous detection using signals that are synchronized with changes in the electric field, only the frequency components that fluctuate in synchronization are extracted and used, completely eliminating the effects of ambient light such as stray light and scattered light due to other impurities. This makes it possible to detect reactions with high precision and sensitivity.

〔Example〕

Examples will be described below with reference to the drawings, taking an antigen-antibody reaction as an example of the reaction.

FIG. 1 is a block diagram showing the basic configuration of reaction measurement according to the present invention, FIG. 2 is a detailed view of the sample cell shown in FIG. 1, and FIG. 3 is a diagram showing an embodiment of the synchronous detection device. In the figure, 1 is a laser light source, 2 is a laser beam,
3 is a condenser lens, 4 is a polarizer, 5 is a sample cell, 6
is a reaction solution, 7 is a condensing lens, 8 is a polarizer, 9 is a photodetector, 10 is an oscillator, 11 is a synchronous detection device, 1
2 is a data processing device, E ₁ and E ₂ are electrodes, Tr ₁ to _{Tr 6}
is a transistor, R ₁ and R ₂ are stabilizing resistors, R ₃ and R ₄
is the output resistance, Ic is the constant current source, and Vcc is the power supply voltage.

Next, the effect will be explained. After the light 2 emitted from the laser light source is focused by the condenser lens 3, the output light from the polarizer 4 is projected onto the sample cell 5. A reaction solution 6 is placed in the sample cell 5, and this reaction solution is used to collect antibodies or antigens that cause a specific antigen-antibody reaction with the antigen or antibody to be measured in the sample, for example, onto a plastic plate with a diameter of about 0.2 μm. It consists of colloidal particles fixed on the surface of a sphere suspended in an appropriate solvent. These colloid particles undergo Brownian motion due to the shock caused by the thermal movement of the solvent molecules. In this state, when a beam of linearly polarized laser light is incident on the sample cell, the plastic sphere emits the laser light. However, because the scatterer is spherical, the forward scattered light maintains the same linear polarization as the incident light. Therefore, the scattered light does not pass through the polarizer 8, which has a polarization plane perpendicular to the polarization plane of the polarizer 4, and does not reach the photodetector 9. However, when an antigen-antibody reaction occurs, the fine particles are aggregated using the antigen or antibody as a mediator, and the scatterer no longer has a spherical shape and exhibits optical anisotropy. As a result, the scattered light has an elliptically polarized wave and a polarization component perpendicular to the plane of polarization of the incident light, so the scattered light passes through the condenser lens 7, the polarizer 8, and the photodetector 9. It will be incident. The output of this photodetector 9 changes when the anisotropy of the scattering medium changes depending on the agglomeration state of the particles in the reaction solution 6, so the immune reaction can be measured by detecting the output change. .

On the other hand, an alternating current electric field modulated at a frequency f is applied to the sample cell 5 from an oscillator 10, and as a result, the aggregated particles are arranged in the direction where the electric energy is minimized. For example, if the aggregated particles are spheroidal, the energy will be minimum and stable when the long axis is parallel to the electric field, which will change the anisotropy of the scattering medium. In this case, due to the influence of Brownian motion, it is not possible to align the directions of all particles in the direction of the electric field, but the greater the electric field strength, the greater the probability that the direction of the aggregated particles will be oriented in the direction of the electric field. Therefore, when the magnitude of the electric field applied from the outside is temporally modulated by the frequency f, the probability that the aggregated particles are oriented in the direction of the electric field is also modulated by the frequency f. Therefore, the anisotropy of the scattering medium will also be modulated with the frequency f. Correspondingly, the intensity of light incident on the photodetector 9 through the condenser lens 7 and polarizer 8 also changes with the frequency f. Therefore, by using the signal from the oscillator 10 as a synchronous input to the synchronous detection device 11 and synchronously detecting the output obtained by the photodetector 9, it is possible to separate and extract only the signal components that change at the frequency f. The state of the antigen-antibody reaction can be detected by this synchronized wave output, and in this case, ambient stray light, light scattered by other impurity particles, etc. will not be modulated by the frequency f. Even if stray light or the like enters the photodetector 9, it can be completely removed, resulting in an improved signal-to-noise ratio and further improved detection sensitivity. Although not shown, if the output light of the laser light source 1 is photoelectrically converted and the synchronous detection output is corrected as a monitor signal for output light intensity fluctuations, the influence of the output light fluctuations can be removed.

Next, the synchronous detection device shown in FIG. 3 will be described. The detection signals from the photodetector 9 are input to the bases of Tr ₁ and Tr ₂ in opposite phases to each other. On the other hand, the synchronization signal from the oscillator 10 is transmitted to the switching transistor.
Added to the base of Tr ₃ to _{Tr 6} . At this time, Tr ₃ ,
The pair of Tr ₆ and the pair of Tr ₄ and Tr ₅ are each in phase,
Moreover, these pairs are switched in opposite phases to each other, and on the output side, Tr ₃ and Tr ₅ and Tr ₄ and Tr ₆ are connected to each other, so that the output resistance R ₃ ,
In _R4 , detection outputs of opposite phases synchronized with the synchronization signal are obtained during the positive and negative periods of the synchronization signal, and by taking the difference between them, the magnitude of the signal generated at one output resistance is twice as large. The output is obtained.

Note that in the above configuration, the anisotropy of the scattering medium is changed by modulating the intensity of the electric field, but instead of this,
The anisotropy of the scattering medium may be changed by using a plurality of electrodes and periodically changing the direction of the applied electric field at a frequency f. Furthermore, if magnetic particles are used instead of plastic particles, a magnetic field can be used instead of an electric field. Furthermore, the polarization of the reactant molecules themselves may be utilized without using plastic particles.

This example can be applied to the measurement of immunoglobulin G (IgG), immunoglobulin A (IgA), IgM, IgD, IgE, and all substances that cause agglutination by the Australian antigen-antibody reaction. Many other modifications and changes are possible. Also,
Although plastic particles such as polystyrene were used as the fine particles, other organic particles and inorganic particles such as glass and metal particles may also be used. Furthermore, in the above-mentioned example, fine particles were present in the antigen-antibody reaction solution from the beginning, but instead of using such fine particles, scattered light from fine particulate products generated as a result of the antigen-antibody reaction was used. You can also. An example of such an antigen-antibody reaction is a reaction using human chorionic gonadotropin (HCG) as an antigen, and the antigen-antibody complexes produced by this reaction can be treated as fine particles.
Furthermore, the antigen itself can also be used as particles. In such an antigen-antibody reaction, Candida albicans (yeast) is used as the antigen and anti-Candida albicans is used as the antibody, and blood cells, cells, microorganisms, etc. can also be used as particles.

FIG. 4 is a block diagram showing an embodiment of the reaction measuring device according to the present invention. In the figure, 21 is a laser light source, 22, 24, and 25 are light beams, 23 is a half mirror, 26 is a condenser lens, 27 is a polarizer, 28 is a cell, 29 is a photodetector, 30 is a reaction liquid, and 31 is a collimator. , 32 is a polarizer, 33 is a photodetector, 34,
37 is an amplifier, 35 is a synchronous detection device, 36 is an oscillator, 38 is a low-pass filter, 39 is a data processing device, and 40 is a display device.

In this example, the light source has a wavelength of 632.8 nm.
A He-Ne gas laser 21 is provided. In addition to such a gas laser, a solid laser such as a semiconductor laser can also be used as a light source that emits coherent light. A laser beam 22 emitted from the light source 21 is separated by a half mirror 23 into a beam 24 and a beam 25. On the other hand, after the light beam 24 is condensed by a condenser lens 26, it is passed through a polarizer 27 made of, for example, a Glan-Thompson prism, and the linearly polarized light is projected onto a transparent cell 28. The other light beam 25 is made incident on a photodetector 29 made of a silicon photodiode and is converted into a monitor signal representing fluctuations in the output light intensity of the light source 21. In the cell 28, a reaction liquid containing fine particles or reactive molecules, for example, a liquid in which spherical fine particles having antibodies or antigens bound to their surfaces are dispersed, and a test liquid containing the corresponding antigen or antibody are mixed. Contains reaction solution 30. As mentioned above, antigen-
When a reaction such as an antibody reaction occurs and interaction occurs between fine particles, the fine particles adhere to each other, thereby changing the state of Brownian motion, thereby changing the polarization state of scattered light. On the other hand, an electric field modulated at a frequency f is applied to the cell 28 by an oscillator 36, and the anisotropy of the scattering medium also changes with the frequency f, as in the case of FIG. The scattered light scattered by the particles in the cell 28 is made incident on a collimator 31 having a pair of pinholes, and a polarizer 3 having a polarization plane orthogonal to the polarization plane of the polarizer 27 is used.
2 and enters a photodetector 33 consisting of a photomultiplier tube. The output of the photodetector 33 is applied to a synchronous detection device 35 which uses the output from an oscillator 36 as a synchronous input, and is subjected to synchronous detection. Only frequency components that fluctuate in synchronization with changes in the applied electric field are extracted, and a low-noise amplifier is used. 37 and a low-pass filter 38 to a data processing device 39;
The monitor signal from 9 is supplied to a data processing device 39 via a low noise amplifier 34 to eliminate the influence of output fluctuations of the laser light source. The data processing device 39 is
It performs signal processing as described below and outputs measurement results of reactions such as antigen-antibody reactions. This measurement result is supplied to the display device 40 and displayed. In this way, a significant relationship is recognized between the state of particle aggregation and the synchronous detection output, and from this it is possible to detect the presence or absence of aggregation, the degree of aggregation, and the like. Note that the polarizer 32
is not in the collimator 31, but in the collimator 31
and the photodetector 33.

In the embodiment described above, the reaction liquid 30 is placed in the cell 28.
However, it is also possible to accommodate a reaction gas containing fine particles or reaction molecules that react with each other.Also, although the batch method was adopted in which the reaction solution is accommodated in a cell and measured, the reaction solution Of course, it is also possible to use a flow method in which measurement is performed while continuously flowing a reaction gas or a reaction gas. Further, although a laser light source that emits coherent light is used as a light source, it is also possible to use a light source that emits incoherent light.

FIG. 5 is a diagram showing a detailed configuration of the collimator 31 shown in FIG. 4. The collimator 31 of this example has a hollow structure, in which a polarizer 32 is disposed, and the hollow 31 has a dark box structure to remove the influence of external light, and its inner surface has an antireflection structure. Pinholes 31b and 31c are provided before and after the cavity 31a.
form.

In the embodiment described above, the angle between the direction of the light beam 24 incident on the cell 28 and the optical axis direction of the collimator 31 was 90 degrees, but as shown in FIG. The angle θ between the collimator 31 and the optical axis can be set arbitrarily. Further, although the homodyne method in which the incident light flux does not directly enter the photodetector 33 is adopted, a heterodyne method in which a part of the incident light flux is made to enter the photodetector 33 may be employed.

Next, a method of estimating the concentration of fine particles that have caused a reaction according to the present invention will be explained.

Assuming that the scattered light intensity Is obtained as the output of the synchronous detection device described above shows a temporal change as shown in FIG. 7, and its average value is Im, the standard deviation of fluctuation from the average value ΔI is: (ΔI) ² =Σ(Is−Im) ² /N. However, N is the number of samples of the scattered light intensity Is. Therefore, when the concentration of fine particles resulting from the reaction is C (g/ml), log [(ΔI) ² /Im] ²
If we draw the relationship between That is, as the density decreases, the relative value of fluctuation increases, and as the density increases, the relative value of fluctuation decreases. Therefore, it is possible to data-process the output of the synchronous detection device described above to obtain (ΔI) ² /Im ² and estimate the concentration C using the calibration curve shown in FIG.

As described above, the present invention can synchronize with changes in the applied electric or magnetic field if the anisotropy of the scattering medium changes due to the reaction and the particles formed as a result of the reaction have electric or magnetic polarization. Since the anisotropy of the scattering medium also changes due to the change in the incident radiation, by detecting the light scattered by the particles of the incident radiation and synchronously detecting the detection output with a signal that synchronizes the changes in the applied electric or magnetic field, it is possible to detect the physical It is possible to measure reactions such as adsorption and chemical reactions.

〔Effect of the invention〕

As is clear from the above description, according to the present invention, the following effects can be obtained.

(1) In principle, it is a zero method, and no signal reaches the photodetector before the reaction occurs, so the sensitivity of the detector can be set sufficiently high, making it possible to use extremely sensitive physical and chemical methods. It becomes possible to detect the reaction and also shorten the measurement time.

(2) Since it uses synchronous detection, it is possible to remove the effects of stray light and scattered light due to impurities, improving detection accuracy, and by using it in conjunction with the zero method, reaction detection sensitivity is further improved. It becomes possible to do so.

(3) If the reaction is an immune reaction, (a) To detect the antigen-antibody reaction, so-called
There is no need to perform BF separation, and there is no need to remove unreacted components from the solution after the reaction.

(b) Since there is no need to use labeling reagents such as enzymes or radioitopes that are expensive and difficult to handle, immunoreaction measurements can be carried out at low cost and easily.

(c) It is more accurate and reproducible than non-labeled immunoanalysis methods such as immunoelectrophoresis, immunodiffusion, and precipitation, so it is possible to obtain highly reliable immune reaction measurement results with high precision.

(d) Compared to methods that quantify antigens or antibodies by determining the average diffusion constant from changes in the spectral width of scattered light, a spectrometer is not required, so the device is smaller and cheaper, and the immune reaction can be performed with high precision and reliability. measurement results are obtained.

[Brief explanation of the drawing]

Fig. 1 is a block diagram showing the basic configuration of the reaction measurement method according to the present invention, Fig. 2 is a detailed view of the sample cell shown in Fig. 1, and Fig. 3 is an example of the synchronous detection device shown in Fig. 1. 4 is a block diagram showing an embodiment of the reaction measuring device according to the present invention, and FIG. 5 is a diagram showing the detailed configuration of the collimator shown in FIG. 4.
FIG. 6 is a diagram showing the configuration of another embodiment of the reaction measuring device of the present invention, FIG. 7 is a graph showing the fluctuation of scattered light obtained as the output of the synchronous detection device, and FIG. 8 is a graph showing the fluctuation of scattered light obtained as the output of the synchronous detection device. FIG. 3 is a diagram showing the relationship between the relative value of fluctuation and the concentration of fine particles resulting from a reaction. DESCRIPTION OF SYMBOLS 1... Laser light source, 2... Laser light, 3... Condensing lens, 4... Polarizer, 5... Sample cell, 6...
...Reaction liquid, 7... Condensing lens, 8... Polarizer, 9
... Photodetector, 10 ... Oscillator, 11 ... Synchronous detection device, 12 ... Data processing device, E ₁ , E ₂ ...
Electrode, Tr ₁ to Tr ₆ ... Transistor, R ₁ , R ₂ ...
Stabilizing resistor, R ₃ , R ₄ ... Output resistance, Ic ... Constant current source, Vcc ... Power supply voltage, 21 ... Laser light source,
22, 24, 25...Light flux, 23...Half mirror, 26...Condensing lens, 27...Polarizer, 28
... Cell, 29 ... Photodetector, 30 ... Reaction liquid,
31... Collimator, 32... Polarizer, 33...
Photodetector, 34, 37... Amplifier, 35... Synchronous detection device, 36... Oscillator, 38... Low pass filter, 39... Data processing device, 40... Display device.

Claims (1)

[Claims] 1. Radiation having a linearly polarized wave is projected onto a reaction liquid or reaction gas containing fine particles or reaction molecules to which an electric field whose size or direction changes over time is applied, and the reaction between the fine particles is induced. Scattered light of the radiation by fine particles aggregated by microparticles or fine particles generated by reactions between reactant molecules is homodyne or heterodyne through a polarizer having a polarization plane orthogonal to the polarization plane of the radiation. A reaction measurement method using synchronous detection of orthogonal polarization components, characterized in that the reaction is measured by detecting the detection output with a signal synchronized with the change in the electric field. 2. A reaction measurement method using synchronous detection of orthogonally polarized components according to claim 1, wherein the reaction is an antigen-antibody reaction. 3. A reaction measurement method using synchronous detection of orthogonal polarization components according to claim 1, wherein the reaction is physical adsorption. 4. Reaction measurement using synchronous detection of orthogonal polarization components according to any one of claims 1 to 3, wherein the reaction liquid or reaction gas is continuously flowing. Method. 5. Claims 1 to 4, characterized in that the fine particles are organic or inorganic particles.
A reaction measurement method using synchronous detection of orthogonal wave components according to any one of the items. 6. Among claims 1 to 5, the reaction is measured based on the ratio of the square of the standard deviation and the square of the average value of the intensity fluctuation of the scattered light obtained from the detection output. A reaction measurement method using synchronous detection of orthogonal polarization components according to any one of the items. 7. Any one of claims 1 to 6, characterized in that the output obtained by the synchronous detection is corrected with a monitor signal of output light intensity fluctuation obtained by photoelectrically converting the projected radiation. A reaction measurement method using synchronous detection of orthogonal polarization components according to item 1. 8 A cell containing a reaction liquid or reaction gas containing fine particles or reaction molecules, a means for applying an electric field whose magnitude or direction changes over time to the reaction liquid or reaction gas in the cell, and a linearly polarized electric field. a light source device that makes the light incident on the cell; and a light source device that makes the linearly polarized radiation incident on the cell; a photodetection device that receives light in a homodyne or heterodyne manner through a polarizer having a polarization plane perpendicular to the polarization plane of the light; and a photodetection device that synchronously detects the output from the photodetection device with a signal synchronized with the change in the electric field. A reaction measurement device using synchronous detection of orthogonal polarization components, comprising a synchronous detection device and a data processing device into which the output of the synchronous detection device is input. 9. A reaction measuring device using synchronous detection of orthogonally polarized components according to claim 8, wherein the reaction is an antigen-antibody reaction. 10. A reaction measuring device using synchronous detection of orthogonal components according to claim 8, wherein the reaction is physical adsorption. 11. Reaction measurement using synchronous detection of orthogonal polarization components according to any one of claims 8 to 10, characterized in that the reaction liquid or reaction gas is continuously flowing. Device. 12. A reaction measuring device using synchronous detection of orthogonally polarized components according to any one of claims 8 to 11, wherein the fine particles are organic or inorganic particles. 13. Claim 8, wherein the data processing device is provided with a calculation means for calculating a ratio between the square of the standard deviation of the intensity fluctuation obtained from the output of the synchronous detection device and the square of the average value. Item to first
A reaction measurement device using synchronous detection of orthogonal polarization components according to any one of the above two items. 14. Any one of claims 8 to 13, characterized in that the data processing device receives a monitor signal of output light intensity fluctuation obtained by photoelectrically converting the output light of the light source device. A reaction measurement device using synchronous detection of orthogonal polarization components as described in 2. 15. Claims 8 to 1, characterized in that the scattered waves are guided to a photodetector through a collimator provided with a pair of pinholes at both ends.
A reaction measuring device using synchronous detection of orthogonally polarized components according to any one of the items 4 to 4. 16. The reaction measurement device using synchronous detection of orthogonal polarization components according to claim 15, wherein the collimator has a hollow dark box structure. 17. A reaction measuring device using synchronous detection of orthogonally polarized components according to claim 15 or 16, wherein the optical axis direction of the collimator and the direction of the incident light beam to the cell are 90 degrees. . 18. The orthogonal polarization component according to claim 15 or 16, wherein the optical axis direction of the collimator and the direction of the light beam incident on the cell are at any angle other than 90°. A reaction measurement device using synchronous detection. 19 Claim 15, wherein the polarizer is disposed within the collimator.
A reaction measuring device using synchronous detection of orthogonally polarized components according to any one of items 1 to 18. 20. Synchronization of orthogonal polarization components according to any one of claims 15 to 18, wherein the polarizer is disposed between the collimator and the photodetector. A reaction measurement device using detection.