Container structure

Abstract

To reduce an influence in which recurrent light of light transmitting through spots and scattering forward becomes background light noise.SOLUTION: A container structure is mounted on an array plate having a top face on which a spot array including tissue-derived substances is formed, and configured to form a container that has the array plate as a bottom plate and can retain liquid. The container structure has a bank part that forms side walls of the container when mounted on the array plate, and a recurrent light extinction part that is arranged opposite to the array plate and reduces a situation where light passing through the spot array makes a recurrence incident on the spot array.SELECTED DRAWING: Figure 2

Description

[Technical Field]

[0001] The present invention relates to a container structure used to supply and hold liquids such as liquid specimens and chemical liquids to an array plate in a specimen analyzer. [Background technology]

[0002] Array plates, such as protein arrays, peptide arrays, and DNA arrays, are known, in which numerous substances, such as proteins, peptides, and nucleic acids, are fixed in the form of spots on a substrate. By using array plates in sample analysis, it is possible to simultaneously observe the interactions between numerous substances fixed on the array plate and substances in the sample. This allows for comprehensive analysis of the interactions between numerous substances and liquid samples of biological origin, such as blood, cell extracts, saliva, and interstitial fluid.

[0003] Another known sample analysis method using an array plate involves selectively fluorescently labeling spots where an interaction of interest has occurred to obtain optical information. A known device for observing fluorescently labeled samples is the confocal laser microscope. The confocal laser microscope has an illumination optical system, a fluorescence detection optical system, and a two-dimensional scanning system. The fluorescence detection optical system has the function of detecting the amount of fluorescent light from spots labeled with fluorescent probes. The two-dimensional scanning system has the function of acquiring a fluorescent image of the spot area on the array plate by two-dimensionally scanning the array plate or the optical system.

[0004] In Patent Document 1, a reaction process is carried out in which a frame is fixed to an array plate to allow liquid storage, and a pipette tip moves over the framed array plate to sequentially supply and drain multiple chemical solutions to cause reactions.Then, a specimen evaluation device is described that, after the reaction process is completed, performs optical scanning measurement while maintaining the liquid retention state, and acquires a fluorescent image of the spot area.

[0005] Patent Document 2 describes a chamber slide in which the bottom plate and the bank portion that forms the side are bonded together to prevent leakage of the chemical solution outside the holding portion. In this chamber slide, the cover is fixed to the bank portion with a snap mechanism, allowing the holding portion to be kept moist and warm. All of the components that make up this covered channel slide are made of materials that allow optical measurement from the outside, so it can also be used directly for fluorescence observation. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Patent Publication No. 2023-12426 [Patent Document 2] US Patent Application Publication No. 2013 / 017143 Summary of the Invention [Problem to be solved by the invention]

[0007] In the device configuration of Patent Document 1, optical scanning measurement involves irradiating fluorescently labeled spots with laser light from the underside of the array plate as excitation light and measuring the fluorescence emitted from them. However, the spots themselves are transparent, and much of the excitation light passes through the spots and scatters upward. Furthermore, because the top of the container that maintains the liquid holding state on the array plate is open, scattered laser light can leak out of the container, reflect off components outside the container, and return to the spots, potentially affecting the measurement light. Furthermore, when excitation light is irradiated around the spots, it can reflect off components outside the container and return to the array plate surface, potentially affecting the measurement of background light around the spots. In reaction measurement using an array plate with multiple fixed spots in an array, the amount of fluorescent light at each spot is calculated from the ratio or difference between the amount of fluorescent light at the spot itself and the amount of background light around it. Therefore, if unwanted reflected light caused by scattered light enters the vicinity of the spots, it may affect the measurement results. Meanwhile, the configuration shown in Patent Document 2 poses similar concerns even when a cover that allows optical measurement is attached to the top of the container. [Means for solving the problem]

[0008] The present invention provides a container structure that is attached to an array plate having an upper surface on which a spot array containing a biological substance is formed, and is configured to form a container that can store a liquid using the array plate as a bottom plate, a bank portion that forms a side wall of the container when attached to the array plate; a retroreflection attenuation unit disposed opposite the array plate and configured to reduce the retroreflection of light passing through the spot array back onto the spot array; The present invention provides a container structure having the above structure, thereby solving the above-mentioned problems. [Effects of the Invention]

[0009] According to the container structure of the present invention, in fluorescence detection measurement after a reaction process on an array plate, unnecessary light caused by scattered light that passes through the array plate when irradiated with laser light can be prevented from entering the array plate during measurement, thereby improving measurement accuracy. [Brief explanation of the drawings]

[0010] [Figure 1] 1A and 1B show a first embodiment of a container structure of the present invention, in which Fig. 1A is a perspective view showing the appearance of a container composed of a container structure and an array plate, and Fig. 1B is an exploded perspective view. [Figure 2] Fig. 2(a) shows a first embodiment of a container structure of the present invention. Fig. 2(a) is a vertical cross-sectional view showing a cross-section of a container composed of a container structure and an array plate, cut along a plane perpendicular to the longitudinal direction of the container. Fig. 2(b) is a vertical cross-sectional view showing a cross-section of the same container, cut along a plane perpendicular to the longitudinal direction of the container. Fig. 2(c) is a horizontal cross-sectional view of the container structure constituting the container, cut horizontally along line 2C-2C in Fig. 2(b), as viewed from below. Fig. 2(d) is a horizontal cross-sectional view of the container structure constituting the container, cut horizontally along line 2D-2D in Fig. 2(b), as viewed from below. [Figure 3] 2(b) shows a second embodiment of the container structure of the present invention, which is a vertical cross-sectional view showing a cross section corresponding to the cross section shown in FIG. [Figure 4] 2(b) shows a third embodiment of the container structure of the present invention, which is a vertical cross-sectional view showing a cross section corresponding to the cross section shown in FIG. [Figure 5] 2(b) shows a fourth embodiment of the container structure of the present invention, which is a vertical cross-sectional view showing a cross section corresponding to the cross section shown in FIG. [Figure 6] 2(b) shows a fifth embodiment of the container structure of the present invention, which is a vertical cross-sectional view showing a cross section corresponding to the cross section shown in FIG. [Figure 7] 7(a) and 7(b) are schematic diagrams showing the internal structure of the housing (enclosure) of a sample analyzer that uses a container composed of the container structure and an array plate of the present invention, where Fig. 7(a) is a schematic diagram of the inside of the device as seen from the front, and Fig. 7(b) is a schematic diagram of the inside of the device as seen from the side. DETAILED DESCRIPTION OF THE INVENTION

[0011] [First embodiment] A first embodiment of a container structure of the present invention will be described with reference to FIGS. FIG. 1(a) is a perspective view schematically illustrating the appearance of a container 1 formed by mounting a container structure 6 of this embodiment on an array plate 2. FIG. 1(b) is an exploded perspective view showing the individual components of the container 1 of FIG. 1(a). The container 1 is configured by engaging an array plate 2 with a container structure 6 including a bank portion 3, an opposing portion 4 facing the array plate 2, and a light-attenuating portion 5. The array plate 2 is a rectangular, flat glass slide, and its upper surface has an array region 2a in which a spot array is formed, each of which includes an array of multiple spots to which a biological substance is fixed. The bank portion 3 forming the side wall of the container structure 6 is formed of a resin material in the shape of a rectangular frame, surrounding a space. Groove-shaped recesses 3a are formed near the lower ends of the inner surfaces of three walls of the bank portion 3 and along the lower ends of the bank portions. The side edges of the array plate 2 slide into and engage with these recesses, thereby fixing the container structure 6 to the array plate 2, thereby forming the container 1. A chemical solution can be stored in the space within the container surrounded by the bank portion 3 and reacted with each spot arranged in the array region 2a on the upper surface of the array plate 2. Therefore, the dimensions of the recess 3a formed in the bank portion 3 are determined so that the side end or side edge of the array plate 2 fits tightly against the recess 3a and the chemical solution does not leak out from the bottom of the container 1. A rectangular flat plate-shaped opposing portion 4 having approximately the same external shape as the bank portion 3 is placed and fixed on the upper end surface of the bank portion 3. A light-attenuating portion 5 is fixed to the lower surface of the opposing portion 4.

[0012] FIG. 2 shows the configuration of the container 1 in more detail. FIG. 2(a) is a vertical cross-sectional view of the container structure 6 constituting the container 1, cut along a plane perpendicular to the longitudinal direction of the container (but excluding the array plate 2), as viewed from the front of the container (front, lower left in FIG. 1(a)). FIG. 2(b) is a vertical cross-sectional view of the container structure 6 constituting the container 1, cut along a plane perpendicular to the longitudinal direction of the container (but excluding the array plate 2), as viewed from the side of the container (longitudinal side, right side in FIG. 2(a)). FIG. 2(c) is a horizontal cross-sectional view of the container structure 6 constituting the container 1, cut horizontally along line 2C-2C in FIG. 2(b), as viewed from below the container structure. FIG. 2(d) is a horizontal cross-sectional view of the container structure 6 constituting the container 1, cut horizontally along line 2D-2D in FIG. 2(b), as viewed from below the container structure.

[0013] As shown in Figures 2(a) and 2(b), the lower surface of the facing portion 4 has a convex shape with a recessed outer periphery, which contacts the upper end surface of the bank portion 3. At the same time, a step fixing portion 4a, a vertical surface formed between the outer periphery and its inner region, contacts and fixes the upper end edge of the inner surface of the bank portion 3, thereby supporting the facing portion 4 on the bank portion 3. The attenuation portion 5 is a rectangular flat plate, and as shown in Figure 2(c), it is formed to be approximately the same size as the inner surface of the bank portion 3. The fixing method between the facing portion 4 and the attenuation portion 5 can be selected depending on the material used, such as shape-based fixing by adhesive bonding or fitting, or welding after installation. The internal space of the container 1 is formed between the attenuation portion 5 and the array plate 2, the inner surface of the bank portion 3, and the upper surface of the array plate 2. The chemical solution is stored in this internal space. As shown in Figure 2(d), an array region 2a is formed on the array plate 2, in which multiple spots 2b are fixed in a two-dimensional array.

[0014] Using the container of this embodiment, laser light is irradiated from below onto multiple spots 2b on the array plate 2, and the fluorescence emitted therefrom is measured. At this time, scattered light that passes through the array plate when the laser light is irradiated is irradiated onto the light-attenuating unit 5 arranged opposite the array plate. The light-attenuating unit 5 uses a resin plate that is absorbent for the wavelengths of the laser excitation light and the fluorescence, and by absorbing the scattered light irradiated onto the light-attenuating unit 5, the amount of light that is reflected and retro-enters the array plate 2 can be reduced. In particular, if light with high energy traveling in the forward direction, such as primary light or secondary light, among the transmitted scattered light, is reflected and retro-enters the array plate 2, it has a significant impact on the amount of fluorescent light to be measured, so the installation of the light-attenuating unit 5 is very effective.

[0015] In this embodiment, the area of ​​the attenuation section 5 is larger than the area of ​​the array region 2a. In addition, the area of ​​the attenuation section 5 projected onto the upper surface of the array plate 2 encompasses the area of ​​the array region 2a. This makes it possible to reliably absorb the transmitted scattered light in the attenuation section 5. Furthermore, even if the light spreads when irradiating the separated attenuation section 5 or if the incident angle of the light irradiated from the back surface of the array plate is slightly shifted, the scattered light can be absorbed.

[0016] As described above, by performing fluorescence detection measurement using a container 1 in which the container structure 6 of this embodiment is engaged with an array plate 2, it is possible to prevent unnecessary light caused by excitation light from entering during measurement, thereby improving measurement accuracy.

[0017] In this embodiment, the attenuation section 5 is preferably a film or a membrane formed by vapor deposition or the like that is absorptive of the wavelengths of the laser excitation light and the fluorescence, as a structure for attenuating the retroreflected light. Alternatively, it may be a metal plate that has been blackened by surface treatment such as black painting or black anodizing, a diffusion film, a plate with a textured surface, a light guide plate, or the like. In other words, any material that can prevent or reduce retroreflected light from reaching the array plate can achieve the same effect. Furthermore, the bank section 3 may have a shape other than a rectangular frame.

[0018] [Second embodiment] Fig. 3 shows a vertical cross-sectional view of a container using a container structure according to a second embodiment of the present invention, which corresponds to Fig. 2(b) in the first embodiment. In the following embodiments, members (parts) having the same functions as those in the first embodiment are given the same reference numerals, and detailed explanations thereof will be omitted.

[0019] In the first embodiment, the light-attenuating section was provided on the lower surface of the facing section 4, but in this embodiment, the light-attenuating section 7 is provided on the upper surface of the facing section 4. The facing section 4 is made of a transparent resin material that does not cause light to return to the array plate, or a glass plate. This configuration reduces the need to consider the resistance to the chemical solution stored in the light-attenuating section 7 and the effect on fluorescence after the labeling reaction, and allows a wide range of materials to be selected that have a high light-attenuating effect.

[0020] [Third embodiment] FIG. 4 shows a vertical cross-sectional view of a container using a container structure according to a third embodiment of the present invention, corresponding to FIG. 2(b) in the first embodiment. In this embodiment, the facing portion 8 combines the functions of the facing portion and the retroreflection attenuation portion of the first and second embodiments. The facing portion 8 is made of a resin material that is absorbent for the wavelengths of the laser excitation light and the fluorescence, and is fixed to the bank portion 3. This configuration integrates the facing portion and the attenuation portion, making it possible to reduce the number of parts.

[0021] [Fourth embodiment] FIG. 5 shows a vertical cross-sectional view of a container using a container structure according to a fourth embodiment of the present invention, corresponding to FIG. 2(b) of the first embodiment. In this embodiment, in addition to the configuration of the first embodiment, a light-attenuating section 9 is added to each inner surface of the frame-shaped bank section 3. Scattered light transmitted through the array plate 2 mainly irradiates the light-attenuating section 5 on the opposing side. Most of the light is absorbed, but some is reflected and scattered by this light-attenuating section 5, and this light may head toward the inner surface of the bank section 3. Reflection by this inner surface may cause light to retro-enter the array plate 2, affecting measurement. Therefore, by fixing a light-attenuating section 9 to each inner surface of the bank section 3 as in this example, the possibility of retro-entering light into the array region can be further reduced.

[0022] [Fifth embodiment] FIG. 6 shows a vertical cross-sectional view of a container using a container structure according to a fifth embodiment of the present invention, corresponding to FIG. 2(b) in the first embodiment. In this embodiment, the container structure 10 is configured by integrating the bank portion and the opposing portion, which function as a retroreflection attenuation portion, in contrast to the configuration of the fourth embodiment. The container structure 10 is configured by processing a resin material that is absorptive of the wavelengths of the laser excitation light and the fluorescence. The configuration of this embodiment allows for a reduction in the number of parts.

[0023] [Example of container structure use] An example of using the container structure of the present invention in a sample analyzer that performs optical measurement of biological materials on an array plate is shown. Figure 7 is a schematic diagram of the measurement system of a sample analyzer that uses the container structure of the present invention. Figure 7(a) is a front view, and Figure 7(b) is a right side view.

[0024] The sample analyzer 101 has an irradiation optical system, a fluorescence detection optical system, and a scanning system, all of which are covered by an enclosure 113. The irradiation optical system generates laser light of a required wavelength via a semiconductor laser (not shown), a collimating lens, and a bandpass filter in the excitation light generator 102. The laser light is reflected by a longpass filter 104 and redirected upward by a mirror 105. It is then focused by an objective lens 106 in the subsequent stage and irradiated as primary light onto the spot array from below the array plate 2 constituting the container 1. At this time, the container 1 is placed on a transport hand 111, which functions as a mounting unit, in the measurement chamber (optical measurement unit) of the sample analyzer 101. That is, the irradiation optical system of the sample analyzer 101 is configured so that the spot array to be observed is irradiated with laser light as primary light from a light irradiation unit located on the opposite side of the retroreflection attenuation unit provided on the container 1. Meanwhile, in the fluorescence detection optical system, the fluorescence emitted as secondary light from the spot fluorescently labeled by the illumination optical system travels along an optical path in the opposite direction to the illumination optical system. The fluorescence then passes through long-pass filter 104 and passes through an imaging lens and pinhole (not shown) in fluorescence receiving unit 103, before being detected by a detection unit equipped with a photomultiplier tube. As described above, in sample analyzer 101, a container including the container structure of the present invention, a mounting unit on which the container is mounted, and a light emitting unit constituting the illumination optical system are housed within enclosure 113.

[0025] The optical system is placed below the container 1 and performs reciprocating scanning in the X direction. A mirror base 107 carrying a mirror 105 and an objective lens 106 is connected to a guide block 108 and configured to be movable in the X direction along a guide rail 109 fixed to a base block 110. A linear motor, a motor rotary-to-linear conversion crank mechanism, or the like (not shown) can be used for driving. Meanwhile, a transport hand 111 carries the container 1 and is fixed to a drive mechanism 112 that is movable in the Y direction. A portion of the mounting portion of the transport hand 111 is open so that laser light can be irradiated from below the array plate 2.

[0026] The above configuration forms a moving unit capable of relative movement within the XY plane by combining the Y-directional movement of the transport hand and the X-directional movement of the optical system. This makes it possible to acquire a fluorescent image by two-dimensional scanning of the observation spot on the array plate 2. Note that inside the container 1 containing the array plate 2, a chemical solution 11 is stored on the upper surface of the array plate 2, and measurements can be performed while maintaining this state.

[0027] At this time, the container structure having a light-attenuating section inside the container reduces the diffused light that has passed through the array region on the array plate, thereby reducing the light that returns to the array plate and preventing the intrusion of unnecessary light that occurs when measuring the amount of fluorescent light, thereby improving measurement accuracy.

[0028] The present invention includes a container structure having the following configuration, as well as a container using the same and a sample analyzer using the same. (Configuration 1) A container structure that is attached to an array plate having an upper surface on which a spot array containing a biological substance is formed, and is configured to form a container that can store liquid using the array plate as a bottom plate, A container structure having a bank portion that forms a side wall of the container when attached to the array plate, and a retroreflection light attenuation portion that is arranged opposite the array plate and reduces the retroreflection of light that has passed through the spot array back into the spot array. (Configuration 2) The container structure of configuration 1, wherein the retroreflecting light attenuating portion is fixed to the bank portion. (Configuration 3) The container structure of configuration 1, wherein the retroreflecting light attenuating portion is supported by an opposing portion fixed to the bank portion. (Configuration 4) The container structure of configuration 1, wherein the retroreflecting light attenuation section is positioned at a distance from the spot array. (Configuration 5) The container structure of configuration 1, wherein the retroreflecting light attenuation section has an area larger than that of the spot array. (Configuration 6) A container structure according to configuration 1, wherein the area obtained by projecting the retroreflection attenuation portion onto the upper surface of the array plate contains the spot array. (Configuration 7) A container constructed from the container structure of any one of Configurations 1 to 6 and the array plate. (Configuration 8) a placement portion on which the container of configuration 7 is placed; a light emitting unit that is located on the opposite side of the retroreflection attenuation unit with respect to the spot array of the container placed on the mounting unit and that irradiates the spot array with primary light; an enclosure that houses the container, the placement portion, and the light emitting portion. (Configuration 9) The device of Configuration 8, further comprising a moving unit that moves the light emitting unit and the placing unit relative to each other. (Configuration 10) 9. The apparatus of configuration 8, further comprising a detector located on the opposite side of the spot array from the retroreflecting light attenuating unit, for detecting secondary light from the spot array. [Explanation of symbols]

[0029] 1 container 2 Array Plates 2a Array region 2b Spot 3 Embankment 3a Recess 4 Opposing part 4a Step fixing part 5 Dimming section 6 Container structure 7 Dimming section 8 Opposing part 9 Dimming section 10 Container structure 11 Chemical Solution 101 Sample analyzer 102 Excitation light generating unit 103 Fluorescence receiving unit 104 Longpass Filter 105 Mirror 106 Objective Lens 107 Mirror base 108 Guide Block 109 Guide Rail 110 Base Block 111 Transport hand 112 Drive mechanism

Claims

1. A container structure that is attached to an array plate having an upper surface on which a spot array containing a biological substance is formed, and is configured to form a container that can store a liquid using the array plate as a bottom plate, a bank portion that forms a side wall of the container when attached to the array plate; a retroreflection attenuation unit disposed opposite the array plate and configured to reduce the retroreflection of light passing through the spot array back onto the spot array; A container structure having:

2. The container structure according to claim 1 , wherein the retro-reflecting light attenuating portion is fixed to the bank portion.

3. The container structure according to claim 1 , wherein the retro-reflecting light attenuating portion is supported by an opposing portion fixed to the bank portion.

4. The container structure according to claim 1 , wherein the retro-light attenuating section is disposed at a distance from the spot array.

5. The container structure according to claim 1 , wherein the retroreflecting light attenuating portion has an area larger than that of the spot array.

6. The container structure according to claim 1 , wherein the area obtained by projecting the retroreflecting light attenuating portion onto the upper surface of the array plate includes the spot array.

7. A container comprising the container structure according to any one of claims 1 to 6 and the array plate.

8. a mounting portion on which the container according to claim 7 is mounted; the container is located on the opposite side of the retroreflection attenuation unit with respect to the spot array of the container placed on the mounting unit, a light emitting unit that irradiates the spot array with primary light; an enclosure that accommodates the container, the placement portion, and the light emitting portion; An apparatus comprising:

9. 9. The device according to claim 8, further comprising a moving section that moves the light emitting section and the placing section relative to each other.

10. The apparatus of claim 8 , further comprising a detector located on the opposite side of the spot array from the retroreflecting light attenuating unit, the detector detecting secondary light from the spot array.

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