JP2003004629A - Microplate and method for measuring emission of light - Google Patents

Abstract

PROBLEM TO BE SOLVED: To realize a microplate which can detect/measure faint emission of fluorescence or chemiluminescence efficiently by suppressing crosstalk effectively. SOLUTION: A plurality of wells Wij for containing a light emitting sample are formed regularly according to a specified arrangement wherein at least the bottom part of each well transmits light and light emitted from each well is measured through the light transmitting bottom part thereof. In such a microplate, a lens face Lzij having a positive power is formed on the bottom part of each well Wij .

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a luminescence measuring microplate and a luminescence measuring method.

[0002]

2. Description of the Related Art A "fluorescent sample that emits fluorescence by excitation light" or "a chemiluminescent sample that emits light by a chemical reaction" is placed in each well of a microplate in which small wells (recesses) are formed to emit light and emit light. Luminescence measurements are known in which light is measured on the bottom side of a microplate. For example,
As a fluorescence measuring device for measuring the fluorescence generated by the excitation light, the one described in JP-A-10-281994 is known.

Since the fluorescence and chemiluminescence generated in the well are generally weak, how to efficiently detect and measure such weak light emission becomes a problem.

Further, since fluorescence and chemiluminescence have no directivity, light from neighboring wells is mixed with light to be detected,
So-called "crosstalk" that reduces the S / N ratio of the detection signal
The problem of cannot be ignored.

[0005]

SUMMARY OF THE INVENTION An object of the present invention is to realize a novel microplate capable of effectively reducing crosstalk. Another object of the present invention is to realize a novel microplate capable of efficiently detecting and measuring weak luminescence such as fluorescence and chemiluminescence.

Another object of the present invention is to realize a luminescence measuring method using the novel microplate.

[0007]

The microplate of the present invention has a plurality of wells for accommodating a luminescent sample, which are regularly formed in accordance with a predetermined arrangement, and at least the bottom of each well is transparent and Microplate for measuring the light emission in the wells through the light transmissive bottom. " The luminescence that is the target of luminescence measurement is “fluorescence, chemiluminescence, or the like”.

The microplate according to claim 1 is characterized in that "a lens surface having a positive power is formed at the bottom of each well". The light emitted in the well is measured as transmitted through the light-transmissive bottom of each well as described above. The lens surface formed on the bottom of each well is
The positive power suppresses "chaotic divergence" and imparts directivity, which enables efficient detection.

In the luminescence measurement microplate according to the first aspect, the "opening edge portion for reducing crosstalk" can be formed on the light emission side surface of each well (the second aspect). The "opening edge" is formed so that its inner wall surface surrounds the effective diameter of the lens surface.

When the opening edge portion is formed in this manner, the inner wall surface of the opening edge portion can be provided with "a taper having a large taper angle such that the opening diameter increases in the light emission direction" (claim). 3). A "taper with a large taper angle" is a taper having a taper angle of 10 degrees or more, preferably 15 degrees or more. For example, in the case where the microplate is formed as a resin molded product, the taper having a large taper angle as described above makes the die cutting “easier”.

The opening edge effectively reduces the "light emitted from the bottom of the well causing crosstalk towards the adjacent well". In the microplate for luminescence measurement according to claim 2 or 3, which has such an opening edge, in order to further enhance the effect of crosstalk reduction by the opening edge, "a reflective film is formed on the inner wall surface of the opening edge. Can be formed ”(claim 4). Such a reflective film also promotes the directivity of light emitted from each well (given by the lens surface).

In the luminescence measurement microplate according to claim 2 or 3 or 4, "a light-shielding layer can be formed on the exit-side end face of the opening edge" in order to further reduce crosstalk. Item 5).

In the microplate for luminescence measurement according to any one of claims 1 to 5, the "lens surface having a positive power" formed on the bottom surface of each well is the inner surface of the well bottom surface. Although it may be formed on the outer side and / or on the outer side, it is preferable that the lens surface is formed on the outer side of the well bottom, that is, on the “light emitting side surface” in view of ease of forming the lens surface.

In the microplate according to claim 7, an opening edge portion for reducing crosstalk is formed on the surface of each well on the light emission side so that the inner wall surface surrounds the emission opening diameter. Is characterized by. That is, in this case, no lens surface is formed.

In the microplate for luminescence measurement according to the present invention, the inner wall surface of the opening edge portion is provided with "a taper having the above-mentioned large taper angle such that the opening diameter increases in the light emission direction". be able to. In addition, claim 7
Alternatively, in the luminescence measurement microplate according to the eighth aspect, a reflection film can be formed on the inner wall surface of the opening edge portion (claim 9).

Since the microplate according to claims 7 and 8 has no lens surface, the light emitted from the well is not given directivity by the lens surface. However, the reflection film formed on the inner wall surface of the opening edge is The light emitted from the well can be given a little directivity, and the efficiency of luminescence detection is promoted.

In particular, when a reflective film is formed on the inner wall surface of the opening edge portion where a taper with a large taper angle is formed, directivity is given to the light that is likely to cause crosstalk, and crosstalk can be effectively reduced. The same applies to the microplate according to claim 4.

Also in the luminescence measuring microplate according to claim 7 or 8 or 9, the crosstalk can be further reduced by forming a light-shielding layer on the exit side end face of the opening edge. it can.

In the microplate for luminescence measurement according to any one of claims 1 to 10, it is needless to say that the plurality of wells can be uniformly formed over the "entire region of the predetermined area" of the microplate. , Claim 1
Multiple wells may be "grouped into a fixed number of wells," such as those described in 1.

That is, in the microplate according to claim 11, a large number of wells for accommodating the luminescence sample
(≧ 2) are divided into groups to make “well groups”.

On the other hand, a plurality of large wells are regularly formed in accordance with a predetermined "large well arrangement", and the bottom of each large well is
The n wells of a 1-well group are regularly formed according to a predetermined "well arrangement".

In the luminescence measuring microplate according to any one of claims 1 to 11, the main part can be formed as a molded product of transparent resin (claim 12). Here, the “main part of the microplate” means a part excluding the reflection film and the light shielding layer.

The luminescence measuring method of the present invention is the above-mentioned claim 1.
The method for measuring luminescence using a microplate according to any one of 1 to 12, wherein an optical fiber that guides light emitted from a well to a detection unit is provided at a predetermined distance from the incident end face to the bottom face of the microplate. The wells are switched while being relatively displaced with respect to the microplate ”(claim 13).

In the case of the luminescence measuring method according to the thirteenth aspect, it is possible to "use a minute positive lens fixed to the incident end face of the optical fiber so as to enhance the light converging property".
4) In this case, it is further possible to use "a microplate in which a lens surface having positive power and an opening edge portion for reducing crosstalk are formed for each well" (claim 15).

Of course, when each of the microplates described above is used, it is needless to say that a plurality of optical fibers may be used to detect the light emission in each well at the same time, and the microplate according to claim 11. In the case of using “a plurality of optical fibers, light emission from n wells in one large well is detected at the same time, and the plurality of optical fibers are integrally displaced relative to the microplate, The wells can be switched ”.

Alternatively, the microplate according to claim 11 is used, one optical fiber is used for each large well, and the light emission from one well is detected for each large well at the same time. It is also possible to perform relative switching with respect to the microplate as a unit so that the wells are switched within each large well.

[0027]

DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 is a diagram for explaining one embodiment of a microplate of the present invention. The microplate 1 for measuring luminescence is the microplate according to claim 11, and the main part is formed as a molded product of transparent resin (claim 12).

FIG. 1A is a plan view of the microplate 1. Since the microplate 1 has a plate shape with four corners rounded and is formed as a molded product, the peripheral portion is tapered at a taper angle of 2 degrees for "die cutting" over the entire circumference. By the way, the size of the microplate 1 is 128 mm in the horizontal direction in the figure and 86 m in the vertical direction.
m, the thickness is 10 mm.

The microplate 1 has a large well GW.
₁₁, GW₁₂, GW_Thirteen... GW ₂₁... GW
_ij,,, are formed in a matrix according to the large well array.
Is made. Each large well GW_ijHave the same shape
Specifically, it is a circular shape with a diameter on the front side of the drawing of 8 mm.
Twelve in the horizontal direction and eight in the vertical direction at an array pitch of mm.
A total of 96 pieces are formed.

The inner wall of the large well GW _ij is tapered at a taper angle of 2 degrees for die cutting. A well group wg _ij is formed at the bottom of each large well GW _ij . FIG. 1B shows an arbitrary large well GW _ij.
FIG. 1 (c) is a plan view of FIG.
-C sectional drawing is shown.

As shown in FIG. 1C, the large well GW
_ij has a depth of 8. from the opening (the left end in the figure) to the bottom.
The inner wall surface is 45 mm and the taper angle is 2 degrees as described above.

As shown in FIG. 1B, the well group wg _ij formed at the bottom of each large well GW _ij is formed by regularly arranging wells W _ij for accommodating a luminescent sample. The wells W _ij have the same shape, and the shape shown in the plan view is circular. Well _{W ij} that make up the well group
The array (well array) is in the form of a matrix as shown in the figure, and 30 wells each have 6 wells in the horizontal direction and 5 wells in the vertical direction.
A total of 30 pieces are formed. These wells _Wij
The arrangement pitch is 1 mm in both the vertical and horizontal directions.

FIG. 1C is an enlarged view of a portion indicated by reference symbol D in FIG. 1C, which shows a cross-sectional shape of each well W _ij . Each well W _ij is a large well GW
drilled in the _ij bottom _{GW ij} (bottom), diameter 0.7mm of the bottom of the well _{W ij} (circular), depth 0.8mm
The taper angle of the taper attached to the inner wall is 5 degrees.

The emission surface side of the bottom of the well W _ij (see FIG. 1)
On the right side surface of (d), a lens surface Lz _ij having the same shape is formed for each well. The lens surface Lz _ij is convex and has positive power, and the one in the description has a radius of curvature of 0.6.
mm, lens effective diameter: 0.8 mm.

An opening edge E for reducing crosstalk is formed so as to surround the effective diameter of each lens surface Lz _ij . The height of the opening edge E is 0.25 mm
On the inner wall surface of the opening edge E, a taper for die cutting is formed with a taper angle of 5 degrees so that the opening becomes larger toward the light exit side. The well groups wg _ij formed in the large well GW _ij are the same. The opening shape of the opening edge E is circular when the state of FIG. 1D is viewed from the right side.

In the embodiment shown in FIG. 1, each well W _ij has a small diameter of about 0.7 mm and a depth of about 0.8 mm. _Therefore , each well W _ij is called a "microwell or microcell" instead of being called a well. Can also be called. Lens surface Lz _ij also has a lens effective diameter: 0.8 mm
Since it is small, it can be called a "microlens surface".

That is, in the microplate 1 of which the embodiment has been described with reference to FIG. 1, a plurality of wells W _ij for accommodating a luminescent sample are regularly formed according to a predetermined arrangement, and at least each well is formed. In a microplate having a light-transmitting bottom and for measuring light emission in each well through the light-transmitting bottom, a lens surface Lz _ij having a positive power is formed for each bottom of each well W _ij . The opening edge portion E for reducing crosstalk is formed on the surface of each well W _{ij on} the light emission side so that the inner wall surface surrounds the effective diameter of the lens surface Lz _ij. It is formed (claim 2). Further, each lens surface Lz _ij is formed on the light emitting side surface (claim 6).

Further, a large number of wells for accommodating the luminescent sample are divided into n (= 30) groups into a well group wg _ij, and a plurality of large wells GW _ij are regularly arranged according to a predetermined large well arrangement. The n wells W _{ij of} one well group are regularly formed at the bottom of each large well GW _ij according to a predetermined well arrangement (claim 11).

2 and 3 show the morphology (type) of wells that can be formed in the microplate of the present invention.
Type: a to type: g shown in FIG. 2 are types of wells belonging to the microplate of claim 6.

Type: a is "a lens surface Lz having a positive power is formed on the light emitting side surface at the bottom of each well"
Is. The type: b is "a lens surface Lz having positive power and an opening edge E formed on the bottom surface of each well on the light emitting side surface" (claim 2). Type: c has an opening edge E1 having a taper with a large taper angle such that the opening diameter increases toward the light emission direction (downward in the drawing) (claim 3). And Figure 1
Each well in the embodiment shown in FIG.

The type: d is a type in which the reflective film Rf is formed by vapor deposition on the inner wall surface of the opening edge E in the type: b (claim 4), and the type: e is the opening edge E in the type: d.
A light-shielding layer S is formed on the end face portion on the emission side of (5)
Is. The light shielding layer S can be formed by, for example, “application or printing of black ink”.

The type: f is one in which a reflective film Rf1 is formed by vapor deposition on the inner wall surface of the opening edge E1 in the type: c (claim 4), and the type: g is the injection of the opening edge E1 in the type: f. The light-shielding layer S is formed on the side end face portion (Claim 5).

The well types shown in FIG. 3 are: h-type:
m is the type of well formed in the microplate according to claims 7-10. Type: h is "a surface on the light emission side of each well, in which an opening edge portion E for reducing crosstalk is formed so that the inner wall surface surrounds the emission opening diameter (claim 7)" And the type: i is "the opening edge portion E1 for reducing crosstalk is formed on the light exit side surface of each well so that the inner wall surface surrounds the exit opening diameter. The inner wall surface has a taper with a large taper angle such that the opening diameter increases in the light emission direction (claim 8).

The type: j is a type in which a reflective film Rf is formed on the inner wall surface of the opening edge E in the type h (claim 9), and the type: k is the opening edge E in the type: i.
The reflective film Rf1 is formed on the inner wall surface of No. 1 (claim 9).

Further, the type: l is a type in which the light shielding layer S is formed on the exit side end surface portion of the opening edge E in the type: j (claim 10), and the type: m is the opening edge in the type: k. The light shielding layer S is formed on the exit side end surface of the portion E1 (claim 10).

FIG. 4 and FIG. 5 are views for explaining an embodiment of the light measuring method. In these figures, reference numeral 10 is a microplate, reference numeral WL is a well, reference numeral F.
B indicates an optical fiber. Further, in FIG. 5, reference numeral ML indicates a “small positive lens”.

As shown in FIGS. 4 and 5, the microplate 10 has wells WL for containing the luminescent sample 0.
Are regularly formed according to a predetermined arrangement, at least the bottom of each well is light-transmissive, and the light emission in each well is measured through the light-transmissive bottom. A lens surface Lz having a positive power is formed on each bottom.

The luminescent sample 0 is contained in each well WL, and the reagent is supplied from the upper part of the well WL. Reagent is well W
Upon contact with the luminescent sample in L, “chemiluminescence or fluorescence” is emitted.

In the luminescence measuring method shown in FIG. 4, the well WL is
By displacing the optical fiber FB that guides the light emitted from the detector to the detection unit, the distance between the incident end surface (the end surface on the microplate side) and the bottom surface of the microplate is maintained at a predetermined distance and the microplate 10 is relatively displaced. , Well WL
Switch.

In the issue measuring method of FIG. 5, the optical fiber FB is used.
A small positive lens M for increasing the light collecting property on the incident end face of
L is fixed and used.

As shown in FIGS. 4 and 5, the optical fiber FB
Thus, by "scanning" the array of the wells WL of the microplate 10 and detecting the light emission, the light emission for each well can be measured.

As an example of measurement, "gene detection by hybridization method" will be described.
In the above, in each well WL of the microplate 10, different oligo probes (DNA having a length of several bases to several tens of bases and a known base sequence) are housed and labeled with a fluorescent molecule as a reagent. The sample solution containing the target nucleic acid "is provided to the well group. When the target nucleic acid in the sample solution binds to the oligo probe, it emits fluorescence.

Therefore, when the well scanning is performed by the optical fiber FB and the well WL emitting fluorescence is specified by the method shown in FIG. 4 or FIG. 5, the base sequence of the oligo probe accommodated in this well becomes the target nucleic acid. You know that you have. In this way, it is possible to efficiently know what base sequence the target nucleic acid in the sample liquid contains.

[0054]

EXAMPLES The inventors conducted a simulation to evaluate the characteristics of luminescence measurement when using the microplate of the present invention. The simulation conditions are as follows. As the microplate, the same one as described with reference to FIG. 1 was assumed. That is, it is assumed that the microplate is made of a resin material (polycarbonate, refractive index for light having a wavelength of 520 nm: 1.562498). As the well, various types shown in FIGS. 2 and 3 were assumed. FIG. 6 illustrates the positional relationship between the four types of wells and the optical fiber FB. The wells labeled Ref are "for comparison", and the types: a, b, and c are all described with reference to FIG.

Well: Ref has a bottom of the well having a diameter of 0.
It has a circular shape of 7 mm, and this bottom surface shape is the same for other types of wells. Further, the thickness of the well bottom portion in the well: Ref (the distance from the well bottom portion to the emission surface) is 0.75 mm.

In the types: b, c and the types: d to g in FIG. 2, the height of the apex of the lens surface Lz and the exit side end face of the opening edges E, E1 are the same. Also in the types: a to g, the distance from the well bottom to the lens surface top or the exit side end face of the opening edge is equal to the thickness of the bottom of the well Ref (0.75 mm).

Also, in the type: h to m having only the opening edge E or E1 without the lens surface shown in FIG. 3, the distance from the well bottom to the exit side end surface of the opening edge is the well: It is equal to the thickness of the bottom of Ref (0.75 mm).

In the types: a to g, the radius of curvature of the lens surface Lz is 0.6 mm, the lens thickness is 0.25 mm, and in the types: a to m, the height of the opening edge is 0.25 mm.
And And for each type of well, the optical fiber F
It is assumed that the end surface of B is located at a distance (distance: DS) of FIG. 6 from the lens surface top portion or the exit side end surface portion of the opening edge portion: 0.3 mm, and the center axis of the optical fiber and the center of the well. The axes were supposed to match.

As the type of light emission, "wavelength: 520 nm fluorescence" is assumed, and it is assumed that this fluorescence is surface-emitted with uniform light intensity at the bottom of the well, and the surface-emitted fluorescence is a complete diffused light. . When the amount of light emitted from the uniform light emission per unit time at the bottom of the well is 100% (because it is completely diffused light, the light that propagates to the optical fiber side is 50%
, And the amount of light detected through the optical fiber FB: X
% Was defined as "light collection efficiency", and this light collection efficiency: X was obtained by simulation.

The amount of light emitted from adjacent wells detected by the optical fiber FB: Y% was determined as "crosstalk". As described with reference to FIG. 1B, the well array is a square matrix, and each well has 8 wells.
Since it is surrounded by the adjacent wells, the light amount: 8Y% is obtained as the maximum value of the crosstalk, and the light amount: X and the light amount:
Ratio with 8Y: X / (8Y) was defined as "S / N ratio".

A representative example of the simulation result is shown in a list. The well type Ref is shown in the leftmost diagram of FIG. 6, and the other types are shown in FIGS. 2 and 3. Further, the taper angle of the inner wall portion of the opening edge portion E1 in the types: c, f, g, i is 22.5 degrees, and the type:
The reflectance of the reflective film Rf1 in f and g was 100%, and the transmittance of the light shielding layer S in type: g was 0%.

The optical fiber has a core diameter of 735 μm,
Outer diameter: 750 μm plastic optical fiber (refractive index of core part: 1.495342, refractive index of clad part: 1.
422831) was assumed.

[0063]   [simulation result] Well type Light collection efficiency: X Crosstalk: Y 8Y S / N     Ref 7.7 0.9 7.2 1.07   Type: a 9.7 0.7 5.6 1.73   Type: b 9.6 0.5 4.0 2.40   Type: c 9.7 0.6 4.8 2.02   Type: h 7.8 0.5 4.0 1.95   Type: i 7.7 0.6 4.8 1.61   Type: f 11.6 0.6 4.8 2.42   Type: g 11.6 0.5 4.0 2.90.

As is clear from these results, by forming the lens surface on the bottom surface of the well as compared with the Ref type well, the light collection efficiency is improved by about 2% (type:
a, b, c) can be seen. As can be seen by comparing Ref type wells and types: b, h, and i, the crosstalk is 0.3 to 0.4% by forming the opening edge portion.
It goes down.

Ref type wells and types: c, i,
As can be seen by comparing f and g, providing a “taper with a large taper angle” on the inner wall surface of the opening edge portion and forming a light shielding layer on the exit side end surface portion of the opening edge portion have the effect of reducing crosstalk. Therefore, it can be seen that forming the reflection film on the inner wall surface has the effect of increasing the light collection efficiency.

The inventors have further described the type: b described above.
Assuming a microplate having a well of, as shown in FIG. 5, a simulation similar to the above is performed for the case where “a small positive lens ML for enhancing the light collecting property is fixed to the incident end surface of the optical fiber FB”. I did. The optical fiber is the same plastic optical fiber as described above.

As the positive lens ML, the paraxial radius of curvature: 0.
4 mm, aspherical coefficient (cone constant): K = -0.43, lens thickness: 0.9 mm, effective diameter: 0.9 mm, material: polycarbonate, assuming that the exit side (flat surface) of this lens is light The simulation was performed by fixing the fiber to the end face of the fiber and setting the distance between the top of the lens ML and the top of the lens surface Lz on the microplate side to 0.3 mm. Well type Light collection efficiency: X Crosstalk: Y 8Y S / N type: b 6.50 0.14 1.14 5.68 The result is the type in the above “when the positive lens ML is not used”: b When compared with the case of, the positive lens M
When L is used, the light collection efficiency: X is 3.1% lower than that when the positive lens ML is not used, but the crosstalk:
Y is greatly reduced to 0.36%, resulting in S / N
The ratio is 2.4 to 5.68, which is 2.4 times higher. That is, it can be seen that fixing the positive lens to the end face of the optical fiber for use has the effect of increasing the S / N ratio.

[0068]

As described above, according to the present invention, a novel microplate and luminescence measuring method can be realized. According to the present invention, it is possible to effectively reduce crosstalk and efficiently detect and measure weak luminescence such as fluorescence or chemiluminescence. The “opening shape of the opening edge portion” is not limited to the circular shape described above, and may be a polygonal shape such as a square shape or a hexagonal shape.

[Brief description of drawings]

FIG. 1 is a diagram for explaining an embodiment of a microplate.

FIG. 2 is a diagram showing types of wells formed in a microplate.

FIG. 3 is a diagram showing types of wells formed in a microplate.

FIG. 4 is a diagram for explaining a luminescence measuring method.

FIG. 5 is a diagram for explaining a luminescence measuring method.

FIG. 6 is a diagram for explaining simulation conditions.

[Explanation of symbols]

1 Microplate GW _ij Large well wg _ij Well group W _ij Well E Opening edge

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Kenichi Ishizuka             109, Ohata 10th District, Hanako, Iwate Prefecture, Rico             -In Optical Co., Ltd. (72) Inventor Yasushi Takahashi             109, Ohata 10th District, Hanako, Iwate Prefecture, Rico             -In Optical Co., Ltd. F-term (reference) 2G043 CA03 DA02 DA06 EA01 EA13                       GA04 GB19 HA01 HA05 KA02                       KA05 MA01                 2G057 AA04 AA14 AC01 BA03 BB06                       DA01 DA03 DA04 DB01 DB05

Claims (15)

[Claims] 1. A plurality of wells for accommodating a luminescent sample are regularly formed in accordance with a predetermined arrangement, at least the bottom of each well is light-transmissive, and the emission of light in each well is defined by the light-transmissive bottom. A microplate for luminescence measurement, wherein a lens surface having a positive power is formed at the bottom of each well in the microplate for measurement via. 2. The microplate for luminescence measurement according to claim 1, wherein an opening edge portion for reducing crosstalk is formed on the surface of each well on the light emission side, and an effective diameter of the lens surface is formed on the inner wall surface thereof. A microplate for luminescence measurement, which is formed so as to surround the microplate. 3. The microplate for luminescence measurement according to claim 2, wherein the inner wall surface of the opening edge portion has a taper with a large taper angle such that the opening diameter increases in the light emission direction. A microplate for luminescence measurement characterized by. 4. The microplate for luminescence measurement according to claim 2 or 3, wherein a reflective film is formed on the inner wall surface of the opening edge portion. 5. The microplate for luminescence measurement according to claim 2, 3 or 4, wherein a light-shielding layer is formed on the exit side end face portion of the opening edge portion. 6. The microplate for luminescence measurement according to any one of claims 1 to 5, wherein each lens surface is formed on a light emitting side surface. 7. A plurality of wells for accommodating a luminescent sample are regularly formed in accordance with a predetermined arrangement, at least the bottom of each well is light-transmissive, and the emission of light in each well is defined by the light-transmissive bottom. In the microplate for measurement via the light emitting side of each well, an opening edge portion for reducing crosstalk is formed so that the inner wall surface surrounds the emitting opening diameter. A microplate for luminescence measurement. 8. The microplate for luminescence measurement according to claim 7, wherein the inner wall surface of the opening edge portion has a taper with a large taper angle such that the opening diameter increases in the light emission direction. A microplate for luminescence measurement characterized by. 9. The microplate for luminescence measurement according to claim 7 or 8, wherein a reflection film is formed on the inner wall surface of the opening edge portion. 10. The microplate for luminescence measurement according to claim 7, 8 or 9, wherein a light-shielding layer is formed on the exit side end face portion of the opening edge portion. 11. The microplate for luminescence measurement according to any one of claims 1 to 10, wherein a number of wells for accommodating a luminescence sample are n (≧ 2).
The individual wells are grouped into well groups, and a plurality of large wells are regularly formed according to a predetermined large well arrangement. At the bottom of each large well, n wells of one well group are formed according to a predetermined well arrangement. A microplate for luminescence measurement characterized by being formed regularly. 12. The microplate for luminescence measurement according to any one of claims 1 to 11, wherein a main part is formed as a molded product of transparent resin. 13. A method for measuring luminescence using a microplate according to any one of claims 1 to 12, wherein an optical fiber for guiding light emitted from a well to a detection part is provided on the incident end face and the bottom face of the microplate. A luminescence measuring method, characterized in that the wells are switched by maintaining a predetermined distance to the microplate and displacing the well relative to the microplate. 14. The luminescence measuring method according to claim 13, wherein a fine positive lens for improving light collecting property is fixed and used on the incident end face of the optical fiber. 15. The luminescence measurement method according to claim 14, wherein a microplate having a lens surface of positive power and an opening edge portion for reducing crosstalk is formed for each well. Luminescence measurement method.