A sample reaction container device and method

Abstract

The present invention discloses a sample reaction accommodating device, comprising a box body; a pressing cover, which is selectively clamped on the upper part of the box body; an opening, which is arranged through the pressing cover; a substrate, on which a sample is arranged, and the substrate is detachably connected between the box body and the pressing cover; wherein, a liquid sealing fence is arranged between the substrate and the pressing cover; the liquid sealing fence is tightly pressed against the upper end of the substrate in a sealed manner under the pressing action of the pressing cover. It can be compatible with all types (transparent, semi-transparent, opaque) of substrates, with convenient operation; saves reaction reagents and effectively eliminates cross-contamination between different samples; can be used not only for manual processing procedures but also for automated instruments; separates the sample area and the waste liquid removal area, solving the problem that the sample is easily punctured during waste liquid removal.

Description

Technical Field

[0001] The present invention belongs to the technical field of biological sample processing, and particularly relates to a sample reaction accommodating device and method. Background Art

[0002] In the field of life science research, it is often necessary to make biological samples into slices and attach them to glass slides for pathological research. During the research, pretreatment operations such as adding reagents, reacting under certain temperature conditions, removing waste liquid, and cleaning are often required for the biological tissue samples attached to the glass slides. Traditional pathological examinations only need to perform operations such as dewaxing, permeabilization, and staining on the slices on the glass slides, and the required reagent costs are relatively low. It only needs to drop an excessive amount of reagent on the glass slide to cover the sample, and there is no precise quantification requirement. In spatial transcriptome sequencing, in addition to dewaxing, permeabilization, staining, etc., the pretreatment process for the slice samples attached to the glass slides also requires blocking reactions, reverse transcription reactions, etc. The reagents required for these reactions are costly, and the requirements for reagent concentration are also high. Moreover, the completely open on-chip reaction environment not only causes the reagents to diffuse to the sample-free area on the glass slide, but also easily leads to reagent evaporation and concentration changes. Therefore, it is necessary to construct a reaction space for the slice samples on the glass slide to meet the requirements of no loss of reagents, addition of reagents, removal of waste liquid, and low contamination during the reaction process.

[0003] The Chinese patent application with the publication number CN116888253A discloses a substrate holder, which includes a base configured to receive a substrate; a cover configured to engage with the base in a matching manner, the cover defining an opening formed by an inner wall; and a removable insert defining a surface, the removable insert being configured to be received within the opening of the cover. The removable insert includes a gasket; a protrusion coupled to the gasket; and at least two insertion tabs extending from opposite sides of the removable insert, each insertion tab being configured to engage with at least one of the inner walls forming the opening of the cover.

[0004] During the use of the above patent solution, it is necessary to first place the glass slide in the limiting groove of the base, and then adjust the gasket (the gasket cannot be moved after being fully pressed) when the upper cover is flipped to the state where it is basically covered but not fully pressed. After the slice sample falls within the range of the gasket, then fully cover and press the base and the upper cover. This operation of aligning the slice sample and the gasket in the state of "seemingly covered but not covered" is not only inconvenient, but also when the slice sample and the gasket are of similar size, it is very difficult to accurately align the sample to the extent that it is completely surrounded by the gasket. There is even a risk that the bottom of the gasket accidentally rubs against the slice sample during the movement, causing partial detachment of the sample. Summary of the Invention

[0005] In order to overcome the technical problem that the relative positions of the sample and the substrate in the prior art are inconvenient to adjust, and even the movement of the substrate may cause partial detachment of the sample; an object of the present invention is to provide a sample reaction accommodating device and method, which adjust the sample position by moving the substrate, and locate the sample through markings or openings on the capping, so that the adjustment of the sample position becomes convenient and reliable, and the position of the sample relative to the accommodating device is fixed, which is convenient for integration into an automated instrument.

[0006] To achieve the above object, the present invention is implemented by the following technical solutions: A sample reaction accommodating device includes a box body; a capping, the capping is selectively snap-connected to the upper part of the box body; an opening, the opening is disposed through the capping; a substrate, a sample is disposed on the substrate, and the substrate is detachably connected between the box body and the capping; wherein, a liquid-sealing fence is disposed between the substrate and the capping; the liquid-sealing fence is hermetically abutted against the upper end of the substrate under the pressing action of the capping.

[0007] Further, the inner cavity of the liquid-sealing fence is divided into a sample area for accommodating the sample and a waste liquid removal area communicating with the bottom of the sample area; the waste liquid removal area is a structure with a wider upper part and a narrower lower part that fits the contour of the pipette tip.

[0008] During the process of sucking the 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 the reaction liquid, the waste liquid removal area is minimized in volume on the premise of ensuring that the pipette tip can be inserted.

[0009] Further, it includes a first temperature control component disposed at the lower end of the substrate and a second temperature control component selectively disposed at the upper end of the liquid-sealing fence; 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 hermetically abutted against the upper end of the liquid-sealing fence; the temperature of the second temperature control component is selectively greater than the temperature of the first temperature control component.

[0010] While the second temperature control component and the first temperature control component provide a reaction temperature for the sample, the temperature of the second temperature control component is greater than the temperature of the first temperature control component, which can prevent the reagent from evaporating and adhering to the isolation pad; the isolation pad is convenient for disassembly; the isolation pad is a disposable item or a material that is easy to clean, preventing mutual interference between different experiments.

[0011] Specifically, the inner bottom end of the box body is a heat-conducting body that abuts against the lower end of the substrate; for the convenience of the operation of taking and placing the substrate, the box body and the first temperature control device are often separable, the heat-conducting body can be fixed on the first temperature control device, and more often it can also be fixed on the box body and is an integral structure with the box body.

[0012] Further, for the inner wall length (Ls) and width (Ws) of the box body, the length (Lb) and width (Wb) of the substrate, and the length (Lg) and width (Wg) of the inner wall of the liquid-sealing fence, the minimum effective sealing width t between the liquid-sealing fence and the substrate in sealed contact satisfies at least one of the following two conditions:

[0013] Condition 1: and W s ≥2W b -W g -2t;

[0014] Condition 2; and L s ≥2L b -L g -2t.

[0015] Since the sample is pre-pasted and its position relative to the substrate is random, the inner wall dimensions of the box body that meet the above conditions ensure that no matter where the sample is located on the substrate, the position of the substrate can be adjusted to align the sample with the opening.

[0016] Optionally, a mark is provided at the lower end of the substrate inside the box body; the liquid-sealing fence can be optionally aligned with the mark.

[0017] When the gland is snap-fitted to the upper part of the box body, the mark is aligned with the opening; a pressing surface is provided on the upper part of the liquid-sealing fence, and the pressing surface can be optionally in tight contact with the lower end of the gland.

[0018] For the transparent substrate, the movement arrangement of the substrate and the movement arrangement of the liquid-sealing fence are positioned by the mark, reducing the operation difficulty and improving the operation convenience and speed.

[0019] Optionally, a groove is provided on the upper part of the outer wall of the liquid-sealing fence; the groove is detachably connected to the gland; a notch for adjusting the position of the substrate is provided on the gland.

[0020] For the non-transparent or semi-transparent substrate (light transmission will affect the sample or the reaction process, so some samples require reactions on the non-transparent or semi-transparent substrate), the position of the substrate is adjusted through the notch. When the geometric centers of the sample and the opening are aligned, the liquid-sealing fence is installed and the gland is closed.

[0021] Specifically, an elastic edge is provided at the upper end of the outer wall of the liquid-sealing fence; a guiding inclined surface is provided at the upper edge of the elastic edge.

[0022] Further, a first buckle is provided on the pressing cover; a second buckle is provided on the base; the first buckle is selectively clamped 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.

[0023] Specifically, the first buckle is slidably connected to the pressing cover, and the elastic body is arranged between the first buckle and the pressing cover; the elastic force of the elastic body is used to make the first buckle clamped with the box body.

[0024] Compared with the opening and closing form of elastic deformation, at the moment of opening and closing, it is easy to cause unsmooth clamping due to improper operation, which will generate vibration on the box body, causing the sample or reagent to overflow or move outwards, and even causing damage to the sample; the pressing cover and the box body are stably connected through a buckle structure, so that the liquid sealing fence is stably clamped. During the process of clamping and loosening the first buckle, the first buckle is driven manually, and the opening and closing process is stable without generating vibration on the box body.

[0025] Further, a clamping mechanism is arranged inside the box body, and the clamping mechanism can selectively press the substrate.

[0026] Specifically, one end of the clamping mechanism is rotatably connected to the box body, and the other end can selectively press the upper end of the substrate.

[0027] The clamping mechanism presses the substrate to prevent the position of the sample or the substrate from changing during the process of moving and adjusting the liquid sealing fence.

[0028] A method for accommodating a sample reaction, which is realized based on the above sample reaction accommodating device, includes the following steps:

[0029] (1) Place the substrate pre - attached with the sample inside the box body and adjust the position of the sample;

[0030] (2) Place the liquid sealing fence on the substrate so that the liquid sealing fence is aligned with the mark, or install the liquid sealing fence on the opening of the pressing cover;

[0031] (3) Close the pressing cover, the lower end of the liquid sealing fence is hermetically abutted against the upper end of the substrate, and the sample is located in the cavity formed by the inner wall of the liquid sealing fence and the upper end of the substrate;

[0032] (4) Add reagent into the liquid sealing fence;

[0033] (5) Press the second temperature control device against the upper end of the liquid sealing fence, and control the first temperature control device and the second temperature control device to appropriate temperatures;

[0034] After the sample and the reagent fully react, remove the second temperature control device;

[0035] (7) Insert the pipette tip into the bottom of the waste liquid removal area to suck away the waste liquid;

[0036] (8) Open the gland and remove the substrate.

[0037] Specifically, in step (2), when the substrate is transparent, it is possible to align the sealing liquid fence with the mark or install the sealing liquid fence on the opening; when the substrate is opaque, install the sealing liquid fence on the opening.

[0038] Particularly for the opaque or semi - transparent substrate (the substrate cannot be positioned through the box body), first close the gland, adjust the geometric centers of the sample and the opening to be directly opposite through the notch, then open the gland to install the sealing liquid fence, and then close the gland again, so that the sample is located within the sealing liquid fence. The adjustment of the substrate is smooth, the sample will not be accidentally touched, and the operation difficulty is reduced.

[0039] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0040] 1. It can be compatible with all types (transparent, semi - transparent, opaque) of substrates. The position of the substrate can be easily adjusted according to the position of the mark / opening, enabling the sample to enter the specified position, reducing the operation difficulty, and improving the convenience.

[0041] 2. By setting the sealing liquid fence, a reaction reagent accommodation space required for sample treatment is directly constructed, saving the reaction reagent and achieving the quantification of the reaction reagent; and the sealing liquid fence is a disposable item or made of easily cleanable material, effectively eliminating cross - contamination between different samples while reducing the test cost.

[0042] 3. By setting the box body, the sample attached to any position of the substrate can be used for treatment and analysis, expanding the applicable range and reducing the requirements for the sample pasting link; and the position of the sample relative to the sample reaction accommodation device is fixed. Therefore, the positions for adding reagents, removing waste liquid, and pressing the heat - resistant cover can be fixed, enabling the device to be used not only for manual processing procedures but also for automated instruments.

[0043] 4. By setting the first temperature control device and the second temperature control device, the temperature conditions and closed space required for sample reaction can be provided, realizing biological reactions such as closed reaction and reverse transcription reaction on the sample on the glass slide. The disposable or easily cleanable isolation pad prevents cross - contamination between different samples.

[0044] 5. The sample area and the waste liquid removal area are separately arranged on the liquid sealing fence, solving the problem that the sample is easily punctured during waste liquid removal.

[0045] 6. The sliding setting of the first buckle not only improves the operation convenience and reliability, but also makes the buckling and opening processes of the box body and the pressing cover smoother, preventing vibration during opening and causing the spillage of samples and reagents.

[0046] 7. By setting the clamping mechanism, the substrate in the adjusted position can be preliminarily positioned, preventing the substrate from being touched when placing the liquid sealing fence, ensuring the position accuracy of the sample, and helping to improve the accuracy of the test. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 、 Figure 2 is a schematic structural diagram of the first embodiment of the present invention;

[0048] Figure 3 、 Figure 4 is a schematic cross-sectional structural diagram of the first embodiment of the present invention;

[0049] Figure 5 is a schematic structural diagram of the liquid sealing fence of the first embodiment of the present invention;

[0050] Figure 6 is a schematic structural diagram of the liquid sealing fence of the second embodiment of the present invention;

[0051] Figure 7 、 Figure 8 is a schematic structural diagram of the second embodiment of the present invention;

[0052] Figure 9 is a top view of the second embodiment of the present invention;

[0053] Figure 10 is a schematic cross-sectional structural diagram of the second embodiment of the present invention;

[0054] Figure 11 is a schematic diagram of the substrate area division of the third embodiment of the present invention;

[0055] Figure 12 is a schematic diagram of the relative position of the sample and the box body of the third embodiment of the present invention;

[0056] Figure 13 is a schematic structural diagram of the fourth embodiment of the present invention;

[0057] Figure 14 is a schematic diagram of the operation steps of the sample reaction accommodation method of the present invention.

[0058] In the figure: 11, substrate; 111, edge area; 112, sample pre-setting area; 1121, first pre-setting area; 1122, second pre-setting area; 1123, overlapping area; 12, sample; 2, box body; 21, accommodating space; 22, mark; 23, heat conductor; 24, clamping mechanism; 25, second buckle; 3, liquid-sealing fence; 31, first sealing surface; 32, surrounding surface; 33, pressing surface; 34, second sealing surface; 35, sample area; 36, waste liquid removal area; 37, groove; 38, guiding inclined surface; 4, pressing cover; 41, opening; 42, upper surface; 43, first buckle; 44, elastic body; 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. Detailed implementation manners

[0059] Next, in combination with the accompanying drawings and the detailed implementation manners, the present invention will be further described. It should be noted that, on the premise of no conflict, the following-described embodiments or technical features can be arbitrarily combined to form new embodiments.

[0060] In the description of the present invention, it should be noted that for the orientation terms, such as the terms "center", "transverse", "longitudinal", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", etc., the indicated orientation and position relationships are based on the orientation or position relationships shown in the accompanying drawings. It is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and cannot be understood as limiting the specific protection scope of the present invention.

[0061] In addition, such terms as "first" and "second" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance or implicitly specifying the quantity of technical features. Thus, the features defined with "first" and "second" may explicitly or implicitly include one or more of such features. In the description of the present invention, the meaning of "several" is two or more, unless otherwise specifically defined.

[0062] In the present invention, unless otherwise clearly specified and limited, such terms as "set", "installed", etc. should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can also be a mechanical connection; it can be directly connected, or connected through an intermediate medium, and can be internally connected and communicated between two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present invention can be understood according to specific situations.

[0063] The Chinese patent application with the publication number CN116888253A discloses a modular determination support device, which constructs a sample reaction cell through a substrate (glass slide) and a gasket fixed on an insert that can move relative to the substrate holder. However, this device has three problems: 1. The position of the gasket surrounding the sliced sample relative to the device base (determined by the pasting position of the slice on the glass slide) is uncertain, resulting in uncertain reagent addition and waste liquid suction positions for different sliced samples. Therefore, it is not suitable to apply this device to automated instruments. 2. When in use, it is necessary to first place the glass slide in the limit groove of the base, and then adjust the gasket (the gasket cannot move after being fully pressed) when the upper cover is flipped to the state of being basically covered but not fully pressed. After the sliced sample falls within the range of the gasket, then fully cover and press the base and the upper cover. This operation of aligning the sliced sample and the gasket in the "almost covered but not fully covered" state is not only inconvenient, but also when the sliced sample and the gasket are of similar size, it is very difficult to accurately align the sample to the extent that it is completely within the surrounding area of the gasket, and there is even a risk that the bottom of the gasket accidentally rubs against the sliced sample during the movement, causing partial detachment. 3) The width of the border of the insert determines that the minimum distance between the pasted sample and the edge of the glass slide cannot be small enough, restricting the application of many pasted samples with relatively edge positions.

[0064] See Figures 1 - 5 , for the first embodiment of the invention, a sample reaction accommodating device, including a box body 2, a pressing cover 4 arranged on the box body 2, and a liquid sealing fence 3 detachably connected inside the box body 2; the pressing cover 4 can be selectively clamped on the upper part of the box body 2; one end of the pressing cover 4 can rotate relative to one end of the box body 2; the pressing cover 4 and the box body 2 are of an integral structure; or two detachable separated structures.

[0065] The upper end of the box body 2 is provided with a rectangular accommodating space 21; a substrate 11 is detachably connected inside the accommodating space 21; a sample 12 is pre-pasted on the upper end of the substrate 11; the accommodating space 21 allows the substrate 11 to move therein to adjust the position of the sample 12.

[0066] The liquid sealing fence 3 is a disposable consumable or an easy-to-clean part, and the shape of the liquid sealing fence 3 is not limited. It can be rectangular, circular, or other irregular shapes. According to the common sample shapes, as a preferred solution, it is rectangular.

[0067] The liquid sealing fence 3 includes a first sealing surface 31 at the lower end of the liquid sealing fence 3, a surrounding surface 32 on the inner wall of the liquid sealing fence 3, and a pressing surface 33 at the upper end of the liquid sealing fence 3; the surrounding area of the surrounding surface 32 is greater than or equal to the projected area of the sample 12.

[0068] The inner cavity of the liquid-sealing fence 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 suck the waste liquid, and the tip part of the pipette tip 7 will not contact the sample 12, thus preventing the sample from being punctured. In order to minimize the volume of the waste liquid removal area 36 as much as possible to reduce the reagent consumption, the cavity of the waste liquid removal area 36 is adapted to the pipette tip 7 to form a structure with a larger upper part and a smaller lower part.

[0069] A clamping mechanism 24 is arranged in the box body 2; 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 adjusting the position of the sample 12, rotate the clamping mechanism 24 to clamp the substrate 11 to prevent the substrate 11 from being moved when aligning and placing the liquid-sealing fence 3 subsequently.

[0070] When the gland 4 is snap-connected to the upper part of the box body 2, the gland 4 acts on the pressing surface 33, so that the first sealing surface 31 closely adheres to the upper surface of the substrate 11, and the surrounding surface 32 and the upper surface of the substrate 11 form a sample reaction accommodation cavity; the sample 12 is located in the sample reaction accommodation cavity; an opening 41 is arranged at the corresponding position of the gland 4 on the liquid-sealing fence 3 to facilitate adding reagents and removing waste liquid.

[0071] In order to make the operation of pressing and releasing the liquid-sealing fence 3 more convenient, a first buckle 43 is arranged on the gland 4; a second buckle 25 is arranged on the box body 2; the first buckle 43 can selectively be snap-connected to 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.

[0072] Specifically, the first buckle 43 is slidably connected to the lower end of the gland 4; an elastic body 44 is arranged between the first buckle 43 and the gland 4; the elastic body 44 can be a spring; when closing the gland 4, the first buckle 43 is hooked and locked 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 gland 4, press the first buckle 43 to separate the first buckle 43 and the second buckle 25, and then turn over the gland 4.

[0073] For the transparent substrate 11, in order to facilitate the adjustment of the position of the sample 12, a mark 22 for indicating the sample placement range is provided at the inner bottom end of the box body 2. The position of the mark 22 relative to the entire box body 2 is fixed. The indication range 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 surrounded by the first sealing surface 31. When adjusting the position of the sample 12, only the substrate 11 needs to be moved so that the sample 12 is completely within the inner frame of the mark 22; when placing the sealing liquid fence 3, the sealing liquid fence 3 is also aligned with the mark 22.

[0074] See Figures 6 - 10 , which 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 positioned through the mark 22 (the indication range of the mark 22 is covered by the substrate 11).

[0075] To solve this problem, a groove 37 is provided at the upper end of the outer wall of the sealing liquid fence 3; an elastic edge is provided on the outer wall of the sealing liquid fence at the upper end of the groove; a guiding inclined surface 38 is provided at the upper edge of the elastic edge; the sealing liquid fence 3 is detachably connected to the opening 41 of the pressing cover 4; the width of the pressing cover 4 is smaller than that of the substrate 11; or a notch 45 for adjusting the substrate 11 is provided on the pressing cover 4.

[0076] First, close the pressing cover 4, adjust the geometric center of the sample 12 to be directly opposite to the opening 41 through the notch 45, then open the pressing cover 4 to install the sealing liquid fence 3, and then close the pressing cover 4, so that the sample 12 can be located within the sealing liquid fence 3.

[0077] See Figure 11 、 Figure 12 , which is the third embodiment of the present invention. Based on the first and second embodiments, the following content is added: The substrate 11 is divided into an edge region 111 located at the outer periphery and a sample pre-positionable region 112 located inside the edge region 111; in order to increase the adaptability of the pre-positioned position of the sample 12, so that any sample within the pre-positionable region on the substrate 11 (excluding the samples outside the edge region) can be adjusted into the sealing liquid fence 3, the length (Ls), width (Ws) of the accommodation space 21, the length (Lb), width (Wb) of the substrate 11, the length (Lg), width (Wg) of the surrounding surface 32, and the minimum effective sealing width t of the first sealing surface 31 at least satisfy one of the following two conditions:

[0078] Condition 1: And W s≥2W b -W g -2t

[0079] Condition 2: and L s ≥2L b -L g -2t

[0080] To reduce the format size of the sample reaction container device, preferably, and W s = 2W b -W g -2t.

[0081] The minimum effective sealing width of the first sealing surface 31 is t, and t mainly depends on the size, material softness and hardness, and pressing degree of the liquid sealing fence 3; the range of t can be 0.1 mm to 10 mm. When t = 0.1 mm, the range of the sample pre-setting area 112 reaches the maximum, achieving the maximum tolerance for the sample pre-setting position; considering the manufacturing cost and sealing effect of the liquid sealing fence 3; preferably, t = 2 mm.

[0082] Any sample 12 located within the sample pre-setting area 112 can be adjusted into the enclosed area of the liquid sealing fence 3. Specifically, the sample 12 is a tissue section with a size of 6 mm × 6 mm, the length Lb of the substrate 11 is 76 mm, and the width Wb is 26 mm; the length Lg of the enclosing surface 32 is 8 mm, and the width Wg is 8 mm. The minimum effective sealing width t of the first sealing surface 31 is 2 mm; the length Ls of the accommodating space 21 is 108 mm, and the width Ws is 40 mm, satisfying and W s = 2W b -W g -2t.

[0083] As Figure 11 shown, the sample pre-setting area 112 includes a first pre-setting area 1121 and a second pre-setting area 1122 located on both sides respectively, and an overlapping area 1123 located in the middle; the overlapping area 1123 is the overlapping area of the first pre-setting area 1121 and the second pre-setting area 1122; the lengths of the first pre-setting area 1121 and the second pre-setting area 1122 are both The width of their overlapping part is equal to Lg, which is 8 mm.

[0084] As Figure 12As shown, the relative positions of the sample 12 and the cartridge body 2 are shown in four extreme positions where the sample 12 is completely located within the sample pre-set area 112. Among them, when the sample is within the first pre-set area 1121 or within the second pre-set area 1122 (when the substrate 11 is horizontally rotated 180°), that is, when the substrate 11 is at any position within the accommodation space 21, the sample 12 can be directly opposite to the opening 41.

[0085] See Figure 13 , which is the fourth embodiment of the present invention. On the basis of the first embodiment, the second embodiment, and the third embodiment, the following content is added: The sample reaction accommodation device further includes a first temperature control device 5 provided at the lower end of the cartridge body 2 and a second temperature control device 6 movably provided at the upper end of the gland 4. The lower end of the cartridge body 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. Thus, the control of the sample reaction temperature can be achieved.

[0086] 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 cartridge body 2 are fixed together. The heat conductor 23 and the first temperature control device 5 are made into one body.

[0087] 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, the evaporation of the reagent and its condensation on the second temperature control device 6 can be prevented.

[0088] The material of the isolation pad 61 is not limited. It can be a soft material such as silica gel, 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. For example, it can be hung through lugs or pasted through an easy-to-peel adhesive.

[0089] The liquid sealing fence 3 further includes a second sealing surface 34. Hanging ears 611 are respectively provided on the two outer walls of the isolation pad 61 facing each other. Hooks 621 respectively engaged with the two hanging ears 611 are provided at the lower end of the heating element 62. The isolation pad 61 and the heating element 62 are easy to fix and separate.

[0090] After the second temperature control device 6 is pressed down, a closed sample reaction accommodating cavity is formed among the upper surface of the substrate 11, the surrounding 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 the reagent in the sample reaction accommodating cavity can react under the required temperature conditions and sealed environment, and the reagent will not evaporate and condense on the surface of the isolation pad 61.

[0091] Both the liquid sealing fence 3 and the isolation pad 61 are disposable consumables or easy-to-clean parts, which prevent cross-contamination between different samples; in order to ensure the sealing effect, after the gland 4 presses the liquid sealing fence 3, the second sealing surface 34 on the liquid sealing fence 3 should not be lower than the upper surface 42 of the gland 4.

[0092] See Figure 14 , a sample reaction accommodating method, realized by the above-mentioned sample reaction accommodating device, for the opaque or semi-transparent substrate, includes the following steps:

[0093] (1) Place the substrate 11 pre-stuck with the sample 12 inside the box body;

[0094] (2) Close the gland 4, adjust the substrate 11 through the notch 45 so that the sample 12 is aligned with the geometric center of the opening 41;

[0095] (3) Rotate the clamping mechanism 24 so that the clamping mechanism 24 presses the substrate 11

[0096] (4) Open the gland 4, and install the groove 37 of the liquid sealing fence 3 into the opening 41;

[0097] (5) Close the gland 4, the lower end of the liquid sealing fence 3 is in sealed contact with the upper end of the substrate 11, and the sample 12 is located inside the liquid sealing fence 3;

[0098] (6) Add the reagent 8 into the liquid sealing fence 3;

[0099] (7) Press the second temperature control device 6 against the upper end of the liquid sealing fence 3, control the first temperature control device 5 and the second temperature control device 6 to a suitable temperature, and the temperature of the second temperature control device 6 is higher than that of the first temperature control device 5;

[0100] (8) After the sample 12 and the reagent 8 have fully reacted, remove the second temperature control device 6;

[0101] (9) Insert the pipette tip 7 into the bottom inside the waste liquid removal area 36 to suck away the waste liquid;

[0102] (10) According to the actual test process, steps (6) to (9) are repeated multiple times;

[0103] (11) Open the gland 4 and remove the substrate 11.

[0104] A method for accommodating a sample reaction, implemented by the above-mentioned sample reaction accommodating device. For the transparent substrate, it includes the following steps:

[0105] (1) Place the substrate 11 pre-stuck with the sample 12 inside the box body;

[0106] (2) Adjust the substrate 11 so that the sample 12 is directly opposite to the mark 22;

[0107] (3) Rotate the clamping mechanism 24 to press the substrate 11 tightly;

[0108] (4) Place the liquid sealing fence 3 on the upper end of the substrate 11 and make the liquid sealing fence 3 directly opposite to the mark 22;

[0109] (5) Close the gland 4. The lower end of the liquid sealing fence 3 is in sealed contact with the upper end of the substrate 11, and the sample 12 is located inside the liquid sealing fence 3;

[0110] (6) Add the reagent 8 into the liquid sealing fence 3;

[0111] (7) Press the second temperature control device 6 against the upper end of the liquid sealing fence 3, and control the first temperature control device 5 and the second temperature control device 6 to appropriate temperatures respectively. The temperature of the second temperature control device 6 is higher than that of the first temperature control device 5;

[0112] (8) After the sample 12 and the reagent 8 have fully reacted, remove the second temperature control device 6;

[0113] (9) Insert the pipette tip 7 into the bottom inside the waste liquid removal area 36 to suck away the waste liquid;

[0114] (10) According to the actual test process, steps (6) to (9) are repeated multiple times;

[0115] (11) Open the gland 4 and remove the substrate 11.

[0116] The above are only specific embodiments 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 device, characterized in that: It includes a box body; a pressing cover, which can be selectively clamped on the upper part of the box body; an opening, which is penetrated and arranged on the pressing cover; a substrate, on which a sample is arranged, and the substrate is detachably connected between the box body and the pressing cover; wherein, the box body allows the substrate to move therein to adjust the position of the sample; a liquid sealing fence is arranged between the substrate and the pressing cover; the liquid sealing fence is hermetically abutted against the upper end of the substrate under the pressing action of the pressing cover; the liquid sealing fence is detachably connected to the pressing cover.

2. The accommodating device according to claim 1, wherein: The inner cavity of the liquid sealing fence is divided into a sample area for accommodating the sample and a waste liquid removal area communicated with the bottom of the sample area.

3. The accommodating device according to claim 1 or 2, wherein: It includes a first temperature control component which can be selectively arranged at the lower end of the substrate and a second temperature control component which can be selectively arranged at the upper end of the liquid sealing fence; the second temperature control component includes a heating body and an isolation pad detachably connected to the lower end of the heating body; the lower end of the isolation pad is hermetically abutted against the upper end of the liquid sealing fence.

4. The accommodating device according to claim 1 or 2, characterized in that: The length (Ls) and width (Ws) of the inner wall of the box body, the length (Lb) and width (Wb) of the substrate, the length (Lg) and width (Wg) of the inner wall of the liquid sealing fence, and the minimum effective sealing width t of the liquid sealing fence hermetically abutted against the substrate satisfy at least one of the following two conditions: Condition 1: and W s ≥ 2W b -W g -2t; Condition 2; and L s ≥ 2L b -L g -2t.

5. The accommodating device according to claim 1 or 2, characterized in that: A mark is arranged at the lower end of the substrate inside the box body; the liquid sealing fence can be selectively aligned with the mark.

6. The accommodating device according to claim 1 or 2, characterized in that: A groove is arranged at the upper part of the outer wall of the liquid sealing fence; the groove is detachably connected to the pressing cover; a notch for adjusting the position of the substrate is arranged on the pressing cover.

7. The accommodating device according to claim 1 or 2, characterized in that: A first buckle is arranged on the pressing cover; a second buckle is arranged on the box body; the first buckle can be selectively clamped 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 accommodating device according to claim 7, characterized in that: A clamping mechanism is arranged inside the box body, and the clamping mechanism can selectively press the substrate.

9. A method for accommodating a sample reaction, characterized in that: Realized based on the sample reaction accommodating device according to claims 1-8, including the following steps: (1) Place the substrate pre-stuck with the sample inside the box body and adjust the position of the sample; (2) A mark is arranged at the lower end of the substrate inside the box body; place the liquid sealing fence on the substrate so that the liquid sealing fence is aligned with the mark, or install the liquid sealing fence on the opening of the pressing cover; (3) Close the pressing cover, the lower end of the liquid sealing fence is hermetically abutted against the upper end of the substrate, and the sample is located in the cavity formed by the inner wall of the liquid sealing fence and the upper end of the substrate; (4) Add a reagent into the liquid sealing fence; (5) The sample reaction accommodating device includes a first temperature control component arranged at the lower end of the substrate and a second temperature control component which can be selectively arranged at the upper end of the liquid sealing fence; press the second temperature control device against the upper end of the liquid sealing fence, and control the first temperature control device and the second temperature control device to appropriate temperatures; (6) After the sample and the reagent react fully, remove the second temperature control device; (7) The inner cavity of the liquid-sealing fence is divided into a sample area for accommodating the sample and a waste liquid removal area communicating with the bottom of the sample area. The pipette tip extends into the bottom of the waste liquid removal area to suck away the waste liquid; (8) Open the gland and remove the substrate.

10. The accommodating method according to claim 9, wherein: In step (2), when the substrate is transparent, it is optional to align the liquid-sealing fence with the mark or install the liquid-sealing fence onto the opening; When the substrate is opaque, install the liquid-sealing fence onto the opening.

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