Sample reaction accommodation device and method

Abstract

Disclosed in the present invention is a sample reaction accommodation device, comprising: a box body; a pressing cover, which is selectively snap-fitted on the upper portion of the box body; an opening, which penetrates through the pressing cover; and a substrate, on which a sample is provided, the substrate being detachably connected between the box body and the pressing cover, a liquid sealing barrier being provided between the substrate and the pressing cover, and the liquid sealing barrier sealingly abutting against the upper end of the substrate under the pressing action of the pressing cover. The present invention is compatible with substrates of all types (transparent, translucent and opaque), is convenient to operate, saves reaction reagents, effectively eliminates cross contamination between different samples, not only can be used for manual processing procedures, but also is applicable to automated instruments, and separately arranges a sample region and a waste liquid removal region, thereby solving the problem that samples are prone to being punctured during waste liquid removal.

Description

A sample reaction containment device and method Technical Field

[0001] This invention belongs to the field of biological sample processing technology, and particularly relates to a sample reaction container and method. Background Technology

[0002] In life science research, biological samples are frequently prepared into slides and mounted on glass slides for pathological studies. During these studies, pretreatment procedures are often required, including adding reagents, reacting under specific temperature conditions, removing waste liquid, and washing. Traditional pathological examinations only require dewaxing, permeabilization, and staining of the slides, requiring relatively low-cost reagents; simply adding excess reagent to the slide until it covers the sample is sufficient, without the need for precise quantification. However, in space transcriptome sequencing, the pretreatment process for slide samples involves not only dewaxing, permeabilization, and staining, but also blocking and reverse transcription reactions. These reactions require expensive reagents and demand high concentrations. A completely open on-slide reaction environment not only leads to reagent diffusion into sample-free areas of the slide but also easily causes reagent evaporation and concentration changes. Therefore, a reaction space needs to be constructed for the slide samples to meet requirements such as no reagent loss during the reaction, the ability to add reagents, the removal of waste liquid, and minimal contamination.

[0003] Chinese invention patent application CN116888253A discloses a substrate holder, including a base configured to receive a substrate; a cover configured to engage with the base and defining an opening formed by an inner sidewall; and a removable insert defining a surface configured to be received within the opening of the cover. The removable insert includes a pad; a protrusion coupled to the pad; and at least two insertion tabs extending from opposite sides of the removable insert, each insertion tab configured to engage with at least one of the inner sidewalls forming the opening of the cover.

[0004] The above-mentioned patented solution requires placing the slide in the limiting groove of the base first, and then adjusting the pad while the top cover is flipped onto the basic cover but not fully pressed down (the pad cannot move after being fully pressed down) so that the slide sample falls within the pad area. Then the top cover is fully closed and the base and top cover are pressed down. This operation of aligning the slide sample and the pad in a "partially closed" state is not only inconvenient, but also difficult to accurately align the slide sample to the point that it is completely surrounded by the pad when the slide sample and the pad are about the same size. There is even a risk that the bottom of the pad may accidentally rub against the slide sample during the movement, causing it to fall off partially. Technical issues

[0005] In order to overcome the technical problems in the existing technology where the relative position of the sample and the substrate is inconvenient to adjust, and even the possibility of moving the substrate causing the sample to fall off locally. Technical solutions

[0006] One object of the present invention is to provide a sample reaction containment device and method, which adjusts the sample position by moving a substrate and positions the sample by marking or openings on a cap, making sample position adjustment convenient and reliable, and fixing the sample position relative to the containment device, which is convenient for integration into automated instruments.

[0007] To achieve the above objectives, the present invention employs the following technical solution: a sample reaction container, comprising a housing; a cap, the cap being selectively snapped onto the upper part of the housing; an opening, the opening being disposed through the cap; a substrate, on which a sample is disposed, the substrate being detachably connected between the housing and the cap; wherein a sealing liquid barrier is provided between the substrate and the cap; the sealing liquid barrier is sealed against the upper end of the substrate under the pressing action of the cap.

[0008] Furthermore, the inner cavity of the sealing barrier is divided into a sample area that can accommodate the sample and a waste liquid removal area that communicates with the bottom of the sample area; the waste liquid removal area has a structure that is wider at the top and narrower at the bottom, conforming to the contour of the pipette tip.

[0009] During the process of removing waste liquid after the reaction is completed, the pipette tip will not touch the sample, ensuring the integrity of the sample. At the same time, in order to reduce the consumption of reaction liquid, the volume of the waste liquid removal area is minimized while ensuring that the pipette tip can be inserted.

[0010] Furthermore, it includes a first temperature control component disposed at the lower end of the substrate and a second temperature control component optionally disposed at the upper end of the sealing liquid barrier; the second temperature control component includes a heating element and an isolation pad detachably connected to the lower end of the heating element; the lower end of the isolation pad is sealed against the upper end of the sealing liquid barrier; the temperature of the second temperature control component can be selectively higher than the temperature of the first temperature control component.

[0011] While the second temperature control component and the first temperature control component provide the reaction temperature for the sample, the temperature of the second temperature control component is higher than that of the first temperature control component, which can prevent the reagent from adhering to the isolation pad after evaporation; the isolation pad is easy to disassemble; the isolation pad is a disposable item or an easy-to-clean material to prevent mutual interference between different experiments.

[0012] Specifically, the inner bottom of the box is a heat conductor that abuts against the lower end of the substrate; in order to facilitate the handling of the substrate, the box and the first temperature control device are often separable, and the heat conductor can be fixed on the first temperature control device, or more often it can be fixed on the box and integrated with the box.

[0013] Furthermore, the inner wall length (Ls) and width (Ws) of the box body, the length (Lb) and width (Wb) of the substrate, the inner wall length (Lg) and width (Wg) of the sealing liquid barrier, and the minimum effective sealing width t of the sealing liquid barrier and the substrate must satisfy at least one of the following two conditions: ; Since the sample is pre-attached and its position relative to the substrate is random, the inner wall dimensions of the box that satisfy the above conditions ensure that, regardless of where the sample is located on the substrate, the position of the substrate can be adjusted to make the sample aligned with the opening.

[0014] Optionally, a mark is provided inside the box at the lower end of the substrate; the sealing liquid barrier may optionally be directly opposite the mark.

[0015] When the cap is snapped onto the upper part of the box, the mark is directly opposite the opening; the upper part of the sealing liquid fence is provided with a pressing surface, which can be selectively pressed against the lower end of the cap.

[0016] For the transparent substrate, the movement of the substrate and the movement of the sealing liquid barrier are positioned by the markings, which reduces the difficulty of operation and improves the convenience and speed of operation.

[0017] Optionally, a groove is provided on the upper part of the outer wall of the sealing liquid enclosure; the groove is detachably connected to the pressure cap; and the pressure cap is provided with a notch for adjusting the position of the substrate.

[0018] For opaque or semi-transparent substrates (light transmission can affect the sample or reaction process, so some samples require reaction on opaque or semi-transparent substrates), the position of the substrate is adjusted through the notch until the geometric center of the sample is aligned with the opening, then the sealing barrier is installed and the cap is closed.

[0019] Specifically, the outer wall of the sealing barrier is provided with an elastic edge at the upper end of the groove; the upper edge of the elastic edge is provided with a guide slope.

[0020] Furthermore, the pressure cap is provided with a first buckle; the base is provided with a second buckle; the first buckle can be selectively engaged with the second buckle; at least one of the first buckle and the second buckle is an elastic structure, or at least one is directly or indirectly connected to an elastic body.

[0021] Specifically, the first buckle is slidably connected to the cover, and the elastic body is disposed between the first buckle and the cover; the elasticity of the elastic body is used to engage the first buckle with the box body.

[0022] Compared to elastic deformation-based opening and closing mechanisms, improper operation during opening and closing can easily lead to uneven engagement, causing vibrations on the box body and resulting in the sample or reagent spilling or moving outwards, or even damaging the sample. The cap and the box body are stably connected by a snap-fit ​​structure, ensuring that the sealing liquid barrier is securely clamped. The first snap-fit ​​is manually driven during the locking and unlocking process, resulting in a smooth opening and closing process without vibration on the box body.

[0023] Furthermore, a clamping mechanism is provided inside the box, which can selectively clamp the substrate.

[0024] Specifically, one end of the clamping mechanism is rotatably connected to the box body, and the other end can selectively press against the upper end of the substrate.

[0025] The clamping mechanism presses the substrate to prevent the position of the sample or the substrate from changing during the adjustment of the sealing liquid fence.

[0026] A sample reaction containment method, implemented based on the above-mentioned sample reaction containment device, includes the following steps:

[0027] (1) Place the substrate with the pre-attached sample inside the box and adjust the position of the sample;

[0028] (2) Place the sealing barrier onto the substrate such that the sealing barrier is aligned with the mark, or install the sealing barrier onto the opening of the cap;

[0029] (3) Close the cap, the lower end of the sealing liquid fence is sealed and pressed against the upper end of the substrate, and the sample is located in the cavity formed by the inner wall of the sealing liquid fence and the upper end of the substrate;

[0030] (4) Add reagents into the sealing liquid enclosure;

[0031] (5) Press the second temperature control device against the upper end of the sealing liquid fence to control the first temperature control device and the second temperature control device to a suitable temperature;

[0032] (6) After the sample and the reagent have reacted sufficiently, the second temperature control device is removed;

[0033] (7) Insert the pipette tip into the bottom of the waste liquid removal area and aspirate the waste liquid;

[0034] (8) Open the cover and remove the substrate.

[0035] Specifically, in step (2), when the substrate is transparent, the sealing liquid barrier can be aligned with the mark, or the sealing liquid barrier can be installed on the opening; when the substrate is opaque, the sealing liquid barrier can be installed on the opening.

[0036] Specifically for opaque or semi-transparent substrates (which cannot be positioned by the box), first close the cap, adjust the sample to be aligned with the geometric center of the opening through the notch, then open the cap to install the sealing liquid fence, and then close the cap again. This will allow the sample to be placed within the sealing liquid fence. The adjustment of the substrate is smooth and will not accidentally touch the sample, reducing the difficulty of operation. Beneficial effects

[0037] 1. It is compatible with all types of substrates (transparent, semi-transparent, opaque). The position of the substrate can be easily adjusted according to the position of the mark / opening, so that the sample can enter the designated position, reducing the difficulty of operation and improving convenience.

[0038] 2. By setting up a sealing barrier, the space for the reaction reagents required for sample processing is directly constructed, saving reaction reagents and realizing the quantitative control of reaction reagents; moreover, the sealing barrier is a disposable item or made of easy-to-clean material, which effectively eliminates cross-contamination between different samples and reduces experimental costs.

[0039] 3. By designing the housing, samples attached to any position on the substrate can be used for processing and analysis, which increases the scope of application and reduces the requirements for sample mounting. Moreover, the position of the sample relative to the sample reaction container is fixed, so the positions for adding reagents, removing waste liquid, and pressing the heating cap can all be fixed, making the device suitable not only for manual processing but also for automated instruments.

[0040] 4. By setting up a first temperature control device and a second temperature control device, the required temperature conditions and enclosed space can be provided for the sample reaction, enabling biological reactions such as closed reactions and reverse transcription reactions on the sample on the slide. The disposable or easy-to-clean isolation pads prevent cross-contamination between different samples.

[0041] 5. The sample area and the waste liquid removal area are separated on the sealing liquid fence, which solves the problem that the sample is easily punctured when removing waste liquid.

[0042] 6. The sliding design of the first buckle improves the convenience and reliability of operation, and also makes the process of locking and unlocking the box and the cap smoother, preventing vibration during opening and causing spillage of samples and reagents.

[0043] 7. By setting up a clamping mechanism, the substrate that has been adjusted can be initially positioned. Placing a sealing liquid barrier prevents the substrate from being touched, ensuring the positional accuracy of the sample and helping to improve the accuracy of the test. Attached Figure Description

[0044] Figures 1 and 2 are schematic diagrams of the structure of the first embodiment of the present invention;

[0045] Figures 3 and 4 are schematic cross-sectional views of the first embodiment of the present invention.

[0046] Figure 5 is a schematic diagram of the liquid sealing fence according to the first embodiment of the present invention;

[0047] Figure 6 is a schematic diagram of the liquid sealing fence according to the second embodiment of the present invention;

[0048] Figures 7 and 8 are schematic diagrams of the structure of the second embodiment of the present invention;

[0049] Figure 9 is a top view of the second embodiment of the present invention;

[0050] Figure 10 is a cross-sectional structural schematic diagram of the second embodiment of the present invention;

[0051] Figure 11 is a schematic diagram of the substrate region division according to the third embodiment of the present invention;

[0052] Figure 12 is a schematic diagram showing the relative positions of the sample and the box in the third embodiment of the present invention;

[0053] Figure 13 is a structural schematic diagram of the fourth embodiment of the present invention;

[0054] Figure 14 is a schematic diagram of the operation steps of the sample reaction containment method of the present invention.

[0055] In the diagram: 11. Substrate; 111. Edge area; 112. Sample pre-positioning area; 1121. First pre-positioning area; 1122. Second pre-positioning area; 1123. Overlapping area; 12. Sample; 2. Box body; 21. Receptacle space; 22. Mark; 23. Heat conductor; 24. Clamping mechanism; 25. Second buckle; 3. Sealing barrier; 31. First sealing surface; 32. Enclosing surface; 33. Pressing surface; 34. Second sealing surface; 35. Sample area; 36. Waste liquid removal area; 37. Groove; 38. Guide slope; 4. Cap; 41. Opening; 42. Upper surface; 43. First buckle; 44. Elastomer; 45. Notch; 5. First temperature control device; 6. Second temperature control device; 61. Isolation pad; 611. Hanging ear; 62. Heating element; 621. Hook; 7. Pipette tip; 8. Reagent. Embodiments of the present invention

[0056] The present invention will now be further described in conjunction with the accompanying drawings and specific embodiments. It should be noted that, without conflict, the various embodiments or technical features described below can be arbitrarily combined to form new embodiments.

[0057] In the description of this invention, it should be noted that the directional terms, such as "center," "lateral," "longitudinal," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," and "counterclockwise," indicate the orientation and positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. They should not be construed as limiting the specific protection scope of this invention.

[0058] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of technical features. Thus, the use of "first" and "second" to define a feature may explicitly or implicitly include one or more of that feature, and in the description of this invention, "a number" means two or more, unless otherwise explicitly specified.

[0059] In this invention, unless otherwise explicitly specified and limited, terms such as "set" and "install" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can also refer to a mechanical connection; they can refer to a direct connection or a connection through an intermediate medium; or they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0060] Chinese invention patent application CN116888253A discloses a modular measurement support device that constructs a sample reaction cell using a substrate (glass slide) and a pad fixed to an insert that can move relative to the substrate holder. However, this device has three problems: 1. The position of the pad surrounding the sample slide relative to the device base (determined by the position of the slide on the glass slide) is uncertain, which leads to uncertainty in the positions for adding reagents and removing waste liquid for different sample slides. Therefore, this device is not suitable for use in automated instruments. 2. When using this device, the slide must first be placed in the limiting groove of the base. Then, the top cover must be flipped up to the basic cover but not fully pressed down. Adjust the pad (the pad cannot move after being fully pressed down) so that the slide sample falls within the pad's range. Then, fully cover the top cover and press down the base and top cover. This operation of aligning the slide sample and pad in a "partially covered" state is not only inconvenient, but also makes it difficult to accurately align the slide sample to the point where it is completely surrounded by the pad when the slide sample and pad are of similar size. There is even a risk that the bottom of the pad may accidentally rub against the slide sample during the movement, causing it to partially fall off. 3) The width of the insert's frame determines that the minimum distance between the sample and the edge of the slide cannot be small enough, which limits the application of many samples that are placed relatively close to the edge.

[0061] Referring to Figures 1-5, a first embodiment of the invention is shown, a sample reaction container device, including a box body 2, a pressure cap 4 disposed on the box body 2, and a sealing liquid fence 3 detachably connected to the inside of the box body 2; the pressure cap 4 can be selectively snapped onto the upper part of the box body 2; one end of the pressure cap 4 can rotate relative to one end of the box body 2; the pressure cap 4 and the box body 2 are an integral structure; or two separate structures that can be detached.

[0062] The upper end of the box body 2 is provided with a rectangular accommodating space 21; a substrate 11 is detachably connected to the accommodating space 21; a sample 12 is pre-attached to the upper end of the substrate 11; the accommodating space 21 allows the substrate 11 to move within it to adjust the position of the sample 12.

[0063] The sealing liquid barrier 3 is a disposable consumable or an easy-to-clean component. The shape of the sealing liquid barrier 3 is not limited; it can be rectangular, circular, or other irregular shapes. Based on common sample shapes, a rectangular shape is preferred.

[0064] The sealing barrier 3 includes a first sealing surface 31 located at the lower end of the sealing barrier 3, a closing surface 32 located on the inner wall of the sealing barrier 3, and a pressing surface 33 located at the upper end of the sealing barrier 3; the closing area of ​​the closing surface 32 is greater than or equal to the projected area of ​​the sample 12.

[0065] The inner cavity of the sealing enclosure 3 is divided into a sample area 35 and a waste liquid removal area 36 communicating with the sample area 35. The waste liquid removal area 36 is separated from the sample area 35 on the projection plane. When removing waste liquid, the pipette tip 7 extends into the bottom of the waste liquid removal area 36 to aspirate the waste liquid. The tip of the pipette tip 7 will not contact the sample 12, thereby preventing puncture of the sample. In order to minimize the volume of the waste liquid removal area 36 and reduce the amount of reagent used, the cavity of the waste liquid removal area 36 is adapted to the pipette tip 7 with a structure that is larger at the top and smaller at the bottom.

[0066] The box body 2 is provided with a clamping mechanism 24; one end of the clamping mechanism 24 is rotatably connected to the box body 2, and the other end can selectively press the substrate 11; after the position of the sample 12 is adjusted, the clamping mechanism 24 is rotated to clamp the substrate 11 to prevent the substrate 11 from being moved when the sealing liquid fence 3 is subsequently aligned and placed.

[0067] When the cap 4 is snapped onto the upper part of the box body 2, the cap 4 acts on the pressing surface 33, so that the first sealing surface 31 is pressed against the upper surface of the substrate 11, and the enclosing surface 32 and the upper surface of the substrate 11 form a sample reaction accommodating cavity; the sample 12 is located in the sample reaction accommodating cavity; the cap 4 is provided with an opening 41 at the corresponding position of the sealing liquid fence 3 to facilitate the addition of reagents and the removal of waste liquid.

[0068] To make the operation of pressing and releasing the sealing liquid fence 3 more convenient, the pressure cap 4 is provided with a first buckle 43; the box body 2 is provided with a second buckle 25; the first buckle 43 can be selectively engaged with the second buckle 25; at least one of the first buckle 43 and the second buckle 25 is an elastic structure, or at least one is directly or indirectly connected to an elastic body 44.

[0069] Specifically, the first buckle 43 is slidably connected to the lower end of the pressure cap 4; an elastic body 44 is provided between the first buckle 43 and the pressure cap 4; the elastic body 44 can be a spring; when the pressure cap 4 is closed, the first buckle 43 hooks and locks with the second buckle 25 under the elastic force of the elastic body 44, so that the liquid sealing fence 3 is pressed; when it is necessary to open the pressure cap 4, pressing the first buckle 43 can separate the first buckle 43 and the second buckle 25, and then the pressure cap 4 can be opened.

[0070] For the transparent substrate 11, to facilitate the adjustment of the sample 12's position, a mark 22 is provided at the bottom of the inner side of the housing 2 to indicate the sample placement area. The position of the mark 22 relative to the entire housing 2 is fixed, and the indicated area of ​​the mark 22 is greater than or equal to the size of the sample 12, and less than or equal to the size of the area enclosed by the first sealing surface 31. When adjusting the position of the sample 12, it is only necessary to move the substrate 11 so that the sample 12 is completely within the inner frame of the mark 22; when placing the sealing liquid barrier 3, the sealing liquid barrier 3 is also aligned with the mark 22.

[0071] Referring to Figures 6-10, this is the second embodiment of the present invention. The difference between this embodiment and the first embodiment is that, for the opaque or semi-transparent substrate 11, after the substrate 11 is placed, the sample 12 and the sealing liquid fence 3 cannot be accurately located by the mark 22 (the schematic range of the mark 22 is covered by the substrate 11).

[0072] To solve this problem, a groove 37 is provided on the upper end of the outer wall of the sealing liquid fence 3; an elastic edge is provided on the upper end of the outer wall of the sealing liquid fence located at the groove; a guide slope 38 is provided on the upper edge of the elastic edge; the sealing liquid fence 3 is detachably connected to the opening 41 of the pressure cap 4; the area of ​​the pressure cap 4 is smaller than that of the substrate 11; or the pressure cap 4 is provided with a notch 45 for adjusting the substrate 11.

[0073] First, close the pressure cap 4, adjust the geometric center of the sample 12 to be aligned with the opening 41 through the notch 45, then open the pressure cap 4 to install the sealing liquid fence 3, and then close the pressure cap 4 again so that the sample 12 is located inside the sealing liquid fence 3.

[0074] Referring to Figures 11 and 12, this is the third embodiment of the present invention. Based on the first and second embodiments, the following additions are made: The substrate 11 is divided into an edge region 111 located at the outer periphery and a sample pre-positioning area 112 located inside the edge region 111; to increase the adaptability to the pre-positioning position of the sample 12, so that any sample located on the substrate 11 (excluding samples outside the edge region) within the pre-positioning area can be adjusted into the sealing liquid enclosure 3, the length (Ls) and width (Ws) of the accommodating space 21, the length (Lb) and width (Wb) of the substrate 11, the length (Lg) and width (Wg) of the enclosing surface 32, and the minimum effective sealing width t of the first sealing surface 31 must satisfy at least one of the following two conditions: To reduce the size of the sample reaction container, preferably, and The minimum effective sealing width of the first sealing surface 31 is t, which mainly depends on the size, material hardness, and compression degree of the sealing liquid enclosure 3. The range of t can be 0.1mm to 10mm. When t = 0.1mm, the sample pre-positioning area 112 reaches its maximum range, achieving maximum containment of the sample pre-positioning location. Considering the manufacturing cost and sealing effect of the sealing liquid enclosure 3, preferably t = 2mm. Any sample 12 located within the sample pre-positioning area 112 can be adjusted into the enclosed area of ​​the sealing liquid enclosure 3. Specifically, the sample 12 is a 6mm × 6mm tissue slice; the substrate 11 has a length Lb = 76mm and a width Wb = 26mm; the enclosed surface 32 has a length Lg = 8mm and a width Wg = 8mm; the minimum effective sealing width of the first sealing surface 31 is t = 2mm; the accommodating space 21 has a length Ls = 108mm and a width Ws = 40mm, satisfying… and As shown in Figure 11, the sample pre-positionable area 112 includes a first pre-positionable area 1121 and a second pre-positionable area 1122 located on both sides, and an overlapping area 1123 located in the middle; the overlapping area 1123 is the overlapping area of ​​the first pre-positionable area 1121 and the second pre-positionable area 1122; the lengths of the first pre-positionable area 1121 and the second pre-positionable area 1122 are both... The width of the overlapping portion is equal to Lg, which is 8mm. As shown in Figure 12, the relative positions of the sample 12 and the box 2 are shown in four extreme positions where the sample 12 is completely located within the sample prepositioning area 112. When the sample is located within the first prepositioning area 1121, or when the sample is located within the second prepositioning area 1122 (when the substrate 11 is rotated horizontally by 180°), that is, when the substrate 11 is located at any position within the accommodating space 21, the sample 12 can be directly opposite the opening 41.

[0075] Referring to Figure 13, which is the fourth embodiment of the present invention, the following additions are made based on the first, second, and third embodiments: the sample reaction container further includes a first temperature control device 5 disposed at the lower end of the housing 2 and a second temperature control device 6 movably disposed at the upper end of the pressure cap 4; the lower end of the housing 2 is a heat conductor 23; the upper surface of the heat conductor 23 is in full contact with the lower surface of the substrate 11; the lower surface of the heat conductor 23 is in full contact with the first temperature control device 5; thereby, the sample reaction temperature can be controlled.

[0076] The material of the heat conductor 23 is not limited; it can be a metal such as aluminum or copper, or a non-metal such as ceramic. The first temperature control device 5 and the box body 2 are fixed together. The heat conductor 23 and the first temperature control device 5 are integrated.

[0077] The second temperature control device 6 includes an isolation pad 61 and a heating element 62 detachably connected to the upper end of the isolation pad 61; by controlling the temperature of the second temperature control device 6 to be higher than the sample reaction temperature, it is possible to prevent the reagent from evaporating and condensing on the second temperature control device 6.

[0078] The material of the isolation pad 61 is not limited; it can be a soft material such as silicone, a hard plastic such as polypropylene, or other materials. The isolation pad 61 and the heating element 62 are easy to fix and separate, and the fixing method is not limited, such as hanging it with an ear piece or sticking it with easy-to-remove adhesive.

[0079] The sealing barrier 3 also includes a second sealing surface 34, and the two outer walls opposite the isolation pad 61 are respectively provided with hanging ears 611; the lower end of the heating element 62 is provided with two hooks 621 that are respectively engaged with the two hanging ears 611; the isolation pad 61 and the heating element 62 are easy to fix and separate.

[0080] After the second temperature control device 6 is pressed down, a closed sample reaction containment cavity is formed between the upper surface of the substrate 11, the enclosure surface 32, and the lower surface of the isolation pad 61. Under the action of the first temperature control device 5 and the second temperature control device 6, the sample 12 and reagent in the sample reaction containment cavity can react under the required temperature conditions and in a sealed environment, and the reagent will not evaporate and condense on the surface of the isolation pad 61.

[0081] Both the sealing liquid barrier 3 and the isolation pad 61 are disposable consumables or easy-to-clean parts, preventing cross-contamination between different samples. In order to ensure the sealing effect, after the cap 4 presses the sealing liquid barrier 3, the second sealing surface 34 on the sealing liquid barrier 3 should not be lower than the upper surface 42 of the cap 4.

[0082] Referring to Figure 14, a sample reaction containment method, implemented using the aforementioned sample reaction containment device, for the opaque or semi-transparent substrate, includes the following steps:

[0083] (1) Place the substrate 11 with the sample 12 pre-attached inside the box;

[0084] (2) Close the pressure cap 4, and adjust the substrate 11 through the notch 45 so that the sample 12 is aligned with the geometric center of the opening 41;

[0085] (3) Rotate the clamping mechanism 24 to press the substrate 11.

[0086] (4) Open the pressure cap 4 and install the groove 37 of the sealing liquid fence 3 into the opening 41;

[0087] (5) Close the cap 4, the lower end of the sealing liquid fence 3 is sealed against the upper end of the substrate 11, and the sample 12 is located inside the sealing liquid fence 3;

[0088] (6) Add reagent 8 into the sealing liquid enclosure 3;

[0089] (7) Press the second temperature control device 6 against the upper end of the sealing liquid fence 3 to control the first temperature control device 5 and the second temperature control device 6 to a suitable temperature, wherein the temperature of the second temperature control device 6 is greater than the temperature of the first temperature control device 5;

[0090] (8) After the sample 12 has fully reacted with the reagent 8, the second temperature control device 6 is removed;

[0091] (9) The pipette tip 7 is inserted into the bottom of the waste liquid removal area 36 to remove the waste liquid;

[0092] (10) According to the actual experimental process, steps (6) to (9) are repeated multiple times;

[0093] (11) Open the cover 4 and remove the substrate 11.

[0094] A sample reaction containment method, implemented using the aforementioned sample reaction containment device, for a transparent substrate, includes the following steps:

[0095] (1) Place the substrate 11 with the sample 12 pre-attached inside the box;

[0096] (2) Adjust the substrate 11 so that the sample 12 is directly aligned with the mark 22;

[0097] (3) Rotate the clamping mechanism 24 to press the substrate 11;

[0098] (4) Place the sealing liquid barrier 3 on the upper end of the substrate 11, and make the sealing liquid barrier 3 face the mark 22;

[0099] (5) Close the cap 4, the lower end of the sealing liquid fence 3 is sealed against the upper end of the substrate 11, and the sample 12 is located inside the sealing liquid fence 3;

[0100] (6) Add reagent 8 into the sealing liquid enclosure 3;

[0101] (7) Press the second temperature control device 6 against the upper end of the sealing liquid fence 3, and control the first temperature control device 5 and the second temperature control device 6 to a suitable temperature respectively, wherein the temperature of the second temperature control device 6 is greater than the temperature of the first temperature control device 5;

[0102] (8) After the sample 12 has fully reacted with the reagent 8, the second temperature control device 6 is removed;

[0103] (9) The pipette tip 7 is inserted into the bottom of the waste liquid removal area 36 to remove the waste liquid;

[0104] (10) According to the actual experimental process, steps (6) to (9) are repeated multiple times;

[0105] (11) Open the cover 4 and remove the substrate 11.

[0106] The above description is only a specific embodiment of the present invention, but the technical features of the present invention are not limited thereto. Any changes or modifications made by those skilled in the art within the scope of the present invention are covered by the patent scope of the present invention.

Claims

1. A sample reaction container, characterized in that: Includes a box; A pressure cap, which can be selectively snapped onto the upper part of the housing; An opening, which extends through the gland; A substrate on which a sample is disposed, and the substrate is detachably connected between the housing and the cover. The box body allows the substrate to move within it to adjust the position of the sample; a sealing liquid barrier is provided between the substrate and the cap; The sealing barrier is sealed against the upper end of the substrate under the pressure of the pressure cap; the sealing barrier is detachably connected to the pressure cap.

2. The receiving device as claimed in claim 1, characterized in that: The inner cavity of the sealing liquid enclosure is divided into a sample area that can accommodate the sample and a waste liquid removal area that communicates with the bottom of the sample area.

3. The receiving device as described in claim 1 or 2, characterized in that: It includes a first temperature control component that can be selectively disposed at the lower end of the substrate and a second temperature control component that can be selectively disposed at the upper end of the sealing liquid barrier; the second temperature control component includes a heating element and an isolation pad that is detachably connected to the lower end of the heating element; the lower end of the isolation pad is sealed against the upper end of the sealing liquid barrier.

4. The receiving device as described in claim 1 or 2, characterized in that: The inner wall length (Ls) and width (Ws) of the box body, the length (Lb) and width (Wb) of the substrate, the inner wall length (Lg) and width (Wg) of the sealing liquid barrier, and the minimum effective sealing width t of the sealing liquid barrier and the substrate must satisfy at least one of the following two conditions: ; .

5. The receiving device as described in claim 1 or 2, characterized in that: A mark is provided inside the box at the lower end of the substrate; the sealing liquid fence can be optionally aligned with the mark.

6. The receiving device as claimed in claim 1 or 2, characterized in that: The upper part of the outer wall of the sealing liquid enclosure is provided with a groove; the groove is detachably connected to the pressure cap; the pressure cap is provided with a notch for adjusting the position of the substrate.

7. The receiving device as claimed in claim 1 or 2, characterized in that: The cover is provided with a first buckle; the box body is provided with a second buckle; the first buckle can be selectively engaged with the second buckle; at least one of the first buckle and the second buckle is an elastic structure, or at least one is directly or indirectly connected to an elastic body.

8. The receiving device as claimed in claim 7, characterized in that: The box is equipped with a clamping mechanism, which can selectively clamp the substrate.

9. A sample reaction containment method, characterized in that: The sample reaction container based on claims 1-8 is implemented by the following steps: (1) Place the substrate with the pre-attached sample inside the box and adjust the position of the sample; (2) A mark is provided at the lower end of the substrate inside the box; the sealing liquid fence is placed on the substrate so that the sealing liquid fence is aligned with the mark, or the sealing liquid fence is installed on the opening of the cap; (3) Close the cap, the lower end of the sealing liquid fence is sealed and pressed against the upper end of the substrate, and the sample is located in the cavity formed by the inner wall of the sealing liquid fence and the upper end of the substrate; (4) Add reagents into the sealing liquid enclosure; (5) The sample reaction container includes a first temperature control component disposed at the lower end of the substrate and a second temperature control component optionally disposed at the upper end of the sealing liquid barrier; the second temperature control device is pressed against the upper end of the sealing liquid barrier to control the first temperature control device and the second temperature control device to a suitable temperature; (6) After the sample and the reagent have reacted sufficiently, the second temperature control device is removed; (7) The inner cavity of the sealing enclosure is divided into a sample area that can accommodate the sample and a waste liquid removal area that communicates with the bottom of the sample area. The pipette tip is inserted into the bottom of the waste liquid removal area to remove the waste liquid. (8) Open the cover and remove the substrate.

10. The accommodating method as described in claim 9, characterized in that: In step (2), when the substrate is transparent, the sealing liquid barrier can be aligned with the mark, or the sealing liquid barrier can be installed on the opening; When the substrate is opaque, the sealing liquid barrier is installed onto the opening.

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