Chamber apparatus, hybridization reaction system, nucleic acid analysis system, and sealing member

Abstract

To provide improved assemblability and operability of a chamber apparatus.SOLUTION: A chamber apparatus 1 includes: a chamber unit 2 including: a sealing member 10 having multiple well-forming sections 20 formed therein that penetrate the sealing member in the thickness direction; a glass slide 11, attached to a first surface of the sealing member 10 where one end of the well-forming section 20 is open, closing one end of the well-forming section 20; a DNA array substrate, attached to a second surface of the sealing member 10 where the other end of the well-forming section 20 is open, closing the other end of the well-forming section 20; and a chamber frame 3 that detachably sandwiches the chamber unit 2 in the direction in which the glass slide 11, sealing member 10, and DNA array substrate overlap.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a chamber apparatus, a hybridization reaction system, a nucleic acid analysis system, and a sealing member. [Background technology]

[0002] Patent Document 1 listed below discloses a chamber apparatus including a chamber frame having an upper integrated gasket and a lower integrated gasket, a substrate, and a substrate frame for positioning and capturing the substrate.

[0003] The following Patent Document 2 discloses a target substance detection device comprising a biochemical reaction cassette having a fixation region for a probe for detecting a target substance, a reaction chamber for reacting a sample with the fixation region, and a first flow path and a second flow path that connect the cassette to the outside, the biochemical reaction cassette being provided with a sealing portion capable of sealing the first flow path and a pressure relief means for maintaining the pressure in the reaction chamber at an approximately constant level. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Special Publication No. 2009-542222 [Patent Document 2] Japanese Patent Application Laid-Open No. 2009-082084 Summary of the Invention [Problem to be solved by the invention]

[0005] The above-mentioned conventional techniques leave room for improvement in terms of ease of assembly and operability of the chamber device.

[0006] An object of the present invention is to improve the assembly and operability of a chamber device. [Means for solving the problem]

[0007] In order to solve the above problem, a chamber device according to a first aspect of the present invention comprises a chamber unit including a sealing member having a plurality of well forming portions formed thereon that penetrate in the thickness direction, a transparent member attached to a first surface side of the sealing member where one end of the well forming portion is open and that closes one end of the well forming portion, and a substrate attached to a second surface side of the sealing member where the other end of the well forming portion is open and that closes the other end of the well forming portion, and a chamber frame that detachably sandwiches the chamber unit in the direction in which the transparent member, the sealing member, and the substrate overlap.

[0008] A chamber device according to a second aspect of the present invention is a chamber device according to the first aspect of the present invention, wherein the chamber frame comprises a first frame abutting the first surface side of the chamber unit, a second frame abutting the second surface side of the chamber unit, a hinge mechanism rotatably connecting the first frame and the second frame, and a locking mechanism locking the first frame and the second frame in a folded state.

[0009] A chamber apparatus according to a third aspect of the present invention is a chamber apparatus according to the second aspect of the present invention, wherein the locking mechanism comprises a latch rotatably mounted on one of the first frame and the second frame, and an engaging portion mounted on the other of the first frame and the second frame to which the latch engages.

[0010] A chamber apparatus according to a fourth aspect of the present invention is the chamber apparatus according to the third aspect of the present invention, wherein a surface of at least one of the latch and the locked portion is plated.

[0011] A chamber device according to a fifth aspect of the present invention is a chamber device according to any one of the first to fourth aspects of the present invention, wherein a ring-shaped convex portion is formed on the peripheral edge of the opening of the well formation portion, which abuts in a compressed state against the substrate and the transparent member when the chamber unit is sandwiched between the chamber frame.

[0012] A chamber device according to a sixth aspect of the present invention is a chamber device according to any one of the first to fifth aspects of the present invention, wherein the sealing member has a first hole portion and a second hole portion extending from the inner wall surface of the well formation portion to the outer surface.

[0013] A chamber apparatus according to a seventh aspect of the present invention is the chamber apparatus according to the sixth aspect of the present invention, wherein a step or wall is formed on the inner wall surface of the well formation portion between the first hole portion and the second hole portion.

[0014] A chamber apparatus according to an eighth aspect of the present invention is the chamber apparatus according to the sixth or seventh aspect of the present invention, further comprising a cover member attached to the seal member and closing the first hole portion and the second hole portion.

[0015] A chamber apparatus according to a ninth aspect of the present invention is a chamber apparatus according to any one of the first to eighth aspects of the present invention, wherein the sealing member has a first engagement groove formed on the first surface side that engages with the peripheral edge of the transparent member, and a second engagement groove formed on the second surface side that engages with the peripheral edge of the substrate.

[0016] A hybridization reaction system according to the tenth aspect of the present invention performs a hybridization reaction using a chamber apparatus according to any one of the first to ninth aspects of the present invention.

[0017] The nucleic acid analysis system according to the eleventh aspect of the present invention analyzes nucleic acids extracted using the hybridization reaction system according to the tenth aspect of the present invention.

[0018] A sealing member according to a twelfth aspect of the present invention comprises a plurality of well forming portions penetrating in the thickness direction, a first hole portion and a second hole portion extending from the inner wall surface of the well forming portion to the outer surface, and a step or wall arranged between the first hole portion and the second hole portion on the inner wall surface of the well forming portion. [Effects of the Invention]

[0019] According to the above aspect of the present invention, the assembly and operability of the chamber device can be improved. [Brief explanation of the drawings]

[0020] [Figure 1] 1 is a perspective view of a chamber apparatus according to a first embodiment of the present invention. [Figure 2] FIG. 2 is a right side view of the chamber apparatus according to the first embodiment of the present invention. [Figure 3] 1 is a perspective view showing a state in which the chamber frame according to the first embodiment of the present invention is opened and a cap is removed. FIG. [Figure 4] FIG. 2 is a perspective view showing a state in which a chamber unit is attached to a chamber frame according to the first embodiment of the present invention. [Figure 5] FIG. 2 is an exploded perspective view of the chamber unit according to the first embodiment of the present invention. [Figure 6] FIG. 2 is a plan view of the chamber unit according to the first embodiment of the present invention. [Figure 7] FIG. 2 is a bottom view of the chamber unit according to the first embodiment of the present invention. [Figure 8] FIG. 8 is a cross-sectional view taken along the line VIII-VIII in FIG. 6. [Figure 9] FIG. 2 is a flow chart of assembling the chamber apparatus according to the first embodiment of the present invention. [Figure 10] 1 is a schematic diagram of a nucleic acid analysis system according to a first embodiment of the present invention. [Figure 11] FIG. 10 is a perspective view of a well forming portion according to a second embodiment of the present invention. [Figure 12] FIG. 10 is a plan view of a chamber apparatus according to a third embodiment of the present invention. [Figure 13] FIG. 10 is a bottom view of a chamber apparatus according to a third embodiment of the present invention. [Figure 14] FIG. 10 is a perspective view of a chamber apparatus according to a fourth embodiment of the present invention. [Figure 15] FIG. 10 is an exploded perspective view of a chamber apparatus according to a fourth embodiment of the present invention. [Figure 16] FIG. 10 is a cross-sectional view of a chamber apparatus according to a fourth embodiment of the present invention. [Figure 17] FIG. 17 is an enlarged view of an area A shown in FIG. DETAILED DESCRIPTION OF THE INVENTION

[0021] Hereinafter, a chamber apparatus, a hybridization reaction system, a nucleic acid analysis system, and a sealing member according to embodiments of the present invention will be described in detail with reference to the drawings. First, an overview of the embodiments of the present invention will be described, followed by a detailed description of each embodiment of the present invention.

[0022] 〔overview〕 The above-mentioned Patent Document 1 discloses a method of fastening and sealing the upper and lower hybridization chamber parts with a full-perimeter snap system. Furthermore, the above-mentioned Patent Document 2 discloses a hybridization chamber technology in which two flow paths for injecting samples are provided, and the sample is stirred to improve reaction efficiency, while a pressure relief means is used to maintain the pressure in the reaction chamber at a substantially constant level, thereby preventing the flow of the sample from being obstructed.

[0023] However, the configuration described in Patent Document 1 requires that the cells be fastened with snaps around the entire circumference after the sample is injected into them, which requires assembly to prevent the injected sample from spilling or contaminating other cells. Furthermore, if disassembly becomes necessary after assembly, the snap fastening around the entire circumference can make disassembly of the hybridization chamber parts difficult. Furthermore, when disassembling the hybridization chamber parts, the enclosed sample, etc., may scatter, requiring cleaning. Furthermore, the configuration described in Patent Document 2 requires bonding the glass and the housing, which can result in high manufacturing costs.

[0024] In an embodiment of the present invention, a chamber unit is assembled by stacking a transparent member, a sealing member, and a substrate, and the chamber unit is then detachably sandwiched between a chamber frame. This configuration allows the chamber device to be constructed using parts that do not have any joints, improving the assembly and operability of the chamber device. Furthermore, by removing and replacing the chamber unit after use, the chamber frame can be reused without cleaning. Furthermore, by disposing of the replaced chamber unit as is, it is possible to prevent the enclosed sample from scattering.

[0025] [First embodiment] Fig. 1 is a perspective view of a chamber apparatus 1 according to a first embodiment of the present invention, and Fig. 2 is a right side view of the chamber apparatus 1 according to the first embodiment of the present invention. As shown in these figures, the chamber device 1 includes a chamber unit 2, a chamber frame 3, and a cap (lid member) 4. The chamber device 1 has a flat, approximately rectangular parallelepiped shape.

[0026] In the following description, an XYZ Cartesian coordinate system is set, and the positional relationship of each component may be described with reference to this XYZ Cartesian coordinate system. The X-axis direction is the width direction (left-right direction) of the chamber apparatus 1. The Y-axis direction is the depth direction (front-rear direction) of the chamber apparatus 1. The Z-axis direction is the thickness direction (up-down direction) of the chamber apparatus 1. The front-rear, up-down, left-right directions of the chamber apparatus 1 are defined by defining the side of the chamber frame 3 where the locking mechanism 60 is located as the front side, and the side where the hinge mechanism 50 is located as the rear side.

[0027] Fig. 3 is a perspective view showing a state in which the chamber frame 3 according to the first embodiment of the present invention is opened and the cap 4 is removed. Fig. 4 is a perspective view showing a state in which the chamber unit 2 is attached to the chamber frame 3 according to the first embodiment of the present invention. As shown in these figures, the chamber device 1 has a chamber unit 2 detachably sandwiched between a chamber frame 3. The chamber unit 2 has a plurality of well formation portions 20 formed therein for accommodating samples.

[0028] Fig. 5 is an exploded perspective view of the chamber unit 2 according to the first embodiment of the present invention. Fig. 6 is a plan view of the chamber unit 2 according to the first embodiment of the present invention. Fig. 7 is a bottom view of the chamber unit 2 according to the first embodiment of the present invention. Fig. 8 is a cross-sectional view taken along the line VIII-VIII shown in Fig. 6. As shown in FIG. 5, the chamber unit 2 includes a seal member 10, a slide glass 11 (transparent member), and a DNA array substrate 12 (substrate).

[0029] The seal member 10 has a flat, rectangular shape in a plan view. The seal member 10 is formed, for example, from elastic silicone rubber. The seal member 10 has a first surface 10A facing upward, a second surface 10B facing downward, a third surface 10C facing forward, a fourth surface 10D facing rightward, a fifth surface 10E facing rearward, and a sixth surface 10F facing leftward.

[0030] The sealing member 10 is formed with a plurality of well-forming portions 20 that penetrate in the thickness direction (Z-axis direction). The well-forming portions 20 are formed in two rows, one in the depth direction (Y-axis direction) and one spaced apart in the width direction (X-axis direction). Note that the number, arrangement, and shape of the well-forming portions 20 are not limited to those shown in the figure.

[0031] The slide glass 11 is attached to the first surface 10A side of the seal member 10, where one end (upper end) of the well formation portion 20 is open. When attached to the seal member 10, the slide glass 11 closes the opening at one end (upper end) of the well formation portion 20. A commercially available standard product can be used as the slide glass 11. Note that the slide glass 11 is not limited to the slide glass 11, and a transparent acrylic plate or the like may also be used as long as it is a transparent member that is rectangular in plan view.

[0032] The DNA array substrate 12 has a plurality of DNA microarrays 12a in which probe carriers for hybridization reactions are arranged in an array. The DNA microarrays 12a are formed in an arrangement corresponding to the well formation portions 20. Note that the number of DNA microarrays 12a does not necessarily have to be the same as the number of well formation portions 20. Furthermore, when a reaction other than a hybridization reaction is to be performed, a substrate having a reaction portion corresponding to that reaction may be used.

[0033] The DNA array substrate 12 is attached to the second surface 10B side of the sealing member 10, where the other end (lower end) of the well forming portion 20 is open. When attached to the sealing member 10, the DNA array substrate 12 closes the opening at the other end (lower end) of the well forming portion 20. The DNA array substrate 12 has a notch 12b formed therein to prevent incorrect assembly (orientation) with respect to the sealing member 10. The notch 12b is formed at one of the four corners of the DNA array substrate 12.

[0034] As shown in Fig. 6, an annular protrusion 21A that abuts against the slide glass 11 is formed on the periphery of the opening of the well-forming portion 20 on the first surface 10A side of the seal member 10. As shown in Fig. 8, the annular protrusion 21A has a semicircular convex shape in cross section. The annular protrusion 21A abuts against the slide glass 11 attached to the seal member 10 in a compressed state in the vertical direction (Z-axis direction), thereby improving the airtightness of the well-forming portion 20.

[0035] Furthermore, a first engagement groove 22 that engages with the peripheral edge of the slide glass 11 is formed on the first surface 10A side of the seal member 10. As shown in Fig. 6, the first engagement groove 22 engages with the peripheral edge of the slide glass 11 over the entire circumference. When the slide glass 11 is engaged with the first engagement groove 22, the seal member 10 is elastically deformed.

[0036] 5, engagement recesses 23 extending in the front-rear direction (Y-axis direction) are formed on the first surface 10A of the seal member 10. The engagement recesses 23 are formed in pairs along the fourth surface 10D and the sixth surface 10F. In addition, engagement recesses 23 extending in the front-rear direction (Y-axis direction) are also formed on the second surface 10B of the seal member 10. These engagement recesses 23 are also formed in pairs along the fourth surface 10D and the sixth surface 10F.

[0037] 6, the sealing member 10 is formed with an inlet 25 (first hole) and an outlet 24 (second hole), which extend in the width direction (X-axis direction) from the inner wall surface of the well-forming portion 20 to the outer side surfaces (fourth surface 10D, sixth surface 10F) of the sealing member 10. An injection needle such as a pipetter is inserted into the inlet 25 to inject a sample into the well-forming portion 20. Air, a sample, etc. are discharged from the well-forming portion 20 through the outlet 24.

[0038] A step 28 is formed on the inner wall surface of the well formation section 20 between the inlet 25 and the outlet 24. In other words, a recess is provided on the inner wall surface of the well formation section 20, and the outlet 24 is formed in the recess. This prevents the reagent injected from the inlet 25 into the well formation section 20 from being short-circuited to the outlet 24 and being discharged immediately thereafter. This makes it possible to efficiently inject a constant amount of reagent into the well formation section 20.

[0039] The outlet side (outer surface side) of the inlet 25 forms a tapered section 25a with an increased diameter. Plate sections 26 that protrude laterally are formed on the outer surfaces (fourth surface 10D, sixth surface 10F) of the seal member 10. The plate sections 26 are provided to separate the inlet 25 and outlet 24 that communicate with the same well formation section 20 from the inlet 25 and outlet 24 that communicate with other well formation sections 20, in order to prevent sample contamination between the well formation sections 20.

[0040] 1 and 2 is attached to the outer surfaces (fourth surface 10D, sixth surface 10F) of the sealing member 10, and closes the inlet 25 and the outlet 24. The cap 4 has two types of protrusions (not shown) that can be inserted into the inlet 25 and the outlet 24, and an accommodating groove (not shown) that accommodates the plate portion 26.

[0041] 8, an annular protrusion 21B that abuts against the DNA array substrate 12 is formed on the periphery of the opening of the well formation portion 20 on the second surface 10B side of the seal member 10. The annular protrusion 21B has a semicircular convex shape in cross section. The annular protrusion 21B abuts against the DNA array substrate 12 attached to the seal member 10 in a compressed state in the vertical direction (Z-axis direction), thereby improving the airtightness of the well formation portion 20.

[0042] Furthermore, a second engagement groove 29 that engages with the peripheral edge of the DNA array substrate 12 is formed on the second surface 10B of the seal member 10. As shown in Fig. 7, the second engagement groove 29 engages with the peripheral edge of the DNA array substrate 12 over substantially the entire periphery. When the DNA array substrate 12 is engaged with the second engagement groove 29, the seal member 10 is elastically deformed.

[0043] Positioning recesses 27a to 27d are formed on the second surface 10B of the sealing member 10 at positions corresponding to the four corners of the DNA array substrate 12. The positioning recesses 27a and 27b are formed at positions corresponding to the two front corners of the DNA array substrate 12. The positioning recesses 27c and 27d are formed at positions corresponding to the two rear corners of the DNA array substrate 12.

[0044] The positioning recesses 27c and 27d communicate with the second engagement groove 29. Therefore, the two rear corners of the DNA array substrate 12 extend from inside the second engagement groove 29 to inside the positioning recesses 27c and 27d. Note that the positioning recesses 27a and 27d do not communicate with the second engagement groove 29, but may communicate with it.

[0045] As shown in Figure 4, the chamber frame 3 includes a first frame 30 that abuts against the first surface 10A side of the chamber unit 2, a second frame 40 that abuts against the second surface 10B side of the chamber unit 2, a hinge mechanism 50 that rotatably connects the first frame 30 and the second frame 40, and a locking mechanism 60 that locks the first frame 30 and the second frame 40 together when they are folded over each other.

[0046] The second frame 40 is formed with a support base 41 that supports the DNA array substrate 12 from below, and positioning protrusions 42a to 42d provided at four locations around the periphery of the support base 41. The support base 41 has a rectangular shape in a plan view and is large enough to be inserted into the second engagement groove 29 shown in FIG.

[0047] The positioning protrusions 42a to 42d have a size that allows them to be inserted into the positioning recesses 27a to 27d shown in Fig. 7. The positioning protrusions 42a to 42d are inserted into the positioning recesses 27a to 27d, thereby positioning the DNA array substrate 12 (chamber unit 2) relative to the second frame 40. Each of the positioning protrusions 42a to 42d has a recess formed therein to avoid interference with the corners of the DNA array substrate 12, as shown in Fig. 4.

[0048] A pair of engagement ribs 43 extending parallel to the front-rear direction (Y-axis direction) are formed on both the left and right sides of the support base 41. The pair of engagement ribs 43 engage with a pair of engagement recesses 23 provided on the second surface 10B side of the seal member 10 described above. The engagement of the pair of engagement ribs 43 with the pair of engagement recesses 23 positions the seal member 10 (chamber unit 2) relative to the second frame 40.

[0049] A pair of clamping protrusions 44 extending parallel to the front-rear direction (Y-axis direction) are formed on both the left and right sides of the pair of engaging ribs 43. The pair of clamping protrusions 44 are protrusions that clamp the cap 4 shown in Figures 1 and 2. This prevents the cap 4 from easily detaching from the seal member 10 (chamber unit 2).

[0050] 4, the first frame 30 is formed with a plurality of window sections 31 for observing the well formation section 20, and a pair of slits 32 arranged on both the left and right sides of the plurality of window sections 31. The plurality of window sections 31 are formed at positions that overlap with the well formation section 20 in the Z-axis direction when the chamber apparatus 1 is assembled as shown in FIG.

[0051] The window portions 31 penetrate the first frame 30 in the thickness direction (Z-axis direction). The window portions 31 are formed in two rows, one in the depth direction (Y-axis direction) and one spaced apart in the width direction (X-axis direction). The number, arrangement, and shape of the window portions 31 are not limited to those shown in the figure. In this embodiment, two well formation portions 20 can be observed from one window portion 31.

[0052] The slit 32 penetrates the first frame 30 in the thickness direction (Z-axis direction). As shown in Fig. 6, the slit 32 is formed at a position that overlaps the middle part of the injection port 25 and the outlet port 24 in the seal member 10 in a plan view. This makes it possible to confirm that the injection needle is securely inserted into the injection port 25 when injecting a sample into the well formation section 20. The seal member 10 is preferably translucent, for example, transparent or translucent.

[0053] 4, a pair of engagement ribs 33 extending parallel to the front-rear direction (Y-axis direction) are formed on both the left and right sides of the pair of slits 32. The pair of engagement ribs 33 engage with a pair of engagement recesses 23 provided on the first surface 10A side of the above-mentioned seal member 10. The engagement of the pair of engagement ribs 33 with the pair of engagement recesses 23 positions the seal member 10 (chamber unit 2) relative to the first frame 30.

[0054] A pair of clamping protrusions 34 extending parallel to the front-rear direction (Y-axis direction) are formed on both the left and right sides of the pair of engaging ribs 33. The pair of clamping protrusions 34 are protrusions that clamp the cap 4 shown in Figures 1 and 2. This prevents the cap 4 from easily detaching from the seal member 10 (chamber unit 2).

[0055] As shown in FIGS. 3 and 4, the hinge mechanism 50 connects the rear ends of the first frame 30 and the second frame 40 to each other so as to be rotatable about an axis extending in the width direction (X-axis direction). As shown in FIG. 1, the locking mechanism 60 releasably fixes the front ends of the first frame 30 and the second frame 40 to each other.

[0056] The locking mechanism 60 includes a latch 61 rotatably provided on the first frame 30, and a latched portion 62 provided on the second frame 40 and engaged with the latch 61. The latch 61 may be rotatably provided on the second frame 40, and the latched portion 62 may be provided on the first frame 30.

[0057] 3, the latch 61 is connected to the front end of the first frame 30 so as to be rotatable around an axis extending in the width direction (X-axis direction). A long hole 61a extending in the width direction is formed in the latch 61. A claw portion 61b protrudes from the inside of the long hole 61a. The claw portion 61b is formed on the inner wall surface of the long hole 61a on the rotation end side (lower side) of the latch 61 and extends in the width direction.

[0058] The locked portion 62 is formed at the front end of the second frame 40. The locked portion 62 is formed in the shape of a plate extending in the width direction. A step portion 62a protrudes from the underside of the locked portion 62. When the locked portion 62 is inserted into the elongated hole 61a of the latch 61, the claw portion 61b climbs over the step portion 62a and is locked to the step portion 62a in the depth direction (Y-axis direction), thereby locking the first frame 30 and the second frame 40 together.

[0059] The locking force of the locking mechanism 60 is preferably 20N to 40N, which is the range of force exerted by an average person's fingers. In this embodiment, the locking force is designed to be, for example, 20N. Furthermore, it is preferable to plate the surfaces of the latch 61 and the locked portion 62 to improve slidability and durability. This allows for smooth opening and closing of the chamber frame 3.

[0060] FIG. 9 is a flow diagram of the assembly of the chamber apparatus 1 according to the first embodiment of the present invention. When assembling the above-described chamber apparatus 1, first, the first frame 30 and the second frame 40 connected by the hinge mechanism 50 are set (placed) in an open state (step S1).

[0061] Next, the slide glass 11 and the DNA array substrate 12 are fitted around their entire circumferences into the first engagement groove 22 and the second engagement groove 29 of the sealing member 10, respectively (step S2). The first and second surfaces 10A and 10B of the sealing member 10 have annular convex portions 21A and 21B formed around the periphery of the opening of the well-forming portion 20, and fitting the slide glass 11 and the DNA array substrate 12 together forms an enclosed space in the well-forming portion 20, i.e., a reaction space between the DNA microarray 12a and the reagent.

[0062] Next, the chamber unit 2, which is an assembly of the sealing member 10, the slide glass 11, and the DNA array substrate 12, is set on the second frame 40 (step S3). The second frame 40 is provided with positioning structures (positioning protrusions 42a to 42d, engaging ribs 33, etc.) for setting the chamber unit 2 in a predetermined position, making it easy to set the chamber unit 2.

[0063] Next, the first frame 30 connected to the second frame 40 is closed, and with the first frame 30 and the second frame 40 folded over each other, the latches 61 are hooked onto the latched portions 62 and fixed in place (step S4).The chamber frame 3 is held by hand or set in a dedicated jig or the like, and a test reagent is injected into the well formation portion 20 from the injection port 25 using a pipetter or the like (step S5).

[0064] Once the injection of the reagent into the well formation portion 20 on one side (for example, the right side) is complete, the inlet 25 and outlet 24 on that side are sealed with the cap 4, and the well formation portion 20 on that side is put into a sealed state (step S6). The same procedure is performed for the well formation portion 20 on the opposite side (for example, the left side) (step S7). Once all of the above steps are completed, the process proceeds to the next step (step S8).

[0065] After completing the processing in the next step, the latch 61 is released from the latched portion 62, and the chamber frame 3 is opened (step S9). Then, the chamber unit 2, which is an assembly of the seal member 10, slide glass 11, and DNA array substrate 12, is removed from the chamber frame 3 with the cap 4 still attached, and is discarded as is (step S10). The slide glass 11 and DNA array substrate 12 are embedded entirely or almost entirely in the seal member 10, and the reagent inside can be retained without leaking for several tens of seconds, so the reagent does not splash onto the chamber frame 3. Therefore, the chamber frame 3 can be used repeatedly without cleaning.

[0066] FIG. 10 is a schematic diagram of a nucleic acid analysis system 600 using the chamber apparatus 1 according to the first embodiment of the present invention. As shown in FIG. 10, a nucleic acid analysis system 600 includes a bacteria recovery system 200 , a nucleic acid extraction system 300 , a hybridization reaction system 400 , and a detection system 500 .

[0067] The microorganism collection system 200 is a system that collects microorganisms (bacteria, fungi, etc.) contained in the sample 100 from the sample 100. For example, in the case of testing a beverage, the sample 100 may be the produced beverage, the water used to produce the beverage, or a liquid in the process of producing the beverage. Alternatively, the sample 100 may be a liquid from which microorganisms have been collected using a cotton swab or the like that has been used to wipe the test environment in order to test for the presence or absence of bacterial contamination in the production environment and the degree of contamination.

[0068] The bacteria can be collected by filtering the collected liquid using a filter or the like, for example, by applying pressure or vacuum to the collected liquid. When collecting bacteria or fungi, the filter preferably has a pore size of 0.22 μm to 0.45 μm. After collecting the bacteria using the filter, the filter is placed in a culture vessel described below and immersed in a culture solution in which the bacteria are cultured, and the bacteria are cultured. Bacterial culture can be performed, for example, by static culture, in which the culture vessel is left stationary, or by shaking culture, in which the culture vessel is shaken. The culture solution in which the bacteria have been cultured is transferred to the next step (nucleic acid extraction system 300). The bacteria may be collected by centrifugation or the like and then transferred to the next step, or the liquid containing the filter may be shaken and the liquid in which the bacteria are suspended may be transferred to the next step.

[0069] The nucleic acid extraction system 300 is a system that destroys (dissolves) the membrane structure of cells in a liquid and extracts nucleic acids from bacterial cells. The sample 100 from which nucleic acids have been extracted may be mixed with a liquid containing other nucleic acids that react with the extracted nucleic acids. Furthermore, the other nucleic acids may be nucleic acids to which a moiety that exhibits fluorescence, luminescence, or quenching effects under specific conditions has been added for detection in the detection step (detection system 500) described below. These may be mixed with the sample 100 before processing with the nucleic acid extraction system 300, or may be mixed with the sample 100 after processing with the nucleic acid extraction system 300.

[0070] The hybridization reaction system 400 is a system that causes a hybridization reaction of nucleic acids in a sample 100. In this process, the chamber apparatus 1 described above is used. In the hybridization reaction, the sample 100 is heated to, for example, 60°C and stirred in the well forming section 20, thereby causing a hybridization reaction that matches with the other nucleic acids described above. In this reaction, for example, a moiety that exhibits fluorescence, luminescence, or quenching effect under specific conditions, which is imparted to the other nucleic acid, reacts with the nucleic acid in the sample 100, thereby causing the fluorescence, luminescence, or quenching effect to be expressed.

[0071] Furthermore, by designing the structure of the other nucleic acid described above to react with a specific nucleic acid, it is possible to make it react only with the nucleic acid contained in a specific bacterium in the sample 100. In other words, in the processing of the hybridization reaction system 400, by using another nucleic acid that reacts with the specific nucleic acid, it is possible to make the fluorescence, luminescence, or quenching effect imparted to the other nucleic acid manifest only when the specific bacterium is contained in the sample 100.

[0072] The detection system 500 detects the presence or absence, and the degree of, fluorescence, luminescence, or quenching that occurs in the sample 100 that has been treated in the hybridization reaction system 400. The detection system 500, for example, excites the fluorescence that occurs in the nucleic acid of the sample 100 with an excitation laser beam, and detects the excited fluorescence with a high-sensitivity camera.

[0073] Alternatively, the detection system 500 uses a high-sensitivity camera to detect the luminescence effect expressed in the nucleic acid of the sample 100. Alternatively, the detection system 500 uses a high-sensitivity camera to detect the degree to which the fluorescence or luminescence imparted near the site to which the quenching effect is imparted is quenched, which is the quenching effect expressed in the nucleic acid of the sample 100. Regarding this detection method, for example, a method such as that described in JP 2020-74726 A may be adopted.

[0074] The nucleic acid analysis system 600 can analyze whether a particular microorganism (bacteria, fungi, etc.) is present in the sample 100 or its concentration by using a series of systems as described above.

[0075] As described above, the chamber apparatus 1 according to this embodiment includes a chamber unit 2 including a seal member 10 having a plurality of well-forming portions 20 formed therethrough in the thickness direction, a slide glass 11 (transparent member) attached to a first surface 10A of the seal member 10 where one end of the well-forming portion 20 is open and blocking one end of the well-forming portion 20, and a DNA array substrate 12 (substrate) attached to a second surface 10B of the seal member 10 where the other end of the well-forming portion 20 is open and blocking the other end of the well-forming portion 20, and a chamber frame 3 that detachably sandwiches the chamber unit 2 in the direction in which the slide glass 11, seal member 10, and DNA array substrate 12 overlap. This configuration enables the chamber apparatus 1 to be constructed using parts that do not have any joints, improving the assembly and operability of the chamber apparatus 1. Furthermore, by removing and replacing the chamber unit 2 after use, the chamber frame 3 can be reused without cleaning. Furthermore, by simply discarding the replaced chamber unit 2, the enclosed sample can be prevented from scattering.

[0076] Furthermore, in the chamber apparatus 1 according to this embodiment, the chamber frame 3 includes a first frame 30 that abuts against the first surface 10A of the chamber unit 2, a second frame 40 that abuts against the second surface 10B of the chamber unit 2, a hinge mechanism 50 that rotatably connects the first frame 30 and the second frame 40, and a locking mechanism 60 that locks the first frame 30 and the second frame 40 in a folded state. With this configuration, the first frame 30 and the second frame 40 are connected by the hinge mechanism 50, so that both frames need only be fixed in one place, simplifying the assembly work of the chamber apparatus 1.

[0077] Furthermore, in the chamber apparatus 1 according to this embodiment, the locking mechanism 60 includes a latch 61 rotatably provided on one of the first frame 30 and the second frame 40, and an engaged portion 62 that is provided on the other of the first frame 30 and the second frame 40 and engages with the latch 61. With this configuration, the chamber frame 3 can be fixed simply by hooking the latch 61 onto the engaged portion 62.

[0078] Furthermore, in the chamber apparatus 1 according to this embodiment, a plating process is applied to the surface of at least one of the latch 61 and the locked portion 62. This configuration improves the slidability and durability of the latch 61 and the locked portion 62, allowing for smoother opening and closing of the chamber frame 3.

[0079] Furthermore, in the chamber device 1 according to this embodiment, annular convex portions 21A and 21B are formed on the periphery of the opening of the well forming portion 20, which come into compressed contact with the DNA array substrate 12 and the slide glass 11 when the chamber unit 2 is sandwiched between the chamber frame 3. With this configuration, the annular convex portions 21A and 21B come into compressed contact with the slide glass 11 and the DNA array substrate 12, thereby forming an enclosed space in the well forming portion 20, i.e., a reaction space between the DNA microarray 12a and the reagent.

[0080] Furthermore, in the chamber apparatus 1 according to this embodiment, the sealing member 10 is formed with an inlet 25 (first hole) and an outlet 24 (second hole) that extend from the inner wall surface of the well formation section 20 to the outer surface (fourth surface 10D, sixth surface 10F). With this configuration, a reagent can be injected from the outer surface while a sealed space is formed in the well formation section 20.

[0081] Furthermore, in the chamber apparatus 1 according to this embodiment, a step 28 is formed on the inner wall surface of the well formation section 20 between the inlet 25 and the outlet 24. This configuration makes it possible to prevent the reagent injected into the well formation section 20 from the inlet 25 from being short-circuited to the outlet 24 and being discharged immediately thereafter.

[0082] Furthermore, the chamber apparatus 1 according to this embodiment is provided with a cap 4 (lid member) that is attached to the sealing member 10 and closes the inlet 25 and the outlet 24. This configuration makes it possible to prevent the reagent injected into the well formation section 20 from leaking out from the inlet 25 or the outlet 24.

[0083] Furthermore, in the chamber apparatus 1 according to this embodiment, the seal member 10 is provided with a first engagement groove 22 formed on the first surface 10A side that engages with the peripheral edge of the slide glass 11, and a second engagement groove 29 formed on the second surface 10B side that engages with the peripheral edge of the DNA array substrate 12. With this configuration, by fitting the slide glass 11 and the DNA array substrate 12 to the seal member 10 all around, the reagent inside can be retained without leaking for several tens of seconds, which prevents the reagent from splashing onto the chamber frame 3 when the chamber frame 3 is discarded.

[0084] Furthermore, the hybridization reaction system 400 according to this embodiment performs a hybridization reaction using the chamber apparatus 1. According to this configuration, the chamber apparatus 1 can be easily assembled and easily operated, thereby improving the workability of the hybridization reaction.

[0085] Furthermore, the nucleic acid analysis system 600 according to this embodiment analyzes nucleic acids extracted using the hybridization reaction system 400. According to this configuration, the ease of assembly and operability of the chamber apparatus 1 is improved, thereby improving the workability of nucleic acid analysis.

[0086] Second Embodiment Next, a second embodiment of the present invention will be described. In the following description, the same or equivalent components as those in the above-described embodiment will be denoted by the same reference numerals, and the description thereof will be simplified or omitted.

[0087] FIG. 11 is a perspective view of a well forming portion 20 according to the second embodiment of the present invention. As shown in FIG. 11, a wall 28a is formed between the inlet 25 and the outlet 24 on the inner wall surface of the well forming portion 20 of the second embodiment.

[0088] Wall 28a extends obliquely from inlet 25 toward outlet 24. Wall 28a is formed in a roughly triangular tongue shape in plan view. Outlet 24 is located closer to the outer surface of seal member 10 than the tip of wall 28a. According to the above configuration, as with step 28 in the first embodiment, it is possible to prevent reagent injected from inlet 25 into well formation section 20 from short-circuiting to outlet 24 and being discharged immediately thereafter.

[0089] Third Embodiment Next, a third embodiment of the present invention will be described. In the following description, the same or equivalent components as those in the above-described embodiment will be denoted by the same reference numerals, and the description thereof will be simplified or omitted.

[0090] Fig. 12 is a plan view of a chamber apparatus 1 according to a third embodiment of the present invention. Fig. 13 is a bottom view of the chamber apparatus 1 according to the third embodiment of the present invention. As shown in these figures, the locking mechanism 60 of the third embodiment has a bayonet structure.

[0091] The locking mechanism 60 of the third embodiment includes a pair of fixing pieces 63 rotatably provided on the first frame 30, and a pair of through holes 64 provided on the second frame 40 and locking the pair of fixing pieces 63 by rotation. Note that the pair of fixing pieces 63 may be rotatably provided on the second frame 40, and the pair of through holes 64 may be provided on the first frame 30.

[0092] As shown in Fig. 12, the fixing piece 63 is attached to the front end of the first frame 30 so as to be rotatable around a rotation axis 63a extending in the thickness direction (Z-axis direction). As shown in Fig. 13, the fixing piece 63 has an insertion end 63b that can be inserted into the through-hole 64. The insertion end 63b has a substantially rectangular shape when viewed from the bottom.

[0093] The through-hole 64 penetrates the front end of the second frame 40 in the thickness direction (Z-axis direction). The through-hole 64 has a generally rectangular shape extending in the front-to-rear direction (Y-axis direction) when viewed from the bottom. The lower end of the fixing piece 63 is inserted into the through-hole 64, and as the fixing piece 63 rotates, the insertion end 63b engages with the opening edge of the through-hole 64, thereby locking the first frame 30 and the second frame 40 together. The above configuration also makes it possible to configure the chamber device 1 using parts that do not have joints, improving the ease of assembly and operability of the chamber device 1.

[0094] [Fourth embodiment] Next, a fourth embodiment of the present invention will be described. In the following description, the same or equivalent components as those in the above-described embodiments will be denoted by the same reference numerals, and the description thereof will be simplified or omitted.

[0095] In each of the above-described embodiments, the chamber frame 3 includes a first frame 30 that abuts against the first surface 10A of the chamber unit 2, a second frame 40 that abuts against the second surface 10B of the chamber unit 2, and a hinge mechanism 50 that rotatably connects the first frame 30 and the second frame 40. The chamber frame 3 in each of the above-described embodiments is intended to be reused, and the hinge mechanism 50 is made of metal. In contrast, the fourth embodiment aims to provide a chamber apparatus 1 that is disposable (throwaway), including the chamber frame 3.

[0096] Fig. 14 is a perspective view of a chamber apparatus 1 according to a fourth embodiment of the present invention. Fig. 15 is an exploded perspective view of the chamber apparatus 1 according to the fourth embodiment of the present invention. As shown in these figures, the chamber apparatus 1 of the fourth embodiment is provided with resin clamp members 70 that clamp the short sides of the first frame 30 and the second frame 40 on both sides in the Y-axis direction.

[0097] The clamp member 70 is formed in a U-shape when viewed from the X-axis direction. Slide grooves 35 extending in the X-axis direction along the short sides of each of the first frame 30 and the second frame 40 are formed on both sides in the Y-axis direction of the first frame 30 and the second frame 40. A protrusion that engages with the slide groove 35 is formed at the tip of the U-shape of the clamp member 70. The clamp member 70 slides in the X-axis direction along the slide groove 35 to engage with the first frame 30 and the second frame 40 and also clamps the first frame 30 and the second frame 40 in the Z-axis direction.

[0098] 15, in the fourth embodiment, a transparent resin member 11A is used as the transparent member instead of the slide glass 11. Examples of the transparent resin member 11A include acrylic, polyethylene terephthalate, polycarbonate, and polyvinyl chloride. The transparent resin member 11A has a plurality of protrusions 11a formed therein and arranged within a plurality of window portions 31.

[0099] This transparent resin member 11A is integrated with the first frame 30 (high-strength resin member) by, for example, two-color molding. By arranging the protrusion 11a in the window portion 31, the strength of the portion that was hollow in each of the above embodiments can be improved. Note that, by integrating the transparent resin member 11A with the first frame 30, the chamber unit 2 is composed of the seal member 10 and the DNA array substrate 12, excluding the transparent member.

[0100] 16 is a cross-sectional view of a chamber apparatus 1 according to a fourth embodiment of the present invention. FIG. 17 is an enlarged view of an area A shown in FIG. 16, an annular convex portion 21B that abuts against the DNA array substrate 12 is formed on the periphery of the opening of the well formation portion 20 on the second surface 10B side (-Z side) of the seal member 10. On the other hand, the annular convex portion 21A of each of the above-mentioned embodiments is not formed on the periphery of the opening of the well formation portion 20 on the first surface 10A side (+Z side) of the seal member 10.

[0101] 17, the sealing performance of well-forming portion 20 on the first surface 10A side (+Z side) of seal member 10 is ensured by flange portion 11b formed on transparent resin member 11A. Flange portion 11b abuts against the inner wall surface of well-forming portion 20 formed by seal member 10 over the entire periphery. This causes the inner wall surface of well-forming portion 20 to be in a compressed state, thereby improving the sealing performance of well-forming portion 20 on the first surface 10A side (+Z side) of seal member 10.

[0102] According to this configuration, by providing a flange portion 11b instead of the above-described annular protrusion 21A and compressing the seal member 10 in the lateral direction (tank direction in the XY plane), the force crushing the seal member 10 in the Z-axis direction can be halved compared to the above-described embodiments. Therefore, even if the metal hinge mechanism 50 is replaced with a resin clamp member 70, sufficient sealing of the well formation portion 20 can be ensured. Therefore, the entire chamber apparatus 1, including the chamber frame 3, can be made disposable.

[0103] While the preferred embodiments of the present invention have been described above with reference to the drawings, the present invention is not limited to the above-described embodiments. The shapes and combinations of the components shown in the above-described embodiments are merely examples, and various modifications can be made based on design requirements, etc., without departing from the spirit of the present invention.

[0104] For example, in the above embodiment, the first frame 30 and the second frame 40 are rotatably connected by the hinge mechanism 50, but the hinge mechanism 50 may be omitted. In this case, a locking mechanism 60 may be provided in the area where the hinge mechanism 50 was located, and the first frame 30 and the second frame 40 may be locked at two locations. [Explanation of symbols]

[0105] 1...chamber device, 2...chamber unit, 3...chamber frame, 4...cap (lid member), 10...sealing member, 10A...first surface, 10B...second surface, 10C...third surface, 10D...fourth surface, 10E...fifth surface, 10F...sixth surface, 11...slide glass (transparent member), 11A...transparent resin member (transparent member), 11a...convex portion, 11b...flange portion, 12...DNA array substrate (substrate), 12a...DNA microarray, 12d...notch portion, 20...well forming portion, 21A...annular convex portion, 21B...annular convex portion, 22...first engagement groove, 23...engagement recess, 24...outlet, 25...inlet, 25a...tapered portion, 26...plate portion, 27a to 17d...positioning recess, 28 ...step, 28a...wall, 29...second engagement groove, 30...first frame, 31...window portion, 32...slit, 33...engagement rib, 34...clamping protrusion, 35...slide groove, 40...second frame, 41...support base, 42a-42d...positioning protrusion, 43...engagement rib, 44...clamping protrusion, 50...hinge mechanism, 60...locking mechanism, 61...latch, 61a...long hole, 61b...claw portion, 62...engaged portion, 62a...step portion, 63...fixing piece, 63a...rotating shaft, 63b...insertion end, 64...through hole, 70...clamp member, 100...sample, 200...bacterial recovery system, 300...nucleic acid extraction system, 400...hybridization reaction system, 500...detection system, 600...nucleic acid analysis system

Claims

1. a sealing member having a plurality of well-forming portions formed therein and penetrating in a thickness direction; a transparent member attached to a first surface side of the sealing member where one end of the well forming portion is open, and which closes the one end of the well forming portion; a substrate attached to a second surface side of the sealing member where the other end of the well formation portion is open, and closing the other end of the well formation portion; a chamber unit comprising: a chamber frame that detachably sandwiches the chamber unit in a direction in which the transparent member, the sealing member, and the substrate overlap, Chamber device.

2. The chamber frame includes: a first frame abutting against the first surface side of the chamber unit; a second frame abutting against the second surface side of the chamber unit; a hinge mechanism that rotatably connects the first frame and the second frame; a locking mechanism that locks the first frame and the second frame together in a folded state. The chamber apparatus according to claim 1 .

3. The locking mechanism is a latch rotatably provided on one of the first frame and the second frame; a latched portion provided on the other of the first frame and the second frame and to which the latch is latched; The chamber apparatus according to claim 2 .

4. At least one surface of the latch and the engaged portion is plated. The chamber apparatus according to claim 3 .

5. an annular convex portion is formed on the periphery of the opening of the well formation portion, the annular convex portion being in compressive contact with the substrate and the transparent member when the chamber unit is sandwiched between the chamber frame; The chamber apparatus according to any one of claims 1 to 4.

6. The sealing member has a first hole and a second hole extending from an inner wall surface of the well forming portion to an outer surface thereof. The chamber apparatus according to any one of claims 1 to 4.

7. a step or a wall is formed on an inner wall surface of the well formation portion between the first hole portion and the second hole portion; The chamber apparatus according to claim 6 .

8. a cover member attached to the sealing member and closing the first hole portion and the second hole portion; The chamber apparatus according to claim 6 .

9. The sealing member is a first engagement groove formed on the first surface side and adapted to engage with a peripheral edge of the transparent member; a second engagement groove formed on the second surface side and adapted to engage with a peripheral edge portion of the substrate; The chamber apparatus according to any one of claims 1 to 4.

10. A hybridization reaction is carried out using the chamber apparatus according to any one of claims 1 to 4. Hybridization reaction system.

11. Analyzing extracted nucleic acids using the hybridization reaction system according to claim 10. Nucleic acid analysis system.

12. a plurality of well forming portions penetrating in the thickness direction; a first hole portion and a second hole portion extending from an inner wall surface to an outer surface of the well forming portion; a step or a wall disposed on an inner wall surface of the well formation portion between the first hole portion and the second hole portion; Sealing material.

Images