JPS6040937A - Immunity-reaction measuring apparatus - Google Patents

Abstract

PURPOSE:To make it possible to perform accurate measurement, by providing a scattered light measuring and fluorescent light measuring device in combination, a transmitted light measuring device, and a data processor, which obtains a ratio between the intensity of transmitted light and the intensity of scatterd light. CONSTITUTION:An electron multiplier tube 5 for receiving projected light is provided on a reflected light path of a beam splitter. A measuring cell 6 is arranged on a transmitted light path. An annular slit 7 is arranged in front of a first lens 8 after the measuring cell. A second lens 11 for conversion is provided at the rear of a reflecting mirror in a coaxial relation with the first lens 8. In the case the apparatus is used also for the measurement of fluorescent light, a lens 12, a filter 16, and the like are further provided. A data processing part 18 is provided in order to obtain the ratio between the intensity of transmitted light and the intensity of the scattered light. By this constitution, the accurate measurement can be performed.

Description

Detailed Description of the Invention (a) Industrial Application Field The present invention relates to an immune reaction measuring device for performing serological tests, and in particular, it is capable of measuring insoluble antigen-antibody complexes generated by antigen-antibody reactions over a wide concentration range. The present invention relates to an immune reaction measuring device that optically detects with high sensitivity.

The present invention also enables optical detection of clusters of particles that are formed by aggregation of latex bound with antibodies (or antigens) using antigens (or antibodies) as bridges,
This relates to an immune reaction measuring device that can measure the amount of antigen or antibody.

The immune reaction measuring device of the present invention is particularly suitable for measuring blood plasma proteins, drugs, hormones, etc., and is widely used in fields such as clinical testing, pharmacy, and biochemistry.

(b) Conventional technology The antigen-antibody precipitate generated by the antigen-antibody reaction in the solution is extremely fine, 0.1 to 1.0 microns, so the amount of the antigen-antibody precipitate is smaller than that of the scattered light or transmitted light. Measured by

Therefore, there is a method (end point method) of measuring the intensity of scattered light or transmitted light after reaction for a certain period of time, but this method has the drawbacks that the reaction time is long and the upper limit of measurable concentration is low. In addition, there is a method Pt1 (rate method) that measures changes in the intensity of scattered light or transmitted light as the immune reaction progresses, but although the measurable upper limit concentration value is higher, the reaction time is shorter and depending on the analysis item. However, since minute changes must be measured, the measurement accuracy deteriorates.

Moreover, in general, measurement using transmitted light has the disadvantage that sensitivity cannot be obtained when the sediment concentration is small, and measurement using scattered light has the disadvantage that when the sediment concentration is large,
The drawback was that it caused a plateauing phenomenon.

(c) Purpose The present invention overcomes the drawbacks of these conventional immune reaction measurement devices, and uses transmitted light to select the optimal measurement method depending on the measurement item (type of antigen or antibody). In addition to providing a detector and a scattered light detector, the ratio of the intensity of scattered light to the intensity of transmitted light (hereinafter referred to as the (scattered light/transmitted light) ratio) can be determined, making immune reaction measurement more sensitive. .

Moreover, it is an object of the present invention to provide an immune reaction measuring device that can perform measurements up to a high concentration range with high accuracy.

(D) Configuration The present invention processes detection signals from a scattered light measuring device/fluorescence measuring device, a transmitted light measuring device, a scattered light detector/fluorescence detector, and a transmitted light detector (scattered light/transmitted light detector). 1. An immunoreaction measuring device characterized by having a data processing device for calculating a light) ratio.

In the present invention, a measuring device refers to a device that represents a physical quantity numerically, and a detector refers to a device that converts a physical quantity into a specific signal.

Therefore, the scattered light measuring instrument/fluorescence measuring instrument of the present invention includes a scattered light detector/fluorescence detector, and converts a detection signal from the scattered light detector/fluorescence detector into a scattered light intensity value or a fluorescence intensity value. It is equipped with a comparison operation-conversion device for converting. Similarly, the transmitted light measuring device includes a transmitted light detector and is equipped with a comparison calculation/conversion device that converts a detection signal from the transmitted light detector into a transmitted light intensity value.

The scattered light measuring device/fluorescence measuring device and the transmitted light measuring device in the immune reaction measuring device of the present invention may be configured in the same manner as the conventionally used scattered light measuring device and transmitted light measuring device. The same applies to the scattered light detector/fluorescence detector and the transmitted light detector, and the configurations of the conventionally used scattered light detector/fluorescence detector and transmitted light detector are sufficient.

In the immune reaction measuring device of the present invention, not only the scattered light measuring device/fluorescence measuring device and the transmitted light measuring device are provided in parallel, but also the detection signals from both detectors are used to determine the difference between the scattered light and the transmitted light. The key point is that a new data processing device was installed that can measure the intensity ratio, that is, the (scattered light/transmitted light) ratio.

The reason why the (scattered light/transmitted light) ratio is particularly important is that it enables highly sensitive measurement, regardless of the concentration of antigen-antibody precipitate in the solution. It is in. Therefore, it is sufficient to have a scattered light detector and a transmitted light detector without providing a scattered light measuring device and a transmitted light measuring device. It is convenient because only one device is needed when measuring scattered light, and the immune reaction time and measurement can be adjusted depending on the item to be measured, such as the type of antigen and antibody. The point is that the most suitable measurement method or calculation method can be selected in consideration of the relationship with sensitivity, the linearity of the measured value with respect to 18 degrees, the measurement range, etc. Furthermore, if monochromatic light is used as the irradiation light and a metal interference filter is installed, the fluorescence intensity can also be measured, and it can also be used for immunoassay measurements that use fluorescent substances as labeling substances. Become.

The data processing device that processes the detection signals from the scattered light detector and the transmitted light detector in the immune reaction measurement device of the present invention 1) processes the detection signals from the scattered light detector and the transmitted light detector. For transmitted light, scattered light, and the (scattered light/transmitted light) ratio, not only the end point method and the rate method are calculated, but also the immune This is a data processing device that can select which calculation method to use in consideration of the relationship between reaction time and measured reaction, the linearity of measured values with respect to concentration, etc., and any conventionally used data processing device can be used.

The immunoreaction measuring device of the present invention not only reduces the amount of particles of antigen-antibody complexes produced by antigen-antibody reactions, but also increases the amount of antigen (or
It is also possible to optically detect the amount of particles produced by agglutination using (or antibody) as a bridge, and thereby measure the amount of antigen.

(e) Actual example The diagram shows an example of the immune reaction measuring device of the present invention.

A light source 1, a spectroscope 2, a chopper 3, and a half-mirror beam splitter 4 are provided in the same manner as in a conventional spectrometer. An electron multiplier tube 5 for receiving irradiated light is provided on the reflected optical path of the beam splitter, and a measurement cell 6 is provided on the transmitted optical path. An annular slit 7 is provided on the front surface of the first lens 8 behind the measurement cell to distinguish the scattered light component from the transmitted light component. Therefore, only the transmitted light component can be collected through the central hole 10 of the annular slit 7. A reflector 1119 is provided behind the first lens 8, and a photomultiplier tube 15 for receiving transmitted light is provided on the reflected optical path.

A second lens 11 for convergence is provided coaxially with the first lens 8 behind the reflecting mirror. If it is to be used also for fluorescence measurement, a lens 12 and a metal filter 16 may be added.
, a condensing lens 13 is provided, and a photomultiplier tube 14 for receiving scattered light and fluorescent light is provided.

Each photomultiplier tube is connected to a synchronous rectification amplification section 17 so that a photodetection signal from each photomultiplier tube is sent to the synchronous rectification amplification section 17. Furthermore, the synchronous rectification amplification section 1
A data processing section 18 for appropriately processing signals from 7 is connected to the synchronous rectification weir IJ section 17. In addition, 19 is a negative high voltage power supply for photomultiplier tubes.

During photometry, the light from the light [1 passes through the spectrometer 2, and light with a designated single wavelength is extracted.

This light is repeatedly illuminated and blocked by a chopper 3 in order to compensate for the dark current of the photomultiplier tube, and the illuminated light is divided into two by a beam splitter 4, one of which receives the irradiated light. The light is received by the electron multiplier tube 5, and the other light is irradiated onto the measurement cell 6.

The alternating current photocurrent ■0 of the photomultiplier tube 5 is converted into a direct current voltage EO by the synchronous rectifier amplifier 17, and fed back to the negative high voltage power supply 19, so that a negative high voltage is generated so that EO always maintains a constant value. This results in a so-called tuple beam photometry method.

On the other hand, the light passing through the measurement cell is divided by the annular slit 7 into a scattered light component (or fluorescent component) passing outside the annular slit 7 and a transmitted light component passing through the hole 10 of the annular slit. The transmitted light component that has passed through the hole 10 of the annular slit 7 passes through the lens 8, is reflected by the reflection 19, and is received by the transmitted light receiving photomultiplier tube 15, and this AC photocurrent It is synchronously rectified. DC voltage Et at the widening part 17
is converted to

On the other hand, the scattered light component (or fluorescence component)
The alternating current photocurrent 1s is received by the synchronous rectifying unit 11 and converted into a direct current voltage ES by the synchronous rectifying unit 17. In the data processing unit 18, in the case of the end point method, Et,
Select either Es 1Es/Et, and in the case of the rate method, change amounts ΔEt, ΔEs, Δ(
Es/Et) and calculate the concentration value from the pre-stored calibration curve.

Of course, the present invention is not limited to this example in any way. For example, a laser beam can be used as the light source, and in this case, there is no need to provide a spectrometer because the light is monochromatic.

Further, for the chopper, DC amplification can be performed, and for the detector, a photoelectric element other than a photomultiplier tube can be used. Furthermore, appropriate changes can be made to the H arrangement of the annular ring, lens, reflecting mirror, etc. in terms of design.

(f) Effects The immunoassay device of the present invention can perform highly accurate measurements by selecting the optimal method depending on the item to be measured. Moreover, by measuring the (scattered light/transmitted light) ratio, it has become possible to perform measurements with higher sensitivity than when measuring scattered light or transmitted light alone.

By providing a metal interference filter, fluorescence measurements can be easily performed and can also be used for immunoassay measurements using fluorescent substances as labeling substances.

The immunoassay device of the present invention can perform such multipurpose analysis.
This was made possible for the first time by combining an optical system with an annular slit and a data processing device, and by adopting a new measurement method of measuring the (scattered light/transmitted light) ratio, it was possible to improve accuracy. Furthermore, fluorescence analysis can be performed simply by inserting a metal interference filter.

The present invention is an excellent immune reaction measuring device that is extremely compact, easy to operate, highly accurate, and highly sensitive compared to conventional immune reaction measuring devices, and is provided thereby. The impact is significant.

[Brief explanation of the drawing]

The figure is a diagram showing a schematic optical arrangement and a 1M yarn system in an embodiment of the immune reaction measuring device of the present invention. 1 is a light source, 2 is a spectrometer, 3 is a chopper, 4 is a beam splitter, 15, 14, 15 are photomultiplier tubes, 6 is a measurement cell, 7 is an annular slit, 8.11.12.13 is a lens, 9 1 is a reflecting mirror, 17+ is a synchronous rectification amplifier, and 18 is a data processing section. Procedural amendment prisoner (released) Commissioner of the Patent Office Kazuo Wakasugi 1, Indication of case Patent application No. 149489 of 1982 @
2. Title of the invention: Immune reaction measuring device 3. Relationship with the amended person's case: Name of patent applicant (199) Shimadzu Corporation 4.
Agent name (750B) Patent attorney Takeshi 1) Masahiko 5, Date of amendment order November 29, 1981 6, Application subject to amendment and Mei 11 I!

Claims (1)

[Claims] Processes the detection signals from the scattered light measurement device, fluorescence measurement device, transmitted light measurement device, scattered light detector/fluorescence detector, and transmitted light detector to calculate the ratio of the intensity of scattered light to the intensity of transmitted light. 1. An immune reaction measuring device comprising: a data processing device for measuring