Container structure

Abstract

This container structure is mounted on an array plate having an upper surface on which a spot array including a biological substance is formed, and the container structure is configured so as to form a container that has the array plate as the bottom plate thereof and that is capable of storing liquid, wherein the container structure includes: an embankment portion that forms a side wall of the container when mounted on the array plate; and a retroreflective light attenuating portion that is disposed facing to the array plate to reduce retroreflective incidence of light that has passed through the spot array onto the spot array. This reduces the impact of retroreflective light that has passed through the spot and is scattered forward becoming background light noise.

Description

container structure

[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.

[0002] Array plates such as protein arrays, peptide arrays, and DNA arrays are known, in which a large number of substances such as proteins, peptides, and nucleic acids are fixed in the form of spots on a substrate. By using an array plate for analyzing a sample, it is possible to simultaneously observe the interactions between the large number of substances fixed on the array plate and substances in the sample. This allows for comprehensive analysis of the interactions between a large number of substances and liquid samples derived from living organisms, 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 a 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 fluorescence 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 performed in which a frame is fixed to an array plate to allow liquid to be stored, and a pipette tip moves above the framed array plate to sequentially supply and drain multiple chemical solutions to cause reactions.Then, a specimen evaluation device is described in which, after the reaction process is completed, optical scanning measurement is performed while maintaining the liquid retention state, and a fluorescent image of the spot area is obtained.

[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 components that make up this covered channel slide are made of materials that allow optical measurements from the outside, so it can be used directly for fluorescence observation.

[0006] JP 2023-12426 A U.S. Patent Application Publication No. 2013 / 017143

[0007] In the device configuration of Patent Document 1, optical scanning measurement involves irradiating fluorescently labeled spots with laser light as excitation light from the underside of an array plate 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.

[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 biological substances is formed, and is configured to form a container that can store liquid using the array plate as a bottom plate, the container structure having: a bank portion that forms the 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 onto the spot array, thereby solving the above-mentioned problems.

[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.

[0010] 2B.

[0024] FIG. 1B is a perspective view showing the appearance of a container composed of a container structure and an array plate, which is a first embodiment of the container structure of the present invention. FIG. 1A is an exploded perspective view of the container shown in FIG. 1A. FIG. 1B is a vertical cross-sectional view showing a cross-section of the container shown in FIGS. 1A and 1B taken along a plane perpendicular to its longitudinal direction ... horizontal cross-sectional view of the container shown in FIGS. 1A and 1B taken horizontally along line 2C-2C in FIG. 2B, viewed from below. FIG. 1B is a horizontal cross-sectional view of the container shown in FIGS. 1A and 1B taken horizontally along line 2D-2D in FIG. 2B, viewed from below.

[0025] FIG. 1B shows a second embodiment of the container structure of the present invention. This figure is a vertical cross-sectional view showing a cross-section corresponding to the cross-section shown in FIG. 2B. FIG. 1C shows a third embodiment of the container structure of the present invention. This figure is a vertical cross-sectional view showing a cross-section corresponding to the cross-section shown in FIG. 2B. FIG. 1D shows a fourth embodiment of the container structure of the present invention. This figure is a vertical cross-sectional view showing a cross-section corresponding to the cross-section shown in FIG. 2B. FIG. 1E shows a fifth embodiment of the container structure of the present invention. This figure is a vertical cross-sectional view showing a cross-section corresponding to the cross-section shown in FIG. 2B. 7A is a schematic diagram showing the internal structure of the housing (enclosure) of a sample analyzer using a container composed of a container structure and an array plate of the present invention, as viewed from the front of the device. 7B is a schematic diagram showing the internal structure of the housing (enclosure) of the sample analyzer shown in FIG. 7A, as viewed from the side of the device.

[0011] [First Embodiment] A first embodiment of a container structure of the present invention will be described with reference to Figs. 1A, 1B and 2A to 2D.

[0012] FIG. 1A 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. 1B is an exploded perspective view individually illustrating the components constituting the container 1 of FIG. 1A. 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 arrangement 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 so as to surround a space in the shape of a rectangular frame. 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 can react 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 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.

[0013] 2A to 2D show the configuration of the container 1 in more detail. FIG. 2A 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. 1A). FIG. 2B 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. 2A). FIG. 2C is a horizontal cross-sectional view of the container structure 6 constituting the container 1, cut horizontally along line 2C-2C in FIG. 2B, as viewed from below the container structure. FIG. 2D is a horizontal cross-sectional view of the container structure 6 constituting the container 1, cut horizontally along line 2D-2D in FIG. 2B, as viewed from below the container structure.

[0014] As shown in Figures 2A and 2B, 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 2C, 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 materials used, such as shape-based fixing by adhesive bonding or fitting, or welding after installation. Furthermore, an 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. A chemical solution is stored in this internal space. As shown in Figure 2D, an array region 2a is formed on the array plate 2, in which multiple spots 2b are fixed in a two-dimensional array.

[0015] 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.

[0016] In this embodiment, the area of ​​the attenuation section 5 is larger than the area of ​​the array region 2a. Furthermore, 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.

[0017] 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.

[0018] 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.

[0019] [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. 2B 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 description thereof will be omitted.

[0020] 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 color development after the labeling reaction, and allows a wide range of materials to be selected that have a high light-attenuating effect.

[0021] [Third embodiment] Figure 4 shows a vertical cross-sectional view, corresponding to Figure 2B in the first embodiment, of a container using a container structure according to a third embodiment of the present invention. 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 fixed to the bank portion 3 by processing a resin material that is absorptive of the wavelengths of the laser excitation light and the fluorescence. This configuration integrates the facing portion and the attenuation portion, making it possible to reduce the number of parts.

[0022] [Fourth Embodiment] Figure 5 shows a vertical cross-sectional view, corresponding to Figure 2B in the first embodiment, of a container using a container structure according to a fourth embodiment of the present invention. 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 from this inner surface may cause light to retro-enter the array plate 2, affecting measurement. Therefore, by fixing the 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.

[0023] [Fifth Embodiment] Figure 6 shows a vertical cross-sectional view corresponding to Figure 2B in the first embodiment of a container using a container structure according to a fifth embodiment of the present invention. In this embodiment, the container structure 10 is configured such that the bank portion and the opposing portion are integrated into one body, and this is given the function of a retroreflection light attenuating portion, in contrast to the configuration of the fourth embodiment. The container structure 10 is configured by processing a resin material that is light-absorbing at the wavelengths of the laser excitation light and the fluorescence. The configuration of this embodiment allows the number of parts to be reduced.

[0024] [Example of Use of Container Structure] 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. Figures 7A and 7B are schematic diagrams of the measurement system of a sample analyzer that uses the container structure of the present invention. Figure 7A is a front view, and Figure 7B is a right side view.

[0025] The sample analyzer 101 has an illumination optical system, a fluorescence detection optical system, and a scanning system, all of which are enclosed by an enclosure 113. The illumination 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. The laser light is then focused by an objective lens 106 in the downstream 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 illumination optical system of the sample analyzer 101 is configured so that the laser light as primary light is irradiated onto the spot array to be observed from a light irradiator 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.

[0026] The optical system is disposed 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-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.

[0027] The above configuration forms a moving unit capable of relative movement within the XY plane by combining the Y-direction movement of the transport hand and the X-direction 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.

[0028] 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.

[0029] The present invention is not limited to the above-described embodiments, and various modifications and variations can be made without departing from the spirit and scope of the present invention. Therefore, the following claims are appended to apprise the public of the scope of the present invention.

[0030] This application claims priority based on Japanese Patent Application No. 2024-021896, filed February 16, 2024, the entire contents of which are incorporated herein by reference.

[0031] REFERENCE SIGNS LIST 1 Container 2 Array plate 2a Array region 2b Spot 3 Bank portion 3a Recess 4 Opposing portion 4a Step fixing portion 5 Light-attenuating portion 6 Container structure 7 Light-attenuating portion 8 Opposing portion (light-attenuating portion) 9 Light-attenuating portion 10 Container structure (light-attenuating portion) 11 Chemical solution 101 Sample analyzer 102 Excitation light generating portion 103 Fluorescence receiving portion 104 Long-pass 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 configured to be attached to an array plate having an upper surface on which a spot array containing biological substances is formed, and to form a container capable of storing liquid with the array plate as a bottom plate, the 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 positioned opposite the array plate and reduces the retroreflection of light that has passed through the spot array back onto the spot array.

2. A container structure according to claim 1, wherein the retroreflecting light attenuating portion is fixed to the bank portion.

3. A container structure according to claim 1 or 2, wherein the retroreflecting light attenuating section is supported by an opposing section fixed to the bank section.

4. A container structure according to any one of claims 1 to 3, wherein the retroreflecting light attenuating section is arranged at a distance from the spot array.

5. A container structure according to any one of claims 1 to 4, wherein the retroreflecting light attenuating section has an area larger than that of the spot array.

6. A container structure according to any one of claims 1 to 5, wherein the area obtained by projecting the retroreflecting light attenuation section 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. An apparatus comprising: a mounting section on which the container described in claim 7 is mounted; a light emitting section located on the opposite side of the spot array of the container placed on the mounting section from the retro-reflecting light attenuating section, and irradiating the spot array with primary light; and an enclosure that houses the container, the mounting section, and the light emitting section.

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

10. The device according to claim 8 or 9, further comprising a detection section located on the opposite side of said spot array from said retroreflecting section, for detecting secondary light from said spot array.

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