JP2010169672A - Vessel for luminescence measurement - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a vessel for luminescence measurement, made of a thermoplastic resin containing light-reflecting component and light-absorbing component so as to enhance signal intensity by reflection on the inner surface of a vessel, reduce background by self light emission from the thermoplastic resin body itself constituting the vessel, and simultaneously attain all light-shielding effects for eliminating unfixed noise. <P>SOLUTION: This vessel for luminescence measurement is made of a molded thermoplastic resin, containing 0.5-10 wt.% light-reflecting component and 0.01-0.5 wt.% light-absorbing component and is colored in a gray or chromatic tint. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a luminescence measurement container used for luminescence measurement. More specifically, the present invention relates to a measurement container such as for chemiluminescence measurement that can simultaneously achieve an excellent signal enhancement effect, a noise reduction effect such as self-luminescence, and an excellent light-shielding effect.

  In the field of today's clinical examinations, for example, by measuring the blood concentration of a marker substance related to a specific disease using various luminescence measurements such as fluorescence measurement, chemiluminescence measurement, bioluminescence measurement, etc. It is used for early detection and treatment of diseases. The luminescence measuring containers used in such fields are generally made of a thermoplastic resin due to ease of mass production, low cost, flexibility such as shape and dimensions, etc. Depending on the case, the uncolored ones, black ones, and white ones are used properly.

  First, when it is more advantageous to improve the measurement sensitivity by reflecting the light emission by reflecting the inner surface of the container, the heat with which the white pigment which is a light reflecting component is blended (kneaded) A white container made of plastic resin is used. Next, when it is necessary to reduce the background due to self-emission emitted from the thermoplastic resin main body constituting the container, it is a white pigment that is a light reflective component to reflect or absorb excitation light, or a light absorbing property. A white or black thermoplastic resin container in which a black pigment as a component is blended (kneaded) is used. And, in order to improve the S / N ratio by eliminating indefinite noise coming from the outside (for example, disturbance light, light emission spilling from an adjacent container, etc.) and avoiding so-called crosstalk, and to achieve high sensitivity, A black thermoplastic resin container having a good light shielding property against ambient light is used.

  For example, Patent Document 1 discloses that, in light emission (fluorescence) measurement in which a sample is irradiated with excitation light, the thermoplastic resin constituting the container is excited by excitation light and emits fluorescence to prevent the background from rising. It discloses the incorporation of white or black pigments. Further, for example, Patent Document 2 discloses that a part of the inner wall is colored in a color other than white in a luminescence measurement container in order to reduce the background.

Japanese Patent Publication No. 3-40818 JP 2006-1119011 A

  In order to increase the measurement sensitivity of luminescence measurement, N (noise) in addition to signal enhancement by reflection on the inner surface of the container and reduction of background caused by self-luminescence emitted from the thermoplastic resin body constituting the container. It is important to improve the S / N ratio of the measurement system by reducing the components, but in order to eliminate indefinite noise from the outside of the measurement system among the N components, the light-shielding property of the luminescence measurement container is improved. It is important to do. Indefinite noise includes, for example, the case where the light emission measurement container is a microplate, and the light emission from the adjacent well is strong, whereas the light emission of the measurement target well is fine. In addition, the strong light emission leaks through the side wall to the target well.

  However, Patent Document 1 only discloses that a black or white pigment is blended, and it is impossible to expect all of reduction of background, enhancement of signal due to reflection on the inner surface of the container, and light shielding properties. When black pigments are kneaded, signal enhancement due to reflection on the inner surface of the container remains as a problem. Conversely, when white pigments are blended, sufficient light-shielding properties cannot be obtained compared with blending black pigments. . In Patent Document 2, since the inner wall of the container is colored in a color other than white, preferably black, reduction of the background can be expected, but signal enhancement due to reflection on the inner surface of the container is insufficient. There is.

  By blending both a black pigment that is a light-absorbing component and a white pigment that is a light-reflecting component, this is achieved by reducing the background and shading achieved by blending the black pigment, and blending the white pigment. Can the reduction of the background and the signal enhancement by reflection on the inner surface of the container be achieved at the same time, and what proportion should be added to increase the signal by reflection on the inner surface of the container and the thermoplastic resin constituting the container It is not known whether it is possible to simultaneously and efficiently achieve the reduction of the background by the self-emission emitted from the main body and the light shielding property to eliminate indefinite noise.

  Therefore, the object of the present invention is to achieve all of the signal enhancement by reflection on the inner surface of the container, the reduction of the background by the self-emission emitted from the thermoplastic resin body constituting the container, and the light shielding property to eliminate indefinite noise at the same time. An object of the present invention is to provide a container for light emission measurement made of a thermoplastic resin in which a light reflecting component and a light absorbing component are blended.

The present invention, which has been made to achieve the above object, contains 0.5 to 10% by weight of a light-reflective component and 0.01 to 0.5% by weight of a light-absorbing component, and is colored gray or chromatic. A molded luminescence measurement container made of a thermoplastic resin. In addition, when the luminescence measurement container has a plurality of openings, the luminescence measurement container of the present invention from the viewpoint of optical characteristics,
The amount of light (A) [cps] from the luminescent solution detected from the opening when the luminescent solution is put into one of the openings of the luminescence measurement container, and one of the openings of the container From the amount of light (B) [cps] from the luminescent solution detected from the opening adjacent to the opening when the luminescent solution is added, equation (1)
[{(A)-(B)} ÷ (A)] × 100 Formula (1)
The shading rate calculated by is 99.99% or more,
The light quantity (A) [cps] is the light quantity [cps] detected from the luminescent solution when the luminescent solution is put into the opening of a container containing 1.0% by weight of black pigment. ], A luminescence measurement container. Hereinafter, the present invention will be described in detail.

  Luminescence in the present invention is visible light generated when a molecule in the ground state by chemical reaction such as chemiluminescence or bioluminescence absorbs energy and transitions to an excited state of a high energy state and returns to the original ground state. And other than fluorescence emission or phosphorescence emission. Luminescence is not limited to visible light emitted directly from the measurement object. For example, once the energy is transferred to a fluorescent substance coexisting in the reaction field, the fluorescent substance becomes excited, and from there It may be visible light emitted when returning to the ground state. Here, fluorescence emission refers to excitation light emitted when an illuminant that absorbs excitation light is electronically excited to enter an intermediate excitation state and return to the ground state by irradiating ultraviolet light or visible light as excitation light. Phosphorescence emission refers to luminescence emitted when electrons that have transitioned to the electronically excited triplet return to the ground state.

  The size of the container is not particularly limited as long as it is made of a thermoplastic resin having an opening for measuring the luminescence generated by the luminescence reaction. Even if an absorptive component is blended, there is a possibility that sufficient light-shielding properties may not be obtained. Therefore, it is preferable to secure a thickness of about 1 mm (0.5 mm to 1.5 mm). As the shape of the container, for example, a tube, a cup, a cuvette, etc., each of which has an individual concave shape (having one opening), or a shape in which a plurality of concave shapes such as a microplate are connected (opening) One having a plurality of parts) can be exemplified. The thermoplastic resin constituting the container is not particularly limited as long as it can be blended with a pigment or dye that is a light-reflecting component or a light-absorbing component, which will be described later. For example, an olefin resin, a styrene resin, a vinyl resin Examples thereof include, but are not limited to, carbonate-based resins, polyester-based resins, and polyamide-based resins. Among the exemplified thermoplastic resins, olefin homopolymers and copolymers having a proven track record as already constituting a container for light emission measurement are particularly preferred thermoplastic resins. More specifically, for example, low density, medium density or high density polyethylene, linear low density polyethylene, polypropylene, polybutene-1, poly-4-methylpentene-1, propylene-ethylene copolymer, ionomer, ethylene- An acrylic copolymer, an ethylene-vinyl acetate copolymer, etc. can be illustrated.

  The light-reflective component blended in the thermoplastic resin that composes the container is designed to reduce the background caused by self-luminescence by preventing signal enhancement by reflection on the inner surface of the container and excitation of the thermoplastic resin body by the reflection. belongs to. Accordingly, examples of the light-reflecting component include white pigments or dyes that reflect light over all wavelengths, and pigments or dyes that reflect a specific wavelength of light emission to be measured. Among these, a pigment is a particularly preferable light reflecting component. More specifically, white pigments such as titanium dioxide, zinc white, lead white, calcium carbonate, talc, and mica can be exemplified. Among these, titanium dioxide is a particularly preferable pigment because it has good dispersibility and concealment properties when molding a thermoplastic resin into a container shape, and has high safety. Of the titanium dioxides, those that are usually selected are those having a small photocatalytic action, or those coated with aluminum hydroxide or alumina to suppress the photocatalytic action. As a pigment of a color that reflects a specific wavelength of light emission to be measured, for example, if the specific wavelength is around 450 nm, a blue pigment such as ultramarine blue, procyanine blue, phthalocyanine blue, or anthraquinone, If the specific wavelength is around 550 nm, a green pigment such as phthalocyanine green, perylene, or nitroso compound is used. If the specific wavelength is around 650 nm, nickel titanium yellow, strontium yellow, yellow lead, chromium yellow, zinc Examples thereof include yellow pigments such as yellow, benzidine yellow, quinophthalone, isoindolinone and imidazolone. The light reflective component demonstrated above may be mix | blended individually by 1 type, and may be mix | blended combining 2 or more types. When two or more types are combined, in addition to combining two or more pigments of the same color, it also includes combining two or more pigments of different colors.

The light-absorbing component blended in the thermoplastic resin that composes the container is intended to reduce the background due to self-emission emitted from the thermoplastic resin body that composes the container, and to block light to eliminate indefinite noise. is there. Accordingly, black pigments or dyes that absorb light over all wavelengths can be exemplified. Among these, a pigment is a particularly preferable light absorbing component. More specifically, examples of the black material include carbon black, titanium black, acetylene black, lamp black, aniline black, iron oxide (Fe ₃ O ₄ ), and reduced iron. These can also be mix | blended independently and can be mix | blended combining 2 or more types.

The light-reflecting component is used for the thermoplastic resin container to reduce the background derived from self-luminescence by preventing signal enhancement by reflection on the inner surface of the container and excitation of the thermoplastic resin body by the reflection. 0.5 to 10% by weight, preferably 1 to 6% by weight is blended. On the other hand, the light-absorbing component is less than the thermoplastic resin container because of the reduction of the background due to self-emission emitted from the thermoplastic resin body constituting the container and the light-shielding property to eliminate indefinite noise. 0.01 to 0.5% by weight, preferably 0.02 to 0.1% by weight. Among the examples exemplified above as the light reflecting component and the light absorbing component, when a white pigment and a black pigment are employed, the luminescence measurement container of the present invention that exhibits gray color is provided. It varies depending on the ratio of white and black pigments to be blended. On the other hand, when a pigment other than white or a pigment other than white and white is employed as the light reflecting component, for example, a blue container (reflects light near 450 nm), a green container (reflects light near 550 nm). ), A yellow container (reflects light near 650 nm) and the like, and a luminescent color measurement container colored in a chromatic color is provided. The amount of the light-reflective component and the light-absorbing component to the thermoplastic resin is based on, for example, a white component containing a white pigment, and a black pigment is compounded in various proportions. Can be determined by evaluating with the intended measurement system. According to the knowledge of the present inventor, the light-shielding property can be achieved by blending a small amount of a light-absorbing component. Therefore, first, after studying the blending amount that can achieve a signal enhancement effect due to reflection on the inner surface of the container with a white, blue, green, or yellow pigment, a small amount of black pigment is blended, and the thermoplastic resin self-luminous. It is possible to exemplify adjusting the composition of the black pigment by examining the background reduction and the light-shielding property.

In the case where the container of the present invention is a container in which a plurality of concave objects are connected (having a plurality of openings), such as the container of FIG.
The amount of light (A) [cps] detected from the luminescent solution when the luminescent solution is put in one of the openings of the container, and the luminescent solution in one of the openings of the container From the amount of light (B) [cps] from the luminescent solution detected from the opening adjacent to the opening when
[{(A)-(B)} ÷ (A)] × 100 Formula (1)
The shading rate calculated by is 99.99% or more,
The light quantity (A) [cps] is the light quantity [cps] detected from the luminescent solution when the luminescent solution is put into the opening of a container containing 1.0% by weight of black pigment. ] Is larger than the container. As a specific aspect of the method for measuring the light shielding rate,
After putting a luminescent solution in one opening and covering the other opening with a light-shielding material, measure the light amount (A) [cps] from the luminescent solution detected from the opening containing the luminescent solution,
After the luminescent solution is put in one opening and the opening is covered with a light-shielding material, the opening adjacent to the opening (if there are a plurality of adjacent openings like a microplate, After measuring the amount of light (B) [cps] from the luminescent solution detected from one of them)
A method of calculating the light shielding rate by the above formula (1) can be mentioned. As the light-shielding material that covers the opening used when measuring the light-shielding rate, a material usually used by those skilled in the art, such as an aluminum foil, may be used.

  The container of the present invention is for luminescence measurement. However, the thermoplastic resin constituting the container has been conventionally used as a reaction container for various reactions such as immune reactions and biochemical reactions. This means that the container of the present invention can be used not only for the stage of luminescence measurement but also as a container for causing various reactions as the previous stage. For example, a trace component in blood or serum is captured on a solid phase as a complex using a biological specific reaction, and a component (Bound) captured on the solid phase and a component (Free) not captured are separated. After performing B / F separation, the container of the present invention can be used as a container for causing the biological specific reaction in a series of operations in which the labeling component in the complex is introduced into the luminescence reaction to measure luminescence. Is possible. Further, if the reagent for the biological specific reaction is sealed in advance with an aluminum seal or the like, the container of the present invention is used as a storage container for the biological specific reaction reagent, and a sample for measurement is added. This can be part of a reagent kit that starts the biological specific reaction and measures the resulting luminescence. Here, the biological specific reaction includes antigen-antibody reaction, biotin-streptavidin reaction, biotin-avidin reaction, nucleic acid-nucleic acid interaction, ligand-receptor reaction, and the like.

  The reagent for the two-step sandwich reaction using the antigen-antibody reaction as a biological specific reaction will be specifically described as an example. The amount of antibody necessary to cause one reaction in the luminescence measurement container of the present invention is described. This corresponds to a solution obtained by dispensing and lyophilizing a solution containing a carrier on which is immobilized. Dispensing and lyophilizing a solution of an antibody labeled with an enzyme such as alkaline phosphatase or an antibody labeled with a luminescent substance such as acridinium in an amount necessary to carry out a single measurement in a container different from this container Keep it. Samples such as blood and serum are dispensed into a container containing a carrier, lyophilized components are dissolved and a first antigen-antibody reaction is performed, and a first B / F separation is performed. Dissolve the solution in another container to dissolve the labeled antibody, and dispense the dissolved labeled antibody into the container that has completed B / F separation to perform the second antigen-antibody reaction. After completion of the second antigen-antibody reaction, second B / F separation is performed, and then an enzyme substrate is dispensed as necessary, and the amount of luminescence is measured and converted to the concentration of the target of the antigen-antibody reaction. In this example, the luminescence measurement container of the present invention is commonly used as a container for storing an immune reaction reagent, an antigen-antibody reaction container, and a luminescence measurement container.

The light emission measuring container of the present invention is a background achieved by blending a black pigment as a light-absorbing component and a white pigment as a light-reflecting component at a certain blending ratio. Reduction, light shielding, and background reduction and signal enhancement by reflection on the inner surface of the container are achieved simultaneously and effectively. The luminescence measurement container of the present invention has a high light shielding property,
(A) Even if there is luminescence derived from a contamination component in the luminescence detector, it can be prevented from taking it in.
(B) Even when the substance to be measured is various unknown samples contained from a very low concentration to a high concentration, if it is measured using the luminescence measurement container of the present invention in the form of a microplate, Can be measured after avoiding crosstalk,
(C) Even if the luminescence measurement container has a multiple tube shape and the reagent that reacts with the luminescent substance held in one container is dispensed into another adjacent tube, the luminescent substance content is measured. It is possible to avoid leakage of noise light that occurs due to splashing at the time of injection,
(D) Compared with the existing black container that realizes a low background, signal enhancement due to reflection on the inner surface of the container can be expected.
It has the effect. In particular, the above (a) to (c) are effective in increasing the reliability of the measured value when measuring a low-concentration measurement object with high sensitivity in luminescence measurement.

FIG. 1 is a diagram for explaining the light shielding rate related to the luminescence measurement container. In the figure, 1 is a fluorescence detector, 2 is a luminescence measuring container, 3 is an enzyme reaction solution, and 4 is an aluminum foil seal. FIG. 2 is a diagram showing a response curve of TSH measured using the black container in Table 1. FIG. 3 is a diagram showing a TSH response curve measured using the # 2 container in Table 1.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, these Examples do not limit this invention.

Example 1
A white pigment (titanium dioxide) is selected as the light reflecting component, and a black pigment (carbon black) is selected as the light absorbing component. As shown in Table 1, the blending ratio with respect to polypropylene is changed in four patterns # 1 to # 4. A two-tank type luminescence measurement container (see FIG. 1) having the same capacity was prepared. As controls, uncolored, black and white containers were also prepared.

  As shown in FIG. 1 (a), one opening of an empty container is sealed with aluminum foil 4, and then 10 ng / mL alkaline phosphatase (ALP) 10 μL and 0.4 mM ALP chemiluminescence are added to the other tank. The enzyme reaction solution 3 consisting of 100 μL of the substrate solution (see JP-A-2005-77142) is dispensed and set in the luminescence detector 1, and the amount of luminescence (A) after the reaction at 37 ° C. for 5 minutes is determined according to the reaction solution. It detected from the opening part upper part of the tank into which it entered.

  Next, as shown in FIG. 1B, a new empty container is prepared, the enzyme reaction solution 3 is dispensed into one tank, the opening is sealed with aluminum foil 4, and the reaction is performed at 37 ° C. for 5 minutes. I let you. After the reaction, a container is set in the luminescence detector 1 so that the light receiving surface is located above the opening of the empty tank, and the empty space passes through a partition wall of about 1 mm at the shortest point from the tank containing the enzyme reaction solution 3. The amount of light (B) leaking into the tank was measured. The light blocking ratio was calculated as follows and summarized in Table 1.

Shading rate [%] =
{(Amount of light on sample side (A) [cps] −Amount of light leaking into empty tank (B) [cps])
÷ Sample-side light intensity (A) [cps]} × 100
Even when using titanium dioxide with good shielding properties as the light-reflective component and using a white container in which the pigment kneading rate is increased to 6% near the molding density limit, the amount of leakage light is as high as 944 and 105 cps, and the light shielding rate is Only 93.6774% is obtained. On the other hand, in black, # 1, # 2, and # 3 (the mixing ratio of carbon black that is a light absorbing component is 1.00, 0.09, 0.048, and 0.024%, respectively) In any case, the amount of light leaked into the empty tank was 30 to 41 cps, which was as low as the dark count of the detector 1, and the light shielding rate showed a good value of 99.997% or more. # 4 in which the blending ratio of carbon black, which is a light absorbing component, was reduced to 0.012% showed a leakage light amount of 1,611 cps, and the light blocking ratio was 99.9662%.

  From the above results, in order to ensure the same level of light shielding rate as that of the black container (that is, 99.99% or more), it is sufficient to incorporate a very small amount of carbon black into the white pigment, and the blending ratio is 0.024%. It turns out that it is above. On the other hand, in the effect of enhancing the amount of luminescence by the enzymatic reaction of ALP, all of # 1, # 2, and # 3 obtained a high signal compared with the black container, and among them, the necessary minimum amount of carbon black was blended. # 3 was the highest. # 4, White 1 and White 2 (especially white containers) have a high signal enhancement effect, but the light shielding properties are poor, light emission leaks, and the measured value becomes false high due to leakage of noise light from outside the container I understand that there is a fear.

実施例２
実施例１において観察された容器外からのノイズ光の漏れ込みが、実際の発光測定においてどの程度の偽高値に相当するのかを評価するために、無着色、黒色、＃２、＃４、白色２のそれぞれの容器を用いて実際に甲状腺刺激ホルモン（ＴＳＨ）の発光測定を行った。

Example 2
In order to evaluate how much false high value the leakage of noise light from outside the container observed in Example 1 corresponds to in actual light emission measurement, no coloring, black, # 2, # 4, white The luminescence measurement of thyroid stimulating hormone (TSH) was actually performed using each of the two containers.

1. Preparation of anti-TSH antibody-immobilized particles Take 10 mg of carboxylated microparticles (microparticles having a carboxy group on the particle surface and a diameter of about 2.5 μm) in a 1.5 mL tube, and use 1 mL of 10 mM MES buffer (pH 6.0). After washing, 1 mL of 10 mM MES buffer (pH 6.0) containing 10% N-ethyl-N ′-(dimethylaminopropyl) carbodiimide (EDC) was added, and the mixture was shaken and stirred at room temperature. After removing the supernatant, it was washed with 1 mL of 10 mM MES buffer (pH 6.0), sonicated, 100 μL of 1 mg / mL antibody solution was added, and the mixture was shaken and stirred at room temperature. After washing with 1 mL of 0.1 M Tris hydrochloric acid buffer (pH 8.0) containing 0.1% sodium azide, 0.1 M Tris hydrochloric acid buffer containing 0.1% BSA and 0.1% sodium azide Blocked at (pH 8.0) and stored in the same buffer.

2. Measurement of TSH Take 10 μL of 0.4% anti-TSH antibody-immobilized particles, 10 μL of TSH sample, and 40 μL of surfactant-containing purified water in a container for luminescence measurement, stir, and then a 2-step sandwich at 37 ° C. for 5 minutes. The first reaction of the method was performed. After the reaction solution was removed by suction, the reaction solution was first treated with 10 mM Tris hydrochloric acid buffer solution (containing 150 mM NaCl, 0.05% Tween-20, 1 mM MgCl ₂ , 0.1% sodium azide) (pH 8.0). Washing was performed. 50 μL of 2.8 μg / mL ALP-labeled anti-TSH antibody was added, and after stirring, the second reaction was performed at 37 ° C. After the second washing, 0.4 μL of a 0.4 mM ALP chemiluminescence substrate solution was added and stirred, reacted at 37 ° C. for 5 minutes, and endpoint photometry was performed.

  Table 2 shows a comparison between the TSH concentration and the count value. The response curves of TSH using the black container and # 2 container are shown in FIGS. 2 and 3, respectively. In this example, a low-concentration TSH sample near the measurement lower limit concentration was used as the sample. In the amount of leakage light in the result of Example 1, 1,611 cps of # 4, which is a container with a light shielding rate of less than 99.99%, has a TSH concentration of about 0.004 μIU / mL or more plus error (0.0021 μIU / mL × 1, 611 / (1,323-523) = 0.0042 μIU / mL), 944,105 cps of the white container 2, which is also a container with a light shielding rate of less than 99.99%, is a positive error of 1.37 μIU / mL or more (0.0021 μIU /ML×944,105/(2,284-839)=1.372 μIU / mL). As a matter of course, there is no upper limit to the noise light that shines outside the container, and the greater the amount of light, the higher the degree of positive error in the measured value. If noise light that cannot be controlled in this way suddenly leaks and the measured value fluctuates, the reliability of measurement in the low concentration region is lowered, which is a serious problem.

  Also, the black color produced in Example 1 by the 2SD method, that is, the standard deviation (SD) calculated by measuring a zero concentration sample a plurality of times and the slope of the calibration curve, are calculated from the TSH detection lower limit concentration (MDC). As a result of using the container and the # 2 container, the slope of the response curve using the # 2 container is about twice as large. In the MDC, the black container is 0.00017 μIU / mL, # When 2 containers are used, 0.000089 μIU / mL is obtained. From this result, it can be seen that the # 2 container can measure the sensitivity twice as high.

DESCRIPTION OF SYMBOLS 1 Luminescence detector 2 Luminescence measuring container 3 Enzyme reaction liquid 4 Aluminum foil (seal)

Claims (9)

A container for light emission measurement made of a thermoplastic resin, containing 0.5 to 10% by weight of a light-reflective component and 0.01 to 0.5% by weight of a light-absorbing component, and colored and molded in gray or chromatic color. The light emission measuring container according to claim 1, wherein the light reflective component is a white pigment. The container for luminescence measurement according to claim 2, wherein the white pigment is titanium dioxide. The container for light emission measurement of Claim 1 whose said light absorptive component is a black pigment. The luminescence measurement container according to claim 4, wherein the black pigment is carbon black. A container for light emission measurement made of a thermoplastic resin, containing 1 to 6% by weight of titanium dioxide and 0.02 to 0.1% by weight of carbon black. The container for luminescence measurement according to any one of claims 1 to 6, comprising a plurality of openings. A luminescence measurement container having a plurality of openings,
The amount of light (A) [cps] from the luminescent solution detected from the opening when the luminescent solution is put into one of the openings of the luminescence measurement container, and one of the openings of the container From the amount of light (B) [cps] from the luminescent solution detected from the opening adjacent to the opening when the luminescent solution is added, equation (1)
[{(A)-(B)} ÷ (A)] × 100 Formula (1)
The shading rate calculated by is 99.99% or more,
The light quantity (A) [cps] is the light quantity [cps] detected from the luminescent solution when the luminescent solution is put into the opening of a container containing 1.0% by weight of black pigment. ], A container for measuring luminescence larger than. A luminescence measurement method, wherein a luminescent solution is placed in one of the openings of the luminescence measurement container according to claim 7 or 8, and light emission from the luminescent solution is detected from above the opening.

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