JPH06201586A - Photometric apparatus of generated light of component of living body - Google Patents

Abstract

PURPOSE:To provide a photometric apparatus equipped with a light shutting structure which can shut an outdoor light effectively in the simple constitution when the light is measured. CONSTITUTION:The apparatus is constituted of an emission detecting means S with a photodetecting part 8 at an end thereof, a hollow shutting member 14 which is movable along the outer peripheral surface of the photodetecting part 8, and a container 2 containing a measuring sample 6. An opening part 4 of the container 2 is set confronting to an end face 8a of the photodetecting part. When the light is to be measured, a gap 12 defined by the end face 8a and an upper end part 2a of the container can be freely shut by the movement of the hollow shutting member 14. Accordingly, the light is shut with high efficiency in the simple shutting structure, and the equipment cost related to the shutting of light is reduced. Moreover, the container can be easily and correctly set at the relative position of the photodetecting part, whereby a plurality of samples can be measured continuously and the working efficiency is improved.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a luminescence metering device for a biological component capable of highly accurately measuring weak luminescence emitted by a biological component in a sample due to bioluminescence, chemiluminescence or the like.

[0002]

2. Description of the Related Art In the fields of medicine, pharmacy, biochemistry and the like, the luminescence method is widely used as a method for optically measuring biological components and detecting specific substances. For example, in the analysis of biological components by the luminescence method, very weak luminescence emitted from a minute amount of substances contained in blood, urine, sputum, stool, etc. is measured, and therefore, when external light enters the device in photometry. Since the photometric accuracy decreases, it is necessary to suppress the intrusion of external light as much as possible.

In conventional photometry, for example, as shown in the schematic sectional view of FIG. 5, the light detecting means S and the sample container 2 are covered with a light-shielding case B, that is, the entire photometric device is covered with a light-shielding body to block outside light. It is known to measure light.

On the other hand, it is also known to locally shield the photometric device. For example, as shown in the schematic sectional view of FIG.
It is known that a light shielding case B for accommodating the container 2 is integrally formed in the light receiving part 8 of the light detecting means S, and the light receiving part 8 of the light detecting means S and the container 2 are covered with a light shield to perform photometry. .

[0005]

However, when photometry is performed by covering the entire photometric device with a light shield, it is relatively difficult to shield external light because of the large light-shielding area, and the structure of the light shield becomes large. Therefore, there was a problem that the facility cost required for shading was high. Further, when the light receiving part of the light detecting means and the container are locally covered with a light shield to perform photometry, the container must be put in and taken out of the light shielding case for each measurement, and the opening part of the container is provided with the light receiving part. It was difficult to set accurately in the relative position of, and the setting was very troublesome and inefficient. Further, when a container for storing a plurality of measurement samples, such as a microplate, is used, stray light from the sample other than the measurement target enters the light receiving section and becomes measurement noise, which causes a problem that the measurement accuracy is reduced. Further, there is a problem in that the apparatus becomes complicated and the cost increases when trying to automate the setting of the container.

SUMMARY OF THE INVENTION An object of the present invention is to solve the problems of the conventional photometric device and to provide a photometric device having a light-shielding structure which can effectively shield external light during photometry with a simple structure.

[0007]

Means for Solving the Problems As a result of a study by the inventors of the present invention, a gap is formed between the light receiving portion of the light detecting means and the container for storing the sample in the conventional photometric device. The present invention has been completed by finding that the above-mentioned object can be achieved by making this gap light-shielding.

[0008] That is, the biological component luminescence photometric device of the present invention contains a luminescence detecting means having a light receiving portion at the tip, a hollow light shielding member movable along the outer peripheral surface of the light receiving portion, and a measurement sample. The container is placed on the front end surface of the light receiving portion with the opening facing the front surface, and covers the gap formed between the front end surface of the light receiving portion and the upper end portion of the container by the movement of the hollow light shielding member during photometry. It is characterized in that it is configured to be free.

The present invention will be described in more detail below. The photometric device of the present invention uses, as a labeling substance, a substance that emits light by itself, a substance that emits light by the action of another substance, or a substance that acts on the reaction of converting a luminescent substrate into a luminescent substance. A specific binding substance for the biological component to which the labeling substance is bound is bound to a measurement sample by a specific binding reaction, and the luminescence amount of the labeling substance of the specific binding substance or a luminescent substance resulting from the action of the labeling substance is measured. It is a device for doing.

The relationship between the measurement sample such as the above-mentioned biological components and the specific binding substance thereto is generally called a ligand-receptor system, which includes an antigen and an antibody, various biological substances and their corresponding receptors, biotin-avidin. Systems based on specific binding proteins such as systems, DNA or RNA and complementary DNA
Alternatively, a nucleic acid-nucleic acid system such as complementary RNA is included.

Examples of the luminescence method include methods utilizing bioluminescence and chemiluminescence. The above-mentioned bioluminescence means the emission of visible light by an organism, and examples of the organism that emits light include luminescent bacteria, fungi such as oyster mushrooms, protozoa such as pearl oysters, and insects such as fireflies. Chemiluminescence refers to a phenomenon in which an atom or molecule is excited by energy generated by a chemical reaction to emit light, and chemiluminescent substances (substrates) include luminol, acridinium ester, stabilized dioxetane, and peroxalic acid. Esters and the like can be mentioned.

As shown in the schematic partial sectional view of FIG. 1, the photometric device of the present invention is equipped with a light emission detecting means S having a light receiving portion 8 at its tip, and is movably attached along the outer peripheral surface of the light receiving portion 8. The hollow light-shielding member 14 and the container 2 that accommodates the measurement sample 6 and is placed with the opening 4 facing the front surface 8a of the light receiving portion. In this photometric device, a gap 1 is provided between the light receiving portion front end surface 8a and the container upper end portion 2a.
2 is formed.

According to the photometric device having the above structure, the hollow light shielding member 14 can be moved in the arrow direction (downward) at the time of photometry to cover at least the gap 12, so that external light can enter the gap 12. Can be prevented, and the luminescence L from the biological component in the measurement sample 6 can be measured with high accuracy.

Light emission detecting means S used in the photometric device of the present invention.
Any material can be used as long as it can detect weak light with high sensitivity, and for example, a luminescence detection device such as a photomultiplier tube is preferably used. In this light emission detecting means S, it is desirable that the light receiving portion 8 at the tip is projected in a cylindrical shape.

The light-shielding member 14 is formed of a light-shielding material in a hollow shape so as to be movable along the cylindrical light-receiving portion 8, and is movable on the outer peripheral surface of the light-receiving portion 8 in the longitudinal direction, preferably. It is slidably fitted. The inner surface of the light shielding member 14 is preferably black, which can absorb light.

As a means for moving the light shielding member 14, a method by a worker's hand or a mechanical method using an external drive source can be used. In addition, in this movement,
It is desirable not to give vibration to the light emission detecting means S.

As the container 2, any container having a hole-shaped opening 4 capable of accommodating a measurement sample may be used, and a container such as a microplate having the above-mentioned opening 4 at least at one place can be preferably used. At least one container 2 may be held in a measurement rack for use. The container 2 and the measurement rack may be transparent or colored, but if they have a plurality of openings in close proximity, it is preferably colored, especially black, in order to suppress stray light from other samples.

The hole-shaped opening 4 of the container 2 accommodates a sample to be measured, a reagent for measuring biological components, and the like. The biological component in the sample reacts with a reagent or the like to emit light, and is emitted to the outside of the container 2 through the opening 4. In the present invention, the light emission is detected and measured by the light emission detection means S via the light receiving unit 8. At this time, the light receiving portion 8 and the container upper end portion 2
Since the gap 12 with respect to a is covered with the hollow light shielding member 14, it is possible to block external light and stray light and measure light emission with high accuracy.

In order to highly shield the outside light,
The photometric device of the present invention preferably has the following configuration. That is, the light shielding member is slidably fitted to the outside of the cylindrical light receiving portion. However, even in this slidable fitting, since a minute gap still exists, it is necessary to block the light entering through this gap when more accurate photometry is required. FIG. 2 is a schematic cross-sectional view showing an example of such a light shielding structure, and FIG.
Is an O-ring 3 made of a light-shielding material on the side surface of the light receiving portion 8.
2 shows a light-shielding structure in which 2 is mounted, and FIG.
Shows a light-shielding structure in which a flexible light-shielding annular synthetic resin member 34 is attached to the outer periphery of the lower end of the light-receiving unit 8.

Further, it is necessary to consider light shielding for a minute gap existing between the container and the light shielding member. Figure 3
FIG. 3A is a schematic cross-sectional view showing an example of such a light-shielding structure, and FIG. 3A shows a light-shielding structure in which a flexible light-shielding annular synthetic resin member 36 is attached to a lower end portion of the light-shielding member 14, FIG. 3B shows a light-shielding structure in which an annular groove 38 is formed on the outer periphery of the opening 4 of the sample container 2 and the light-shielding member 14 is loosely fitted in the annular groove 38. In particular, the latter light-shielding structure is preferable because the light detecting means and the sample container can be more reliably positioned and the measurement accuracy is further improved. The annular groove may be formed on the outer circumference of the sample container on the upper surface of the measurement rack holding the sample container.

Further, in the apparatus of the present invention, when the container or the measurement rack holding the container is mounted on a moving table such as a conveyor, the photometry of the sample can be continuously performed, It is preferable because the work efficiency can be improved.

[0022]

According to the structure of the photometric device of the present invention, a high degree of light shielding can be achieved with a simple light shielding structure, and the cost related to light shielding can be reduced. Further, since the light shielding member can be moved, the container can be easily replaced except when the light is measured, and the mounting position of the container is determined, so that the container can be easily and accurately set at the relative position of the light receiving unit. Like Therefore, when the containers are automatically moved to predetermined positions in sequence, it is possible to continuously and accurately measure a plurality of samples.

[0023]

EXAMPLES Examples of the present invention will be shown below for more specific description. Needless to say, the present invention is not limited to this. Example 1 Stabilized dioxetane catalyzed by alkaline phosphatase was used as a measurement sample, and its chemiluminescence was measured by a photon counting method using a photomultiplier as a luminescence detection means. In this example, the photometric device having the configuration shown in FIG. 4 was used. In the figure, A is a photometric device, a photomultiplier tube S having a cylindrical light receiving portion 8 having an outer diameter of φ14 mm at its tip, and a photomultiplier tube S movably fitted to the outer peripheral surface of the light receiving portion 8 for external driving. The tubular light-shielding member 14 that slides in the axial direction by the device D, the microplate 2 that accommodates the measurement sample 6 and has the hole-shaped opening 4 having a diameter of 7 mm facing the tip surface 8a of the light-receiving section 8; The measurement rack 16 for holding the microplate 2 of FIG. An annular groove 38 concentric with the opening 4 of the microplate 2 to be held is formed on the upper surface of the measurement rack 16. The measurement rack 16 is placed on the belt conveyor B that moves in the longitudinal direction.

In the measurement of the chemiluminescence, the hole-shaped opening 4 of the microplate 2 is set so as to be coaxially aligned with the light receiving portion 8 of the photodetection multiplier S, and then the light shielding member 14 is set by the external driving device D. Move it in the direction of the arrow, and
The gap 12 formed between the tip surface 8a and the microplate upper end portion 2a was covered, and the lower end portion of the light shielding member 14 was loosely fitted so as to be in close contact with the bottom surface of the annular groove 38. Then, in this state, the photomultiplier tube S is operated to introduce the chemiluminescence L of the stabilized dioxetane in the measurement sample 6 housed in the hole-shaped opening 4 of the microplate 2 into the photodetector 8 and It was measured by the counting method. After the measurement is completed, the light shielding member 14 is attached to the external drive device D.
After moving to the original position of the light receiving unit 8 by the, the belt conveyor B on which the measurement rack 16 is placed is moved in the direction of the arrow so that the hole-shaped opening 4a of the microplate accommodating the next measurement sample receives light. It was set so as to be coaxial with the portion 8. Then, the above operation was repeated to continuously measure the chemiluminescence of the measurement sample in the plurality of microplates held in the measurement rack 16. In addition, as comparative examples, one having a structure in which the gap is not shielded (Comparative Example 1), one having a structure for locally shielding light shown in FIG. 6 (Comparative Example 2) and the entire photometric device shown in FIG. The same measurement sample as the above was photometrically measured with the one having a structure covered with (Comparative Example 3). The results are shown in Table 1.

[0025]

[Table 1]

As is apparent from Table 1 above, according to the method of the embodiment, it is possible to block external light from entering the gap and stray light from the sample that is not the object of measurement, which is equivalent to that of the photometric device of Comparative Example 2, or It showed extremely high measurement accuracy. Further, the workability was higher than that of the photometric device of Comparative Example 3.

[0027]

Since the present invention has the configuration described above,
The following effects are achieved. According to the configuration of the photometric device of the present invention, since a high degree of light shielding is performed with a simple light shielding structure, it is possible to reduce the facility cost related to light shielding. In addition, since the light shielding member can be moved, the container can be easily moved, and the mounting position of the container can be determined.
The container can be easily and accurately set at the relative position of the light receiving section. Therefore, when the container is automatically moved to a predetermined position, it is possible to continuously measure a plurality of samples with high accuracy, and it is possible to improve work efficiency and reduce work cost related to measurement.

[Brief description of drawings]

FIG. 1 is a schematic partial cross-sectional view showing the basic configuration of a photometric device of the present invention.

FIG. 2 is a schematic cross-sectional view showing a structure that shields light from a gap between a light receiving portion and a light shielding member.

FIG. 3 is a schematic cross-sectional view showing a structure that shields a gap between a container and a light blocking member.

FIG. 4 is a schematic partial cross-sectional view showing the configuration of a photometric device according to an embodiment of the present invention.

FIG. 5 is a schematic cross-sectional view showing a light shielding structure of a conventional photometric device.

FIG. 6 is a schematic cross-sectional view showing a light shielding structure of another conventional photometric device.

[Explanation of symbols]

 A Photometer S Luminescence detection means 2 Container 2a Upper end 4 Opening 6 Measurement sample 8 Light receiving part 8a Tip surface 12 Gap 14 Light-shielding member

Claims (1)

[Claims] 1. A light emission detecting means having a light receiving portion at its tip,
It consists of a hollow light-shielding member that is movable along the outer peripheral surface of the light-receiving section, and a container that holds the measurement sample and is placed on the tip of the light-receiving section with its opening facing the light-shielding section. 1. A luminescence photometric device for a biological component, characterized in that it is configured such that a gap formed between the light receiving portion front end surface and the container upper end portion can be freely covered by movement of a member.