In-vitro preparation instrument for biological organ structure

Abstract

The invention discloses a biological organ structure in-vitro preparation instrument, which is a precise device for preparing a biological organ structure model for in-vitro culture, relates to the technical field of biomedical experiment research, and comprises a slicing part, a micro-motion part and a linkage part, the micro semi-automatic device creatively combines the biological structure characteristics of a microelectronic control system, peripheral nervous nerves, central nervous nerves, various solid visceral organs and tumor tissues, and adopts a spiral micrometer micrometer order stepping device as an organ-like tissue thickness control structure. And the organoid tissue structure with the thickness of 100-500 microns can be intuitively, conveniently and efficiently prepared under the in-vitro sterile condition.

Description

Technical Field

[0001] The present invention relates to the field of biomedical science experimental research technology, and specifically to a precision device for preparing biological organoid structures cultured in vitro, which is applied to the preparation of organoid models of the peripheral and central nervous systems, various solid organs and tumor tissues. Background Art

[0002] Over the past 20 years, the development of neural tissue engineering technology has received widespread attention from researchers in multiple disciplines and fields, including neurobiology, biomaterials science, and clinical neurological disease. It uses standardized in vitro microscopy methods to prepare neural tissue slices with a thickness of 200-500um. Under a special in vitro culture environment, it observes the various neurobiological behavioral changes of neural cells at the tissue level in isolated neural tissue, including various peripheral and central nervous tissues, under the influence of various research factors. Through immunocytochemistry, immunohistochemistry, immunoelectron microscopy, and various molecular biology techniques, it explores the cellular and molecular biological mechanisms of neural tissue repair and regeneration. In particular, it provides an extremely convenient intervention and observation method for observing the interaction between grafts and host tissues after various cell transplants, including stem cell transplantation. It also provides efficient and precise in vitro research methods for in-depth study of the interaction between various new biomaterials and host neural tissues after implantation. These are the most eye-catching research directions in neurobiology and neural tissue engineering.

[0003] The international 3R principles (Reduction, Replacement, and Refinement) for the use of experimental animals in biological research are currently being advocated worldwide. These principles emphasize the adoption of various scientific approaches to replace the use of experimental animals, optimize research protocols, and minimize the number of experimental animals used. Countries are promoting the establishment of animal welfare systems aligned with international standards. In recent years, organoid model technology has become the most direct and powerful embodiment of the 3R principles for experimental animals. Organoid models utilize specialized, high-tech equipment to minimize the use of experimental animals, optimize study designs, obtain target organ tissue, and utilize in vitro culture techniques under specialized conditions to reconstruct a three-dimensional spatial structure similar to the target organ. This approach maximizes the preservation and reproduction of the biological properties of the target organ, including the structural composition of major cellular components, the spatially ordered organization of tissue structures, key biological features, and specific cellular biological activities. This technique is currently the most authentic in vitro model for observing the cell-tissue biological properties of the target organ, allowing researchers to closely mimic the in vivo state.

[0004] Nervous system injuries encompass both peripheral and central nervous system damage. Mechanical injury to the spinal cord or brain is the most challenging central nervous system injury among various high-energy traumas, and has long been a major challenge plaguing clinical medicine, neurobiology, rehabilitation medicine, and other life science research fields. Vast research funds are invested globally (primarily in developed Western countries) annually in neuroprotection and neuroregeneration research. However, to date, few breakthroughs have been achieved. Research on spinal cord injury typically relies on various in vivo models. Currently, commonly used models include heavy impact, ventral and dorsal clamping, hemisection, and total resection. Following spinal cord injury, animals experience random loss of sensory and motor function below the level of injury, severe impairment of bowel and bladder function, and visceral sympathetic and parasympathetic nervous system function. In particular, severe pain and secondary infections can lead to devastating consequences for the quality of life of experimental animals. For these animals, this type of neurological trauma is tantamount to the most severe form of physical torture. Therefore, constructing an in vitro research model of spinal cord injury is an urgent direction of research on spinal cord injury in order to achieve the current requirements of experimental animal ethics and animal welfare protection.

[0005] Undoubtedly, the ideal in vitro model for spinal cord or brain injury is to observe spinal cord or brain tissue at the organ-level in vitro. However, current research and preparation techniques are unable to fully preserve the blood supply and cerebrospinal fluid circulation system of spinal cord tissue in vitro. Currently, organoid models that can be prepared are generally based on thick slices of spinal cord or brain tissue. With the continuous advancement of cutting technology and in vitro culture technology, the thickness of neural tissue slices is increasing, which has brought the concepts of in vitro central nervous system tissue blocks and central nervous system organoid models closer and closer, thus making the preparation of spinal cord organoid models more and more perfect. This in vitro model can more completely observe the various specific neurobiological behaviors of spinal cord tissue after injury, such as the interaction between key central nervous system components such as neurons, glial cells, extracellular matrix, and axons. At the level of classic neurobiological research, spinal cord organoid models can simulate the degenerative characteristics of injured neural components at different time stages in vivo, the interactions and respective outcomes of various neurons and glial cells, and various changes in sensory and motor axons. This will greatly facilitate in-depth research on neuroactive drug screening, neurotoxicity testing, neuroprotection, and regeneration, particularly in molecular and cell biology studies of neural stem cell differentiation and axonal regeneration, by simulating an in vivo experimental environment. This technology also provides a highly effective in vitro model for observing the effects of various novel biomaterials or tissue-engineered complexes on neuroprotection and neuroregeneration.

[0006] It is noteworthy that the spinal cord organoid model has unique and major research value in the development and design application of neurobiomaterials. The implantation of biomaterials in the body will involve complex microsurgery. The various physiological activities of experimental animals after surgery will produce various physical strains on the material-host tissue interface, which will bring many adverse factors to the changes of biomaterials in the body, increase the complexity of the interpretation of research results, and is not conducive to the early judgment of the key biological effects of materials and host tissues in the research stage, and is not conducive to the targeted improvement of the physical and chemical properties of biomaterials, which has a significant adverse effect on the tissue engineering design optimization of neurobiomaterials.

[0007] Therefore, the preparation and optimization technology of spinal cord organoid models will bring more and more efficient research observation methods and technical solutions to the regeneration and repair of spinal cord injury. It is especially conducive to observing the application of exogenous neural repair strategies such as neural biomaterial scaffolds and various stem cell transplantations in the regeneration of injured spinal cord and the exploration of related molecular-cellular biological mechanisms.

[0008] The current commercial organoid tissue slicing technology prepares thick tissue slices through a semi-automated mechanical repetitive action. The mechanical vibration generated is large, and the various mechanical parameters during the cutting process cannot be fine-tuned and optimized according to the actual situation. As a result, vibration-induced mechanical damage to fresh tissue in vitro, destruction of tissue microstructure, and uneven thickness of slices resulting in poor sample uniformity may occur during the cutting process. These shortcomings have brought significant disadvantages to the in vitro application of organoid models, and there is an urgent need to improve related technologies to optimize the preparation of organoid tissue. Summary of the Invention

[0009] In response to the above-mentioned problems existing in the prior art, the present invention provides a precision device for preparing biological organoid structures cultured in vitro.

[0010] In order to achieve the above technical objectives and the above technical effects, the present invention is implemented through the following technical solutions: A precision device for preparing biological organoid structures cultured in vitro, comprising a slicing part, a micro-motion part and a linkage part, the slicing part comprising a cutter, a knife holder and a sliding seat, the cutter being arranged on the knife holder, one end of the knife holder being fixedly connected to a rotating shaft, the sliding seat sliding along the vertical direction of the cutter, the micro-motion part comprising a precision displacement tool and a chimeric frame, the chimeric frame and the sliding seat cooperating, the precision displacement tool being provided with a linkage part, the linkage part comprising an engaging component cooperating with the rotating shaft, a fitting component cooperating with the moving tool, and a driving component that enables the fitting component to work unidirectionally, the driving component being driven by the engaging component.

[0011] The top end of the tool holder is fixedly provided with a toothed connecting strip which is cooperatively connected with the toothed connecting strip, and the lower end of the tool holder is connected with the toothed connecting strip.

[0012] Furthermore, the tool holder is provided with an upper groove, an upper frame is provided on one side of the tool holder, the upper frame is slidably connected in the upper groove, and an arc frame is fixedly connected to the upper end of the sliding seat.

[0013] Furthermore, the micro-motion part includes a limit frame, a connecting frame, a mosaic frame, a clamping frame, and a micrometer. The limit frame is fixedly connected to the base on the rear side of the fixed seat. The limit frame is provided with a lower groove. A lower frame is provided on the lower side of the connecting frame. The lower frame is slidably connected in the lower groove. One end of the connecting frame is fixedly connected to the mosaic frame. One side of the limit frame is fixedly connected to the clamping frame. A micrometer is detachably connected in the clamping frame. The micrometer is a screw micrometer. A digital display component of the screw micrometer is placed in the clamping frame. One end of the micrometer is in contact with the connecting frame.

[0014] Furthermore, the linkage part includes a main frame, a swivel, a slip ring, a tightening button and a one-way tooth, the main frame is fixedly connected to the base, the swivel is rotatably connected to the main frame through a stepped ring frame, the slip ring is detachably connected to the swivel, one side of the slip ring is threadedly connected to the tightening button, the tightening button abuts against the micrometer, and the swivel and slip ring are fixedly connected to the one-way tooth; The linkage part also includes a sub-frame, a reciprocating ring, a moving ring, a weight, a fixed column, an inclined block, a reference frame and a limit seat. The auxiliary frame is fixedly connected to one side of the main frame, the reciprocating ring is rotatably connected to the auxiliary frame, the moving ring is slidably connected to the reciprocating ring, the reciprocating ring is connected to a weight, the moving ring is fixedly connected to a fixed column, the reciprocating ring is slidably connected to an inclined block, the inclined block cooperates with a one-way tooth, one side of the auxiliary frame is fixedly connected to the reference frame, the reference frame is slidably connected to the limit seat, and the limit is fixedly connected to a pointing block; A pull rope is connected to the fixed column, one end of the rotating shaft is rotatably connected to the support, one end of the rotating shaft is fixedly connected to the main shaft, the main shaft is fixedly connected to the main gear, the support is rotatably connected to the secondary shaft, the secondary shaft is fixedly connected to the secondary gear, the main gear is meshed with the secondary gear, rope grooves are opened on the main shaft and the secondary shaft, and the pull rope is wound around the rope grooves.

[0015] Beneficial effects of the present invention: This micro-precision device innovatively combines a microelectronic control system with the biological structural characteristics of central nervous system tissue. Using a micrometer-level stepper as the thickness control mechanism for organoid tissue slices, it can intuitively, conveniently, and efficiently prepare organoid tissue structures with a thickness of 100-500 microns in vitro under sterile conditions. Of course, any product implementing the present invention does not necessarily need to achieve all of the advantages described above at the same time. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0017] Figure 1 To prepare the mechanical diagram of the instrument Panorama-1; Figure 2 To prepare the mechanical diagram of the instrument Panorama-2; Figure 3 A mechanical stage for the preparation instrument; Figure 4 A sample tray for the mechanical stage of the preparation instrument; Figure 5 Mechanical sample cutter for the preparation instrument; Figure 6 For the preparation of instrument mechanical stepping and transmission structure-1; Figure 7 For the preparation of instrument mechanical stepping and transmission structure-2; Figure 8 For the preparation of instrument mechanical stepping and transmission structure-3; Figure 9 For the preparation of instrument mechanical stepping and transmission structure-4; Figure 10 For the preparation of instrument mechanical stepping and transmission structure-5; Figure 11 For the preparation of instrument mechanical stepping and transmission structure-6; Figure 12 For the preparation of instrument mechanical stepping and transmission structure-7; Figure 13For the preparation of instrument mechanical stepping and transmission structure-8; Figure 14 For the preparation of instrument mechanical stepping and transmission structure-9; Figure 15 For the preparation of instrument mechanical stepping and transmission structure-10; Figure 16 For the preparation of instrument mechanical stepping and transmission structure-11; Figure 17 For the preparation of instrument mechanical stepping and transmission structure-12; Figure 18 For the preparation of instrument mechanical stepping and transmission structure-13; Figure 19 For the preparation of instrument mechanical stepping and transmission structure-14; Figure 20 For the preparation of instrument mechanical stepping and transmission structure-15; Figure 21 To prepare the mechanical chassis of the instrument; Figure 22 To prepare the mechanical part chassis-2 of the instrument; Figure 23 To prepare the mechanical part chassis-3 of the instrument; Figure 24 For the preparation instrument sample thickness precision setting device-1; Figure 25 For the preparation instrument sample thickness precision setting device-2; Figure 26 For the preparation instrument sample thickness precision setting device-3; Figure 27 For the preparation instrument sample thickness precision setting device-4; Figure 28 For the preparation instrument sample thickness precision setting device-5; Figure 29 For the sample thickness precision setting device of the preparation instrument-6; Figure 30 For the preparation instrument sample thickness precision setting device-7; Figure 31 For the sample thickness precision setting device of the preparation instrument-8; Figure 32 It is the device for setting the sample thickness precision of the preparation instrument-9.

[0018] In the accompanying drawings, the components represented by the reference numerals are as follows: 1-base, 110-fixed seat, 111-end seat, 112-lower nail, 113-slide, 120-slide seat, 121-arc frame, 122-upper nail, 123-slide, 130-bracket, 131-rotating shaft, 132-knife holder, 133-upper slot, 134-control handle, 135-fixed frame, 136-upper frame, 137-adjustment frame, 138-cutter, 2-limiting frame, 201-lower slot, 202- Connecting frame, 203-lower frame, 204-mosaic frame, 205-card frame, 3-micrometer, 310-main frame, 311-swivel, 312-step ring frame, 313-one-way tooth, 314-step groove, 315-slip ring, 316-step block, 317-tightening button, 318-screw, 319-top tightening washer, 320-sub-frame, 321-reciprocating ring, 322-fixed rope, 323-weight, 324-transfer ring, 3 25-fixed column, 326-oblique block, 327-spring, 330-reference frame, 331-size mark, 332-concave frame, 333-tightening nail, 334-limiting seat, 335-pointing block, 340-rope winding, 341-pull rope, 350-support, 351-main shaft, 352-main gear, 353-sub-gear, 354-sub-shaft, 355-rope groove, 101-bottom pad, 102-extension ring, 103-pressure Spring, 104-fixing pad, 360-displacement motor, 361-output shaft, 362-driving gear, 363-support frame, 364-limiting groove, 365-half ring, 366-step teeth, 367-side groove, 368-side ring, 369-side frame, 370-drive motor, 380-control frame, 381-display part, 382-adjustment key, 383-on / off key, 384-lower connecting wire, 385-upper connecting wire. DETAILED DESCRIPTION

[0019] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts shall fall within the scope of protection of the present invention.

[0020] Example 1 like Figure 1-19 shown A precision device for preparing a biological organoid structure cultured in vitro, comprising a slicing part, a micro-motion part and a linkage part, wherein the slicing part comprises a cutter (138), a knife holder (132) and a sliding seat (120), wherein the cutter (138) is arranged on the knife holder (132), and one end of the knife holder (132) is fixedly connected to a rotating shaft (131), and the sliding seat (120) slides in a direction perpendicular to the cutter (138), wherein the micro-motion part comprises a precision displacement tool and a chimeric frame (204), wherein the chimeric frame (204) cooperates with the sliding seat (120), and wherein a linkage part is provided on the precision displacement tool, wherein the linkage part comprises a meshing component cooperating with the rotating shaft (131), a fitting component cooperating with the moving tool, and a driving component for making the fitting component work in one direction, wherein the driving component is driven by the meshing component; The slicing part comprises a base (1), a fixed base (110), an end base (111), a slide (113), a sliding base (120), a tension spring (124), a bracket (130) and a rotating shaft (131), wherein the fixed base (110) is fixedly connected to the front end of the base (1), one end of the fixed base (110) is fixedly connected to the end base (111), a slide (113) is provided on the upper side of the fixed base (110), the upper side of the fixed base (110) is slidably connected to the sliding base (120), a sliding groove (123) is provided on the lower side of the sliding base (120), the sliding groove (123) cooperates with the slide (113), and one end of the slide (113) is fixedly connected to the upper nail (122 ), one end of the fixing seat (110) is fixedly connected to a lower nail (112), a tension spring (124) is fixedly connected between the upper nail (122) and the lower nail (112), a bracket (130) is fixedly connected to the rear side of the base (1), a rotating shaft (131) is rotatably connected to the bracket (130), a knife holder (132) is fixedly connected to the rotating shaft (131), a fixing frame (135) is detachably connected to the knife holder (132), a direction adjustment frame (137) is detachably connected to the lower side of the fixing frame (135), a cutter (138) is detachably connected to the lower side of the direction adjustment frame (137), and a control handle (134) is fixedly connected to the upper end of the knife holder (132); The tool holder (132) is provided with an upper groove (133), an upper frame (136) is provided on one side of the tool holder (132), the upper frame (136) is slidably connected in the upper groove (133), and an arc frame (121) is fixedly connected to the upper end of the sliding seat (120); The micro-motion part comprises a limit frame (2), a connecting frame (202), a chimeric frame (204), a card frame (205), and a micrometer (3), wherein the limit frame (2) is fixedly connected to the base (1) on the rear side of the fixed seat (110), the limit frame (2) is provided with a lower groove (201), the lower side of the connecting frame (202) is provided with a lower frame (203), the lower frame (203) is slidably connected in the lower groove (201), one end of the connecting frame (202) is fixedly connected to the chimeric frame (204), one side of the limit frame (2) is fixedly connected to the card frame (205), the card frame (205) is detachably connected to the micrometer (3), the micrometer (3) is a screw micrometer (3), the digital display component of the screw micrometer (3) is placed in the card frame (205), and one end of the micrometer (3) is in contact with the connecting frame (202); The linkage part includes a main frame (310), a rotating ring (311), a slip ring (315), a tightening button (317) and a one-way tooth (313), wherein the main frame (310) is fixedly connected to the base (1), the rotating ring (311) is rotatably connected to the main frame (310) by being provided with a stepped ring frame (312), the slip ring (315) is detachably connected to the rotating ring (311), and a tightening button (317) is threadedly connected to one side of the slip ring (315), the tightening button (317) abuts against the micrometer (3), and the rotating ring (310) is fixedly connected to the base (1), the rotating ring (311) is rotatably connected to the main frame (310), and the slip ring (315) is detachably connected to the rotating ring (311). The ring (311) and the slip ring (315) are fixedly connected to the one-way tooth (313); the linkage part also includes a sub-frame (320), a reciprocating ring (321), a shift ring (324), a weight (323), a fixed column (325), an inclined block (326), a reference frame (330) and a limit seat (334), the sub-frame is fixedly connected to one side of the main frame (310), the reciprocating ring (321) is rotatably connected to the sub-frame, the shift ring (324) is slidably connected to the reciprocating ring (321), and the reciprocating ring (321) is fixedly connected to the one side of the main frame (310). ) is connected to a weight (323), a fixed column (325) is fixedly connected to the moving ring (324), an inclined block (326) is slidably connected to the reciprocating ring (321), the inclined block (326) cooperates with the one-way tooth (313), one side of the auxiliary frame is fixedly connected to a reference frame (330), a limited position seat (334) is slidably connected to the reference frame (330), and a pointing block (335) is fixedly connected to the limit position; a pull rope (341) is connected to the fixed column (325), and one end of the rotating shaft (131) is connected to the fixed column (325). The rotating shaft (131) is rotatably connected to the support (350), one end of the rotating shaft (131) is fixedly connected to a main shaft (351), a main gear (352) is fixedly connected to the main shaft (351), a secondary shaft (354) is rotatably connected to the support (350), a secondary gear (353) is fixedly connected to the secondary shaft (354), the main gear (352) is meshed with the secondary gear (353), a rope groove (355) is provided on the main shaft (351) and the secondary shaft (354), and the pull rope (341) is wound around the rope groove (355).

[0021] In manual control, the operator mainly controls the lifting and pressing of the cutter. During the lifting process of the cutter, if Figure 17 In the figure, the main shaft rotates clockwise and the secondary shaft rotates counterclockwise to release the rope. Figure 13As shown, the reciprocating ring rotates counterclockwise under the traction of the weight, and the compression spring contracts after the inclined surface of the inclined block cooperates with the inclined surface of the one-way tooth. At this time, the knife is completed and enters the downward pressing process. The main shaft and the secondary shaft reverse and wrap the pull rope. The pull rope pulls the fixed column to restore the fixed column to the direction of the rope cutting point on the secondary shaft. The rope slides and cooperates with the position change of the fixed column to make the reciprocating ring rotate clockwise. At this time, the one-way teeth of the inclined block are stuck, thereby driving the rotating ring and the slip ring to rotate, completing the control of the micrometer, and achieving the effect of precise tissue displacement along with the cutting knife action; before operation, open the slip ring and place the micrometer, the slip ring and the rotating ring are buckled and fixed to the position of the micrometer by tightening the knob, and the position of the concave frame on the reference frame is adjusted by the tightening pin. The limit seat limits the travel space of the fixed column, adjusts the displacement distance, and controls the slice thickness.

[0022] Example 2 like Figure 20-32 shown A buffer base (1) is provided at the four corners of the lower side of the base (1), and the buffer base (1) includes a bottom pad (101) and a fixed pad (104), the fixed pad (104) is fixedly connected to the base (1), the bottom pad (101) is provided on the lower side of the fixed pad (104), a middle groove is provided in the middle of the bottom pad (101) and the fixed pad (104), the bottom pad (101) and the fixed pad (104) are fixedly connected with a compression spring (103) at the middle groove position, and an extension ring (102) is fixedly connected to the upper side of the bottom pad (101) at the middle groove position, and the extension ring (102) is slidably connected in the middle groove of the fixed pad (104); The base (1) is fixedly connected to a support frame (363) at the control part of the micrometer (3), and a limiting groove (364) is opened on the support frame (363). A semi-ring (365) is slidably connected to the support frame (363). The outer side of the semi-ring (365) is fixedly connected to a plurality of stepped teeth (366), and the stepped teeth (366) cooperate with the limiting groove (364). The cross section of the semi-ring (365) is provided with a side groove (367). The cross section of the semi-ring (365) is detachably connected to a side ring (368), and the cross section of the side ring (368) is fixedly connected to a side frame (369), and the side frame (369) is slidably connected to the side frame (369). Connected to the side groove (367), the outer side of the side ring (368) is fixedly connected to a plurality of stepped teeth (366), the inner sides of the half ring (365) and the side ring (368) are provided with elastic pads, which are fitted with the control component of the micrometer (3) through the elastic pads, and one side of the support frame (363) is fixedly connected to a displacement motor (360), and the displacement motor (360) is provided with an output shaft (361) on the side close to the stepped teeth (366), and a driving gear (362) is fixedly connected to the output shaft (361), and the driving gear (362) is meshed with the stepped teeth (366) on the half ring (365) and the side ring (368); The base (1) is fixedly connected to a control frame (380) on one side of the displacement motor (360), and the control frame (380) includes a prior art control chip and functional elements. A display unit (381) is provided on the upper side of the control frame (380), and the display unit (381) includes an interval time display screen and a movement distance display screen. The control frame (380) is provided with adjustment keys (382) corresponding to adjustment parameters on the lower side of the display unit (381). The interval time adjustment range is "500ms, 1s, 2s", and the displacement distance adjustment range is "100 -500μm", the control frame (380) is fixedly connected to a drive motor (370) at a position close to the rotating shaft (131), the drive motor (370) drives the rotating shaft (131) to rotate, a lower connecting line (384) is fixedly connected between the control frame (380) and the displacement motor (360), an upper connecting line (385) is fixedly connected between the control frame (380) and the drive motor (370), and a switch key (383) for controlling the operation of the displacement motor (360) and the drive motor (370) is also provided on the control frame (380); The control frame (380) achieves the following effects through the control chip of the prior art: the working amplitude of the displacement motor (360) is controlled by the adjustment key (382), and the moving distance of the tissue is achieved by controlling the movement of the micrometer (3) under the meshing transmission of the driving gear (362) and the stepped teeth (366), and finally displayed on the display element (381) of the moving distance display screen; the adjustment key (382) controls the drive motor (370) to do reciprocating work, completes the falling and lifting of the cutter (138), and displays the interval time between the two working times of the drive motor (370) on the display element (381) of the interval time display screen. After turning on the switch key (383), the control chip of the control frame (380) adjusts the displacement motor (360) and the drive motor (370) to work alternately.

[0023] The representative of the existing conventional fresh tissue slicing technology is the McIlwain tissue chopper tissue slicer, which is prone to tissue slice adhesion and excessive mechanical vibration during the slicing process, resulting in tissue slice failure. The cutter part and sliding seat structure of the present application have been prepared and have been applied to the cutting of various central nervous tissues and in vitro organoid model cultivation in university neurobiology laboratories. The automated control system of the present application performs automated work through chip control, and the system quickly completes slicing according to the set cutting program through parameter adjustment. The human central nervous system organoids cultured in vitro targeted by the present application are embedded in low-melting-point agarose, are small and soft, and the vibrations generated during the operation of the machine affect the stability of the tissue, resulting in unsatisfactory final slicing results or even slicing failure. Among them, the patent: Method and device for cutting fresh tissue slices - US20100175520A1 - A method and device for cutting fresh tissue slices, discloses the vibration slicing technology of related fresh tissues; The usage method of this application is divided into manual and automated operations. During the contact between the cutter and the tissue, the operator can manually perform gentle cutting, which is suitable for timely intervention in problems that occur during the slicing process and avoid mechanical vibration affecting the slicing results; the control frame can also use microelectronic technology to coordinate various parameters to achieve a complete and efficient cutting control system automation, avoiding fresh tissue from being exposed to the air for too long and affecting tissue vitality, which is crucial for the preparation of organoids of central nervous tissue.

[0024] This neural tissue organoid model preparation instrument can be widely used to prepare organoid models of peripheral and central nervous tissues, such as the brain and spinal cord, and has broad application prospects in key neurobiological research fields such as neuroprotection, nerve injury repair, stem cell transplantation, research on the biological effect mechanisms of neuroactive molecules, and neural regeneration. Compared with similar preparation instruments at home and abroad, this design has the following advantages: 1. The thickness and cutting stability of the tissue slices are basically comparable to those of foreign electric products through the high-precision control of the movement distance by the micrometer, and the subsequent preparation cost is 1 / 6 of similar equipment; 2. The preparation method of tissue slices is more flexible than existing products. By adjusting the position of the culture dish or the tissue between the arcs of the sliding base, sagittal and coronal sections of spinal cord, brain, and peripheral nerve tissue can be prepared at any time. Cross-sections and longitudinal sections of neural tissue can be prepared at various angles, and tissue protection is better implemented during the slicing process, demonstrating the unique advantages of this design device; 3. The unobstructed sliding base based on zero notch allows operators to observe and perform slicing without blind spots, and the entire slicing process is visualized without obstacles, which greatly saves precious animal tissue.

[0025] Current domestic neurobiology research lacks standardized cutting tools in the preparation of central nervous system organoid models. Through the precision device for preparing biological organoid structures cultured in vitro in this application, the reliable effect of accurately controlling the thickness of the slices is achieved, and the slicing work is stably completed under the transmission cooperation between components; the preparation cost of this precision equipment in this application is about 1 / 6 of similar electric equipment abroad. The low-cost advantage of this application will help to promote its application in the corresponding scientific research field.

[0026] The device of the present application can be used by an operator to manually control the cutter to perform precise slicing operations on demand, or it can be controlled by adding an automation module to adjust various parameters to perform various precise automatic slicing operations. This greatly overcomes the disadvantages of existing similar foreign equipment such as excessive vibration and sample disturbance during use, which can cause adverse consequences such as uneven sample thickness and microstructural damage to fragile fresh tissue. For example, during the electric slicing process, certain foreign tissue slicers often cause significant mechanical vibration due to excessive cutting force from the cutter, resulting in damage to fragile central nervous system tissue or microstructural damage. In addition, improper vibration of the stage causes unstable tissue cutting, resulting in uneven slice thickness and uneven results, which significantly affects sample quality. In view of this, the present application can not only slice tissue blocks more gently through manually operated mechanical devices, but also significantly improve the relevant disadvantages of competing products by scientifically setting the mechanical force and scientifically designing the shock-absorbing stage. While effectively achieving precise slicing, it greatly protects the fragile microstructure of central nervous tissue, allowing the preparation of organoid structures of various tissues more scientifically and reasonably through this precision device, effectively improving the scientific quality of related biological research.

[0027] The invention can prepare various axially oriented tissue structures according to the actual biological observation direction, and is particularly suitable for the preparation of organoid structures of central and peripheral nervous tissues with highly ordered functional structures. The scope of application can be extended to the preparation of in vitro culture models of various solid tumors and various solid organ organoids, providing a relatively economical, efficient and reliable organoid preparation tool for the visualization and observation of biological characteristics of various three-dimensional structures. The finished product of this patent has been prepared and has been used by research groups at Sun Yat-sen University and Wuhan University with smooth progress, providing great convenience for the study of organoids of central nervous tissues such as spinal cord injuries. Subsequent work will cooperate with bio-mechanical companies to complete the relevant market promotion and industrialization process, providing an important scientific research means and practical tool for the in vitro culture and scientific research of three-dimensional biological structures of micro-organs and organoids with great research potential.

[0028] In summary, this micro-precision device innovatively combines the microelectronic control system with the biological structural characteristics of central nervous system tissue, and uses a spiral micrometer micrometer-level stepping device as the thickness control structure of the organoid tissue slice. It can intuitively, conveniently and efficiently prepare organoid tissue structures with a thickness of 100-500 microns under sterile conditions in vitro.

[0029] Throughout this specification, references to terms such as "one embodiment," "example," or "specific example" indicate that the specific features, structures, materials, or characteristics described in conjunction with that embodiment or example are included in at least one embodiment or example of the present invention. In this specification, schematic representations of these terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

[0030] The preferred embodiments of the present invention disclosed above are intended only to help illustrate the present invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the present invention to the specific embodiments described. Obviously, many modifications and variations are possible based on the content of this specification. These embodiments are selected and described in detail in this specification to better explain the principles and practical applications of the present invention, thereby enabling those skilled in the art to better understand and utilize the present invention. The present invention is limited only by the claims and their full scope and equivalents.

Claims

1. An in vitro preparation apparatus for biological organoid structures. This invention is a precision device for preparing in vitro cultured biological organoid structures, characterized by: It includes a slicing part, a micro-motion part and a linkage part. The slicing part includes a cutter, a knife holder and a sliding seat. The cutter is set on the knife holder. One end of the knife holder is fixedly connected to a rotating shaft. The sliding seat slides along the vertical direction of the cutter. The micro-motion part includes a precision displacement tool and a fitting frame. The fitting frame cooperates with the sliding seat. A linkage part is provided on the precision displacement tool. The linkage part includes an engaging component that cooperates with the rotating shaft, a fitting component that cooperates with the moving tool, and a driving component that enables the fitting component to work unidirectionally. The driving component is driven by the engaging component.

2. The precision device for preparing in vitro cultured biological organoid structures according to claim 1, characterized in that: The slicing part comprises a base, a fixed base, an end base, a slide, a sliding base, a tension spring, a bracket and a rotating shaft, the fixed base is fixedly connected to the front end of the base, one end of the fixed base is fixedly connected to the end base, the upper end of the fixed base is provided with a slide, the upper end of the fixed base is slidably connected to the sliding base, the lower end of the sliding base is provided with a slide, and the slide is matched with the slide, one end of the slide is fixedly connected to an upper nail, one end of the fixed base is fixedly connected to a lower nail, and the upper nail and the lower nail are fixedly connected with a tension spring between the upper nail and the lower nail. The rear end of the base is fixedly connected to the bracket, the bracket is rotatably connected to the rotating shaft, the rotating shaft is fixedly connected to the knife holder, the knife holder is detachably connected to the fixed base, the lower side of the fixed base is detachably connected to the adjustment bracket, and the lower side of the adjustment bracket is detachably connected to the cutting knife 3. The precision device for preparing in vitro cultured biological organoid structures according to claim 2, characterized in that: The tool holder is provided with an upper groove, an upper frame is provided on one side of the tool holder, the upper frame is slidably connected in the upper groove, and an arc frame is fixedly connected to the upper end of the sliding seat.

4. The precision device for preparing in vitro cultured biological organoid structures according to claim 3, characterized in that: The micro-motion part includes a limit frame, a connecting frame, a mosaic frame, a clamping frame, and a micrometer. The limit frame is fixedly connected to the base on the rear side of the fixed seat. The limit frame is provided with a lower groove. A lower frame is provided on the lower side of the connecting frame. The lower frame is slidably connected in the lower groove. One end of the connecting frame is fixedly connected to the mosaic frame. One side of the limit frame is fixedly connected to the clamping frame. A micrometer is detachably connected in the clamping frame. The micrometer is a screw micrometer. A digital display component of the screw micrometer is placed in the clamping frame. One end of the micrometer abuts against the connecting frame.

5. The precision device for preparing in vitro cultured biological organoid structures according to claim 4, characterized in that: The linkage part includes a main frame, a swivel, a slip ring, a tightening button and a one-way tooth. The main frame is fixedly connected to the base. The swivel is rotatably connected to the main frame through a stepped ring frame. The slip ring is detachably connected to the swivel. A tightening button is threadedly connected to one side of the slip ring. The tightening button abuts against the micrometer. The swivel and the slip ring are fixedly connected to the one-way tooth. The linkage part also includes a sub-frame, a reciprocating ring, a moving ring, a weight, a fixed column, an inclined block, a reference frame and a limit seat. The auxiliary frame is fixedly connected to one side of the main frame, the reciprocating ring is rotatably connected to the auxiliary frame, the moving ring is slidably connected to the reciprocating ring, the reciprocating ring is connected to a weight, the moving ring is fixedly connected to a fixed column, the reciprocating ring is slidably connected to an inclined block, the inclined block cooperates with a one-way tooth, one side of the auxiliary frame is fixedly connected to the reference frame, the reference frame is slidably connected to the limit seat, and the limit is fixedly connected to a pointing block; A pull rope is connected to the fixed column, one end of the rotating shaft is rotatably connected to the support, one end of the rotating shaft is fixedly connected to the main shaft, the main shaft is fixedly connected to the main gear, the support is rotatably connected to the secondary shaft, the secondary shaft is fixedly connected to the secondary gear, the main gear is meshed with the secondary gear, rope grooves are opened on the main shaft and the secondary shaft, and the pull rope is wound around the rope grooves.

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