A mold device for organoid embedding

Abstract

The application relates to the field of organoid embedding technology, and particularly relates to a mold device for organoid embedding, which solves the problems of the inability to simplify the whole process of 'dehydration-embedding', the risk of organoid damage and position deviation in the prior art, and comprises a bottom plate and a top cover, the bottom plate and the top cover are detachably buckled and form an embedding groove; a plurality of dehydration channels are formed in the mold device to enable the embedding groove to communicate with the outside; the top cover is provided with a wax liquid flow guide mechanism, and the mold device further comprises a demolding mechanism, the demolding mechanism comprises a knob, a threaded rod and a demolding unit; the knob is rotationally connected to the bottom surface of the bottom plate, the top surface of the knob is fixedly connected with one end of the threaded rod, the demolding unit is sleeved on the threaded rod, and the threaded rod is rotated to lift or lower the demolding unit. The device has the dual functions of dehydration treatment and paraffin embedding, realizes in-situ operation of the whole process from dehydration to molding of the organoid sample, completely eliminates the manual transfer link, and avoids structural damage and position deviation of the pre-embedded sample.

Description

Technical Field

[0001] This invention relates to the field of organoid embedding technology, and more specifically, to a mold device for organoid embedding. Background Technology

[0002] In the fields of organoid pathological analysis, histomorphological studies, and drug sensitivity testing, organoid samples are typically prepared into high-quality paraffin-embedded blocks for ultrathin sectioning (usually 3-5 micrometers thick) and subsequent hematoxylin-eosin (H&E) staining, immunohistochemistry (IHC), or in situ hybridization (ISH). During the paraffin embedding process, to protect the fragile three-dimensional structure of the organoid from damage by chemical reagents and mechanical stress during subsequent dehydration, clearing, and paraffin impregnation steps, the organoid is usually pre-embedded in agarose gel for fixation (i.e., agarose pre-embedding). After pre-embedding, a multi-container transfer process is required: first, the agarose-embedded organoid sample is placed in a dedicated porous dehydration box for gradient dehydration, clearing, and paraffin impregnation; then, the dehydrated sample is manually removed from the dehydration box and transferred to the metal forming tank of an independent paraffin embedding mold, where molten paraffin is re-injected. After cooling and solidification, the paraffin-embedded block attached to the bottom of the metal forming tank is separated from the metal forming tank by utilizing the inherent low interfacial adhesion between the metal forming tank surface and the solidified paraffin. At this time, the bottom of the dehydration box forms a raised area of ​​paraffin-embedded block due to the groove structure of the metal forming tank. The organoid is then positioned in this raised area. Finally, the dehydration box is fixed on the microtome so that the paraffin-embedded block is close to the slicing area for subsequent slicing operations.

[0003] The traditional operating method described above has significant drawbacks. After dehydration, samples must be exposed to air for manual transfer across containers. This process is highly susceptible to damage from tweezers or collisions, which can break the pre-embedded agarose gel, leading to organoid structural damage or loss of spatial orientation. This severely impacts the quality of subsequent sections and the accuracy of microscopic observation. Therefore, it is necessary to develop a device that integrates dehydration and embedding functions and features a controllable, active demolding structure. This would eliminate the sample transfer step and ensure stable, non-destructive molding of the embedded block, thereby guaranteeing the integrity of organoid samples and the quality of sections, and meeting the requirements of high-precision pathological analysis.

[0004] A search revealed that existing embedding molds share similar main structures, primarily optimizing demolding by changing the material of the positioning grooves. For example, Chinese Patent (CN216899865U) discloses a novel organoid tissue embedding box. This box utilizes the elastic and plastic properties of silicone to achieve press-type demolding by setting silicone positioning grooves and matching positioning blocks. While this design improves operational comfort and demolding convenience, it is essentially still an independent embedding container, lacking integrated dehydration processing. Samples still need to be manually transferred from the dehydration box to this embedding box, failing to simplify the entire "dehydration-embedding" process and still not addressing the risks of organoid damage and orientation shift. Therefore, to address these issues, an integrated "dehydration-embedding" organoid processing device is proposed. Summary of the Invention

[0005] The purpose of this invention is to provide a mold device for embedding organoids. This device integrates the dual functions of dehydration treatment and paraffin embedding, enabling in-situ operation of organoid samples from dehydration to molding, completely eliminating the manual transfer step, and avoiding structural damage and orientation shift of the pre-embedded sample.

[0006] The present invention is implemented as follows: a mold device for embedding organoids includes a base plate and a top cover. The base plate and the top cover are detachably fastened together to form an embedding groove. The mold device has multiple dehydration channels to connect the embedding groove with the outside. The top cover is provided with a wax liquid guiding mechanism. The wax liquid guiding mechanism can guide molten paraffin into the embedding groove.

[0007] Furthermore, it includes a demolding mechanism that allows the complete paraffin-embedded block to protrude from the surface of the base plate.

[0008] Furthermore, the demolding mechanism includes a knob, a threaded rod, and a demolding unit; the knob is rotatably connected to the bottom surface of the base plate, the top surface of the knob is fixedly connected to one end of the threaded rod, the demolding unit is sleeved on the threaded rod, and the rotation of the threaded rod causes the demolding unit to rise and fall.

[0009] Furthermore, the embedding groove is opened on the top surface of the base plate; the demolding unit is a sleeve with a top sealing end, the sleeve is sleeved on the threaded rod and the outer side wall slides in contact with the inside of the base plate; when the sleeve is in the lower limit position, the top surface of the sleeve is flush with the bottom surface of the embedding groove.

[0010] Furthermore, the embedding groove is opened on the bottom surface of the top cover, and the demolding unit is a positioning cylinder with an open top. The positioning cylinder is sleeved on the threaded rod and its outer wall slides in contact with the inside of the base plate. When the positioning cylinder is in the lower limit position, the top surface of the positioning cylinder is flush with the top surface of the base plate; the top surface of the threaded rod is flush with the top surface of the base plate.

[0011] Furthermore, the wax liquid guiding mechanism is a cover body, which is detachably installed on the top cover, and multiple dehydration channels are also opened inside the cover body.

[0012] Furthermore, the wax liquid guiding mechanism includes a storage cavity, multiple release channels, and a heat exchange unit; both the storage cavity and the release channels are located inside the top cover, and the multiple release channels connect the storage cavity to the embedding tank; the heat exchange unit is embedded inside the top cover, and the two heat exchange ends of the heat exchange unit are located in the storage cavity and the embedding tank, respectively.

[0013] Furthermore, the heat exchange unit includes multiple cooling semiconductors, two heat sinks, and multiple heat sinks; the two ends of the multiple cooling semiconductors are respectively connected to the two heat sinks, the multiple heat sinks are fixedly installed on the surface of the heat sinks in the storage cavity, and the heat dissipation surface of another heat sink is located in the embedding groove.

[0014] Furthermore, the wax guiding mechanism consists of multiple sealing blocks, all of which are slidably disposed on the top cover, and the bottom of each sealing block is fixedly connected to the top surface of the positioning cylinder. When the positioning cylinder is at the upper limit position, the sealing blocks are connected to the top cover; when the positioning cylinder is at the lower limit position, the sealing blocks are located inside the bottom plate and flush with the top surface of the bottom plate.

[0015] Furthermore, the embedding groove is opened on the top surface of the base plate, the surface of the embedding groove is a smooth surface, and the bottom surface of the top cover is a rough surface.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] 1. This device integrates the dual functions of dehydration and paraffin embedding, enabling in-situ operation of organoid samples from dehydration to shaping, completely eliminating the manual transfer step and avoiding structural damage and orientation shift of the pre-embedded samples. At the same time, the built-in demolding mechanism in the base plate, combined with the low adhesion characteristics of the mold surface, achieves zero-contact and controllable demolding of the embedding block, ensuring the structural integrity of the embedding block and the positioning accuracy of the organoid, significantly improving the pass rate of slides and the reliability of pathological analysis.

[0018] 2. The wax liquid guiding mechanism is set as a cover. When closed, the agarose pre-embedded block in the embedding groove is constrained by the inner wall of the cover to prevent it from falling out. The dehydration channel through the surface of the cover can ensure the penetration of dehydration reagent. When the cover is opened, the opening of the embedding groove can be exposed, so that the molten paraffin can be directly injected into the embedding groove to realize the in-situ embedding of organoids without the need for sample transfer throughout the process.

[0019] 3. This device designs the top surfaces of the embedding groove and the sleeve rod as coplanar, forming a continuous support plane to ensure that the paraffin-embedded block can adhere to the top surface of the sleeve rod after solidification. The demolding mechanism drives the sleeve rod to lift, achieving zero-contact demolding. After demolding, the solidified paraffin-embedded block adheres to the top surface of the sleeve rod, allowing the demolded embedding block to be directly sliced ​​while maintaining its original positioning, completely eliminating organoid structural damage caused by traditional manual transfer. This simplifies the operation steps and significantly improves the reliability of pathological analysis.

[0020] 4. The top cover is designed as an inverted U-shaped structure, with the embedding groove located at the bottom of the top cover. This creates a three-dimensional spatial constraint during the dehydration stage. Combined with the positioning cylinder's limiting effect, this controls the displacement of the agarose pre-embedded block, completely preventing sample displacement caused by dehydration reagent rinsing. During demolding, the positioning cylinder sinks down along the threaded rod into the bottom plate, exposing a flat cutting surface with a uniform height for the paraffin-embedded block. This allows for direct slicing without secondary trimming, further improving operational efficiency.

[0021] 5. The sealing block and positioning cylinder are rigidly connected, which enables automatic switching between dehydration and wax injection modes: when the positioning cylinder is at the upper limit, the sealing block closes the top cover to form a zero-displacement anti-fall barrier, ensuring the stability of the dehydration treatment of the agarose pre-embedded block; when the positioning cylinder is lowered to the lower limit, the sealing block sinks simultaneously to fully expose the wax injection opening, supporting direct pouring of molten paraffin, eliminating the traditional opening operation, and further improving work efficiency.

[0022] 6. By integrating a storage cavity, multiple vertically arranged release channels, and a dual-mode heat exchange unit, an automatic paraffin block embedding structure is constructed. Before dehydration, molten paraffin can be pre-injected into the storage cavity of the top cover and rapidly solidified through phase change via the heat exchange unit. After dehydration, the heating mode is activated to efficiently melt the paraffin in the storage cavity. The molten paraffin is then injected into the embedding tank through the release channel. At the same time, the end of the heat exchange unit closest to the embedding tank is the cooling end, enabling rapid solidification of the paraffin in the embedding tank area and quickly forming paraffin embedding blocks, further improving embedding efficiency. Attached Figure Description

[0023] Figure 1 This is a schematic diagram of the structure of an organoid embedding mold device provided in Embodiment 1 of the present invention;

[0024] Figure 2 This is a schematic diagram of the top cover and bottom plate separated during demolding of an organoid embedding mold device provided in Embodiment 1 of the present invention;

[0025] Figure 3 This is a schematic diagram of the structure of an organoid embedding mold device provided in Embodiment 2 of the present invention;

[0026] Figure 4 This is a schematic diagram of the base plate provided in Embodiment 2 of the present invention during demolding;

[0027] Figure 5 This is a bottom view of the base plate provided in Embodiment 2 of the present invention;

[0028] Figure 6 This is a schematic diagram of the structure of an organoid embedding mold device provided in Embodiment 3 of the present invention;

[0029] Figure 7 yes Figure 6Enlarged view of point A in the middle;

[0030] Figure 8 This is a schematic diagram of the structure of the bottom plate and top cover of the organoid embedding mold device provided in Embodiment 3 of the present invention when they are separated;

[0031] Figure 9 This is a top view of the top cover provided in Embodiment 3 of the present invention;

[0032] Figure 10 This is a bottom view of the top cover provided in Embodiment 3 of the present invention;

[0033] Figure 11 This is a schematic diagram of the structure of an organoid embedding mold device provided in Embodiment 4 of the present invention;

[0034] Figure 12 This is a schematic diagram of the structure of an organoid embedding mold device provided in Embodiment 5 of the present invention during the dehydration of agarose pre-embedded blocks;

[0035] Figure 13 This is a top view of the top cover provided in Embodiment 5 of the present invention;

[0036] Figure 14 This is a schematic diagram of the structure of an organoid embedding mold device provided in Embodiment 5 of the present invention during the injection of molten paraffin;

[0037] Figure 15 This is a schematic diagram of the structure of an organoid embedding mold device provided in Embodiment 6 of the present invention during the dehydration of agarose pre-embedded blocks;

[0038] Figure 16 This is a schematic diagram of the structure of the base plate during slicing provided in Embodiment 6 of the present invention.

[0039] Reference numerals used in the above figures:

[0040] 1. Top cover; 2. Bottom plate; 3. Agarose pre-embedded block; 4. Embedding groove; 5. Paraffin wax; 6. Cover body; 7. Dehydration channel; 8. Threaded rod; 9. Sleeve rod; 10. Knob; 11. Release channel; 12. Storage cavity; 13. Heat sink; 14. Refrigeration semiconductor; 15. Heat sink plate; 16. Positioning cylinder; 17. Sealing block. Detailed Implementation

[0041] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0042] The implementation of the present invention will be described in detail below with reference to specific embodiments.

[0043] In the accompanying drawings of this embodiment, the same or similar reference numerals correspond to the same or similar components. In the description of this invention, it should be understood that if terms such as "upper," "lower," "left," and "right" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the 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. Therefore, the terms used to describe positional relationships in the drawings are only for illustrative purposes and should not be construed as limiting this invention. For those skilled in the art, the specific meaning of the above terms can be understood according to the specific circumstances.

[0044] Reference Figures 1-16 The image shows a preferred embodiment of the present invention.

[0045] Example 1: A mold device for organoid embedding. The mold device in this example is a basic model provided in this application, such as... Figure 1 and Figure 2 As shown, it mainly consists of a top cover 1 and a bottom plate 2, which are connected by a snap-fit ​​mechanism. An embedding groove 4 is formed on the top surface of the bottom plate 2. To facilitate demolding of the paraffin wax 5 embedded block, the bottom plate 2 and the embedding groove 4 in this embodiment are made of smooth metal, while the bottom surface of the top cover 1 is made of non-smooth plastic. Multiple vertically penetrating dehydration channels 7 are formed on the top cover 1 directly above the embedding groove 4. These dehydration channels 7 not only allow the dehydrating agent to enter the embedding groove 4 to dehydrate the agarose pre-embedded block 3, but also make the bottom surface of the top cover 1 porous and rough, thereby improving the adhesion of the solidified paraffin wax 5 to the top cover 1 and making the entire paraffin wax 5 embedded block easier to demold. A wax liquid guiding mechanism is also installed on the top cover 1. In this embodiment, the wax liquid guiding mechanism is a cover body 6, with an opening on the top cover 1 for injecting molten paraffin wax 5. During dehydration, the cover body 6 closes at this opening. In order to make it easier to fix the cover 6 at the opening, the cover 6 in this embodiment is preferably a rubber cover.

[0046] In order to allow the dehydrating agent to enter the embedding tank 4 better and improve the dehydration effect, this embodiment also provides multiple through dehydration channels 7 on the cover body 6.

[0047] like Figure 2 As shown in this embodiment, since the paraffin 5 embedding block will adhere to the bottom surface of the top cover 1 after demolding, the top cover 1 will be fixed on the slicer during subsequent slicing. Therefore, the cross-sectional shape of the top cover 1 is not limited. It can be set as a circle or a rectangle. The size and dimensions can be designed according to the existing slicer fixing method.

[0048] To avoid interference between the slicer and the top cover 1 during slicing, this embodiment sets the slot inside the top cover 1 and sets the snap block on the top surface of the bottom plate 2. In this way, there is no protrusion on the bottom surface of the top cover 1, and the slicer can normally slice the protruding paraffin 5 embedded block.

[0049] Working principle: Place the agarose pre-embedded block 3 into the embedding tank 4, and then snap the top cover 1 and bottom plate 2 together. The entire mold device is sequentially immersed in 70% ethanol, 80% ethanol, and 90% ethanol for gradient dehydration, washing at least once for at least 45 minutes each time, followed by washing at least twice with anhydrous ethanol for at least 60 minutes each time. After dehydration, remove the cover 6, and inject molten paraffin 5 into the embedding tank 4 using a paraffin 5 embedding machine. During injection, the liquid level of the molten paraffin 5 should be positioned within the dehydration channels 7. After cooling and solidification, the paraffin 5 embedding block will stably adhere to the bottom surface of the top cover 1, and the experimenter can then directly remove the top cover 1. Figure 2 As shown in the figure, demolding is completed. Finally, the top cover 1 is fixed on the slicer, so that the raised paraffin 5 embedded block faces the blade of the slicer, and the slicing operation can be performed.

[0050] Example 2: A mold device for embedding organoids, such as Figures 3-5 As shown, in order to make the paraffin 5 embedded block more stable and controllable in demolding and achieve no contact throughout the process, this embodiment sets a demolding mechanism on the base plate 2 on the basic model provided in embodiment 1.

[0051] The demolding mechanism in this embodiment mainly consists of a knob 10, a threaded rod 8, and a demolding unit. The knob 10 is attached to the bottom surface of the base plate 2, and its top surface is connected to the bottom surface of the base plate 2 by a snap-fit ​​ring. A threaded rod 8 is installed at the top axis of the knob 10. The demolding unit uses a sleeve 9 with a top-sealed end. The sleeve 9 is threaded onto the threaded rod 8, and the sleeve 9 rises and falls when the threaded rod 8 rotates. Figure 3 As shown, when the sleeve rod 9 is at the lower limit, all the threaded teeth of the sleeve rod 9 are fully engaged with the threaded rod 8, and the top surface of the sleeve rod 9 is flush with the bottom surface of the embedding groove 4. In order to facilitate the demolding operation of the paraffin 5 embedding block when the sleeve rod 9 rises, the top surface of the sleeve rod 9 in this embodiment is a rough surface, the bottom surface of the top cover 1 and the surface of the embedding groove 4 are smooth metal surfaces, and the bottom surface of the cover 6 is a smooth surface.

[0052] like Figure 4As shown, after demolding, the paraffin 5 embedding block adheres to the top surface of the sleeve 9 and protrudes from the top surface of the base plate 2. Therefore, in this embodiment, the base plate 2 needs to be fixed to the slicer during subsequent slicing operations. Thus, the cross-sectional shape of the base plate 2 can be circular or rectangular, and its size and dimensions can be designed according to the existing slicer fixing method. Furthermore, to avoid interference between the base plate 2 and the slicer during slicing, a slot is provided inside the base plate 2, and a locking block is provided on the bottom surface of the top cover 1. To prevent the slicer from contacting the knob 10 during fixing, combined with… Figure 4 and Figure 5 As shown, an annular protrusion is provided on the bottom surface of the base plate 2. The annular protrusion is integrally formed with the base plate 2. The annular protrusion can not only protect the knob 10 when the slicer is fixed, but also take advantage of the integral design to allow the cold energy of the freezing stage to be quickly transferred to the embedding tank 4 when the paraffin 5 embedding block cools and solidifies, so that the molten paraffin 5 solidifies quickly.

[0053] Working principle: After the agarose pre-embedded block 3 has completed dehydration, the cover 6 is removed, and molten paraffin 5 is injected into the embedding tank 4 using a paraffin 5 embedding machine. To facilitate demolding, the liquid level of the molten paraffin 5 should be lower than the top surface of the base plate 2 during injection, avoiding contact between the molten paraffin 5 and the bottom surface of the top cover 1 as much as possible. After cooling and solidification, the paraffin 5 embedding block will stably adhere to the top surface of the sleeve 9. The top cover 1 is opened, and the experimenter rotates the knob 10 to move the sleeve 9 upwards, thus allowing the entire paraffin 5 embedding block to protrude from the top surface of the base plate 2. Figure 4 (As shown). Finally, fix the base plate 2 onto the microtome, so that the raised paraffin 5 embedded block faces the direction of the microtome blade, and then the sectioning operation can be performed.

[0054] Example 3: A mold device for organoid embedding, such as Figures 6-10 As shown, this embodiment provides a new wax guiding mechanism compared to Embodiment 2. The wax guiding mechanism of this embodiment mainly consists of a storage chamber 12, multiple release channels 11, and a heat exchange unit. The storage chamber 12 and the release channels 11 are located inside the top cover 1. The storage chamber 12 is used to store paraffin wax 5. The tops of the multiple release channels 11 are simultaneously connected to the storage chamber 12, and when the top cover 1 and the bottom plate 2 are closed, the bottoms of the release channels 11 are connected to the embedding groove 4. The heat exchange unit is also located inside the top cover 1, with its heat exchange ends located in the storage chamber 12 and the embedding groove 4, respectively.

[0055] The design of the dehydration channel 7 needs to be adjusted according to the design dimensions of the storage chamber 12, for example... Figure 6As shown, when the cross-section of the storage cavity 12 is larger than the cross-section of the embedding tank 4, not only is a vertically penetrating dehydration channel 7 required inside the top cover 1, but an L-shaped dehydration channel 7 also needs to be correspondingly provided inside the bottom plate 2. When the top cover 1 and the bottom plate 2 are closed, the two sets of dehydration channels 7 are connected. To improve the dehydration effect and allow the dehydrating agent to enter the embedding tank 4 as much as possible, such as... Figure 9 As shown, the dehydration channel 7 is arranged along the circumference of the storage cavity 12. When the cross-section of the storage cavity 12 is smaller than that of the embedding tank 4, the dehydration channel 7 does not need to be provided in the bottom plate 2. The dehydration channel 7 in the top cover 1 is the same as in embodiment 2 and can be directly connected to the embedding tank 4.

[0056] Combination Figure 6 and Figure 7 As shown, the heat exchange unit in this embodiment mainly consists of multiple NP cooling semiconductors 14, two heat sinks, and multiple heat sinks 13. The two ends of the cooling semiconductors 14 are respectively connected to two heat sinks 15. In order to prevent the paraffin wax 5 from solidifying and adhering to the heat sinks 15, multiple heat sinks 13 are installed on the surface of the heat sinks 15 near the storage cavity 12, and the bottom surface of the heat sinks 15 near the embedding groove 4 is a smooth metal surface.

[0057] In order to control the operation of the cooling semiconductor 14, a battery (not shown in the figure) and a control component (not shown in the figure) are embedded in the top cover 1. The test personnel can switch the cooling semiconductor 14 to cooling or heating by adjusting the control component.

[0058] The demolding mechanism in this embodiment is the same as that in Embodiment 2, and will not be described again here.

[0059] Working principle: Before the agarose pre-embedded block 3 completes dehydration, with the bottom surface of the top cover 1 facing upwards, molten paraffin 5 is injected into the storage cavity 12 through the release channel 11 using a paraffin 5 embedding machine. After the storage cavity 12 is filled, the cooling semiconductor 14 is adjusted by the control component, so that one end of the cooling semiconductor 14 near the storage cavity 12 generates cold energy, and the other end generates heat. Under the action of the cooling semiconductor 14, the molten paraffin 5 in the storage cavity 12 solidifies rapidly. The cooling semiconductor 14 is turned off, the agarose pre-embedded block 3 is placed into the embedding tank 4, and the top cover 1 and the bottom plate 2 are closed for dehydration treatment. After dehydration, the researchers then adjust the cooling semiconductor 14 through the control components. The cooling semiconductor 14 generates heat at one end near the storage cavity 12 and generates cold at the other end. The heat generated can melt the solid paraffin 5 in the storage cavity 12 and flow into the embedding tank 4 through the release channel 11. The paraffin 5 in the embedding tank 4 is then rapidly solidified by the cold generated at the other end of the cooling semiconductor 14 to form a paraffin 5 embedding block.

[0060] The wax flow guiding mechanism of this embodiment directly skips the traditional cryostat cooling process, seamlessly connecting organoid dehydration with paraffin 5 embedding, thus improving experimental efficiency. It also eliminates the power consumption generated by continuous cryostat cooling, meeting green laboratory standards. Furthermore, this solution fully utilizes the cold and heat generated by the cooling semiconductor 14, reducing energy loss while increasing the melting and solidification speed of paraffin 5, further enhancing experimental efficiency.

[0061] Example 4: A mold device for embedding organoids, such as Figure 11 As shown, this embodiment provides a new demolding mechanism compared to embodiment 2, and the embedding groove 4 is provided on the bottom surface of the top cover 1. The top cover 1 and the bottom plate 2 are also connected by a snap-fit ​​method, forming a closed embedding groove 4 when the top cover 1 and the bottom plate 2 are fastened together. The dewatering channel 7 is provided through the top surface of the top cover 1. The cover body 6 is also provided on the top surface of the top cover 1, and the installation method is the same as in embodiment 2.

[0062] The demolding mechanism in this embodiment differs from that in Embodiment 2 in that the demolding unit has a different structure. In this embodiment, the demolding unit is a positioning cylinder 16 with an open top, which is sleeved on the threaded rod 8. In this embodiment, the cross-section of the threaded rod 8 is larger than that of the threaded rod 8 in Embodiment 2, and is greater than the length of the agarose pre-embedded block 3. When dehydrating the agarose pre-embedded block 3, the positioning cylinder 16 is at its upper limit position. At this time, the positioning cylinder 16 limits the agarose pre-embedded block 3, ensuring that the agarose pre-embedded block 3 can be located in the middle position of the paraffin 5 embedding block. When the positioning cylinder 16 is at its lower limit position, the top surface of the positioning cylinder 16, the top surface of the threaded rod 8, and the top surface of the base plate 2 are flush, avoiding interference between the slicer and the mold device when slicing. In addition, the top surface of the base plate 2, the top surface of the threaded rod 8, and the top surface of the positioning cylinder 16 in this embodiment are all rough planes, which allows the paraffin 5 embedding block to better adhere to the plane, while the surface of the embedding groove 4 is a smooth metal surface.

[0063] Working principle: Before the agarose pre-embedded block 3 completes dehydration, the experimenter rotates knob 10 to move the positioning cylinder 16 to its upper limit position under the action of the threaded rod 8, thereby positioning the agarose pre-embedded block 3. After dehydration, the cover 6 is removed, and knob 10 is rotated in the opposite direction to lower the positioning cylinder 16 to its lower limit position. Then, molten paraffin 5 is injected into the embedding tank 4 using a paraffin 5 embedding machine. To facilitate better demolding, the surface of the molten paraffin 5 is kept away from the inner top surface of the top cover 1 during injection. After cooling and solidification, the top cover 1 is removed, and the paraffin 5 embedded block adheres to the top surface of the base plate 2. Finally, the base plate 2 is fixed to the microtome, with the protruding paraffin 5 embedded block facing the direction of the microtome blade, and the slicing operation can be performed.

[0064] Example 5: A mold device for embedding organoids, such as Figures 12-14As shown, this embodiment provides a new wax guiding mechanism compared to embodiment 4. The remaining structure is the same as that of embodiment 4, and will not be described again here. The wax guiding mechanism in this embodiment consists of four sealing blocks 17. In the dehydrated state, the sealing blocks 17 are located at the four openings of the top cover 1, and the four openings are used to inject molten paraffin wax 5. The bottom of the sealing block 17 is connected to the top surface of the positioning cylinder 16 through a rod. When the positioning cylinder 16 is in the lower limit position, both the sealing block 17 and the positioning cylinder 16 move into the bottom plate 2, and the top surface of the sealing block 17, the top surface of the threaded rod 8, and the top surface of the bottom plate 2 are flush.

[0065] The structure of this embodiment can further reduce the workload of experimenters, eliminating the need to open the wax injection opening separately, thus further improving experimental efficiency.

[0066] Working principle: Before the agarose pre-embedded block 3 completes dehydration, the experimenter rotates knob 10 to move the positioning cylinder 16 to its upper limit position under the action of the threaded rod 8, thereby positioning the agarose pre-embedded block 3. At the same time, the sealing block 17 blocks the wax injection opening. After dehydration, simply rotate knob 10 in the opposite direction to lower the positioning cylinder 16 to its lower limit position. At this time, the sealing block 17 is also located inside the base plate 2, and the wax injection opening is opened. Then, molten paraffin 5 is injected into the embedding tank 4 through the paraffin 5 embedding machine. After cooling and solidification, the top cover 1 is removed, and the paraffin 5 embedded block is attached to the top surface of the base plate 2. Finally, the base plate 2 is fixed on the microtome, with the raised paraffin 5 embedded block facing the direction of the microtome blade, and the sectioning operation can be performed.

[0067] Example 6: A mold device for embedding organoids, such as Figures 15-16 As shown, the main difference between this embodiment and embodiments 4 and 5 is the wax flow guiding mechanism, while the rest of the structure is the same. The wax flow guiding mechanism in this embodiment is the same as that in embodiment 3, and will not be described again here.

[0068] It should be clearly stated that the scope of protection claimed in this application covers all technical variations based on the core architecture of "dehydration-embedding integration": regardless of whether the embedding tank 4 is set on the top cover 1 or the bottom plate 2, and regardless of whether the wax liquid guiding mechanism adopts a cover type, sealing block type or storage cavity type structure, as long as the mold device achieves in-situ zero transfer operation of organoid samples from dehydration treatment to paraffin embedding through structural innovation, it shall be deemed to fall within the scope of protection of the claims of this application.

[0069] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A mold device for embedding organoids, characterized in that, The device includes a base plate (2) and a top cover (1). The base plate (2) and the top cover (1) are detachably fastened together to form an embedding groove (4). The mold device has multiple dehydration channels (7) to connect the embedding groove (4) with the outside. The top cover (1) is equipped with a wax liquid guiding mechanism. The wax liquid guiding mechanism can guide molten paraffin (5) into the embedding groove (4). The wax liquid guiding mechanism includes a storage cavity (12), multiple release channels (11) and a heat exchange unit; the storage cavity (12) and the release channels (11) are both opened in the top cover (1), and the multiple release channels (11) connect the storage cavity (12) to the embedding tank (4); the heat exchange unit is embedded in the top cover (1), and the two heat exchange ends of the heat exchange unit are located in the storage cavity (12) and the embedding tank (4) respectively; The heat exchange unit includes multiple cooling semiconductors (14), two heat sinks (15) and multiple heat sinks (13); the two ends of the multiple cooling semiconductors (14) are respectively connected to the two heat sinks (15), and the multiple heat sinks (13) are fixedly installed on the surface of the heat sinks (15) in the storage cavity (12), and the heat dissipation surface of the other heat sink (15) is located in the embedding groove (4). Before dehydration, the end of the cooling semiconductor (14) near the storage cavity (12) generates cold energy to cool the molten paraffin injected into the storage cavity (12) and solidify it. After dehydration, the end of the cooling semiconductor (14) near the storage cavity (12) generates heat and the other end generates cold energy. The heat generated can melt the solid paraffin in the storage cavity (12) and flow into the embedding tank through the release channel (11). The paraffin in the embedding tank is then rapidly solidified by the cold energy generated at the other end of the cooling semiconductor (14) to form a paraffin embedding block.

2. The organoid embedding mold device according to claim 1, characterized in that, Includes a demolding mechanism that allows the complete paraffin (5) embedded block to protrude from the surface of the base plate (2).

3. The organoid embedding mold device according to claim 2, characterized in that, The demolding mechanism includes a knob (10), a threaded rod (8), and a demolding unit; the knob (10) is rotatably connected to the bottom surface of the base plate (2), and the top surface of the knob (10) is fixedly connected to one end of the threaded rod (8). The demolding unit is sleeved on the threaded rod (8), and the rotation of the threaded rod (8) causes the demolding unit to rise and fall.

4. The organoid embedding mold device according to claim 3, characterized in that, The embedding groove (4) is opened on the top surface of the base plate (2); the demolding unit is a sleeve (9) with a top sealing opening. The sleeve (9) is sleeved on the threaded rod (8) and its outer side wall slides in contact with the inside of the base plate (2); when the sleeve (9) is in the lower limit position, the top surface of the sleeve (9) is flush with the bottom surface of the embedding groove (4).

5. The organoid embedding mold device according to claim 3, characterized in that, The embedding groove (4) is opened on the bottom surface of the top cover (1). The demolding unit is a positioning cylinder (16) with an open top. The positioning cylinder (16) is sleeved on the threaded rod (8) and its outer side wall slides in contact with the inside of the bottom plate (2). When the positioning cylinder (16) is in the lower limit position, the top surface of the positioning cylinder (16) is flush with the top surface of the bottom plate (2). The top surface of the threaded rod (8) is flush with the top surface of the bottom plate (2).

6. The organoid embedding mold device according to claim 1, characterized in that, The embedding groove (4) is located on the top surface of the base plate (2). The surface of the embedding groove (4) is smooth, while the bottom surface of the top cover (1) is rough.

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