JP2000241708A - Light-condensing device for emission analysis and emission analysis device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a light-condensing device which is same as or smaller than a conventional light-condensing device in size and which does not cause an increase in the production cost by forming such a structure that the light emitted from a sample housed in a housing portion enters a polyhedron and that part of the light entering the polyhedron reaches a reflection plane is then outgoes as condensed light from the light-emitting face. SOLUTION: This device is a polyhedron made of a material which is translucent for the light emitted from a specified material and composed of a light-reflecting face 2 and a translucent face. The light-reflecting face 2 has a curved form with the cross section being almost a part of an ellipse, and has a light-reflecting film 11 formed on the outer face. The light-outgoing face 3 is disposed facing the light- reflecting face 2 and the light reflected by the light-reflecting face 2 enters the face 3 at an angle not larger than the angle of total reflection and outgoes from the polyhedron. A housing part to house a sample or a container holding a sample is formed at one focal point near the light-reflecting face 2 of the two focal points of the ellipse. Part of the light emitted from the sample housed in the housing part and entering the polyhedron reaches the light-reflecting face 2 and then outgoes as condensed light from the light-outgoing face 3.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a light-collecting element for efficiently collecting light such as fluorescence, chemiluminescence, and bioluminescence from a specific substance contained in a sample. The present invention also relates to an emission spectrometer including the light-collecting element, which detects fluorescence, chemiluminescence, bioluminescence, and the like from a specific substance contained in a sample, and performs analysis such as quantification of the specific substance from the amount of luminescence. It is.

[0002]

2. Description of the Related Art Light emitted from a specific substance contained in a sample, such as fluorescence, chemiluminescence, or bioluminescence, is generally emitted radially. On the other hand, a photodetector provided in a general emission analyzer has only one light receiving surface oriented in one direction. For this reason, a light-collecting element that efficiently condenses the radially emitted light on the light-receiving surface of the light-detecting element is required. In particular, the smaller the amount of light emitted from a specific substance contained in the sample, the more the light-collecting element becomes necessary. Increases.

Conventionally used light-collecting elements include optical elements such as optical lenses and concave reflecting mirrors, and optical waveguides represented by optical fibers. For chemiluminescence or bioluminescence, the light-receiving surface of the photodetector may be brought close to the test sample without using a light-collecting element.

FIG. 13 shows a conventional fluorescence analyzer using an optical lens as a condensing element. In this apparatus, after the excitation light whose wavelength is selected by an optical filter (27) or the like is adjusted by a converging lens (26), it is irradiated on a test sample in a sample container, and fluorescence from a specific substance contained in the sample is emitted. , Condensing with an optical lens, and an optical filter (2
After performing the wavelength selection of the light according to 4), the light detection element (2)
Detect in 2). When analyzing chemiluminescence or bioluminescence, there is basically no excitation light source and no wavelength selection means on the excitation light side, and the light-emitting light condensing element has the same form. When an optical lens is used as a light-collecting element as in the conventional example of FIG. 13, the solid angle extending from one point of the sample to the effective diameter of the optical lens determines the light-collecting efficiency. FIG. 14 shows a conventional apparatus using a concave reflecting mirror as a light collecting element. Also in the conventional example of FIG. 14, the solid angle extending from one point of the sample to the effective surface of the concave reflecting mirror determines the light collection efficiency.

Among the conventional examples shown in FIG. 14, a large light-collecting efficiency is expected especially in a light-collecting element using a reflecting mirror having a cross section that is a part of an elliptical shape (hereinafter simply referred to as an elliptical concave reflecting mirror). it can. An ellipse has the geometric property that an angle consisting of a line segment connecting a point on the ellipse and two focal points is bisected by a normal passing through the point on the ellipse. This is because light radiated from one focal point (first focal point) can be collected at another focal point (second focal point).

FIG. 15 shows a conventional emission analyzer using an optical waveguide typified by an optical fiber as a condensing element. In this device, one of the end faces of the optical waveguide is disposed on a sample (a sample container holding the sample), and the other is disposed close to a photodetector (22), and light emitted from the sample is transmitted to the photodetector. When wavelength selection is performed, a wavelength selection element such as an optical filter is interposed between the optical waveguide and the photodetector or between the sample and the optical waveguide. This example is a device for analyzing chemiluminescence or bioluminescence, but in the case of fluorescence analysis, only an excitation light source and a means for selecting the excitation light are added,
The fluorescent light collecting element has a similar form. When the optical waveguide is used as a condensing element as in the conventional example of FIG.
The smaller of the solid angle extending from one point of the sample to the core end surface of the optical waveguide or the allowable light receiving angle calculated from the refractive indices of the core portion and the cladding portion of the optical waveguide, determines the light collection efficiency.

FIG. 16 shows a conventional apparatus for analyzing chemiluminescence or bioluminescence by disposing a light-receiving surface of a light-detecting element close to a sample without using a light-collecting element. When wavelength selection of light emission is performed, a wavelength selection element such as an optical filter is interposed between the sample and the light detection element. In the conventional example of FIG. 16, the solid angle extending from one point of the sample to the light receiving surface of the photodetector determines the light collection efficiency.

[0008]

SUMMARY OF THE INVENTION When detecting light emission such as fluorescence, chemiluminescence, bioluminescence, etc. from a specific substance contained in a sample by a detection element having a light receiving surface oriented in one direction, the light-collecting element described above is used. Has the following issues.

An optical lens (FIG. 13) and a concave reflecting mirror (FIG. 1)
The light collection efficiency when an optical element such as 4) is used is determined by a solid angle extending from one point of the sample to the effective diameter of the optical element.
When the optical waveguide is used (FIG. 15), the light-collecting efficiency is determined by the solid angle extending from one point of the sample to the core end face of the optical waveguide or the allowable light receiving angle calculated from the refractive indexes of the core and the cladding of the optical waveguide. Is determined by the smaller one. In the case where the light-receiving surface of the light-detecting element is disposed close to the test sample and chemiluminescence or bioluminescence is detected (FIG. 16) without using the light-collecting element, the light-collecting efficiency of the light-detecting element is measured from one point of the sample. It is determined by the solid angle formed on the light receiving surface. In either case, each light-collecting element exists only in a certain direction when viewed from the sample, and only light that happens to be emitted from the sample in that direction is collected, and light detection is performed. The light is guided to the light receiving surface of the element. Therefore, when applied to luminescence emitted radially, such as fluorescence from a specific substance contained in the sample, chemiluminescence, bioluminescence, etc., only a part of the light is collected,
No detection is possible and efficient luminescence analysis is not possible. In particular, when the amount of emitted light is very small, there is a problem that analysis cannot be performed. For example, in the immunoassay method, alkaline phosphatase (ALP) is frequently used as a label, but in recent years, the sensitivity has been increased to 1 × 10 ⁻²⁰ mol.
An emission spectrometer capable of realizing the following ALP detection is required. However, it is extremely difficult to detect 1 × 10 ⁻²⁰ mol of ALP with a conventional analyzer using a light-collecting element, and even if it can be detected, deterioration of accuracy is unavoidable at present.

In order to solve such a problem, a combination of a plurality of light-collecting elements may be used. For example, it is conceivable to use a combination of an optical lens and a concave reflecting mirror or a combination of an optical waveguide and a concave reflecting mirror. However, there are problems such as an increase in the size of the analyzer and an increase in the manufacturing cost of the analyzer. Newly occurs.

As described above, a light-collecting element using an elliptical concave reflecting mirror is ideal in terms of light-collecting efficiency, and thus has the possibility of realizing efficient light emission analysis. . However, in a light-collecting element using an elliptical concave reflecting mirror, light emitted radially at one focal point (first focal point) is also radially collected at another focal point (second focal point). Would. On the other hand, the photodetector usually has only one light-receiving surface, and can only detect light traveling toward the light-receiving surface. In fact, the efficient light-gathering power of an elliptical concave reflector is sufficient. There is a problem that it cannot be used. In addition, in order to use an elliptical concave reflecting mirror as a light-collecting element, it is necessary to three-dimensionally surround the sample with a reflecting mirror. Since the manufacturing cost of a three-dimensional reflecting mirror called a mirror is high, there has been a problem that the manufacturing cost of the analyzer increases as a result.

As described above, in order to realize a small-sized emission spectrometer capable of realizing high-sensitivity and high-accuracy luminescence analysis, a large amount of light that a specific substance contained in a sample radially diverges is used as a whole. As a light-collecting element to be a light beam going in one direction, equivalent to or smaller than the conventional light-collecting element,
Moreover, it is required to provide a device that does not cause an increase in manufacturing cost in order to perform high-sensitivity emission analysis using a photodetector having only one light receiving surface.

[0013]

SUMMARY OF THE INVENTION The present invention relates to a light-collecting element which converts a large amount of light radiated from a specific substance contained in a sample into a light beam as a whole in one direction. To provide a light emission analyzer that is equal to or smaller than that and does not cause an increase in manufacturing cost, and to provide a small-sized light emission analyzer capable of realizing high-sensitivity and high-accuracy light emission analysis using such a light-collecting element. It was done for the purpose.

That is, the invention of claim 1 of the present invention, which has been made to achieve the above object, is a light-collecting element for condensing light emitted from a specific substance contained in a sample. A polyhedron formed of at least a light-reflecting surface and a light-transmitting surface, which is formed of a material that transmits light.
(B) the light-reflecting surface has a curved surface shape whose cross section is substantially a part of an ellipse, and a light-reflecting film is formed outside; and (c) the light-emitting surface is arranged to face the light-reflecting surface. The light reflected by the light reflecting surface can enter at an angle not exceeding the total reflection angle and can be emitted outside the polyhedron, and (d) the focal point on the side closer to the light reflecting surface among the two focal points of the ellipse A storage section for storing the sample or a container holding the sample, and (e) collecting, among the light emitted from the sample stored in the storage section and incident on the polyhedron, reaching the reflection surface; A light-collecting element, wherein the light is emitted from the light-emitting surface in a lighted state.

According to a seventh aspect of the present invention, which is provided to achieve the above object, there is provided a light-collecting element according to the present invention having the above-mentioned features, and a light-emitting element for detecting light emitted from a light exit surface of the light-collecting element. An apparatus for detecting and analyzing luminescence from a specific substance contained in a sample, comprising: a detection element. Hereinafter, the present invention will be described in detail.

The light-collecting device according to the present invention can emit light from a specific substance contained in a sample contained in a polyhedral container or a specific substance contained in a sample contained in a container contained in a container (hereinafter simply referred to as “ Light emission). The housing section, for example, provided a hole in the polyhedron,
What is necessary is just to accommodate a sample and a container in this hole. In the light-collecting element of the present invention, light emission enters the inside of the polyhedron from the inner surface of the accommodation hole.

The polyhedron, which is the light-collecting element of the present invention, is not limited as long as it is made of a material transmissive to light emission and composed of two or more surfaces. Examples of the dihedron include a shape obtained by cutting an ellipsoidal sphere at a plane that intersects perpendicularly with the major axis, and a shape obtained by forming a spherical surface protruding toward the inside of the ellipsoidal sphere. As the trihedron, for example, a polyhedron obtained by cutting a pointed portion of a cone having a curved bottom whose cross section becomes a part of an ellipse can be exemplified. Other than these, the polyhedron may be a tetrahedron or more polyhedron as long as the above requirements are satisfied.

The polyhedron has at least a light reflecting surface for reflecting light emission, and a light emitting surface facing the light reflecting surface such that the light reflected by the light reflecting surface is incident at an angle not exceeding the total reflection angle. Having. The light reflecting surface has a curved surface shape such that an arbitrary cross-sectional shape is substantially a part of an ellipse, and a light reflecting film is formed outside in order to efficiently reflect light. On the other hand, the light exit surface is a surface on which light incident thereon can exit to the outside of the polyhedron.

When the polyhedron of the present invention is a dihedron, one surface is a light reflecting surface and the other surface is a light emitting surface.
In the case of a polyhedron having more than surfaces, each surface is configured so that one or more surfaces function as a waveguide for guiding light incident thereon to a light reflecting surface or a light emitting surface.

Since the light reflecting surface has a curved shape such that an arbitrary cross-sectional shape substantially becomes a part of an ellipse, the light reflected there is collected at two focal points of the ellipse. In the polyhedron of the present invention, the accommodation hole is provided at a focal point closer to the light reflecting surface among the two focal points. This allows
Light emitted from one of the two focal points and entering the inside of the polyhedron arrives at the light exit surface after passing through another focal point or on the way to another focal point. Also, it goes without saying that a part of the light emitted to the polyhedron directly reaches the light emitting surface. Then, of the light emitted to the polyhedron, the light emitted to a surface other than the light reflecting surface or the light emitting surface finally reaches the light emitting surface by the above-described waveguide function of these surfaces.

As described above, in the light-collecting device of the present invention, of the light emitted to the inside of the polyhedron, the light that reaches the light-emitting surface via reflection on the light-reflecting surface is condensed from the light-emitting surface to the polyhedron. It can be emitted to the outside.

The analyzer of the present invention is an apparatus having the above-described light-collecting element of the present invention and an excitation light source for fluorescently exciting a sample in a receiving hole provided in a polyhedron which is a light-collecting element. Such an apparatus also has a detection element for detecting light emission emitted from the light emission surface of the light collection element.

The detecting element is arranged on or away from the light emitting surface of the light-collecting element. Considering the maintenance of the detection element, etc., it is preferable to arrange them separately.However, if they are arranged separately, they should be arranged in parallel, so that a wavelength selecting means such as an optical filter is sandwiched between them. Becomes possible. In the case of disposing them separately, the light emitting surface of the polyhedron may be a curved surface that is convex toward the outside of the polyhedron, or the light receiving surface area of the detection element may be larger than the light emitting surface area. It is possible to improve the light collection efficiency of the light emission on the detection element.

Further, when the analyzer of the present invention is applied to emission analysis in which a sample such as a fluorescence analyzer needs to be irradiated with excitation light, the sample accommodated in the accommodation hole or the specimen accommodated in the accommodation hole is used. What is necessary is just to add an excitation light source for inducing fluorescence in the sample in the container thus set.

[0025]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the drawings and examples, but these show only one embodiment of the present invention and do not limit the present invention.

FIG. 1 shows an example of the light-collecting element of the present invention which is an eight-sided polyhedron. FIG. 2 is a top plan view (a) and a cross-sectional view of the light-collecting element shown in FIG. (B). In the figure, reference numeral 1 denotes an accommodation hole for accommodating a sample container for holding a sample;
Is a light reflecting surface whose arbitrary cross section (see FIG. 2) is a part of an ellipse, 3 is a light emitting surface facing the light reflecting surface, and 4 to 9 are light reflecting surfaces which reflect light incident inside the polyhedron. This is another surface having the function of a waveguide for guiding to the light emitting surface, and 11 is a light reflection film formed outside the light-collecting element.

The size and depth of the housing hole 1 can be appropriately determined in consideration of the amount of the sample to be provided to the light-collecting element, but the volume is made as small as possible so as not to cut off the optical path inside the polyhedron. Is preferred.

In this embodiment, in addition to the light reflecting surface 2, a light reflecting film is formed outside the surfaces 6 and 7 functioning as an optical waveguide. The light reflection film 11 can be formed outside all surfaces except the surface forming the accommodation hole 1 which is the light incident surface where light emission enters the inside of the polyhedron and the light exit surface where light emission exits outside the polyhedron. From the viewpoint of reducing the manufacturing cost, it is sufficient to form at least the light reflecting surface 2 outside. In the example of FIG. 1, the light reflecting films are formed on the surfaces 6 and 7 because, as shown in FIG. 1B, the light emission on the surfaces 6 and 7 is hardly guided to the light emitting surface 3 by total reflection. It is particularly preferable to form a light reflecting surface here. The reflecting surface can be formed by, for example, aluminum evaporation or silver plating on the outer surface, or by bonding an aluminum foil to the outer surface.

The light-collecting element of the present invention is formed of a material that is transparent to light emitted from a specific substance as a whole. As such a material, for example, among transparent materials such as transparent glass, transparent acrylic resin, and transparent polystyrene resin, light emission wavelength, light absorption characteristics of the material, and ease of processing for forming a polyhedron are considered. It is sufficient to select and use as appropriate. From the viewpoint of ease of processing and low cost, a transparent acrylic resin can be exemplified as a preferable material.

The light reflecting surface 2 has an arbitrary cross section (surface 8) as shown in FIG.
The cross section in a plane parallel to is 60 mm long axis and 39.2 m short axis
m is a curved surface that becomes a part of the ellipse, and its height is 8 mm
It is. As can be easily understood from FIG. 2, the cross section of the light reflecting surface 2 is an elliptical arc formed by each 20 degrees from the major axis of the ellipse, and has a shape symmetric to the major axis of the ellipse.
The light reflecting surface 2 has two focal points on the long axis at positions of 10 mm and 50 mm from the reflecting surface, respectively.

The focal point closer to the light reflecting surface 2 (hereinafter referred to as the first focal point) of the two focal points coincides with the center of the cylindrical housing hole 1 and is located inside the same. in this way,
It is preferable that the first focal point of the light reflecting surface be present in the sample liquid when the sample liquid is actually stored in the storage hole, more preferably in the storage hole. However, the first focal point may be located outside the accommodation hole (that is, inside the polyhedron) and very close to the accommodation hole. In the example shown in the figure, since the polyhedron is a flat plate, the first focal point of the light reflecting surface continuously exists on an 8 mm line corresponding to the thickness (8 mm), but the first focal point exists inside the accommodation hole. It is not necessary that all of them exist inside the accommodation hole even if the configuration is made to perform the above. For example, in the configuration shown in the figure, for example, the depth of the accommodation hole may be set to about 5 mm.

The example shown in the figure is an embodiment in which a container holding a sample is accommodated in the accommodation hole 1. However, the sample can be directly accommodated in the accommodation hole 1. Further, for example, a flow type sample container can be accommodated in the accommodation hole. For this reason, for example, a container made of the same material as the material constituting the polyhedron after arranging the accommodation holes of an appropriate size on the polyhedron so that the center thereof coincides with the first focus, For example, it is possible to prepare a container having the same external dimensions but differing only in the content of the container, hold the sample in the container, and then store the sample in the storage hole.

When the sample or the container is accommodated in the accommodation hole, in order to improve the light-collecting efficiency by the light-collecting element of the present invention,
Determining the dimensions of the receiving hole and the sample container to be stored in the receiving hole, and the amount of the sample liquid to be analyzed so that the liquid level of the sample is substantially equal to the upper surface of the receiving hole (that is, the upper surface of the polyhedron). Is preferred.

In the example shown in the figure, the receiving hole is formed in the direction perpendicular to the surface 8 which is the upper surface of the polyhedron which is the light condensing element. However, if the container is a closed type or a flow cell type container is used, For example, the opening of the accommodation hole can be formed in the surface 6 or the surface 7.

The light exit surface 3 is opposed to the light reflection surface 2, and the light reflected by the light reflection surface enters at an angle not exceeding the total reflection angle and exits outside the polyhedron. As can be easily understood from FIG. 2, this surface has an arbitrary cross-section (a surface in which a cross-section that becomes a part of an ellipse in the light reflection surface 2, in this example, a cross-section in a plane parallel to the surface 8). , Symmetric to the major axis of the ellipse, in other words parallel to the minor axis of the ellipse.

The focal point farther from the light reflecting surface 2 (hereinafter referred to as a second focal point) is located at the light reflecting surface 2 with the light emitting surface 3 interposed therebetween.
, That is, outside the polyhedron. In the light-collecting element of the present invention, besides positioning the second focal point outside the polyhedron, it is also possible to position the second focal point on the surface (outer surface) of the light exit surface 3 or to position the second focal point inside the polyhedron. is there.

Normally, when the condensed light emission is detected using a light detection element, the light emission surface area of the light detection element is reduced by condensing the light emission in as narrow a range as possible. It is preferable in preventing light emission from decay. On the other hand, when a wavelength selecting means such as an optical filter is provided, it is necessary to arrange the means between the light detecting element and the light emitting surface. Therefore, in the analyzer according to the present invention, the second focal point is located outside the polyhedron as in the example of FIG. 2, and the light emitting surface and the detecting element are spaced apart from each other, and the wavelength selecting means and the like are interposed therebetween. Is preferred.

With respect to the embodiment of the light-collecting device of the present invention shown in FIG. 1, the dimensions of each part and the actual manufacturing process will be described with reference to FIG. The polyhedron was a flat plate of transparent acrylic resin having a thickness of 8 mm, and was formed by shaving from a plate having a thickness of 8 mm. First, an accommodation hole having a diameter of 8 mm is formed so that a container having an outer diameter of 7.7 mm can be accommodated. The depth and shape of the storage hole were determined so that when 200 μl of the liquid sample was placed in the sample container, the liquid level of the sample was almost equal to the upper surface of the storage hole.

Next, the receiving hole was cut out so as to be surrounded by a surface composed of the light reflecting surface 2 and five surfaces (a total of seven surfaces including the upper surface 8 and the lower surface 9). At this time, the first focal point of the light reflecting surface 2 was located at the center of the receiving hole. The light reflecting surface 2 and the other five surfaces are all flat plate upper surfaces 8.
And the lower surface 9. The light exit surface 3 is parallel to the minor axis of the ellipse partially including the light reflection surface 2 in a cross section parallel to 8, and the distance from the center of the ellipse is 12
mm.

It should be noted that the side surface 4 connected perpendicularly to the light exit surface 3
(Hereinafter referred to as the SL1 surface) and the side surface 5 (hereinafter referred to as the SR1 surface) are parallel, and the sensation is 18 mm. Further, the side surface 6 (hereinafter, referred to as SL2 surface) and the side surface 7 (hereinafter, referred to as SR2 surface) connected to the light reflection surface 2 form an angle of 20 degrees with the major axis of the ellipse, and are parallel to the upper surface 8. At a position 4 mm away from the short axis of the ellipse partially including the light reflecting surface 2 in the cross section toward the first focal point, the SL1 surface or the SR
Connected to one side.

After the cutting, each surface was roughly polished with diamond particles of φ10 μm, and further, φ1 μm, φ0.3
Precision polishing was sequentially performed using alumina particles of μm and φ0.06 μm to obtain a mirror finish. Light reflective surface 2 for finishing, SL
The light reflection film 11 was formed on the two surfaces and the SR2 surface by performing aluminum evaporation.

FIG. 3 is a view showing an embodiment of the light-collecting device according to the sixth aspect, and FIGS. 4A and 4B are a top sectional view (a) and a cross-sectional view of the light-collecting device shown in FIG. b).

In the light-collecting element of this embodiment, the light exit surface 3 is formed as a curved surface that is convex toward the outside of the polyhedron.
In the figure, the light emitting surface 3 is a curved surface having a radius of curvature of 8.5 mm. By forming the light emitting surface as a curved surface protruding toward the outside of the polyhedron, the light-collecting element is provided in the emission analyzer. In this case, the light collection efficiency on the light receiving surface of the photodetector can be further improved.

FIG. 4 illustrates the operation of the light-collecting device shown in FIG. 1 or FIG. Housing hole (light reflection surface 2
Out of the light emitted from the sample or the like accommodated in the first light-emitting surface of the light-emitting surface 3
Out of the polyhedron (optical path a). The light emitted toward the light reflecting surface 2 is reflected toward the second focal point by the light reflecting film formed on the outer surface of the light reflecting surface 2 and is emitted from the light emitting surface 3 to the outside of the polyhedron (optical path b).

On the other hand, the light emitted toward the SL1 surface or the SR1 surface is totally reflected and goes to the light emitting surface 3 because the incident angle to the SL1 surface and the SR1 surface exceeds the total reflection angle, and the light is emitted from the surface to the outside of the polyhedron. (Optical path c). SL2 surface or SR2
Light emitted toward the surfaces is reflected by the light reflecting films formed on these surfaces, and a part of the light is guided to the light emitting surface 3 by repeated reflection on the light reflecting surface 2 and other surfaces, from which the polyhedron exterior is exposed. (Optical path d). Most of the light emitted toward the surface 8 or 9, which is the upper surface and the lower surface of the light-collecting element, is guided to the light emitting surface 3 while repeating total reflection, and is emitted from the surface to the outside of the polyhedron, as in the optical path e.

Although a combination of the above-mentioned reflections may occur, the light emitted from the receiving hole and incident on the inside of the polyhedron, which is a light-collecting element, is transmitted in the direction of the light emitting surface 3 as a whole. 3, the light is emitted to the outside of the polyhedron. Therefore, in the light-collecting device shown in the figure, the light-receiving surface of the light-detecting device is arranged on the light-emitting surface 3 and both are brought into close contact with each other, or
If both the vertical and horizontal sizes of the light receiving surface are sufficiently larger than the size of the light emitting surface 3, all of the light emitted from the light emitting surface 3 can be incident on the light receiving surface.

However, since the light receiving surface of the photodetector is housed inside the vacuum tube or inside the protective case, it is often difficult to bring it into close contact with the light emitting surface 3. For this reason, when it is necessary to dispose them both as described above, as described above, a light detection element (light receiving surface) having a larger area than the light emitting surface 3, particularly, both the vertical and horizontal sizes of the light emitting surface 3 are used.
It is preferable to use a light-receiving surface sufficiently larger than the size of the light-emitting surface. However, as shown in FIG. When the emitted light is emitted from the polyhedron, the light is refracted in the direction of the central axis, and is made incident on the light receiving surface to increase the detection efficiency.

As described above, in the light-collecting device of the present invention shown in the figure, the surface functioning as a concave reflecting mirror having a columnar shape with an arbitrary cross-section substantially forming a part of an ellipse and functioning as an optical waveguide. Since it is a polyhedron having both surfaces and surfaces, it is a compact, low-cost, high-efficiency light-collecting element that can make the majority of the emitted light emitted radially as a light flux directed toward the light-emitting surface. .

FIGS. 5 and 6 are views showing an embodiment of an emission spectrometer according to claim 7 equipped with the light-collecting element of the present invention shown in FIG.

In this example, 8 × 2
A side-on type photomultiplier tube having a light receiving surface 23 of 4 mm and the light condensing element 21 shown in FIG. 3 are provided, and the light emitting surface of the light condensing element and the light receiving surface of the photomultiplier tube are separated and parallel. They are arranged facing each other.

The size of the light exit surface of the light-collecting device shown in FIG.
It is a cylindrical surface with a radius of curvature of 8.5 mm based on a plane of × 18 mm, and most of the light emitted from the light emitting surface is incident on the light receiving surface of the photomultiplier and detected. Further, since the light-collecting element and the light-detecting element are spaced apart from each other, as shown in FIG. 6, it is possible to arrange the optical filter 24, which is a means for selecting the wavelength of light, between them. Emission analysis can be performed on the emission from the specific substance after wavelength selection.

FIG. 7 is also a diagram showing an embodiment of an emission spectrometer according to claim 7 equipped with the light-collecting element shown in FIG. In this example, 8 × 24 mm
3, a side-on type photomultiplier tube having a light receiving surface, a condensing element 21 shown in FIG. 3, an excitation light source 25 for inducing fluorescence of a specific substance contained in a sample, a converging lens 26 for excitation light, Equipped with excitation light wavelength selection means 27,
The light emitting surface of the light condensing element and the light receiving surface of the photomultiplier tube are spaced apart and opposed to each other in parallel, and an optical filter 24 serving as light wavelength selecting means is arranged between them. The emission spectrometer of FIG. 7 shows an example in which the incident direction of the excitation light for inducing the fluorescence is above the opening of the accommodation hole. Excitation light can also be incident from the bottom.

FIG. 8 shows an embodiment of a light-collecting element (a) obtained by partially improving the polyhedron in FIG. 3, and an emission analyzer equipped with the light-collecting element and irradiating excitation from the lateral direction of the polyhedron. FIG. According to such a configuration, for example, the light-collecting element and the emission spectrometer of the present invention can be applied by using a flow-type sample container for liquid chromatography, flow injection, electrophoresis, or the like, or a column itself in column chromatography as a container. Can be.

FIG. 8 specifically illustrates a light-collecting element for performing fluorescence analysis using a flow type sample container 30 of a liquid chromatograph, and an emission analyzer using the light-collecting element. ing. The accommodation hole in the light condensing element 21 of FIG. 3 is a through hole of φ8 mm, and further, light passing holes 28 and 29 connecting the light reflection surface 2, the accommodation hole and the SR2 surface are formed, and the excitation light source is passed through the light passage hole. The excitation light 25 that has passed through the excitation light converging lens 26 and the excitation light wavelength selecting means 27 can excite the sample in the accommodation hole. In such a configuration, in order to prevent scattering of the excitation light as much as possible, it is preferable to design the irradiation direction of the excitation light so as to be perpendicular to the direction in which the fluorescence signal is collected. It is easy to change the design such that the excitation light enters from the SR2 surface and exits from the light reflection surface 2.

This example shows an embodiment for performing fluorescence analysis. For example, in the case of detecting chemiluminescence or bioluminescence, an excitation light source is arranged or light for excitation light is used. It is not necessary to arrange a passage hole. Further, when the light-collecting element and the emission spectrometer of the present invention are applied using a column in a liquid chromatograph as a sample container, the thickness of the light-collecting element shown in FIG. 3 can be approximately equal to the column length. However, in order to reduce the size of the light-collecting element, and consequently the size of the analyzer, and to reduce the manufacturing cost, for example, a light-collecting element, an excitation light source, a photodetector, and the like illustrated in FIG. For example, it is possible to add a mechanism for performing the analysis and perform the analysis while moving these.
This also makes it possible to sequentially analyze each substance separated along the axis of the column.

FIGS. 9 and 10 show another embodiment of the light-collecting device of the present invention. In the figure, 2 indicates a light reflecting surface, 3 indicates a light emitting surface, and 11 indicates a light reflecting film. FIG.
Is the SL2 surface or SR2 in the light-collecting device shown in FIG.
FIG. 10 shows a light condensing element having a shape in which one surface is added to the left and right surfaces between the light reflecting surface 2 and the light reflecting surface 2. FIG. It is a light collecting element. Both examples have a flat plate shape, and in a cross section in a plane parallel to a plane constituting an upper surface or a lower surface of the polyhedron,
The cross sections of the light reflecting surface 2 and the light emitting surface 3 are symmetric with respect to the major axis of the ellipse, and the first focal point of the light reflecting surface exists inside the receiving hole.

FIG. 11 is a diagram showing an embodiment of the light-collecting element of claim 2, that is, a dihedral light-collecting element. The polyhedron main body is a dihedron in a form in which an elliptical sphere is cut at a plane perpendicular to the major axis thereof. In this example, the cut surface is the light emitting surface 3, and the other curved surface 2 is the light reflecting surface. Although not shown in the figure, a light reflecting film is formed on the curved surface 2 (other than the cut surface 3) outside the polyhedron.

The light reflecting surface 2 has a curved shape whose cross section is a part of an ellipse as is apparent from the figure, and has two focal points on the long axis. Of these focal points, an accommodation hole 1 for accommodating the sample or a container for holding the sample is formed at the focal point closer to the light reflection surface 2.

FIG. 12 is a view showing another embodiment of the light-collecting device according to the third aspect, that is, a dihedral light-collecting device. 11 is that the light emitting surface 3 in the example of FIG. 10 is replaced by an ellipsoidal sphere 3 which is convex toward the inside. The center of the spherical surface 3 as the light emitting surface coincides with the focal point on the side farther from the outer surface among the two focal points of the light reflecting surface 2. The light emitting surface 3 is formed as a spherical surface that is convex toward the inside of the elliptical sphere in order to improve the light collection efficiency of light emitted from the light emitting surface.

As described above, the light-collecting element of the present invention has been described based on the embodiment. In this embodiment, the light-receiving surface size of the photodetector used, the sample capacity, and the size of the light-collecting element are taken into consideration. At the same time, the design can be appropriately changed for the purpose of improving the light collection efficiency.

The improvement of the light collecting efficiency will be described below based on the light collecting element shown in FIG. In the light-collecting element having a different form from that of FIG.
The design change can be facilitated by reading the description regarding the surface and the SR2 surface as the surface corresponding thereto.

For example, if the light emitted from the light emitting surface 3 to the outside of the polyhedron deviates from the range of the light receiving surface of the light detecting element, a detection loss occurs. Therefore, the size of the light emitting surface 3 is made smaller than the light receiving surface of the light detecting element. Is generally preferred. In this case, it is particularly preferable to consider the refraction of light emission on the light emitting surface, such as making the light emitting surface a curved surface convex toward the outside of the polyhedron.

Also, for example, if light emission is incident on the light exit surface at an angle exceeding the total reflection angle and is totally reflected, a condensing loss will occur. Therefore, the light is emitted from the first focal point, enters the inside of the polyhedron, and then strikes the light exit surface. Among the arriving light, at least the light that reaches the light emitting surface directly, the light that reaches the light emitting surface through one or more reflections on the surface corresponding to the SL1 surface or the SR1 surface, and the light that reaches the light reflecting surface. It is preferable that the angle of incidence of the light arriving after one reflection on the light exit surface does not exceed the total reflection angle.

For example, it is preferable that the light emitted from the first focal point, incident on the inside of the polyhedron, and reaches the surface corresponding to the SL1 surface or the SR1 surface is designed to be totally reflected on each surface. This makes it unnecessary to form a light reflection film on these surfaces. If light that does not undergo total reflection is included, and the light has an angle that allows the light to pass through the light exit surface, it is preferable to form a light reflection film on the column surfaces corresponding to the SL1 surface and the SR1 surface.

For example, the light reflected on the light reflecting surface is S
When the light is incident on a plane corresponding to the L2 plane or the SR2 plane, a light-collecting loss is caused. Therefore, it is preferable to design so that a line segment connecting the second focal point and a point on the light reflection surface does not cover the surface corresponding to the SL2 surface or the SR2 surface. However, this does not apply when the entire light reflecting surface is not used as an effective surface for condensing light. Further, in addition to the design in which the light reflected by the light reflecting surface directly reaches the light emitting surface as in the above example, SL1
It is also possible to design such that the light is reflected at least once on the surface corresponding to the surface or the SR1 surface and then reaches the light emitting surface. In this case, it is preferable to design so as to satisfy the condition of total reflection on the surface corresponding to the SL1 surface or the SR1 surface.

The planes corresponding to the SL1 plane and the SR1 plane,
In some cases, it is difficult to optimize both light reflection surfaces from the viewpoint of the amount of reflected light. For example, when optimization is performed on the surfaces corresponding to the SL1 surface and the SR1 surface, the amount of reflection on the light reflecting surface is smaller than the optimum value. Therefore, it is preferable to design the light-collecting element by deciding the allocation of the contribution rates of the two depending on the constraints such as the total light-collecting amount and the overall size, the intended use, and the like. In addition, the surfaces 8 and 9 constituting the upper and lower surfaces of the flat plate in the example of FIG. 3 are surfaces on which total reflection transmission is expected, and it is not always necessary to form a light reflection film on both surfaces.

Example 1 Chemiluminescence generated by the reaction of alkaline phosphatase (ALP) and adamantyl methoxyphosphoryl phenyl dioxetane solution (AMPPD) was analyzed using the emission spectrometer shown in FIG. AMPPD is ALP
Releases a phosphate group by the action of the compound to generate an excited singlet-state methyloxymetabenzoate via intramolecular electron transfer and cleavage of a dioxetane bond, and emits light having a peak at 470 nm. Therefore, a sufficient amount of AMPPD
It is possible to quantify the ALP by measuring the light emission quantity in the presence of a conventional 10 ^-19 to 10 ^-20
It is known as a chemiluminescent substrate capable of detecting mol ALP.

First, 100 mM TES solution (pH 8.0) containing 0.1% BSA was used as a diluent to prepare ALP solutions of various concentrations. 10 μl of this ALP solution was dispensed into a sample container made of polystyrene, and 3.3
100 μl of a × 10 ⁻⁴ M AMPPD solution was added, and the mixture was allowed to stand for 15 minutes. Then, the chemiluminescence for 2 seconds was measured by the emission analyzer shown in FIG.

FIG. 17 shows the average value obtained by repeating the measurement six times for each of the ALP solutions having various concentrations, as a calibration curve for ALP analysis. Note that FIG. 17 also shows, for comparison, results measured by an emission spectrometer using the conventional light-collecting device shown in FIGS. 15 and 16.

As can be seen from FIG. 17, in the luminescence analyzer equipped with the light-collecting device of the present invention, the amount of luminescence measurement clearly increases as compared with the luminescence analyzer equipped with the conventional light-collecting device. An improvement in light collection efficiency is observed. Also, ALP
The linearity in the low-concentration region is also improved, and it can be seen that 1 × 10 ⁻²⁰ mol of ALP can be accurately detected, which was difficult to detect with the conventional device.

Example 2 Using the emission spectrometer shown in FIG. 7, the fluorescence generated when the Eu ³⁺ chelate was excited by ultraviolet rays was measured in a time-resolved manner.

First, a commercially available solution (Phar
Eucia ⁺ chelate solutions of various concentrations were prepared using an enhancer solution (trade name: DELFIA system, manufactured by Macia Corporation). 200 μl of this Eu ³⁺ chelate solution was dispensed into a polystyrene sample container and analyzed by the emission spectrometer shown in FIG. For the analysis, a half-width of 1μ
Using a Xe flash lamp as a pulse light source for sec, light of 370 nm was selected by an optical filter and used as excitation light. A 560 nm cut-off filter is inserted between the light-collecting element and the light-detecting element as a means for selecting the wavelength of light emission,
In addition to preventing the excitation light from being incident on the photodetector,
The fluorescence of the Eu ³⁺ chelate centered at 5 nm was transmitted and detected by the photodetector. In addition, a side-on type photomultiplier tube is used as a photodetector, and 400 to 800
μsec was selected, and signals for 1000 pulses of excitation light were integrated to obtain a measured value.

For Eu ³⁺ chelate solutions of various concentrations,
The average value obtained by repeating six measurements is shown in FIG. 18 as a calibration curve for Eu ³⁺ chelate analysis. FIG.
FIG. 8 also shows, for comparison, the results of measurement using an apparatus using an optical lens as a light-collecting element as shown in FIG. As is clear from FIG. 18, the emission spectrometer equipped with the light-collecting element of the present invention has an emission measurement amount increased by one digit or more as compared with the conventional light-emission analyzer equipped with the light-collecting element. In addition, it can be seen that the light collection efficiency has been improved.

[0074]

As described above, the light-collecting device of the present invention is a small-sized, low-cost light-collecting device having the features of a concave reflecting mirror, which is a part of an elliptic cylinder, and an optical waveguide. Can be efficiently transmitted to the light-receiving surface of the photodetector element. Also,
An emission spectrometer equipped with this light-collecting element can improve the detection efficiency of luminescence as compared with a conventional analyzer, and can lower the lower detection limit concentration of a target specific substance.

The light-collecting device of the present invention can be used not only for a sample or the like held in a sample container, but also for a flow-type sample container for liquid chromatography, flow injection analysis, electrophoresis, etc.
Furthermore, a column itself such as a column chromatograph can be used as a sample container, and can be applied to a wide range of uses, such as when fluorescence, chemiluminescence or bioluminescence is used as a detection means in these analyzers. It is.

[Brief description of the drawings]

FIG. 1 is a diagram showing an embodiment of a polyhedron that is a light-collecting element of the present invention.

FIG. 2 is a diagram showing dimensions of each part of the polyhedron shown in FIG. 1;

FIG. 3 is a diagram showing another embodiment of the polyhedron that is the light-collecting element of the present invention.

FIG. 4 is a view for explaining an optical path of light emission in the light-collecting device shown in FIG.

FIG. 5 is a diagram showing an embodiment of an emission spectrometer of the present invention.

FIG. 6 is a view showing another embodiment of the emission spectrometer shown in FIG. 5;

FIG. 7 is a diagram showing another embodiment of the emission analyzer of the present invention.

FIG. 8 is a diagram showing another embodiment (a) of a polyhedron that is a light-collecting element of the present invention and another embodiment of an emission analyzer of the present invention using the same.

FIG. 9 is a diagram showing another embodiment of the polyhedron that is the light-collecting element of the present invention.

FIG. 10 is a view showing another embodiment of the polyhedron which is the light-collecting element of the present invention.

FIG. 11 is a view showing an embodiment of a dihedral body which is a light condensing element of the present invention.

FIG. 12 is a view showing another embodiment of a dihedral body which is a light condensing element of the present invention.

FIG. 13 is an explanatory diagram of a configuration of a conventional emission analyzer using an optical lens.

FIG. 14 is an explanatory diagram of a configuration of a conventional emission analyzer using a concave reflecting mirror.

FIG. 15 is an explanatory diagram of a configuration of a conventional emission analysis device using an optical waveguide.

FIG. 16 is an explanatory diagram of a configuration of a conventional emission spectrometer in which a photodetector is brought close to a sample container.

FIG. 17 is a diagram showing the results of Example 1. In the figure, the vertical axis shows the count number, and the horizontal axis shows the ALP amount, respectively. The case and the small black square show the result when the photodetector is brought close to the case, respectively.

FIG. 18 is a diagram showing the results of Example 2. In the figure, the vertical axis indicates the count number, and the horizontal axis indicates the Eu ³⁺ amount. The large black square in the figure indicates the case where the light-collecting element of the present invention is used, and the small black square indicates the conventional optical lens type light-collecting element. The results when the element was used are shown.

[Explanation of symbols]

1; accommodation hole; 2; light reflecting surface; 3; light emitting surface; 4; surface (S
L1 plane), 5; plane (SR1 plane), 6; plane (SL2 plane),
7; surface (SR1 surface), 8, 9; upper or lower surface of flat plate, 1
DESCRIPTION OF SYMBOLS 1; Light reflection film, 21; Condensing element, 22;
3; the light receiving surface of the photodetector; 24; wavelength selecting means; 25;
Excitation light source, 26; excitation light converging lens, 27; excitation light wavelength selection means, 28, 29; excitation light light passage hole, 30;
Flow type container

Claims (7)

[Claims] 1. A light-collecting element for condensing light emitted from a specific substance contained in a sample, comprising: (a) a light-reflective material formed of a material transmissive to light emitted from the specific substance; (B) the light reflecting surface has a curved surface shape whose cross section is substantially a part of an ellipse;
A light reflection film is formed outside, and (c) the light emission surface is disposed to face the light reflection surface, and light reflected by the light reflection surface enters at an angle not exceeding the total reflection angle. And (d) forming an accommodation portion for accommodating a sample or a container holding the sample at a focal point on the side closer to the light reflecting surface among the two focal points of the ellipse, and (e) A light condensing element wherein, of the light emitted from the sample accommodated in the accommodation section and incident on the polyhedron, the light reaching the reflection surface is emitted from the light exit surface in a condensed state. 2. The light-collecting element is a dihedron in which an elliptical sphere is cut at a plane perpendicular to the major axis thereof, the cut surface being the light-emitting surface, and the other curved surface being the light-reflecting surface. The light-collecting device according to claim 1, wherein 3. The light-collecting element is a dihedron in which an ellipsoidal sphere is formed with a spherical surface protruding toward the inside thereof, the spherical surface being the light-emitting surface, and another curved surface being the light-reflecting surface. age,
2. The light-collecting device according to claim 1, wherein the center of the spherical surface coincides with a focal point far from the light reflecting surface among the two focal points of the elliptical sphere. 4. The light-collecting element is a polyhedron having three or more surfaces, and a surface other than the light reflecting surface and the light emitting surface reflects light incident inside the polyhedron to form a light reflecting surface or a light emitting surface. The light-collecting device according to claim 1, wherein the light-collecting device has a function as a waveguide for guiding to a surface. 5. The light-collecting device according to claim 1, wherein the light-collecting device is a polyhedron in which the cross sections of the light reflecting surface and the light emitting surface are symmetric with respect to the major axis of the ellipse. 6. The light-collecting device according to claim 1, wherein said light-collecting device is a polyhedron having a curved light-emitting surface protruding toward the outside of the polyhedron. 7. A light emission from a specific substance contained in a sample, comprising: the light collection element according to claim 1; and a detection element for detecting light emission emitted from a light emission surface of the light collection element. For detecting and analyzing