Matrix gel wrapping organoid dehydration mechanism and its degumming method

Abstract

The application relates to a matrix glue wrapped organoid dehydration mechanism and a glue removing method thereof and relates to the technical field of biological tissue engineering. The application solves the problems that, in the existing matrix glue embedding method for culturing organoids, the environment cannot be guaranteed to be constant in temperature due to uneven hot gas flow, the activity of the organoids is affected, the dehydration degree of the organoids in different dehydration modules is uncontrollable, and the organoids are deactivated due to long dehydration time. A longitudinal channel is arranged in the center of the base, two gas inlet hole sections are arranged on the top of the longitudinal channel in a tangential center-symmetrical mode, a horizontal gas inlet channel is arranged in the base and communicates with the gas inlet ends of the two gas inlet hole sections, a center distribution channel is arranged at the bottom of the longitudinal channel, a plurality of horizontal gas outlet channels are arranged outside the center distribution channel, and the center distribution channel communicates with a plurality of butt joint channels through the plurality of horizontal gas outlet channels. The application guarantees that the hot gas flow is stable, the hot gas flow is uniform, and the dehydration degree is controllable during the dehydration of the matrix glue wrapped organoids.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological tissue engineering, in particular to a matrix gel wrapped organoid dehydration mechanism and a gel removal method thereof. BACKGROUND

[0002] In the existing organoid culture system, at least the following problems exist in the dehydration process: because a whole cavity is opened in the dehydration platform as a heat source cavity, the heat source cavity is directly communicated with the dehydration channels of multiple dehydration modules, and the heated pressure air enters the heat source cavity from one side of the air inlet hole and then flows out from the other side of the air outlet hole, which cannot guarantee that the flow of hot gas into each dehydration channel of the dehydration module is uniform. The uneven flow of hot gas cannot guarantee a constant temperature environment, which affects the activity of the organoid. In addition, the uneven flow of hot gas also causes the dehydration degree of the organoid in multiple dehydration modules to be uncontrollable, which brings great difficulty to monitoring, and the dehydration time is too long, which will cause the organoid to be inactivated. Because the nutrients in the container are limited and do not have the function of additional oxygen supply, if the dehydration time of the organoid is too long and is not discovered in time, not only the cell activity will be affected, but also the possibility of cell contamination will be greatly increased, which will eventually lead to the inactivation of the organoid due to dehydration failure. SUMMARY

[0003] The purpose of the present application is to solve the problems that the existing matrix gel embedding method for culturing organoids cannot guarantee a constant temperature environment due to uneven flow of hot gas, affects the activity of the organoid, and the dehydration degree of the organoid in different dehydration modules is uncontrollable, and has the risk of inactivation of the organoid due to too long dehydration time, and to provide a matrix gel wrapped organoid dehydration mechanism and a gel removal method thereof.

[0004] The technical solution of the present application is:

[0005] The present application provides a matrix gel wrapped organoid dehydration mechanism, which comprises:

[0006] One or more dehydration seats, the dehydration seat is provided with a dehydration channel and a moisture removal channel;

[0007] A base, the top end of the base is provided with a plurality of dehydration seat mounting grooves for inserting the dehydration seat, the bottom of the dehydration seat mounting groove is provided with a butt joint channel communicated with the dehydration channel and the moisture removal channel, the inside of the base is provided with a longitudinal channel in the center, the top of the longitudinal channel is provided with two tangential center symmetrically arranged air inlet hole sections, the inside of the base is further provided with two transverse air inlet channels communicated with the air inlet ends of the two air inlet hole sections, the bottom of the longitudinal channel is provided with a center distribution channel, the outside of the center distribution channel is provided with a plurality of transverse air outlet channels, and the center distribution channel is communicated with the plurality of butt joint channels through the plurality of transverse air outlet channels.

[0008] Further, the dehydration seat is used for bearing the array holder, and water absorption filter paper is arranged above the dehydration channel and the moisture removal channel of the dehydration seat.

[0009] Further, the base comprises an upper seat plate, an intermediate seat plate and a lower seat plate which are horizontally stacked from top to bottom.

[0010] Further, the longitudinal channel is a cylindrical hole structure penetrating through the upper and lower surfaces of the intermediate seat plate, the bottom of the longitudinal channel is provided with a cylindrical air outlet expansion hole structure, the center lines of the two air inlet hole sections are located on the same horizontal plane, and the center lines of the two air inlet hole sections are arranged in parallel and staggered and tangent to the longitudinal channel.

[0011] Further, the dehydration seat comprises a bottom block, a pressing block, two elastic locking mechanisms and two pressing block connecting pieces, the bottom block is in a cylindrical structure, the bottom block is detachably installed in the dehydration seat installation slot of the base, the middle part of the top end of the bottom block is provided with an integrally formed cuboid structure, the center of the bottom block is provided with a dehydration seat channel penetrating vertically through the upper and lower surfaces of the bottom block, air outlet notches are arranged on both sides of the dehydration seat channel, the bottom of the pressing block is provided with a pressing block installation through slot matched with the bottom block, the pressing block cover is arranged on the cuboid structure at the top of the bottom block, the pressing block is connected with the bottom block through the two pressing block connecting pieces, there is an air outlet gap between the outer wall of the cuboid structure and the inner wall of the pressing block installation through slot, the top end of the pressing block is provided with an array block installation slot penetrating through the pressing block installation through slot, the two end faces of the pressing block are processed into vertical faces matched with the array holder, the middle parts of the vertical faces on both sides of the pressing block are provided with locking piece installation holes vertically penetrating through the pressing block installation through slot, the two elastic locking mechanisms are respectively installed in the two locking piece installation holes, and the array holder is detachably fixedly connected with the pressing block through the two elastic locking mechanisms.

[0012] Further, the dehydration mechanism further comprises a plurality of silica gel ring flat pads, the docking channel is a cylindrical hole structure penetrating through the upper and lower surfaces of the intermediate seat plate, the top end of the docking channel is provided with an elastic flat pad installation slot arranged coaxially, the silica gel ring flat pad is embedded in the elastic flat pad installation slot, the depth of the elastic flat pad installation slot is less than the thickness of the silica gel ring flat pad, the dehydration seat installation slot is a cylindrical hole structure penetrating through the upper and lower surfaces of the upper seat plate, the inner wall of the dehydration seat installation slot is provided with a plurality of base clamping ports uniformly distributed in the circumferential direction, the base clamping port is a rectangular insertion slot penetrating through the upper and lower surfaces of the upper seat plate, the side bottom of the rectangular insertion slot is provided with a smoothly transitioned clamping groove, the side bottom of the bottom block is provided with a plurality of rectangular protrusions uniformly distributed in the circumferential direction, the width of the rectangular protrusion is consistent with the width of the rectangular insertion slot of the base clamping port, and the thickness of the rectangular protrusion is less than the depth of the clamping groove of the base clamping port.

[0013] Further, the dehydration mechanism further comprises two air inlet joints, the openings of the two lateral air inlets are located on the lower surface of the middle seat plate, the two lateral air inlets are away from the air inlet hole section at one end as the base air inlets, the upper half of the air inlet hole section is a cylindrical hole structure penetrating through the upper and lower surfaces of the upper seat plate, the lower half of the air inlet hole section is a cylindrical hole structure penetrating through the upper and lower surfaces of the middle seat plate, and the two air inlet joints are respectively installed in the upper half of the air inlet hole section of the two base air inlets.

[0014] Further, the dehydration mechanism further comprises a calibration block, the calibration block is a cylindrical structure, and the calibration block is detachably installed in the dehydration seat installation groove located at the upper left corner.

[0015] Further, the central distribution channel is a cylindrical hole structure penetrating through the upper and lower surfaces of the lower seat plate, the inner wall of the central distribution channel is provided with a plurality of distribution channel outlets uniformly distributed in the circumferential direction, the lateral air outlet channel is a linear slot structure, the opening of the lateral air outlet channel is located on the upper surface of the lower seat plate, a plurality of lateral air outlet channels are arranged in a diverging manner with the same cross section, one end of each of the plurality of lateral air outlet channels is in communication with the plurality of distribution channel outlets of the central distribution channel, the other end of each of the plurality of lateral air outlet channels is respectively provided with a plurality of second circular interfaces with smooth transitions, and the plurality of second circular interfaces are respectively in communication with the plurality of butt joint channels.

[0016] The application also provides a dehydration method based on a matrix glue wrapping organoid dehydration mechanism, the dehydration method comprising:

[0017] Assembling the array holder and the dehydration seat: using a mechanical arm to clamp the array holder with the agarose microarray block from the culture dish, and installing the array holder in the array block installation groove of the dehydration seat, while the elastic locking mechanism arranged on both sides of the dehydration seat automatically locks and fixes the array holder;

[0018] Absorbing water in the agarose microarray block: using dry water-absorbing filter paper to quickly absorb the water in the micropores of the agarose microarray block installed on the array holder;

[0019] Evaporating water in the water-absorbing filter paper: the two air inlet joints are connected with the constant-temperature gas source in advance, the constant-temperature gas source is turned on, two constant-temperature gas flows enter the two lateral air inlets from the two air inlet joints, and then converge in the longitudinal channel, the converged constant-temperature gas flow forms a constant-temperature rotational flow gas under the action of the longitudinal channel, the constant-temperature rotational flow gas is evenly distributed under the action of the central distribution channel and then flows into the plurality of lateral air outlet channels, and the plurality of constant-temperature gas flows flow into the dehydration seat channels of the plurality of dehydration seats through the butt joint channels, so as to take away the water on the water-absorbing filter paper, and make the agarose microarray block installed on the plurality of array holders continuously dehydrate;

[0020] Adding matrix glue to the agarose microarray block: using an electric dispenser to add matrix glue to the array region of the agarose microarray block, so that the matrix glue covers the sinking array region;

[0021] Separation of the surface of the matrix glue and the agarose microarray block: a new culture dish is taken by the mechanical arm, and the culture dish is inverted on the array holder, then the mechanical arm is used to pick up the constant temperature heat block, and the constant temperature heat block is pasted on the outer bottom of the culture dish, the constant temperature of the constant temperature heat block is maintained, the matrix glue is solidified and adsorbed on the bottom of the culture dish, and due to the continuous dehydration, the agarose microarray block will shrink slightly, and the array holes on the agarose microarray block will slightly enlarge under the shrinkage, so that the surface of the matrix glue and the agarose microarray block are separated;

[0022] Culture of organoids: the matrix glue wrapped cell ball solidified on the bottom of the culture dish is taken off by the mechanical arm, and the culture medium is added in the culture dish for organoid culture.

[0023] Compared with the prior art, the present application has the following effects:

[0024] 1. Two plastic gas inlet joints are installed on the upper seat plate of the matrix glue wrapped organoid dehydration mechanism, constant temperature dry gas is introduced into the two horizontal gas inlet channels of the middle seat plate, and after spiral mixing at the longitudinal channel of the upper end surface of the middle seat plate, the gas is introduced into the central distribution channel of the lower seat plate. The spiral mixed gas flows into six horizontal gas outlet channels with the same cross section, and then uniformly flows into six butt joint channels and the dehydration seat channel of the bottom block, and then flows out from the gas outlet gap and the gas outlet gap between the pressing block and the bottom block, so that the gas flow in each dehydration seat channel is uniform. The present application can ensure that the constant temperature gas flowing into the multiple dehydration seats is uniform, thereby ensuring the constant temperature environment and the activity of the organoids, and the dehydration degree of the organoids in different dehydration modules is controllable, which effectively avoids the risk of inactivation of the organoids due to too long dehydration time.

[0025] 2. A layer of water absorption filter paper is provided at the array contact position of the dehydration mechanism and the agarose microarray block, so that the water in the agarose array area can quickly penetrate from the loose and porous structure of the agarose to the water absorption filter paper. The bottom of the filter paper is supplied with constant temperature dry gas at 37℃, which can take away the water in the water absorption filter paper through air flow, so that the water absorption filter paper can maintain a certain water absorption and can be reused.

[0026] 3. Two elastic bumping beads are installed on the pressing block, which can press and fix the array holder with a certain force after loading. The array holder and the dehydration mechanism are matched in size, which can be easily clamped on the array block mounting groove of the dehydration seat and fixed by the elastic bumping beads. Therefore, the present application can cooperate with the mechanical arm to complete the automatic operation. The calibration block can be detachably installed in the dehydration seat mounting groove at the upper left corner, which plays a visual calibration role and can cooperate with the mechanical arm to complete the adaptive automatic operation.

[0027] 4, The matrix glue wrapped organoid degelation method of the application takes into account the high throughput and consistency of organoid culture by glue droplet embedding method, is conducive to the establishment of organoid standardization, and has very important significance for accelerating drug development and screening, promoting personalized medicine, and improving the depth and breadth of disease research. BRIEF DESCRIPTION OF DRAWINGS

[0028] Figure 1 is a matrix glue wrapped organoid dehydration mechanism isometric view of the application;

[0029] Figure 2 is an isometric view of the dehydration seat and array holder assembly of the application;

[0030] Figure 3 is an isometric view of the dehydration seat of the application;

[0031] Figure 4 is an exploded view of the dehydration seat of the application;

[0032] Figure 5 is an exploded view of the base and air inlet gas path joint assembly of the application;

[0033] Figure 6 is a schematic diagram of the gas flow in the base of the application;

[0034] Figure 7 is a schematic diagram of the array holder in a matrix glue wrapped organoid degelation method of the application;

[0035] Figure 8 is a schematic diagram of the agarose array area sinking in an array holder in a matrix glue wrapped organoid degelation method of the application;

[0036] Figure 9 is a schematic diagram of the mechanical arm clamping inverted culture dish in a matrix glue wrapped organoid degelation method of the application.

[0037] In the figure: 1, base; 109, upper seat plate; 110, middle seat plate; 111, lower seat plate; 101, dehydration seat mounting slot; 102, docking channel; 103, air inlet; 104, longitudinal channel; 105, transverse air inlet channel; 106, central distribution channel; 107, transverse air outlet channel; 108, base bayonet; 2, dehydration seat; 208, bottom block; 209, pressing block; 210, elastic locking mechanism; 211, pressing block connecting piece; 212, water absorption filter paper; 201, dehydration seat channel; 202, air outlet gap; 203, pressing block mounting slot; 204, air outlet gap; 205, array block mounting slot; 206, locking piece mounting hole; 207, rectangular protrusion; 3, array holder; 4, air inlet gas path joint; 5, calibration block; 6, mechanical arm; 7, culture dish. DETAILED DESCRIPTION

[0038] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work fall within the scope of the present application.

[0039] It should be noted that the embodiments in the present application and the features in the embodiments can be combined with each other without conflict.

[0040] In the existing organoid culture system, at least the following problems exist in the dehydration process: because a whole cavity is opened in the dehydration platform as a heat source cavity, the heat source cavity is directly communicated with the dehydration channels of a plurality of dehydration modules, and the heated pressure air enters the heat source cavity from one side of the air inlet hole and then flows out from the other side of the air outlet hole, which cannot guarantee that the flow of hot gas flowing into the dehydration channel of each dehydration module is uniform. The uneven flow of hot gas cannot guarantee a constant temperature environment, which affects the activity of the organoids. In addition, the uneven flow of hot gas also causes the dehydration degree of the organoids in the plurality of dehydration modules to be uncontrollable, which brings great difficulty to monitoring, and the dehydration time is too long, which will cause the organoids to be inactivated. Because the nutrients in the container are limited and do not have the function of additional oxygen supply, if the dehydration time of the organoids is too long and is not discovered in time, not only the cell activity will be affected, but also the possibility of cell contamination will be greatly increased, which will eventually lead to the inactivation of the organoids due to dehydration failure. In addition, because the air inlet hole and the air outlet hole of the heat source cavity are located below the dehydration module, and the dehydration module above is a sealed space, the amount of upward airflow is small or even no hot gas flows upward, which causes the dehydration of the organoids to fail, or the limited oxygen in the container causes the organoids to be inactivated because the dehydration time is too long.

[0041] Therefore, the present application provides a matrix glue wrapped organoid dehydration mechanism and a glue removal method to solve the above technical problems. The present application will be further described below in combination with the drawings and specific embodiments, but is not limited thereto.

[0042] Specific implementation method one: combination Figures 1 to 9 In this embodiment, a matrix glue wrapped organoid dehydration mechanism is described. The dehydration mechanism comprises:

[0043] One or more dehydration seats, the dehydration seat is provided with a dehydration channel and a moisture removal channel;

[0044] The base 1 is provided with a plurality of dehydration seat installation grooves 101 at the top end of the base 1, the dehydration seat installation grooves 101 are used for inserting the dehydration seat 2, the bottom of the dehydration seat installation groove 101 is provided with a butt joint channel 102 communicated with the dehydration channel and the wet exhaust channel, the inside of the base 1 is provided with a longitudinal channel 104 in the center, the top of the longitudinal channel 104 is provided with two tangential center symmetrically arranged air inlet hole sections, the inside of the base 1 is further provided with two transverse air inlet channels 105 respectively communicated with the air inlet ends of the two air inlet hole sections, the bottom of the longitudinal channel 104 is provided with a center distribution channel 106, the outside of the center distribution channel 106 is provided with a plurality of transverse air outlet channels 107, the center distribution channel 106 is communicated with the plurality of butt joint channels 102 through the plurality of transverse air outlet channels 107.

[0045] In the embodiment, the plurality of dehydration seat installation grooves 101 are arranged on the upper surface of the base 1 in the form of a rectangular array. The longitudinal channel 104 adopts a longitudinal spiral channel, the inner wall of the longitudinal spiral channel is provided with a continuous spiral line-shaped protrusion, two routes of constant temperature gas entering the two transverse air inlet channels 105 from the two air inlet gas joints 4 are collected in the longitudinal spiral channel, and the collected constant temperature gas flow forms constant temperature rotational flow gas under the action of the longitudinal spiral channel; the plurality of transverse air outlet channels 107 are distributed in a radial and circumferential manner outside the center distribution channel 106.

[0046] Specific implementation method two: combined with Figures 1 to 9 It is explained that the dehydration seat 2 of the embodiment is used for carrying the array holding frame 3, and the dehydration seat 2 is provided with the water absorption filter paper 212 above the dehydration channel and the wet exhaust channel. In this way, there is a layer of water absorption filter paper 212 at the array contact position of the dehydration mechanism and the agarose microarray block, so that the water in the agarose array region is quickly penetrated from the loose and porous structure of the agarose to the water absorption filter paper 212, the bottom of the filter paper is communicated with the constant temperature drying gas at 37℃, and the water in the water absorption filter paper 212 is taken away by the gas flow. The other components and connection relationships are the same as those of the specific implementation method one.

[0047] In the embodiment, the water absorption filter paper 212 is horizontally arranged at the intersection of the dehydration seat channel 201 and the array block installation groove 205, and the upper and lower surfaces of the water absorption filter paper 212 are respectively in contact with the lower surface of the pressing block 209 and the upper surface of the bottom block 208.

[0048] Specific implementation method three: combined with Figures 1 to 9 It is explained that the base 1 of the embodiment includes an upper seat plate 109, an intermediate seat plate 110 and a lower seat plate 111 which are horizontally stacked in sequence from top to bottom.

[0049] In this way, the base 1 is divided into a three-layer structure, which facilitates the processing of various channels in the base 1. The other components and connection relationships are the same as those of the specific implementation method one or two.

[0050] In the embodiment, the upper seat plate 109 is made of metal material to ensure the mechanical strength and wear resistance of the base socket 108, and the middle seat plate 110 and the lower seat plate 111 are made of plastic material to enhance the heat insulation performance. The upper seat plate 109, the middle seat plate 110 and the lower seat plate 111 are all rectangular plate structures, the upper surface of the upper seat plate 109 is provided with four seat plate counterbores, the upper surface of the middle seat plate 110 is provided with four seat plate light holes, and the upper surface of the lower seat plate 111 is provided with four seat plate screw holes. The upper seat plate 109, the middle seat plate 110 and the lower seat plate 111 are connected by four seat plate screws. In order to ensure the sealing of the butt joint of the three seat plates, the machining precision of the surfaces of the three seat plates needs to be ensured, so that the channel formed between the adjacent layers of seat plates has good sealing performance to avoid gas leakage. In order to further ensure the sealing, the periphery of each channel can be processed into a continuous sealing groove with an embedded rubber sealing ring.

[0051] Specific implementation four: combination Figures 1 to 9 In this embodiment, the longitudinal channel 104 is a cylindrical hole structure penetrating through the upper and lower surfaces of the middle seat plate 110, the bottom of the longitudinal channel 104 is provided with a cylindrical gas outlet expansion hole structure, the center lines of the two gas inlet hole sections are located on the same horizontal plane, and the center lines of the two gas inlet hole sections are arranged in parallel and staggered and tangent to the longitudinal channel 104.

[0052] In this way, the two constant temperature gases flowing into the middle seat plate 110 pass through the two transverse gas inlet channels 105, are mixed spirally at the upper end surface of the middle seat plate 110, and then enter the central distribution channel 106 of the lower seat plate 111. The other components and connection relationships are the same as those of the first, second or third embodiment.

[0053] Further, the gas inlet hole section is a straight line type gas inlet hole section, two gas inlet hole sections are arranged on both sides of the longitudinal channel 104, the gas outlet of each gas inlet hole section communicates with the gas inlet of the longitudinal channel 104, and the gas outlet of each gas inlet hole section enters the longitudinal channel 104 tangentially, and the gas outlet centers of the two gas inlet hole sections are symmetrically arranged.

[0054] Specific implementation five: combination Figures 1 to 9In this embodiment, the dehydration seat 2 comprises a bottom block 208, a pressing block 209, two elastic locking mechanisms 210 and two pressing block connecting members 211. The bottom block 208 is in a cylindrical structure at the bottom, and is detachably installed in the dehydration seat installation slot 101 of the base 1. A cuboid structure is integrally formed in the middle of the top end of the bottom block 208. A dehydration seat passage 201 is vertically formed through the upper and lower surfaces of the bottom block 208. Gas outlets 202 are arranged on both sides of the dehydration seat passage 201. The bottom of the pressing block 209 is provided with a pressing block installation through slot 203 matched with the bottom block 208. The pressing block 209 is arranged on the cuboid structure at the top of the bottom block 208. The pressing block 209 is connected with the bottom block 208 through the two pressing block connecting members 211. The outer wall of the cuboid structure and the inner wall of the pressing block installation through slot 203 form a gas outlet gap 204. The top end of the pressing block 209 is provided with an array block installation slot 205 penetrating through the pressing block installation through slot 203. The two end faces of the pressing block 209 are processed into vertical faces matched with the array holder 3. The middle part of the vertical faces on both sides of the pressing block 209 is provided with a locking member installation hole 206 vertically penetrating through the pressing block installation through slot 203. The two elastic locking mechanisms 210 are respectively installed in the two locking member installation holes 206. The array holder 3 is detachably fixedly connected with the pressing block 209 through the two elastic locking mechanisms 210.

[0055] In this way, two elastic bumping beads are installed on the pressing block 209, and the array holder 3 is tightly fixed after being loaded. The array holder 3 and the dehydration mechanism are matched in size, and can be conveniently clamped on the array block installation slot 205 of the dehydration seat 2 and fixed through the elastic bumping beads. The bottom block 208 is hollow. The constant-temperature drying gas enters the bottom block 208 through the base 1, and flows out from the gas outlets 202 and the gas outlet gap 204 between the pressing block 209 and the bottom block 208, so as to take away the water on the water-absorbing filter paper 212, so that the water-absorbing filter paper 212 maintains a certain water-absorbing property and can be repeatedly used. The other components and connection relationships are the same as those in the first, second, third or fourth embodiments.

[0056] In this embodiment, the pressing block connecting member 211 is a countersunk screw. Two pressing block connecting circular holes are arranged on both sides of the upper surface of the pressing block 209. Two bottom block connecting screw holes are arranged on the upper surface of the bottom block 208. The pressing block 209 is installed on the bottom block 208 through the two countersunk screws, and the water-absorbing filter paper 212 is tightly pressed therebetween. The pressing block 209 and the bottom block 208 can also be installed through a rotating clamping manner.

[0057] Further, the elastic locking mechanism 210 adopts elastic impact beads, the elastic impact beads are inserted into the locking piece mounting hole 206, the inner side surfaces of the two side arms of the array holder 3 are respectively provided with arc-shaped grooves corresponding to the locking piece mounting hole 206, when the array holder 3 is installed, the steel beads of the elastic impact beads are matched with the arc-shaped grooves of the two side arms of the array holder 3 under the action of the spring, so that the array holder 3 is fixed on the bottom block 208.

[0058] Specific embodiment six: in combination with Figures 1 to 9 In this embodiment, the dehydration mechanism further includes a plurality of silica gel ring flat pads, the docking channel 102 is a cylindrical hole structure penetrating through the upper and lower surfaces of the middle seat plate 110, the docking channel 102 is provided with an elastic flat pad mounting groove coaxially arranged at the top end, the silica gel ring flat pad is embedded in the elastic flat pad mounting groove, the depth of the elastic flat pad mounting groove is less than the thickness of the silica gel ring flat pad, the dehydration seat mounting groove 101 is a cylindrical hole structure penetrating through the upper and lower surfaces of the upper seat plate 109, the inner wall of the dehydration seat mounting groove 101 is provided with a plurality of base clamping ports 108 uniformly distributed in the circumferential direction, the base clamping port 108 is a rectangular insertion slot penetrating through the upper and lower surfaces of the upper seat plate 109, the side surface of the rectangular insertion slot is provided with a smoothly transitioned clamping groove at the bottom, the side surface of the bottom block 208 is provided with a plurality of rectangular protrusions 207 uniformly distributed in the circumferential direction, the width of the rectangular protrusion 207 is consistent with the width of the rectangular insertion slot of the base clamping port 108, and the thickness of the rectangular protrusion is less than the depth of the clamping groove of the base clamping port 108.

[0059] In this way, the four rectangular protrusions 207 are arranged on the outer side of the bottom block 208, so that the dehydration seat 2 can be clamped and rotated on the base clamping port 108. The silica gel ring flat pad has a certain elasticity and can play a sealing and compression role on the base clamping port 108. The thickness of the rectangular protrusion 207 is slightly smaller than the depth of the clamping groove of the base clamping port 108, and under the action of the silica gel flat pad, the rectangular protrusion 207 can be tightly pressed with the base clamping port 108 without loosening. The other components and connection relationships are the same as those in the first, second, third, fourth or fifth specific embodiments.

[0060] In this embodiment, four base clamping ports 108 are arranged around each dehydration seat mounting groove 101, and four rectangular protrusions 207 matched with the base clamping port 108 are arranged on the outer side of each bottom block 208.

[0061] Specific embodiment seven: in combination with Figures 1 to 9In this embodiment, the dehydration mechanism further comprises two air inlet joints 4, the openings of the two lateral air inlets 105 are located on the lower surface of the middle seat plate 110, the ends of the two lateral air inlets 105 away from the air inlet hole section serve as the base air inlets 103, the upper half of the air inlet hole section of the air inlet 103 is a cylindrical hole structure penetrating through the upper and lower surfaces of the upper seat plate 109, the lower half of the air inlet hole section of the air inlet 103 is a cylindrical hole structure penetrating through the upper and lower surfaces of the middle seat plate 110, the two air inlet joints 4 are respectively installed in the upper half of the air inlet hole section of the two base air inlets 103, and the air inlet joints 4 are made of plastic.

[0062] In this way, the air inlet joints 4 are made of plastic, and the upper seat plate 109 is provided with two air inlet joints 4 made of plastic, and the constant-temperature dry gas is introduced into the channel structure between the middle seat plate 110 and the lower seat plate 111. The other components and connection relationships are the same as those in the first, second, third, fourth, fifth, or sixth embodiment.

[0063] In this embodiment, the lateral air inlets 105 are L-shaped channel structures, one end of the lateral air inlets 105 is provided with a first circular interface with a smooth transition, the first circular interface is in communication with the air inlet 103, and the other end of the lateral air inlets 105 is in communication with the air inlet hole section in the longitudinal channel 104.

[0064] Specific embodiment eight: in combination Figures 1 to 9 In this embodiment, the dehydration mechanism further comprises a calibration block 5, the calibration block 5 is a cylindrical structure, and the calibration block 5 is detachably installed in the dehydration seat installation groove 101 located at the upper left corner.

[0065] In this way, the calibration block 5 plays a visual calibration role and can cooperate with the mechanical arm to complete adaptive automatic operation. The other components and connection relationships are the same as those in the first, second, third, fourth, fifth, sixth, or seventh embodiment.

[0066] Specific embodiment nine: in combination Figures 1 to 9 In this embodiment, the central distribution channel 106 is a cylindrical hole structure penetrating through the upper and lower surfaces of the lower seat plate 111, the inner wall of the central distribution channel 106 is provided with a plurality of distribution channel outlets uniformly distributed in the circumferential direction, the lateral air outlets 107 are channel structures, the openings of the lateral air outlets 107 are located on the upper surface of the lower seat plate 111, a plurality of lateral air outlets 107 have the same cross section and are arranged in a diverging manner, one end of each of the plurality of lateral air outlets 107 is in communication with the plurality of distribution channel outlets of the central distribution channel 106, the other end of each of the plurality of lateral air outlets 107 is provided with a second circular interface with a smooth transition, and the plurality of second circular interfaces are respectively in communication with the plurality of butt joint channels 102.

[0067] In this way, the mixed gas is evenly distributed into the six abutment channels 102 and the dehydration seat channels 201 of the bottom block 208 through the six lateral gas outlet channels 107 with the same cross section. The gas flow in each dehydration seat channel 201 is uniform. The other components and connection relationships are the same as those in the first, second, third, fourth, fifth, sixth, seventh or eighth embodiments.

[0068] Organoids, as three-dimensional cell models in vitro, have important value in disease research, drug screening and regenerative medicine. Traditional organoid culture relies on manual operation, and the cell clusters are required not to adhere to the wall during 3D organoid culture. The commonly used matrix glue droplet method in array culture of organoids is to add treated source cells into a multi-well plate (such as a 96-well plate or a 384-well plate) with a U-shaped bottom. After the cells form clusters, a pipette gun is used to transfer them out of the well, mix them with matrix glue, and then drop them into the well plate for solidification and culture. This culture method is to culture multiple organoids in the same glue droplet, and the position of the organoids in the glue droplet greatly affects the consistency of the development of the organoids. Currently, there is also a method of using a pipette gun to suck the cultured cell clusters and seed them in the matrix glue for solidification. Although this method can ensure the approximate uniform distribution of organoids in the horizontal plane, the position of the organoids in the vertical direction is also difficult to unify, and the exchange rate of nutrients during the growth of the organoids also has a large difference. Moreover, this method is highly dependent on the operation level of the biological experimenters, and the culture efficiency is very low. The currently available matrix glue embedding method for culturing organoids generally has the problems of low throughput and poor consistency of organoid growth. There is no public method that can balance the high throughput and consistency of matrix glue embedding organoid culture.

[0069] Specific embodiment ten: combination Figures 1 to 9 To illustrate the present embodiment, the present embodiment is a gel removal method based on a matrix gel wrapping organoid dehydration mechanism, which comprises the following steps:

[0070] Assembling the array holder 3 and the dehydration seat 2: using a mechanical arm 6 to pick up the array holder 3 with the agarose microarray block from the culture dish 7, and installing the array holder 3 in the array block installation groove 205 of the dehydration seat 2, while the elastic locking mechanism 210 arranged on both sides of the dehydration seat 2 automatically locks and fixes the array holder 3;

[0071] Absorbing water in the agarose microarray block: using dry water-absorbing filter paper 212 to quickly absorb the water in the microwells of the agarose microarray block installed on the array holder 3;

[0072] Evaporative water absorption filter paper 212 moisture: in advance, two sides of the air inlet gas path joint 4 and the constant temperature gas source is connected, open the constant temperature gas source, two constant temperature gas respectively from two air inlet gas path joint 4 into two horizontal air inlet channel 105, then in the vertical channel 104, after the collection of constant temperature gas flow in the vertical channel 104 under the action of form constant temperature cyclone gas, constant temperature cyclone gas under the action of the center distribution channel 106 after the average distribution flow into multiple horizontal air outlet channel 107, multiple constant temperature gas flow again through the docking channel 102 flow into the dehydration seat 2 of the dehydration seat channel 201 will absorb water filter paper 212 on the water, so that the agarose microarray block installed on the array holder 3 on the agarose microarray block sustained dehydration;

[0073] Add Matrigel to agarose microarray block: use electric dispenser, add Matrigel in the array area of agarose microarray block, so that Matrigel covers the sinking array area;

[0074] Separate Matrigel surface from agarose microarray block: use mechanical arm 6 to pick up new culture dish 7, and then use mechanical arm to pick up constant temperature heat conducting block and paste it on the outer bottom of culture dish 7, so as to keep the constant temperature of constant temperature heat conducting block, assist Matrigel curing, and make Matrigel cure and adsorb on the bottom of culture dish 7. Due to the continuous dehydration effect, the agarose microarray block will shrink slightly, and the array hole on the agarose microarray block will slightly enlarge under the shrinking effect, so that the Matrigel surface and the agarose microarray block are separated.

[0075] Culture organoids: use mechanical arm 6 to take down the culture dish 7, and then add culture medium in the culture dish 7 for organoid culture.

[0076] In this way, the agarose microarray block is provided with a plurality of array holes arranged uniformly, and each array hole of the agarose microarray block separately contains a Matrigel wrapping cell ball. Compared with the method of culturing multiple organoids in the same gel droplet adopted by the existing array type culture organoids, all the organoids in the gel droplet are consistent in position, so that the development consistency of the organoids can be ensured. In addition, during the dehydration process of the Matrigel wrapping organoids, due to the same cross section of the horizontal air outlet channel 107 in the base 1 for connecting the dehydration channel, the constant temperature cyclone gas flowing out of the center distribution channel 106 uniformly flows into the six docking channels 102 and the dehydration seat channels 201 of the bottom block 208 through the six horizontal air outlet channels 107 with the same cross section, so that the gas flow in each dehydration seat channel 201 is uniform, and the high throughput and consistency of the Matrigel embedding organoid culture can be considered. The other components and connection relationships are the same as those of the first, second, third, fourth, fifth, sixth, seventh, eighth or ninth embodiment.

[0077] In this embodiment, the constant temperature heat conducting block can maintain a constant temperature of 37℃. In addition, the surface of the separation matrix glue and the agarose microarray block can also be cooled at 37℃. The constant temperature air flow in the constant temperature air source is introduced by the air duct, so that the constant temperature air flow at 37℃ continuously blows the bottom of the culture dish, and under the action of the constant temperature air flow, the heat is conducted to the matrix glue to solidify it.

[0078] Working principle

[0079] Combination ​ The working principle of the matrix glue wrapped organoid dehydration mechanism is described as follows: after culturing the cell clusters on the agarose microarray block, the agarose microarray block is transferred to the matrix glue wrapped organoid dehydration mechanism by manual operation or mechanical arm 6. The array holder 3 and the dehydration mechanism are matched in size, which can be easily clamped on the array block mounting groove 205 of the dehydration seat 2, and fixed by elastic beads. There is a layer of water absorption filter paper 212 at the contact between the dehydration mechanism and the agarose microarray block, so that the water in the agarose array area quickly penetrates from the loose and porous structure of the agarose to the water absorption filter paper 212. The bottom of the filter paper is provided with a constant temperature drying gas at 37℃, which carries away the water in the water absorption filter paper 212 through the air flow. After the dehydration of the agarose microarray block is completed, a certain amount of matrix glue is added in the agarose array area, and a new culture dish 7 is inverted to cover the agarose microarray block. The other side of the culture dish 7 is continuously blown by 37℃ sterile gas, so that the matrix glue is heated and solidified. The solidified matrix glue is separated from the agarose array and adheres to the culture dish 7, and the cell clusters are solidified in the matrix glue microcolumn. The culture medium is added in the culture dish 7 for subsequent organoid culture.

[0080] The above examples are only used to illustrate the technical solutions of the present application, but not to limit them; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that they can still modify the technical solutions recorded in the foregoing examples, or make equivalent replacement for part of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A matrix gel-encapsulated organoid dehydration mechanism, characterized in that, The dehydration mechanism includes: One or more dehydration seats (2), the dehydration seats (2) are provided with dehydration channels and dehumidification channels; The base (1) has multiple dehydration seat mounting slots (101) at the top. The dehydration seat mounting slots (101) are used to insert the dehydration seat (2). The bottom of the dehydration seat mounting slots (101) is provided with a docking channel (102) that communicates with the dehydration channel and the dehumidification channel. The center of the base (1) has a longitudinal channel (104). The top two sides of the longitudinal channel (104) are provided with two air inlet sections symmetrically arranged tangentially. The base (1) also has two transverse air inlet channels (105) that communicate with the air inlet ends of the two air inlet sections respectively. The bottom of the longitudinal channel (104) has a central distribution channel (106). The outer side of the central distribution channel (106) has multiple transverse air outlet channels (107). The central distribution channel (106) is connected to multiple docking channels (102) through multiple transverse air outlet channels (107). The base (1) includes an upper base plate (109), a middle base plate (110) and a lower base plate (111) that are horizontally stacked from top to bottom. The longitudinal channel (104) is a cylindrical hole structure that runs through the upper and lower surfaces of the intermediate seat plate (110). The bottom of the longitudinal channel (104) is provided with a cylindrical air outlet expansion hole structure. The center lines of the two air inlet sections are located on the same horizontal plane. The center lines of the two air inlet sections are arranged in parallel and intersecting and are tangent to the longitudinal channel (104). The dehydration mechanism also includes two air inlet connectors (4). The opening of the transverse air inlet channel (105) is located on the lower surface of the middle seat plate (110). The two transverse air inlet channels (105) are located at the end away from the air inlet section as the base air inlet (103). The upper half of the air inlet (103) is a cylindrical hole structure that penetrates the upper and lower surfaces of the upper seat plate (109). The lower half of the air inlet (103) is a cylindrical hole structure that penetrates the upper and lower surfaces of the middle seat plate (110). The two air inlet connectors (4) are respectively installed in the upper half of the two base air inlets (103). The central distribution channel (106) is a cylindrical hole structure that runs through the upper and lower surfaces of the lower seat plate (111). The inner wall of the central distribution channel (106) is provided with multiple distribution channel outlets evenly distributed in the circumference. The transverse air outlet channel (107) is a groove structure. The opening of the transverse air outlet channel (107) is located on the upper surface of the lower seat plate (111). The multiple transverse air outlet channels (107) have the same cross-section. One end of the multiple transverse air outlet channels (107) is connected to the multiple distribution channel outlets of the central distribution channel (106). The other end of the multiple transverse air outlet channels (107) is provided with multiple smoothly transitioned second circular interfaces. The multiple second circular interfaces are connected to multiple docking channels (102).

2. The matrix gel-encapsulated organoid dehydration mechanism according to claim 1, characterized in that, The dehydration seat (2) is used to support the array holder (3), and the dehydration channel and the dehumidification channel of the dehydration seat (2) are provided with absorbent filter paper (212).

3. The matrix gel-encapsulated organoid dehydration mechanism according to claim 2, characterized in that, The dehydration seat (2) includes a base block (208), a pressure block (209), two elastic locking mechanisms (210), and two pressure block connectors (211). The bottom of the base block (208) is a cylindrical structure. The bottom of the base block (208) is detachably installed in the dehydration seat mounting groove (101) of the base (1). The top center of the base block (208) has an integrally formed cuboid structure. The center of the base block (208) has a dehydration seat channel (201) that runs vertically through the upper and lower surfaces of the base block (208). The dehydration seat channel (201) has air outlets (202) on both sides. The bottom of the pressure block (209) has a pressure block mounting groove (203) that matches the bottom block (208). The pressure block (209) covers the cuboid structure on the top of the base block (208). The pressure block (209) is connected to the bottom block (208) through two pressure block connectors (211). There is an air vent gap (204) between the outer wall of the cuboid structure and the inner wall of the pressure block mounting groove (203). The top of the pressure block (209) is provided with an array block mounting groove (205) that is connected to the pressure block mounting groove (203). The two ends of the pressure block (209) are machined into vertical surfaces that match the array retainer (3). The middle of the vertical surfaces on both sides of the pressure block (209) is provided with locking component mounting holes (206) that are vertically connected to the pressure block mounting groove (203). Two elastic locking mechanisms (210) are respectively installed in the two locking component mounting holes (206). The array retainer (3) is detachably fixed to the pressure block (209) through the two elastic locking mechanisms (210).

4. The matrix gel-encapsulated organoid dehydration mechanism according to claim 3, characterized in that, The dehydration mechanism also includes multiple silicone ring flat pads. The docking channel (102) is a cylindrical hole structure that penetrates the upper and lower surfaces of the intermediate seat plate (110). The top of the docking channel (102) is provided with a coaxially arranged elastic flat pad mounting groove. The silicone ring flat pads are embedded in the elastic flat pad mounting grooves. The depth of the elastic flat pad mounting grooves is less than the thickness of the silicone ring flat pads. The dehydration seat mounting groove (101) is a cylindrical hole structure that penetrates the upper and lower surfaces of the upper seat plate (109). The inner wall is provided with multiple base slots (108) evenly distributed around the circumference. The base slots (108) are rectangular slots that penetrate the upper and lower surfaces of the upper base plate (109). The bottom side of the rectangular slot is provided with a smoothly transitioned snap-fit ​​groove. The bottom side of the base block (208) is provided with multiple rectangular protrusions (207) evenly distributed around the circumference. The width of the rectangular protrusions (207) is the same as the width of the rectangular slot of the base slot (108), and the thickness of the rectangular protrusions is less than the depth of the snap-fit ​​groove of the base slot (108).

5. The matrix gel-encapsulated organoid dehydration mechanism according to claim 4, characterized in that, The dehydration mechanism also includes a calibration block (5), which is a cylindrical structure and is detachably installed in the dehydration seat mounting groove (101) located in the upper left corner.

6. The degumming method based on the matrix adhesive-encapsulated organoid dehydration mechanism according to claim 5, characterized in that, include: Assemble the array holder (3) and the dehydration seat (2): Use a robotic arm (6) to grip the array holder (3) with agarose microarray blocks (8) and install the array holder (3) in the array block mounting slot (205) of the dehydration seat (2). At the same time, the elastic locking mechanism (210) set on both sides of the dehydration seat (2) automatically locks and fixes the array holder (3). The water in the micropores of the agarose microarray block (8) is quickly absorbed by the absorbent filter paper (212); Evaporation of moisture from absorbent filter paper (212): First, connect the two air inlet connectors (4) to the constant temperature gas source. Turn on the constant temperature gas source. The two constant temperature gases enter the two transverse air inlet channels (105) from the two air inlet connectors (4) respectively. Then they converge in the longitudinal channel (104). The converged constant temperature airflow forms a constant temperature swirling gas under the action of the longitudinal channel (104). The constant temperature swirling gas is evenly distributed under the action of the central distribution channel (106) and flows into multiple transverse air outlet channels (107). The multiple constant temperature airflows then flow into the dehydration seat channels (201) of multiple dehydration seats (2) through the docking channel (102) to carry away the moisture on the absorbent filter paper (212) and make the agarose microarray block (8) continuously dehydrated. Using an electric dispenser, add matrix gel to the array area of ​​the agarose microarray block (8) so that the matrix gel covers the sinking array area. Separating the surface of the matrix gel from the agarose microarray block (8): Use the robotic arm (6) to pick up the new culture dish (7) and invert it onto the array holder (3). Then use the robotic arm to pick up the constant temperature heat conduction block and attach it to the bottom of the culture dish (7) so that the matrix gel can be solidified and adsorbed on the bottom of the culture dish (7). The continuous dehydration action separates the surface of the matrix gel from the agarose microarray block (8). Organoid culture: The robotic arm (6) removes the culture dish (7), and the matrix gel encapsulates the cell spheres and solidifies them at the bottom of the culture dish (7). Culture medium is added to the culture dish (7) for organoid culture.

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