JP2936973B2 - Luminescence measuring device - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an optimal luminescence measuring apparatus in a method for measuring a biological component in a sample using bioluminescence, chemiluminescence, etc., and is used in the fields of medicine, pharmacy, biochemistry and the like. Is done.

[0002]

2. Description of the Related Art In recent years, a luminescence method has been widely used as a method for detecting a biological component or detecting a specific substance with high sensitivity. For example, when analyzing a biological component by a luminescence method,
It measures the weak luminescence emitted by trace substances contained in blood, urine, sputum and the like. At this time, in the photometry, if external light enters the device, the photometric accuracy is reduced. Therefore, it is necessary to suppress the entry of external light as much as possible.

[0003] As a device capable of preventing the penetration of external light,
For example, as shown in the schematic cross-sectional view of FIG. 5, there is known a device in which the light detecting means S and the sample container 6 are covered with a light-shielding case B, that is, the entire light emission measuring device is completely covered with a light-shielding body and external light is shielded. . However, when measuring luminescence with the device having the above configuration,
It is relatively difficult to block external light due to the large light-blocking area,
In addition, there is a problem that the light-shielding structure becomes large and the equipment cost is high.

[0004] It is also known to locally shield the luminescence measuring device from light. For example, as shown in the schematic cross-sectional view of FIG. 6, there is known a luminescence measuring device in which a light shielding case B is integrally formed with a light detecting means S and the light receiving section 1 and the container 6 are shielded by a light shielding body. However, in any of the luminescence measuring devices that require the light-shielding case, when the container 6 is put in and out of the light-shielding case, external light enters the light-shielding case through the opening, and the light-receiving element of the light detection means S is directly As a result, the light receiving element may be deteriorated and its function may be lost. In addition, in the case of the above-described device in which light is locally covered with a light-shielding body, the container must be taken in and out of the light-shielding case each time measurement is performed, and the opening of the container is positioned at a relative position of the light-receiving portion. It was difficult to set accurately, and setting was very troublesome and inefficient.

[0005]

In order to prevent the above-mentioned inconvenience due to external light, an apparatus having a double structure of a light shielding case can effectively shield external light. When performing a plurality of measurement samples by using, for example, light emission (stray light) generated from a sample outside the measurement target
However, there is a problem in that the measurement noise enters the light receiving portion of the light emission detecting means and becomes measurement noise, thereby lowering the measurement accuracy. An object of the present invention is to solve the above-mentioned conventional luminescence measuring device at a glance, and to prevent external light and stray light from entering the light receiving portion of the luminescence detecting means regardless of luminescence measurement or non-luminescence measurement. To provide a luminescence measuring device having high measurement accuracy.

[0006]

In order to achieve the above object, the present invention provides a luminescence measuring device comprising: a luminescence detecting means; a cylindrical light receiving portion for introducing luminescence of a sample into the luminescence detecting means; A hollow light-blocking member movable along the outer peripheral surface of the light-receiving section, an openable and closable shutter that normally closes an optical path of the cylindrical light-receiving section and opens at the time of emission measurement, and a cylindrical light-receiving section that accommodates a sample to be measured. And a container arranged with the opening facing the portion, and desirably, the light emission detecting means comprises means for photoelectrically converting light emission, and means for measuring the amount of light emission from the photoelectric conversion value. Things.

[0007]

Since the light emission measuring device of the present invention is provided with an openable and closable shutter which always closes the light path of the light receiving portion and opens at the time of light emission measurement, when no light emission measurement is performed, external light is detected. Intrusion into the means is prevented. Therefore, the light receiving element does not encounter the external light, and the deterioration can be prevented. In addition, a light-shielding case that shields the entire device from light or that is formed integrally with the device and that locally shields light is not required, thereby facilitating replacement of the container containing the measurement sample. Further, since the light shielding case is not required, the mounting position of the container can be easily determined, and the container can be easily and accurately set at the relative position of the light receiving section.

Further, a hollow light-shielding member movable along the outer peripheral surface of the light-receiving portion is provided as light-shielding means, and the hollow light-shielding member is moved during light emission measurement to cover a gap between the light-receiving portion and the container. With this configuration, it is possible to effectively prevent external light from invading the optical path at the time of measuring light emission. Further, even when a plurality of measurement samples are performed using a microplate-shaped container or the like, stray light is prevented from entering the light receiving portion.

Furthermore, the light emission measuring device of the present invention forms a light shielding structure having a simple structure comprising a shutter for blocking an optical path of a light receiving part and a hollow light shielding member for shielding a gap between the light receiving part and the container. As a result, a light-shielding case can be eliminated,
The device itself can be miniaturized.

[0010]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail with reference to the drawings showing embodiments. Needless to say, the present invention is not limited to these examples. FIG. 1 is a schematic cross-sectional view showing one embodiment of a light-emitting photometer according to the present invention. In FIG. 1, reference symbol A denotes a light emission measuring device, which is a light emission detecting means S, a light receiving section 1 for introducing light emission of a sample into the light emission detecting means, and a hollow light shielding member movable along the outer peripheral surface of the light receiving section 1. 3, an openable and closable shutter 2 that always closes the optical path 10 of the light receiving unit and opens at the time of emission measurement, and a container 6 that houses the measurement sample 4 and is arranged with the opening 5 opposed to the optical conduction path. It has. Shutter 2 above
Are connected to the shutter driving mechanism 8 and the hollow light shielding member 3 is connected to the light shielding member driving mechanism 9, respectively.

The luminescence detecting means S may be any one which can detect the luminescence of the sample and measure its amount, and a conventionally known device can be used. In the present invention, as the light receiving element 11 that can detect weak light with high sensitivity, for example, a device that combines a photoelectric conversion device using a photomultiplier tube or the like and a device that measures the amount of light from this conversion value, for example, a photon counter is used. It can be suitably used.

The light receiving section 1 is formed of a hollow member such as a cylindrical body, and is integrally formed with one end face of the light emission detecting means S or joined to another member to project outward. The hollow portion is used as an optical path 10 to guide light emission of the sample to the light receiving element 11. The cross-sectional shape of the light receiving portion is not particularly limited, and a circular shape, a triangular shape, a polygonal shape such as a quadrangle, or the like can be used. In the present invention, it is preferable that the light receiving portion has a shape similar to the shape of the opening of the container, and a cylindrical member is usually used. The material of the hollow member is not particularly limited as long as it does not absorb the luminescence of the sample. For example, aluminum, stainless steel, or the like is preferably used.

The shutter 2 is, in the above embodiment,
The optical path 10 of the light receiving unit 1 can be opened and closed by being inserted into a gap formed at the junction between the light emission detecting unit S and the light receiving unit 1. In the present invention, the optical path 10 can be opened and closed. Any configuration may be used. For example, a structure such as a shutter or an aperture mechanism of a camera may be used. Further, the gap for inserting the shutter may be formed in the light receiving section. As such a shutter 2, for example, a plate-shaped shutter having substantially the same size as the gap and having a width and a thickness that can be inserted into the gap is exemplified. In this shutter, a hole having a shape similar to the cross-sectional shape of the light receiving portion and having substantially the same size is formed. This shutter is connected to the light path 1
0 in the direction perpendicular to the light path 10
When positioned above, the light path can be opened. The material of the shutter 2 is not particularly limited as long as it can shield light. For example, stainless steel, aluminum,
A metal such as an iron plate is preferably used. Note that the surface of the shutter 2 is preferably black so as to absorb light.

The shutter 2 has a shutter driving mechanism 8
Are connected. By the operation of the shutter driving mechanism 8, the shutter 2 is moved in the axial direction (the direction of the arrow) with respect to the light path 10. The shutter drive mechanism 8 does not apply vibration to the light emission detection means S.
For example, it is desirable to provide a mechanism for controlling the number of steps by a stepping motor.

In the present invention, in order to prevent external light from entering the light receiving element 11 through a gap provided for inserting the shutter 2, a seal by an O-ring 12 is provided, or the shutter and the entire driving mechanism are shielded from light. preferable. With this configuration, the shielding of external light is more effectively performed.

The light shielding member 3 is made of a light shielding hollow member,
The outer peripheral surface of the light receiving unit 1 is fitted so as to be movable in the longitudinal direction, preferably slidably. The material of this light shielding member is
There is no particular limitation as long as it can shield light, and stainless steel, aluminum and the like are preferably used. It is preferable that the inner surface of the light-shielding member 3 be black, which can absorb light. The hollow light shielding member 3 is connected to a light shielding member driving mechanism 9. By the operation of the light-shielding member driving mechanism 9, the light-shielding member 3 is moved in the axial direction (the direction of the arrow) along the outer peripheral surface of the light receiving section.

The container 6 has only to have a hole-shaped opening 5 for accommodating a measurement sample, and a container having the above-mentioned opening 5 at at least one place can be used. For example, if the container is a microplate having a plurality of hole-shaped openings, it can be suitably used because the trouble of replacing the container can be omitted. The container may be used by holding at least one container in a measurement rack. The container and the measurement rack may be transparent or colored, but when a plurality of openings are provided close to each other, it is preferable that the container and the measurement rack be colored, particularly black in order to suppress stray light from other samples.

A sample to be measured, a reagent for measuring a biological component, and the like are accommodated in the hole-shaped opening 5 of the container. Biological components in the sample react with reagents and the like to emit light, and the emitted light is emitted to the outside of the container through the opening.

In the luminescence measuring apparatus of the present invention, when luminescence measurement is not performed, the shutter 2 closes the light path 10 of the light receiving section 1 of the light detecting means S as shown in FIG. Even when the secondary light shielding of the entire apparatus is insufficient, external light is prevented from entering the light receiving section 1 of the light detecting means S from the gap 7, and the light receiving element is prevented from being deteriorated or broken.

When the luminescence measurement is performed from the above state, the container 6 containing the measurement sample 4 is placed in the light receiving section 1 of the luminescence detection means S.
And the opening 5 of the container are placed at a predetermined position facing each other. At this time, a gap 7 is formed between the light receiving unit 1 and the container 6. When the light-shielding member driving mechanism 9 is operated at the time of light emission measurement, the hollow light-shielding member 3 held on the outer peripheral surface of the light receiving unit 1 slides axially downward (in the direction of the arrow), as shown in FIG. In addition, at least the gap 7 between the light receiving unit 1 and the container 6 is covered with the hollow light-blocking member 3 so that the gap 7 is shielded, and external light enters the light receiving unit 1 of the light detecting means S. Is prevented.

On the other hand, when the shutter driving mechanism 8 is operated, the shutter 2 is moved in the direction of the arrow, the light path 10 of the light receiving section 1 is opened, and the light emission L of the measurement sample 4 is reduced as shown in FIG. The light emission detecting means S is introduced.

Light emitted to the light emission detecting means S is:
A photomultiplier tube device (not shown) which is detected by a light receiving element,
The light quantity is measured by a photon counter device that measures the light quantity from this conversion value.

In the above luminescence measurement, even if a microplate having a plurality of hole-shaped openings is used as a container, the gap 7 is shielded from light, so that luminescence (stray light) from a sample other than the object to be measured is transmitted into the light receiving unit. The measurement noise is prevented.

As described above, it is possible to prevent external light from invading the light receiving portion of the light emission detecting means regardless of whether light emission is measured or light emission is not measured. Further, at the time of light emission measurement, stray light can be prevented from entering the light receiving portion of the light emission detecting means. Therefore, even if the secondary light shielding of the entire apparatus is insufficient, external light and stray light can be prevented from entering the light receiving section of the light emission detecting means, and deterioration of the light receiving element and generation of measurement noise are eliminated. Also, since no light shielding case is required,
Container can be set accurately. Therefore, the measurement accuracy of light emission can be greatly improved. Further, since a light-shielding structure having a simple structure is formed to eliminate the need for a light-shielding case, the size of the device can be reduced, and the manufacturing cost of the device can be reduced.

If the shutter drive mechanism 8 is operated first during the above-mentioned light emission measurement, the light detecting means S is operated in a state where the gap 7 between the light receiving section 1 and the container 6 is not shielded from light.
Since the light path of the light receiving section 1 is opened, external light or stray light may enter the light path, deteriorating the light receiving element or causing measurement noise, thereby lowering the measurement accuracy. Therefore, in the present invention, it is preferable that the shutter drive mechanism 8 be operated simultaneously with or after the operation of the light shielding member drive mechanism 9.

In the luminescence measuring device of the present invention,
In order to highly shield the external light, the following configuration is preferable. That is, the light shielding member is slidably fitted to the outside of the cylindrical light receiving portion. However,
Even in this slidable fitting, there is still a minute gap, so if more accurate photometry is required, it is necessary to block light entering through this gap.

FIG. 3 is a schematic sectional view showing an example of such a light shielding structure. FIG. 3A shows a structure in which a flexible light shielding annular synthetic resin member 14 is attached to the lower end of the light shielding member 3. FIG. 3B shows an opening 5 of the container 6.
A light-shielding structure is shown in which an annular groove 15 is formed on the outer periphery of the light-emitting device and the light-shielding member 3 is loosely fitted into the annular groove 15. In particular, the latter light-shielding structure is preferable because the positioning between the light detection means and the container is more reliable, and the measurement accuracy is further improved. The annular groove 15
May be formed on the outer periphery of the container on the upper surface of the measurement rack that holds the sample container.

Further, in the apparatus of the present invention, if the container or the measuring rack holding the container is configured to be mounted on a moving table such as a conveyor, the sample can be continuously measured. It is preferable because the working efficiency can be improved.

(Experimental Examples) Experimental examples of the present invention are shown below.
This will be described more specifically. Experimental Example 1 Using the luminescence measuring device having the configuration shown in FIG. 1, the degree of penetration of external light when luminescence was not measured was measured by the photon counting method. At this time, in the luminescence measuring device, the light path 10 was closed by the shutter 2 and the light blocking member 3 was positioned above the light receiving section. As external light, the light-shielding effect of the shutter was confirmed under various conditions when a 40 W indoor fluorescent lamp was lit at a distance of 4 m, when a 10 W bulb was lit at a distance of 2 m, and when both were lit at the same time. . The results were as shown in Table 1.

[0030]

[Table 1]

As is clear from Table 1, according to the apparatus of the embodiment, the external light can be completely blocked by the shutter, so that the secondary light shielding can be performed by keeping the shutter closed except for the emission measurement. There is no need to do it.

Experimental Example 2 Stabilized dioxetane using alkaline phosphatase as a catalyst was used as a measurement sample, and the chemiluminescence was measured by a photon counting method using a photomultiplier tube as a light receiving element.

In this experimental example, the luminescence measuring device shown in FIG. 4 was used. Although the details of the shutter structure are not shown in FIG. 4, the same structure as the shutter structure shown in FIG. 1 is formed. The luminescence measuring device A1 is movably fitted on the outer peripheral surface of the light receiving unit S1 having a cylindrical light receiving unit 1 having an outer diameter of 14 mm and an inner diameter of 7 mm at the end, and is driven by a light shielding member driving mechanism 9. Axial direction (arrow direction)
A light-blocking cylindrical member 3, a 1-well microplate 6 for accommodating the measurement sample 4, and having a hole-shaped opening (diameter 7 mm) 5 opposed to the light-receiving section 1, and a measurement rack for holding the microplate 13.
Note that an annular groove 15 concentric with the opening 5 of the microplate 6 to be held is formed on the upper surface of the measurement rack 13. The measuring rack 13 is placed on a belt conveyor C that moves in the longitudinal direction.

The measurement of the chemiluminescence was carried out as follows. The hole-shaped opening 5 of the microplate 6 is set so as to be coaxially coincident with the light-receiving portion 1 of the light-emission detecting means S, and the light-shielding member driving mechanism 9 holds the light-shielding portion held over the outer periphery of the light-receiving portion 1. The member 3 is moved in the direction of the arrow, and the distal end 1a of the light receiving section 1 and the opening 5 of the microplate are moved.
And the lower end of the light shielding member 3 is loosely fitted so as to be in close contact with the bottom surface of the annular groove 15. Then, the shutter drive mechanism 8
After the shutter 2 is moved to the open state by
Activating the photomultiplier tube used in the light receiving element to cause the chemiluminescence L of stabilized dioxycetane in the measurement sample 4 contained in the microplate 6 to be introduced into the light receiving unit 1;
Measured by the photon counting method.

After the measurement, the shutter driving mechanism 8
The shutter 2 is moved to the original position by the
The light path is closed, and the light shielding member 3 is moved to the original position of the light receiving section 1 by the light shielding member driving mechanism 9. On the other hand, the belt conveyor C on which the measurement rack 13 is placed is moved in one direction (the direction of the arrow), and the container of the microplate containing the next measurement sample is opened in the same manner as described above. Were set so that they coincided with each other, and the chemiluminescence of the sample was measured sequentially.

As a comparative example, the light emitting device was constructed so as not to shield the gap (Comparative Example 1), and the structure having the structure shown in FIG. 6 for shielding light locally (Comparative Example 2). Was measured respectively. The results were as shown in Table 2.

[0037]

[Table 2]

[0038]

According to the apparatus of the present invention, a simple light shielding structure for blocking the light receiving portion of the light emission detecting means is formed by a shutter when light emission measurement is not performed, and by a shielding member when light emission measurement is performed. By doing so, it is possible to effectively prevent external light or stray light from a sample other than the measurement target from entering the light emission detecting means. Since the external light is prevented from entering the light emission detection means, the light emission detection element is not deteriorated by the external light that enters. In addition, at the time of light emission measurement, the gap between the light receiving portion of the light detection means and the container can be sufficiently shielded, so that measurement noise does not occur.

As described above, it is possible to prevent external light and stray light from entering the light path of the light receiving section during the light emission measurement or when the light emission measurement is not performed, and to eliminate the deterioration of the light receiving element of the light detecting means and the measurement noise. Since the container can be set accurately, the measurement accuracy of light emission can be greatly improved.

Further, since the light shielding case can be dispensed with, the size of the apparatus itself can be reduced, and the price can be reduced. Further, according to the apparatus of the present invention, a microplate having a plurality of hole-shaped openings can be used as a container, and further, the sample mounted in each container mounted on a moving means can be successively measured. The measurement efficiency is improved.

[Brief description of the drawings]

FIG. 1 is a schematic partial sectional view showing a luminescence measuring device according to one embodiment of the present invention.

FIG. 2 is a schematic partial cross-sectional view illustrating operations of a shutter and a light shielding member at the time of measuring light emission.

FIG. 3 is a schematic cross-sectional view showing a structure for shielding a gap between a container and a light shielding member. (A) shows an example in which a flexible light-shielding annular synthetic resin member is attached to the lower end of the light-shielding member, and (b) shows an example in which the light-shielding member is loosely fitted into an annular groove formed on the outer periphery of the container. .

FIG. 4 is a schematic partial sectional view showing a luminescence measuring device according to another embodiment of the present invention.

FIG. 5 is a schematic sectional view showing a light shielding structure of a conventional luminescence measuring device.

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

[Explanation of symbols]

 Reference Signs List A light emission measuring device S light emission detection means 1 light receiving unit 2 shutter 3 hollow light shielding member 4 measurement sample 5 opening 6 container 7 gap 8 shutter driving mechanism 9 light shielding member driving mechanism 10 light path

──────────────────────────────────────────────────続 き Continuation of the front page (56) References JP-A-61-116647 (JP, A) JP-A-5-157699 (JP, A) JP-A-5-209830 (JP, A) 506433 (JP, A) (58) Fields investigated (Int. Cl. ⁶ , DB name) G01N 21/75 G01N 21/64

Claims (2)

(57) [Claims] 1. A light emission detecting means, a cylindrical light receiving portion for introducing light emission of a sample into the light emission detecting means, a hollow light shielding member movable along an outer peripheral surface of the cylindrical light receiving portion, A luminescence measuring device comprising: a shutter which can be opened and closed, which always closes an optical path of a light receiving portion and is opened at the time of luminescence measurement, and a container which accommodates a measurement sample and which is arranged with the opening facing the cylindrical light receiving portion. . 2. The luminescence measuring device according to claim 1, wherein the luminescence detecting means comprises means for photoelectrically converting the luminescence, and means for measuring the amount of luminescence from the photoelectric conversion value.