JPS6165141A - Method and instrument for measuring immune reaction - Google Patents

Abstract

PURPOSE:To improve S/N and measurement accuracy by detecting parallel the scattered light by the pulverous particles formed as a result of antigen- antibody reaction by plural channels and averaging the power spectral density of each channel. CONSTITUTION:The antigen-antibody reaction arises in a cell 8 and the scattered light scattered by the pulverous particles generated by such reaction is made incident in parallel through an optical fiber array 10 to a photodiode array 11. The output from each photodiode of the photodiode array 11 is supplied through low-noise amplifiers 12-1-12-N, A/D converters 13-1-13-N, memories 14-1-14-N and multiplexers 15 to a divider 16. The output from the divider 16 is supplied to a high-speed Fourier transform device 20 by which the power spectral density of the intensity fluctuation of the scattered light is determined. The power spectral is further averaged by a demultiplexer 21, memories 22-1-22-N, normalizer 23 and arithmetic processing part 24.

Description

DETAILED DESCRIPTION OF THE INVENTION (Prior Art) The present invention relates to a method and apparatus for measuring an immune reaction based on an antigen-antibody reaction using intensity fluctuations of light scattered by fine particles.

There is an immunoassay method that utilizes a specific selective reaction of the immune reaction as a method for measuring in-vivo microcomponents such as immune substances, phorin, pharmaceuticals, and immunomodulators.They can be roughly divided into labels that use enzymes or radioactive isotopes as labeling substances. Two well-known methods are immunoassay and non-labeled immunoassay, which directly measures antigen-antibody complexes.

The former labeled immunoassay is radioimmunoassay (
RIA), It? The elementary immunoassay (EIA) and the fluorescence immunoassay (FIA) are well known, and although they are highly sensitive, they have various limitations such as handling of isotopes and waste disposal, and also require a long time for measurement. However, there are drawbacks such as high testing costs due to the expensive labeling reagents.

The latter non-labeled immunoassay methods include immunoelectrophoresis, immunodiffusion, and precipitation, and these methods are either simple or insufficient for precise J11 determination in terms of sensitivity, quantitative performance, and reproducibility. be. Regarding this type of immunoassay method, please refer to the "Recommendation of Clinical Laboratory Test Methods".
(Written by Izumihara Kanai, edited by Masamitsu Kanai, Aftershock Publishing), "Clinical Testing J Vol. 22° No. 5 (1978), ? 4
71-487u1. :Details L<Explained.

Also, “l mmunochemistry, V
oλ, 12° No. 4 (1975), No. 349-35
On page 1, 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 using the scattered light of laser light. Disclosed is a method for quantitatively analyzing antigens or antibodies by determining from changes in the spectral width of . This analysis method has the advantage of not using labeled reagents, but 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, so the disadvantage is that the equipment is large and expensive. In addition, errors occur when the spectrometer is mechanically driven, 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.

As mentioned above, conventional immunoassay methods use expensive labeling reagents, resulting in high analysis running costs, troublesome liquid handling and processing, long processing times, and high costs. However, this method requires a large spectrometer, has poor accuracy and reproducibility, and has the disadvantages of providing only a small amount of information.

- In order to eliminate these drawbacks, the above-mentioned conventional drawbacks can be overcome by measuring the antigen-antibody reaction by utilizing the fact that the intensity fluctuation of light scattered by fine particles is closely related to the antigen-antibody reaction. It is possible to perform measurements with high precision and reproducibility without using expensive labeling reagents or expensive large spectrometers, and also shortens measurement time.
Japanese Patent Application No. 148878-1987 discloses a method for measuring an immune reaction and an apparatus for carrying out such a method, which allows self-LJ measurement of an antigen-antibody reaction and obtains a lot of useful information about the antigen-antibody reaction. proposed in No.

This immune reaction measurement method involves projecting radiation onto an antigen-antibody reaction solution containing at least an antigen and an antibody. The scattered light generated by the antigen-antibody reaction is detected in a homodyne or heterodyne manner, and the antigen-antibody reaction is determined as 1 (III) based on the power spectrum density of the intensity fluctuation of this detection output.

In such an immune reaction measurement method, the intensity of scattered light generated by microparticles as a result of an antigen-antibody reaction, or the intensity of scattered light generated by an antigen-antibody reaction of microparticles on which antibodies or antigens are immobilized, is determined by light interference. By focusing on the fact that the power spectral density of this intensity fluctuation depends on the shape and size of the particle, we can detect the presence or absence of an antigen-antibody reaction, the antigen or antibody, by detecting the power spectral density of the intensity fluctuation. It is possible to obtain a lot of useful information, such as the determination of the amount of microparticles involved in the antigen-antibody reaction, and the state of aggregation of microparticles (particle size indicator 5). In this method, the scattered light is received by a photodetector and fluctuations in the output signal intensity are detected, so there is no need to use a labeling reagent, and since the method does not involve spectrum analysis of the scattered light, a spectrometer is used. There is no need to use . In addition, by utilizing the fact that the relaxation frequency of the power spectral density of the intensity fluctuation of scattered light depends on the size of the particle, the ratio of the relaxation frequencies before and after the antigen-antibody reaction is determined, and from the value of this ratio, the antigen-antibody reaction By measuring the antibody reaction, or by using the fact that the integral unit of the low-frequency side of the power spectral density of the intensity fluctuation of scattered light depends on the size of the particle, we can calculate the integration box before and after the antigen-antibody reaction. The ratio can be determined and the antigen-antibody reaction can be measured from the field of this ratio.

However, when measuring antigen-antibody reactions based on the power spectral density of intensity fluctuations of such scattered light, the energy of the scattered light is small, so the signal representing the power spectral density is easily affected by noise, and its S /N is small, which makes it difficult, for example, to accurately determine the relaxation frequency, resulting in a disadvantage of low measurement accuracy.

(Objective of the Invention) The object of the present invention is to maintain the advantages of the above-mentioned immune reaction measurement method using the intensity fluctuation of scattered light, effectively eliminate the drawbacks, and improve the S/N of the signal. The object of the present invention is to provide a measuring method and a measuring device that can improve measurement accuracy.

(Summary of the Invention) The measurement method of the present invention involves projecting radiation onto a reaction solution containing an antigen and an antibody, and fixing the antibody or antigen added to the reaction solution or by scattering light from fine particles generated by an antigen-antibody reaction. Light scattered by particles is received in parallel on multiple channels, the photoelectric conversion output of each channel is processed, the power spectrum density of the intensity fluctuation of the scattered light is determined for each channel, and these power spectrum densities are averaged. This method is characterized by measuring an antigen-antibody reaction based on the obtained power spectral density.

In addition, the measurement device of the present invention projects light onto a reaction solution containing an antigen and an antibody, and detects scattered light by fine particles generated by an antigen-antibody reaction or scattered light by fine particles on which antibodies or antigens are immobilized added to the reaction solution. The device detects the antigen-antibody reaction and measures the antigen-antibody reaction based on the power spectral density of the intensity fluctuation of the detection output, which comprises a cell containing a reaction solution for performing the antigen-antibody reaction, and a cell that emits coherent light, a light source device that makes light enter the cell; a plurality of photodetection devices that receive scattered light from different locations of the cell in parallel; and a first photodetection device that stores output signals from the plurality of photodetection devices.
storage means, means for sequentially reading out the output signals of each photodetector from the first storage means and determining the power spectral density of the intensity fluctuation, and a second storage for storing the plurality of power spectral densities. means for reading out and averaging a plurality of power spectral densities from the second storage means;
The method is characterized by comprising a means for measuring an antibody reaction.

(Example) FIG. 1 is a diagram showing the configuration of an example of the immune reaction measuring device according to the present invention. In this example, a He--Ne gas laser 1 with a wavelength of 632.8 nm is provided as a light source that emits coherent light. In addition to such a gas laser, a solid laser such as a semiconductor laser can also be used as a light source capable of emitting coherent light. A laser beam 2 emitted from a light source 1 is separated into a beam 4 and a beam 5 by a semi-transparent mirror 3. One of the light beams 4 is condensed by a condensing lens 6 and then projected onto a transparent cell 8 as a thin parallel beam by a collimator lens 7. The other light beam 5 is made incident on a photodetector 9 made of a silicon photodiode and converted into a Tanabata signal representing fluctuations in the output light intensity of the light source 1.

The cell 8 contains an antigen-antibody reaction Si, which is a mixture of a buffer solution in which fine particles having antibodies or antigens bound to their surfaces are dispersed, and a test liquid containing the antigen or antibodies. Therefore, an antigen-antibody reaction occurs in the cell 8, and interaction occurs between fine particles, and the LA powder particles adhere to each other, resulting in a change in the state of the brush movement. cell 8
Scattered light scattered by particles at different locations inside is made to enter a photodiode array 11 in parallel through an optical fiber array 10. The number of optical fibers in the optical fiber array 10 and the number of photodiodes in the photodiode array 11 do not necessarily have to match; for example, if scattered light transmitted by two adjacent optical fibers is It can also be made incident on a photodiode. In the present invention, a plurality of N channels are provided, and scattered lights from different positions of the cell 8 are received in parallel by these channels.

The charge conversion outputs output from each photodiode of the photodiode array 11 are amplified by low noise amplifiers 12-1 to 12-N, respectively, and then the A/D converter 1
3-1 to 13-N convert it into an M-bit digital signal. The A/D converters 13-1 to +3-iN sample P digital signals during each measurement cycle time,
What is the primary means of remembering these? +'lf J are stored in the respective first memories 74-1 to 14-N. Therefore, each memo J-no-tani (6) requires PXM dots or more.

Next, the PIIMI digital signal stored in each of these first memories 14-1 to 14-N is transferred to the multiplexer 7.
5 are sequentially read out and supplied to the divider 16.

The output of the photodetector 9 is connected to the divider 18 by a low noise amplifier? 7. It is supplied via an A/D converter 18 and a memory 19. The problems of these A/D converters 18 and memory 19 are exactly the same as those of the above-mentioned A/D converters 13-1... and memories 14-1... P pieces of M-bit digital signals are created and stored in the memory 19. The divider 16 provides a signal in which output fluctuations of the light source 1 have been corrected. Next, this signal is supplied to the fast Fourier transformer 20, where fast Fourier transform is performed to obtain the power spectrum density of the scattered light intensity fluctuation.

In this way, the photoelectric conversion output signals from the sequential channels are processed to obtain the power spectral density, and these are sent via the demultiplexer 21 to the second memories 22-1 to 22-N constituting the second storage means. Memorize one after another. The capacity of each of these second memories may also be PxM bits or more. After processing the signals of all channels in this manner, they are supplied to the normalizer 23 to be averaged, and the averaged power spectrum density is supplied to the arithmetic processing section 24. The arithmetic processing unit 24 performs @W98 theory based on the averaged power spectrum density, and determines whether or not there is an aggregation reaction.
Obtain measurement results such as the concentration of antigen or antibody in the sample,
This is supplied to the printer 25 to display the measurement results.

In the present invention, 1. l:) The power spectral density of the intensity fluctuation of the scattered light is detected as if it were a blur, but this power spectral density is composed of a term due to the fluctuation of the interference component due to the diffusion of the fine particles over a distance of the wavelength fertility ~t. , and a term due to fluctuations in the number of particles caused by the movement of particles into and out of the scattering volume.

Among these, the fluctuation of scattered light due to interference is observed as spatial fluctuation of the speckle pattern, but if this is directly incident on a single optical pressure detector with a wide light receiving surface,
Since spatial smoothing is performed over the area of the light-receiving surface, the detected fluctuation will be small, but as mentioned above, by using multiple channels to limit the field of view of each photodiode, the fluctuation can be reduced. Since detection can be performed with high sensitivity and the power spectral densities obtained from the outputs of multiple channels are averaged, the S/N ratio can be significantly improved. Generally, when the number of channels is N, S/, N is IFJ IB.

Therefore, for example, if 100 channels are provided, the S/N will be 1.
It becomes 0 times.

In the embodiment described above, the direction of the light beam 4 incident on the cell 8 and the optical axis direction of the optical fiber array 10 are set at 90 degrees, and a homodyne method is adopted in which one unit of the incident light beam does not directly enter the photodiode array 11. It is also possible to employ a heterodyne method in which a part of the incident light beam is also incident on the photodiode array 11. When detecting scattered light in a homodyne manner, the output signal of the photodiode array 11 consisting of a photomultiplier tube is proportional to the average value E2 of the square of the electric field strength of the scattered light. In the case of heterodyne detection in which scattered light and incident light are detected together, the electric field strength of the direct input light is E8.
Then, the output signal of the photodiode array 11 is -] (E8+E8) 2 =E6+2E8・Es+Es
・(1) becomes. Here, 4 has no fluctuation (even if there is, it is made loose compared to the fluctuation of the scattered light), so the fluctuation component of the output of the photodiode array 11 is almost equal to the second term 2 ES . In other words, an output signal approximately proportional to the electric field strength ξ of the scattered light can be obtained.

Next, a description will be given of how the power spectrum density of the intensity fluctuation of scattered light was obtained by fast Fourier transforming the output signal of the photodiode array 11 using the above-mentioned apparatus.

Here, the power spectral density S([) of the steady-state establishment process x(shi) can be expressed as follows.

Based on this equation (2), the power spectrum density is calculated using fast Fourier transform. However, the power spectral density obtained from the output from one channel is S
/N is low, as shown in Figure 2A, and it is difficult to accurately determine the relaxation frequency, which is the frequency at the shoulder of the curve. When the power spectral densities obtained are averaged, a power spectral density with significantly reduced noise is obtained as shown in FIG. 2B, and from this, the relaxation frequency "," can be accurately determined.

Next, the measuring method of the present invention will be explained based on numerical examples. In this embodiment, data is processed in real time, and a waveform representing the averaged power spectral density can be monitored in real time on the screen of the cathode ray tube 26.

First, the sampling rate in the A/'D converters 13-1 to 13-N is set to be at least twice the signal frequency based on the numbering theorem, but since the frequency component of the intensity fluctuation of scattered light is several hundred Hz or less, Set the sampling frequency to 2K
Let it be Hz. Therefore, the period is 500 μs. This time becomes the cycle time TO. Also, assuming that the number of samples P in each measurement cycle is 1024 points, each measurement cycle time TC is 500 μs × 102
It will be 4 works and 500m5. In other words, the time required to capture all data is approximately sooms.

Since the fast Fourier transformer 20 requires high-speed processing,
Completely hardware configuration. cycle time 20
If 4 cycles of butterfly X ti are performed at 0 nS, 800 n3 is required per butterfly.

On the other hand, the number of butterfly operations required for fast Fourier transform of P = 1024 points is P / 2 X to (I z P = 512
10=5120. The time required for fast Fourier transform is 800 nS per channel.
5120-: 4mS. Therefore, if the data acquisition cycle time TC is 500 m3 as shown in Fig. 3, the processing time T for each channel is 4 m after data is acquired during 500 m3 and the time required for the next data to be acquired.
Real-time processing is possible if Fourier transform processing, which requires S, is performed on all N channels, so at least 1
Channels around 00 can be processed in real time.

In Figures 4 and 5, the particle size is 0.188 μm, respectively.
This shows the averaged power spectral density obtained when a liquid in which 0.305 μm latex particles are dispersed is placed in cell 8. This represents the Lorentzian power spectral density, and is the This is due to interference effects in the power spectrum density of intensity fluctuations. It can be seen that the relaxation frequency of these power spectral densities is inversely proportional to the particle diameter. In other words,
As mentioned above, the intensity fluctuation of the scattered light is a combination of the component due to the interference of coherent light based on the movement of fine particles and the component due to the change in the number of particles in the scattering volume C).
In this example, interference components are mainly detected, and the relaxation frequency of the power spectral density is the reciprocal of the time it takes a particle to travel a distance of the wavelength of light, so the larger the particle size, the longer the travel time, and the relaxation frequency will decrease. As described above, the relaxation frequency of the power spectral density is inversely proportional to the particle size, so the presence or absence of antigen-antibody aggregation and the degree of aggregation can be detected from changes in this relaxation frequency.

As mentioned above, the intensity fluctuation of scattered light is the sum of the coherent component due to the Brownian motion of particles and the incoherent component due to changes in the number of particles within the scattering volume, but as the number of particles within the scattering volume decreases. When the amount of interfering components decreases to the same level as non-interfering components, components other than changes in scattered light intensity due to Brownian motion of particles will also be detected, making it impossible to accurately detect antigen-antibody reactions. . therefore,
The concentration of particles must be selected in a range that is low enough to obtain a sufficient intensity of incident light within the scattering volume and in which the coherent component is larger than the incoherent component, but the particle size of the scatterer If it is constant, the relative fluctuation will be constant over a fairly wide range of 13 degrees.

Figures 6 and 7 show latex particles with a diameter of 0.3 μm on which immunoglobulin G antibodies are immobilized.
10-4 g/m℃ and io-
9g/m, f! An antigen-antibody reaction solution to which immunoglobulin G of 1 degree is added is housed in a cell, and the power spectral density before and after the start of the antigen-antibody reaction is shown. In the case of the original concentration 10-' (1/m fl) shown in FIG. 6, the relaxation frequency before the reaction is about 50, whereas the relaxation frequency after the reaction has changed to 10. On the other hand, when the antigen concentration is 10-9 g/m℃, the relaxation frequency at the beginning of the reaction σ■ is about 95 Hz, and the relaxation frequency after the reaction is about 40 Hz.
The ratio F of the relaxation frequencies before and after the antibody reaction is left constant as follows, and when this one cycle is calculated for several antigen concentrations and plotted in a graph, it becomes as shown in FIG. In other words, in Fig. 8, the horizontal axis represents the antigen concentration, and the vertical axis represents the value of the relaxation frequency ratio F, but the antigen concentration can be detected by determining the relaxation frequency ratio F. I can do it.

On the other hand, in FIGS. 6 and 7, it can also be seen that the ratio (R) of relative fluctuation before and after the antigen-antibody reaction has a certain relationship with the antigen degree. Of course, the relative fluctuation can be calculated by finding the relaxation frequency "1" from the graph of the paraspectral density. At this time, the relative fluctuation ratio R can be expressed by the following equation.

The relative fluctuation ratio R was determined using this equation (3), and the relationship between this and the antigen concentration was determined in a graph as shown in FIG.

As is clear from this graph, the unknown antigen concentration can be determined by determining the ratio R of relative fluctuation before and after the antigen-antibody reaction. That is, prior to measurement, the relative fluctuation ratio R is determined for standard samples with different known antigen concentrations to obtain a calibration curve as shown in FIG. 9, and the relative fluctuation ratio R is determined for a test sample with an unknown antigen concentration. The antigen concentration can be determined based on the previously determined calibration curve.

In normal measurement, 10-8 to 10-9 g/mf:l
Although it is necessary to carry out accurate measurements near the antigen concentration of , the present invention fully satisfies such requirements.

On the other hand, the relative fluctuation ratio R according to equation (3) can also be determined as a ratio of changes in the integral value in the low frequency band of the power spectral density shown in FIGS. 6 and 7. That is, the relative fluctuation ratio R can be determined based on the following.

Here, the integral value A of the power spectral density before the antigen-antibody reaction and the integral 1 axis B after the reaction are TO-1-10'H
It is the integral value that is applied to the low frequency band of z.
L (assuming that the pass filter passes frequencies below 10'fiz).

When the particle size is constant, the power spectral density is Lorentzian, and decreases in inverse proportion to the square of the frequency at frequencies J5 greater than the relaxation frequency.

On the other hand, if grains (¥) are distributed, the observation is a superposition of Lorentzian spectra with relaxation frequencies corresponding to each grain size, so the power spectral density in the high frequency part no longer depends on the frequency. Therefore, from the shape of this part, it is possible to know the particle size distribution of the particles aggregated by the reaction.Such data has not been previously available, and it is important to understand the nature of the antigen-antibody reaction. This is useful information when analyzing the state.

FIG. 10 shows another embodiment of a channel portion that receives scattered light from the cell 8 in parallel.

In this example, a one-dimensional multi-channel image intensifier 30 is arranged between the optical fiber array 10 and the seven-dimensional diode array 11, and the optical fiber array 1
The weak scattered light, which normally radiates from 0, is multiplied and applied to the photodiode array 11. The image intensifier 30 has a configuration in which a photocathode surface 32 is disposed on the incident side of a one-dimensional multichannel plate 31, a fluorescent surface 33 is disposed on the output side, and an accelerating voltage is applied to these from a power source 34. Since it has an extremely large amplification factor, it is possible to multiply extremely weak scattered light, and the S/N ratio can be greatly increased.

FIG. 11 shows another embodiment of the channel portion, and in this example, scattered light from the cell 8 is transmitted to a large number of photomultiplier tubes 41- through a large number of optical fibers 40-1 to 40-N.
1-4l-N1. : Configure to guide.

Since the photomultiplier tube has very high sensitivity, this embodiment is effective when the scattered light is extremely weak.

FIG. 12 shows yet another embodiment of the channel portion. In this example, an imaging lens 50 is disposed between the cell 8 and the optical fiber array 10 so that the image of the surface on which the light beam 4 in the cell 8 is incident is formed on the incident end surface of the optical fiber array 10. In this case, the imaging lens 50 does not need to have imaging performance in the direction perpendicular to the paper plane of FIG. 12, so it can be constructed using a cylindrical lens.

The present invention is not limited to the embodiments described above, but can be modified and changed in many ways. The above explanation was given as an example for immunoglobulin G (+ (JG)), but immunoglobulin A (I (IA>).

[It can be applied to the measurement of all substances that cause agglutination due to antigen-antibody reactions, such as OM, IQ D, IQ E, Australian antigen, syphilis antigen, and insulin. Furthermore, in the above-mentioned example, antibodies were immobilized on the surface of microparticles to detect antigens in the specimen, but it is also possible to immobilize antigens on the surface of microparticles and detect antibodies in the specimen. can. Generally speaking, although polystyrene latex particles were used as the fine particles in the above embodiments, other organic particles or inorganic particles such as crow can 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, the scattered light by the fine particulate products generated as a result of the antigen-antibody reaction could be absorbed. You can also use it. An example of such an antigen-antibody reaction is a reaction using human chorionic gonadotropin (HCG) as an antigen and anti-human chorionic gonadotropin (anti-HCG>) as an antibody, and the antigen-antibody complex generated by this reaction. The body can be treated as microparticles.Furthermore, the antigen itself can also be used as a particle.For such an antigen-antibody reaction, Candida albicans is used as the antigen.
Examples of using anti-Candida albicans as an antibody,
In addition, blood cells, cells, microorganisms, etc. can also be used as particles. In the first embodiment shown in Figure 1, a batch method was used in which the antigen-antibody reaction solution was stored in a cell and measurements were performed, but a flow type method was adopted in which measurements were performed while the antigen-antibody reaction solution was continuously flowing. Of course, this is also possible.

(Effect of the invention) The effects of the present invention described above are summarized as follows.

(1) Since there is no need to use expensive and 1 mO'll sample labeled with enzymes or radioisotopes, the process can be carried out at low cost and easily.

(2) It has a higher viscosity and higher reproducibility than non-labeled immunoanalytical methods such as immunoelectrophoresis, immunodiffusion, and precipitation, so it is possible to obtain highly reliable measurement results with high precision.

(3) Since the method detects intensity fluctuations of scattered light based on the Brownian motion of fine particles, highly accurate measurement can be performed using an ultra-trace amount of sample, and the measurement time can be shortened.

(4) Compared to methods that orient 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 small and safe, and measurement is highly accurate and reliable. Get results.

(5) Since measurements are performed based on the power spectral density of optical fluctuations, a lot of useful information about antigen-antibody reactions can be obtained.

(6) Multiple channels are provided to separately receive scattered light from different positions, and the power spectrum density of the intensity fluctuation of the scattered light is calculated from the photoelectric conversion output signal of each channel, and then averaged. Since the antigen-antibody reaction is measured based on this, the S/N can be increased and the measurement accuracy can be improved.

7) Since it is a spatial multi-channel system, real-time processing is possible.

[Brief explanation of the drawing]

FIG. 1 is a diagram showing the configuration of an embodiment of the immune reaction measuring device according to the present invention; FIGS. 2A and B are diagrams showing its effects; FIG. 3 is a diagram showing its operation; Figures 4 and 5 are graphs showing the power spectral densities for fine particles with particle diameters of 0.488 μm and 0.305 μm, respectively, and Figures 6 and 7 are graphs showing the power spectrum densities for fine particles with particle sizes of 0.488 μm and 0.305 μm, respectively.
Graph showing the power spectral density before and after antigen-antibody reaction for mλ and 10-9g 7mi, Figure 8 is a graph showing the relationship between antigen concentration and relaxation frequency ratio, Figure 9 is antigen concentration and relative fluctuation. Graphs showing the relationship with the ratio, FIGS. 10, 11, and 12 are one-line diagrams showing other examples of the structure of the channel portion. 1... Laser light source 2. 4. 5... Luminous flux 3... Semi-transparent mirror 6... Condensing lens 7...
・Collimator lens 8...Cell 9...Photodetector 10...Nine fufufuiha array 11...Futaiode array 13-1~13-N...A/D converter 14-1~ 14
-N...First memory 15...Multiplexer 16...Divider 20.
...Fast Fourier transformer 21...Demultiplexer 22-1 to 22-N...Second memory 23...Normal +f 24...Arithmetic processing unit 25...Printer 26...Cathode ray tube 30・
・Image intensifier 40-1 to 40-N・
...Optical fibers 41-1 to 41-N...Photomultiplier tube 50...Imaging lens. Patent Applicant: Olympus Optical Industry Co., Ltd. Figure 2 A B Figure 3 Figure 5 ND liquid@CHnno Figure 7 Figure 7 Number of eyebrow tears (H2) Figure 8 Figure 9 Proverbs & Concentration (612

Claims (1)

[Scope of Claims] 1. Radiation is projected onto a reaction solution containing an antigen and an antibody, and scattered light is generated by fine particles generated by an antigen-antibody reaction, or scattered light by fine particles on which antibodies or antigens immobilized are added to the reaction solution. is received in parallel by multiple channels, the photoelectric conversion output of each channel is processed, the power spectrum density of the intensity fluctuation of the scattered light is determined for each channel, and the power spectrum obtained by averaging these power spectrum densities. An immune reaction measurement method characterized by measuring an antigen-antibody reaction based on density. 2. Project light onto the reaction solution containing antigen and antibody, and
Detects scattered light by particles generated by antibody reaction or scattered light by particles immobilized with antibody or antigen added to the reaction solution, and measures antigen-antibody reaction based on the power spectrum density of intensity fluctuation of this detection output. The device includes: a cell containing a reaction solution for performing the antigen-antibody reaction; a light source device that emits coherent light and makes it enter the cell; and a parallel reception of scattered light from different locations in the cell. a plurality of photodetecting devices, a first storage means for storing output signals from the plurality of photodetection devices; and a first storage means for sequentially reading out the output signals of each photodetection device from the first storage means, means for determining power spectral density of fluctuation; second storage means for storing the plurality of power spectral densities; means for reading and averaging the plurality of power spectral densities from the second storage means; 1. An immune reaction measuring device comprising means for measuring an antigen-antibody reaction based on the determined power spectral density.