CN219951074U - Organoid spheroid preparation facilities - Google Patents
Organoid spheroid preparation facilities Download PDFInfo
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- CN219951074U CN219951074U CN202321538568.XU CN202321538568U CN219951074U CN 219951074 U CN219951074 U CN 219951074U CN 202321538568 U CN202321538568 U CN 202321538568U CN 219951074 U CN219951074 U CN 219951074U
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Abstract
The utility model discloses an organoid sphere preparation device, which comprises a platform, a microfluidic chip, a switching mechanism, a first pushing/absorbing mechanism and a second pushing/absorbing mechanism, wherein the platform is connected with the microfluidic chip; the microfluidic chip comprises a chip body arranged at the top end of the platform, and a liquid inlet channel, a liquid storage channel, a liquid outlet channel, a ball outlet channel and a continuous phase liquid inlet channel which are arranged in the chip body, wherein the first end of the liquid inlet channel is connected with the first pushing/absorbing mechanism, the first end of the liquid storage channel is connected with the first pushing/absorbing mechanism, the first end of the ball outlet channel is respectively communicated with the first end of the liquid outlet channel and the first end of the continuous phase liquid inlet channel, and the second end of the ball outlet channel extends to the top end of the chip body and is used for being connected with a ball outlet connecting pipe. The utility model can realize high-flux preparation of organoid spheres, has high degree of automation, and can realize large-scale production and commercial use.
Description
Technical Field
The utility model relates to the field of medical equipment, in particular to a device for preparing organoid spheres.
Background
The cancer threatens the health of all human beings, the heterogeneity of the cancer determines that the same medicament has different response rates aiming at different individuals, the traditional 2D culture method has lower modeling success rate, and the factors such as tumor microenvironment which determines medicament sensitivity are greatly limited in clinical application, and the establishment of a xenograft model (PDX) aiming at a patient can reconstruct the tumor microenvironment and be used for testing the effectiveness of different medicaments, but the high cost and the long period limit the wide clinical application. Tumor organoids are prepared by extracting cancer cells and their stromal cells that make up their microenvironment from a patient's tumor tissue and performing 3D culture in vitro, and have been demonstrated to be highly capable of reconstructing the tumor microenvironment and maintaining the phenotype of the patient's cancer cells, and can be used to predict drug sensitivity for the patient.
The preparation of tumor organoids is mainly divided into the preparation of organoid spheres, the maturation of organoid spheres and the cultivation of organoid spheres. The existing organoid sphere is usually prepared by manually adding a cell mixed solution into a liquid-transferring gun, then forming a matrix spherical drop of the mixed cells by utilizing a liquid-transferring gun head, and controlling the size of the drop by controlling factors such as injection speed, contact angle, caliber of the liquid-transferring gun head and the like so as to form uniform liquid drops as the organoid sphere. The preparation method relies on manual operation, has low automation degree, is difficult to realize high-flux preparation of organoid spheres, and is not suitable for large-scale production and commercial use.
Disclosure of Invention
In order to overcome the defects of the prior art, the utility model provides the organoid sphere preparation device which can realize high-throughput preparation of organoid spheres, has high automation degree and can realize large-scale production and commercial use.
The technical scheme adopted for solving the technical problems is as follows:
the utility model provides a organoid sphere preparation device, which comprises a platform, a microfluidic chip, a switching mechanism, a first pushing/absorbing mechanism and a second pushing/absorbing mechanism, wherein the platform is connected with the microfluidic chip; the microfluidic chip comprises a chip body arranged at the top end of the platform, and a liquid inlet flow channel, a liquid storage flow channel, a liquid outlet flow channel, a ball outlet flow channel and a continuous phase liquid inlet flow channel which are arranged in the chip body, wherein the first end of the liquid inlet flow channel is connected with the first pushing/absorbing mechanism, the first pushing/absorbing mechanism is used for being connected with a sample injection pipe, the first end of the liquid storage flow channel is connected with the first pushing/absorbing mechanism, the first end of the ball outlet flow channel is respectively communicated with the first end of the liquid outlet flow channel and the first end of the continuous phase liquid inlet flow channel, the second end of the ball outlet flow channel extends to the top end of the chip body and is used for being connected with a ball outlet connecting pipe, and the second end of the continuous phase liquid inlet flow channel is connected with the second pushing/absorbing mechanism which is used for being connected with a liquid storage container; the switching mechanism is arranged at the top end of the chip body and is provided with a connecting channel, the switching mechanism can rotate between a first position and a second position relative to the chip body, when the switching mechanism is positioned at the first position, the first end of the connecting channel is communicated with the second end of the liquid storage flow channel, the second end of the connecting channel is communicated with the second end of the liquid inlet flow channel, and when the switching mechanism is positioned at the second position, the first end of the connecting channel is communicated with the second end of the liquid storage flow channel, and the second end of the connecting channel is communicated with the second end of the liquid outlet flow channel.
As the preferable technical scheme, be equipped with disperse phase feed liquor passageway, first stock solution passageway and continuous phase feed liquor passageway in the chip body, disperse phase feed liquor passageway's first end with the first end intercommunication of feed liquor runner, disperse phase feed liquor passageway's second end extends to the top of chip body, first stock solution passageway's first end with the first end intercommunication of stock solution runner, first stock solution passageway's second end extends to the top of chip body, continuous phase feed liquor passageway's first end with the second end intercommunication of continuous phase feed liquor runner, continuous phase feed liquor passageway's second end extends to the top of chip body.
As the preferable technical scheme, the top of chip body is equipped with disperse phase feed liquor joint, stock solution joint, continuous phase feed liquor joint, play ball joint, disperse phase feed liquor joint with disperse phase feed liquor passageway's second end intercommunication, disperse phase feed liquor joint through first connecting pipe with first push/imbibition mechanism is connected, first push/imbibition mechanism is used for being connected with the sample injection pipe through the second connecting pipe, the stock solution joint with first stock solution passageway's second end intercommunication, the stock solution joint through the third connecting pipe with first push/imbibition mechanism is connected, continuous phase feed liquor joint with continuous phase feed liquor passageway's second end intercommunication, continuous phase feed liquor joint through the fourth connecting pipe with second push/imbibition mechanism is connected, second push/imbibition mechanism is used for being connected with the stock solution container through the fifth connecting pipe, play ball joint with go out ball runner's second end intercommunication, play ball joint is used for with play ball connection.
As the preferred technical scheme, be equipped with disperse phase drain passageway, second stock solution passageway and play liquid connecting channel in the chip body, disperse phase drain passageway's first end with the second end intercommunication of feed liquor runner, disperse phase drain passageway's second end extends to the top of chip body, second stock solution passageway's first end with the second end intercommunication of stock solution runner, the second end of second stock solution passageway extends to the top of chip body, go out liquid connecting channel's first end with go out liquid runner's second end intercommunication, go out liquid connecting channel's second end extends to the top of chip body.
As the preferred technical scheme, the switching mechanism includes knob and switch piece, the top of chip body is formed with the ring portion, disperse looks drain channel's second end, second stock solution passageway's second end, go out the second end and all be located the inboard of ring portion, the bottom of knob holding in the ring portion and can be relative the chip body rotates between first position and second position, the bottom of knob is equipped with the installation cavity, the switch piece sets up in the installation cavity, the bottom of switch piece with the top in close contact with of chip body has the connecting channel, the rotation of knob can drive the switch piece is relative the chip body rotates between first position and second position, when knob and switch piece are located the first position, the first end of connecting channel with the second end intercommunication of second stock solution passageway, and the second end of connecting channel with disperse looks drain channel, when the knob and switch piece are located the second position, the second end of connecting channel with the second end intercommunication of second stock solution passageway and second end intercommunication.
As the preferable technical scheme, the rotary table further comprises a driving mechanism arranged at the top end of the table, wherein the top end of the knob is connected with the driving mechanism, and the driving mechanism is used for driving the knob to rotate between a first position and a second position.
As a preferable technical scheme, the liquid storage flow channel is a serpentine flow channel.
As a preferable technical scheme, the size of the ball outlet flow channel is larger than that of the continuous phase liquid inlet flow channel.
As a preferable technical scheme, the first pushing/sucking mechanism and the second pushing/sucking mechanism are injection pumps.
The beneficial effects of the utility model are as follows: according to the utility model, the liquid inlet flow channel, the liquid storage flow channel, the liquid outlet flow channel, the ball outlet flow channel and the continuous phase liquid inlet flow channel which are arranged in the chip body, and the first pushing/absorbing mechanism, the second pushing/absorbing mechanism and the switching mechanism which are arranged, so that the high-throughput preparation of the organoid sphere can be realized, the automation degree is high, the preparation efficiency is improved, and the large-scale production and the commercial use can be realized.
Drawings
The utility model will be further described with reference to the drawings and examples.
FIG. 1 is a schematic diagram of an apparatus for preparing organoid spheres according to an embodiment of the present utility model;
FIG. 2 is a schematic illustration of the organoid sphere preparation apparatus of FIG. 1 with the refrigeration mechanism removed;
FIG. 3 is a schematic top view of the microfluidic chip and switching mechanism of the organoid sphere preparation device of FIG. 1;
FIG. 4 is a schematic cross-sectional view of a microfluidic chip of the organoid sphere preparation device shown in FIG. 3;
FIG. 5 is an exploded schematic view of a first angle of the microfluidic chip and switching mechanism of the organoid sphere preparation device of FIG. 3;
FIG. 6 is an exploded schematic view of a second angle of the microfluidic chip and switching mechanism of the organoid sphere preparation device of FIG. 3;
fig. 7 is a flow chart of a method for preparing organoid spheres based on the organoid sphere preparation apparatus shown in fig. 1.
Detailed Description
The conception, specific structure, and technical effects produced by the present utility model will be clearly and completely described below with reference to the embodiments and the drawings to fully understand the objects, features, and effects of the present utility model. It is apparent that the described embodiments are only some embodiments of the present utility model, but not all embodiments, and that other embodiments obtained by those skilled in the art without inventive effort are within the scope of the present utility model based on the embodiments of the present utility model. In addition, all the coupling/connection relationships referred to in the patent are not direct connection of the single-finger members, but rather, it means that a better coupling structure can be formed by adding or subtracting coupling aids depending on the specific implementation. The technical features in the utility model can be interactively combined on the premise of no contradiction and conflict.
Referring to fig. 1 and 2, an apparatus for preparing organoid spheres according to an embodiment of the present utility model includes a platform 10, a microfluidic chip 20, a switching mechanism 30, a first pushing/sucking mechanism 60, a second pushing/sucking mechanism 50, a driving mechanism 70 and a refrigerating mechanism 80.
Platform 10 is disposed at the top end of refrigeration mechanism 80. The refrigeration mechanism 80 is of a conventional construction. The refrigeration mechanism 80 is mainly composed of a custom thermoelectric refrigeration (peltier semiconductor refrigeration) module, a radiator, and a temperature sensor. The microfluidic chip 20 and the driving mechanism 70 are both arranged at the top end of the platform 10. The switching mechanism 30 is disposed at the top end of the microfluidic chip 20. The first push/suction mechanism 60 and the second push/suction mechanism 50 are provided on one side of the platform 10.
As shown in fig. 3 to 6, the microfluidic chip 20 includes a chip body 21 disposed at the top end of the platform 10, and a liquid inlet channel 22, a liquid storage channel 23, a liquid outlet channel 24, a ball outlet channel 262, a disperse phase liquid inlet channel 263, a disperse phase liquid outlet channel 266, a first liquid storage channel 264, a second liquid storage channel 267, a liquid outlet connection channel 268, a continuous phase liquid inlet channel 25, and a continuous phase liquid inlet channel 265 disposed in the chip body 21.
The chip body 21 has a plate-like structure, and is made of, for example, PC (polycarbonate) material. In this embodiment, the top end of the platform 10 is provided with a mounting groove, and the chip body 21 is disposed in the mounting groove.
In this embodiment, two microfluidic chips 20 are arranged at intervals along the length direction of the platform 10, and the number of the switching mechanisms 30 and the number of the mounting grooves of the platform 10 correspond to the number of the microfluidic chips 20, and are also two. It is understood that the number of microfluidic chips 20 may be set according to practical situations.
The liquid inlet channel 22, the liquid storage channel 23, the liquid outlet channel 24, the ball outlet channel 262 and the continuous phase liquid inlet channel 25 respectively have a first end and a second end. The disperse phase liquid inlet channel 263, the disperse phase liquid outlet channel 266, the first liquid storage channel 264, the second liquid storage channel 267, the liquid outlet connecting channel 268 and the continuous phase liquid inlet channel 265 respectively have a first end and a second end.
The first end and the second end of the dispersed phase liquid inlet channel 263, the dispersed phase liquid outlet channel 266, the first liquid storage channel 264, the second liquid storage channel 267, the liquid outlet connecting channel 268, the continuous phase liquid inlet channel 265, and the ball outlet channel 262 are respectively closed end and open end.
The first end of the dispersed phase liquid inlet channel 263 communicates with the first end of the liquid inlet channel 22, and the second end of the dispersed phase liquid inlet channel 263 extends to the top end of the chip body 21. The first end of the dispersed phase liquid outlet passage 266 communicates with the second end of the liquid inlet passage 22, and the second end of the dispersed phase liquid outlet passage 266 extends to the top end of the chip body 21. The first end of the first liquid storage channel 264 is communicated with the first end of the liquid storage channel 23, and the second end of the first liquid storage channel 264 extends to the top end of the chip body 21. The first end of the second liquid storage channel 267 is communicated with the second end of the liquid storage channel 23, and the second end of the second liquid storage channel 267 extends to the top end of the chip body 21. The first end of the liquid outlet connecting channel 268 communicates with the second end of the liquid outlet channel 24, and the second end of the liquid outlet connecting channel 268 extends to the top end of the chip body 21. The first end of the ball outlet flow channel 262 is respectively communicated with the first end of the continuous phase liquid inlet flow channel 25 and the first end of the liquid outlet flow channel 24, and the second end of the ball outlet flow channel 262 extends to the top end of the chip body 21. The first end of the continuous phase liquid inlet channel 265 communicates with the second end of the continuous phase liquid inlet channel 25, and the second end of the continuous phase liquid inlet channel 265 extends to the top end of the chip body 21.
In this embodiment, the liquid inlet channel 22, the liquid storage channel 23, the liquid outlet channel 24 and the continuous phase liquid inlet channel 25 are horizontally arranged. The disperse phase liquid inlet passage 263 and the disperse phase liquid outlet passage 266 are arranged perpendicular to the liquid inlet passage 22. The first liquid storage channel 264 and the second liquid storage channel 267 are arranged perpendicular to the liquid storage channel 23. The liquid outlet connecting channel 268 is perpendicular to the liquid outlet channel 24. The ball outlet flow passage 262 is perpendicular to the liquid outlet flow passage 24 and the continuous phase liquid inlet flow passage 25, and a T-shaped structure is formed between the ball outlet flow passage 262 and the liquid outlet flow passage 24 and between the ball outlet flow passage 262 and the continuous phase liquid inlet flow passage 25. The continuous phase liquid inlet passage 265 is disposed perpendicular to the continuous phase liquid inlet passage 25.
ChipThe top end of the body 21 is provided with a disperse phase liquid inlet connector 212, a liquid storage connector 213, a continuous phase liquid inlet connector 215 and a ball outlet connector 214. The dispersed phase feed connector 212 communicates with a second end of the dispersed phase feed passage 263. The dispersed phase liquid inlet connector 212 is connected to the first push/liquid suction mechanism 60 through a first connecting pipe 92. The first push/pull mechanism 60 is configured to be coupled to the sample tube via a second coupling tube. The sample inlet tube is used for containing a disperse phase which is wrapped with cells, namely a cell mixed solution. The cells are primary tumor tissue cells extracted from primary tumor tissue, and it is understood that the cells may be stem cells, cell lines, etc. and the type and number of the cells may be set according to practical situations. The density of cells packed in the dispersed phase was 1X 10 7 It will be appreciated that the density of cells may be set according to the circumstances. The dispersion phase is preferably matrigel, and it is understood that the dispersion phase may be hydrogel, collagen, hyaluronic acid, alginic acid, and the like, and may be set according to practical situations.
The reservoir connection 213 communicates with a second end of the first reservoir channel 264. The liquid storage joint 213 is connected to the first push/suction mechanism 60 through a third connection pipe. The first pushing/sucking mechanism 60 is used for sucking the dispersed phase in the sample tube through the second connecting tube and pushing the sucked dispersed phase into the liquid storage flow channel 23 through the first connecting tube 92, the dispersed phase liquid inlet connector 212, the dispersed phase liquid inlet channel 263, the liquid inlet channel 22, the dispersed phase liquid outlet channel 266, the connecting channel 331 of the switching mechanism 30 and the second liquid storage channel 267, so as to store the dispersed phase through the liquid storage flow channel 23. Continuous phase feed connector 215 communicates with a second end of continuous phase feed passage 265. The continuous phase liquid inlet connector 215 is connected to the second push/liquid suction mechanism 50 through a fourth connecting pipe 93. The second push/pull mechanism 50 is adapted to be connected to the first port 41 of the reservoir 40 via a fifth connecting tube 94. The second pushing/sucking mechanism 50 is used for sucking the continuous phase in the liquid storage container 40 through the fifth connecting pipe 94 and pushing the sucked continuous phase into the continuous phase liquid inlet channel 25 through the fourth connecting pipe 93, the continuous phase liquid inlet joint 215, and the second end of the continuous phase liquid inlet channel 265. The reservoir 40 may be disposed between the first push/wick 60 and the second push/wick 50. The reservoir 40 is adapted to hold a continuous phase, preferably an electronic fluorinated liquid, which is preferably capable of shearing the cell-encapsulated dispersed phase into organoid spheres of uniform size. It will be appreciated that the continuous phase may be a material which is compatible with cells and immiscible with water, such as vegetable oil, and may be provided according to the circumstances.
The ball outlet fitting 214 communicates with a second end of the ball outlet flow passage 262. The ball outlet joint 214 is adapted to connect with the ball outlet connection tube 95. One end of the ball outlet connecting pipe 95, which is far away from the ball outlet joint 214, is wound into a plurality of circles and then is accommodated in the accommodating cavity of the heating and curing device.
The first push/pull mechanism 60 and the second push/pull mechanism 50 are both syringe pumps, which are conventional.
In this embodiment, the angle between the second end of the dispersed phase liquid outlet channel 266 and the second end of the second liquid storage channel 267 and the second end of the liquid outlet connecting channel 268 and the second end of the second liquid storage channel 267 is 90 degrees, and it is understood that the angle of the included angle may be other.
The switching mechanism 30 is provided at the top end of the chip body 21.
Specifically, the switching mechanism 30 includes a knob 32 and a switching piece 33. The tip of the chip body 21 is formed with an upwardly extending ring portion 2111. The second end of the dispersed phase liquid outlet passage 266, the second end of the second liquid storage passage 267, and the second end of the liquid outlet connection passage 268 are all located inside the ring portion 2111. The bottom end of knob 32 is fitted with ring portion 2111. The bottom end of the knob 32 is accommodated in the ring portion 2111 and the bottom end of the knob 32 is in close contact with the top end of the chip body 21. The knob 32 is rotatable relative to the chip body 21 between a first position and a second position. The bottom of knob 32 is equipped with installation cavity 322, and switch piece 33 sets up in installation cavity 322, and the bottom of switch piece 33 is parallel and level with the bottom of knob 32. The bottom end of the switching piece 33 is in close contact with the top end of the chip body 21 and has a connection passage 331. The connection passage 331 has a first end 331a and a second end 331b, the first end 331a of the connection passage 331 is located at a center position of the bottom end of the switch piece 33, and the second end 331b of the connection passage 331 is located at one side of the center of the bottom end of the switch piece 33. The first end 331a of the connection passage 331 communicates with the second end of the second reservoir passage 267. Rotation of the knob 32 may rotate the switch 33 between the first position and the second position relative to the chip body 21. Since the first end 331a of the connection channel 331 is located at the center of the bottom end of the switch plate 33, the first end 331a of the connection channel 331 can be always kept in communication with the second end of the second liquid storage channel 267 during rotation of the switch plate 33, that is, when the knob 32 and the switch plate 33 are located at the first position, the first end 331a of the connection channel 331 is in communication with the second end of the second liquid storage channel 267, so that the first end 331a of the connection channel 331 can be in communication with the second end of the liquid storage channel 23 through the second liquid storage channel 267, and the second end 331b of the connection channel 331 is in communication with the second end of the dispersion liquid channel 266, so that the second end of the connection channel 331 can be in communication with the second end of the liquid inlet channel 22 through the dispersion liquid channel 266, and when the knob 32 and the switch plate 33 are located at the second position, the first end 331a of the connection channel 331 can be in communication with the second end of the second liquid storage channel 267, so that the first end 331a of the connection channel 331 can be in communication with the second end of the liquid storage channel 23 through the second liquid storage channel 267, and the second end 331b of the connection channel 331 can be in communication with the second end of the liquid storage channel 268 through the second end of the second liquid storage channel 24.
The outer circumference of the ring portion 2111 is fixedly sleeved with a knob cover 2112, the knob 32 is accommodated in the knob cover 2112, the outer circumference of the knob 32 is sleeved with an elastic member 34, and the elastic member 34 is preferably a spring. One end of the elastic member 34 is connected to the knob 32, and the other end is connected to the top inside the knob cover 2112. The elastic piece 34 is in a compressed state, the elastic piece 34 is used for pressing the knob 32, tight contact between the knob 32, the switching piece 33 and the top end of the chip body 21 is guaranteed to realize sealing, and the length of the elastic piece 34 can be dynamically adjusted in the follow-up rotation operation, so that interference friction and extrusion damage among the knob 32, the switching piece 33 and the chip body 21 are prevented.
A drive mechanism 70 is provided at the top end of the platform 10. Specifically, two mounting posts 12 are arranged at the top end of the platform 10 at one side of the microfluidic chip 20, mounting plates 13 are sleeved on the peripheries of the two mounting posts 12, and the mounting plates 13 are positioned above the microfluidic chip 20 and the switching mechanism 30. The driving mechanism 70 is, for example, a rotary cylinder, a motor, or the like. The top end of the knob 32 has a groove 323, the top end of the knob cover 2112 has an opening corresponding to the groove 323, and the end of the output end of the driving mechanism 70 passes through the through hole of the mounting plate 13, the opening of the knob cover 2112, and is engaged with the groove 323 of the knob 32. The driving mechanism 70 is used for driving the knob 32 to rotate between a first position and a second position relative to the chip body 21, so as to drive the switching piece 33 to rotate between the first position and the second position relative to the chip body 21. The number of mounting posts 12 may be set according to the actual circumstances. Through the actuating mechanism that sets up for the rotation of knob 32 and switch piece 33 between first position and second position can realize automaticly, convenient to use.
With the above structure, in practical application, the knob 32 and the switch 33 are driven by the driving mechanism 70 to rotate to the first position, at this time, the first end 331a of the connection channel 331 is communicated with the second end of the liquid storage channel 23 through the second liquid storage channel 267, and the second end 331b of the connection channel 331 is communicated with the second end of the liquid inlet channel 22 through the dispersed phase liquid outlet channel 266. Then the first pushing/absorbing mechanism 60 is used for absorbing the disperse phase in the sample tube through the second connecting tube and pushing the absorbed disperse phase into the liquid storage flow channel 23 through the first connecting tube 92, the disperse phase liquid inlet joint 212, the disperse phase liquid inlet channel 263, the liquid inlet channel 22, the disperse phase liquid outlet channel 266, the connecting channel 331 and the second liquid storage channel 267 so as to store the disperse phase through the liquid storage flow channel 23, and at the moment, the cooling mechanism 80 is used for cooling the stored disperse phase in the platform 10 and the liquid storage flow channel 23 so as to ensure that the liquid temperature of the disperse phase is between 0 ℃ and 10 ℃, thereby preventing the disperse phase from solidifying and ensuring the low viscosity physical state of the disperse phase so as to further perform droplet forming. Then, the knob 32 and the switch 33 are driven to rotate to the second position by the driving mechanism 70, at this time, the first end 331a of the connection channel 331 is communicated with the second end of the liquid storage channel 23 through the second liquid storage channel 267, and the second end 331b of the connection channel 331 is communicated with the second end of the liquid outlet channel 24 through the liquid outlet connection channel 268. Then, the dispersed phase stored in the liquid storage channel 23 is pushed into the liquid outlet channel 24 by the first pushing/sucking mechanism 60 through the third connecting pipe via the second liquid storage channel 267, the connecting channel 331 and the liquid outlet connecting channel 268, and the dispersed phase in the liquid outlet channel 24 can enter the first end of the ball outlet channel 262. Meanwhile, the second pushing/sucking mechanism 50 sucks the continuous phase in the liquid storage container 40 through the fifth connecting pipe 94 and pushes the sucked continuous phase into the continuous phase liquid inlet channel 25 through the fourth connecting pipe 93, the continuous phase liquid inlet joint 215 and the second end of the continuous phase liquid inlet channel 265, when the continuous phase in the continuous phase liquid inlet channel 25 enters the first end of the ball outlet channel 262, the continuous phase can shear the dispersed phase entering the first end of the ball outlet channel 262 into organoid spheres with uniform size under the pressure of the first pushing/sucking mechanism 60 and the second pushing/sucking mechanism 50, and the sheared organoid spheres can enter the ball outlet channel 262, so that the organoid spheres are prepared. Then the organoid sphere can enter the ball outlet connecting pipe 95 through the second end of the ball outlet flow passage 262 and the ball outlet joint 214, and then the organoid sphere can be heated and cured through the heating curing device, and then the cured organoid sphere can be cultured.
The utility model can realize high-flux preparation of organoid spheres by arranging the liquid inlet flow channel 22, the liquid storage flow channel 23, the liquid outlet flow channel 24, the ball outlet flow channel 262, the continuous phase liquid inlet flow channel 25 and the first pushing/sucking mechanism 60, the second pushing/sucking mechanism 50 and the switching mechanism 30 in the chip body 21, has high automation degree and can realize large-scale production and commercial use. The liquid inlet channel 22, the liquid storage channel 23, the liquid outlet channel 24, the ball outlet channel 262 and the continuous phase liquid inlet channel 25 are arranged in the chip body 21, the size is small, the cost is low, the liquid inlet channel 22 is communicated with the liquid storage channel 23 or the liquid storage channel 23 is communicated with the liquid outlet channel 24 by utilizing the rotation of the switching mechanism 30, so that the disperse phase can be transferred into the liquid storage channel 23 for storage under the action of the first pushing/sucking mechanism 60, and then transferred into the liquid outlet channel 24 under the action of the first pushing/sucking mechanism 60, the disperse phase transfer is performed on the same carrier, the disperse phase loss is relatively low, the preparation efficiency is improved, meanwhile, the disperse phase is stored through the liquid storage channel 23, the volume of the liquid storage channel 23 is small, and the disperse phase waste can not be caused. The utility model is especially suitable for the condition of small volume of disperse phase, and can realize micro-preparation of organoid spheres.
In this embodiment, the liquid storage channel 23 is preferably a serpentine channel, so as to reduce the size of the chip body 21. The cross-sectional shapes of the liquid inlet channel 22, the liquid storage channel 23, the liquid outlet channel 24 and the continuous phase liquid inlet channel 25 are all square, and the side length is 1 mm, for example. The cross-sectional shapes of the disperse phase liquid inlet channel 263, the disperse phase liquid outlet channel 266, the first liquid storage channel 264, the second liquid storage channel 267, the liquid outlet connecting channel 268, the ball outlet flow channel 262 and the continuous phase liquid inlet channel 265 are all round. The inner diameter of the ball outlet flow channel 262 is, for example, 0.45 mm, and the size of the ball outlet flow channel 262 is larger than that of the continuous phase liquid inlet flow channel 25, so that the sheared organoid ball can smoothly enter the ball outlet flow channel 262, and the ball blocking phenomenon is avoided. The size of the continuous phase liquid inlet channel 25 is the same as the size of the liquid outlet channel 24.
Referring to fig. 7, the present utility model further provides a method for preparing organoid spheres based on the organoid sphere preparation device, comprising the following steps:
s2, the switching mechanism 30 is rotated to a first position, so that the first end 331a of the connecting channel 331 is communicated with the second end of the liquid storage channel 23, and the second end of the connecting channel 331 is communicated with the second end of the liquid inlet channel 22.
Specifically, the knob 32 and the switching piece 33 are driven to rotate to the first position by the driving mechanism 70, so that the first end 331a of the connection channel 331 is communicated with the second end of the liquid storage channel 23 through the second liquid storage channel 267, and the second end 331b of the connection channel 331 is communicated with the second end of the liquid inlet channel 22 through the dispersed phase liquid outlet channel 266.
S3, sucking the disperse phase in the sample tube through the first pushing/sucking mechanism 60, and pushing the sucked disperse phase into the liquid storage flow channel 23 through the first connecting pipe 92, the disperse phase liquid inlet connector 212, the disperse phase liquid inlet channel 263, the liquid inlet channel 22, the disperse phase liquid outlet channel 266, the connecting channel 331 and the second liquid storage channel 267 so as to store the disperse phase through the liquid storage flow channel 23. The cooling mechanism 80 can cool the dispersed phase stored in the platform 10 and the liquid storage flow channel 23.
And S4, rotating the switching mechanism 30 to a second position, so that the first end of the connecting channel 331 is communicated with the second end of the liquid storage flow channel 23, and the second end of the connecting channel 331 is communicated with the second end of the liquid outlet flow channel 24.
Specifically, the knob 32 and the switch 33 are driven by the driving mechanism 70 to rotate to the second position, so that the first end 331a of the connection channel 331 is communicated with the second end of the liquid storage channel 23 through the second liquid storage channel 267, and the second end 331b of the connection channel 331 is communicated with the second end of the liquid outlet channel 24 through the liquid outlet connection channel 268.
S5, the disperse phase stored in the liquid storage flow channel 23 is pushed into the liquid outlet flow channel 24 through the second liquid storage channel 267, the connecting channel 331 and the liquid outlet connecting channel 268 by the first pushing/sucking mechanism 60, and the disperse phase in the liquid outlet flow channel 24 can enter the first end of the ball outlet flow channel 262. Simultaneously, the second pushing/sucking mechanism 50 sucks the continuous phase in the liquid storage container 40 and pushes the sucked continuous phase into the continuous phase liquid inlet channel 25 through the fourth connecting pipe 93, the continuous phase liquid inlet connector 215 and the second end of the continuous phase liquid inlet channel 265, when the continuous phase in the continuous phase liquid inlet channel 25 enters the first end of the ball outlet channel 262, the continuous phase can shear the disperse phase entering the first end of the ball outlet channel 262 into organoid spheres with uniform size under the pressure of the first pushing/sucking mechanism 60 and the second pushing/sucking mechanism 50, and the sheared organoid spheres can enter the ball outlet channel 262, so that the preparation of the organoid spheres is completed.
Then the organoid sphere can enter the ball outlet connecting pipe 95 through the second end of the ball outlet flow passage 262 and the ball outlet joint 214, and then the organoid sphere can be heated and cured through the heating curing device, and then the cured organoid sphere can be cultured.
While the preferred embodiment of the present utility model has been described in detail, the present utility model is not limited to the embodiments, and those skilled in the art can make various equivalent modifications or substitutions without departing from the spirit of the present utility model, and the equivalent modifications or substitutions are included in the scope of the present utility model as defined in the appended claims.
Claims (9)
1. The organoid sphere preparation device is characterized by comprising a platform, a microfluidic chip, a switching mechanism, a first pushing/absorbing mechanism and a second pushing/absorbing mechanism;
the microfluidic chip comprises a chip body arranged at the top end of the platform, and a liquid inlet flow channel, a liquid storage flow channel, a liquid outlet flow channel, a ball outlet flow channel and a continuous phase liquid inlet flow channel which are arranged in the chip body, wherein the first end of the liquid inlet flow channel is connected with the first pushing/absorbing mechanism, the first pushing/absorbing mechanism is used for being connected with a sample injection pipe, the first end of the liquid storage flow channel is connected with the first pushing/absorbing mechanism, the first end of the ball outlet flow channel is respectively communicated with the first end of the liquid outlet flow channel and the first end of the continuous phase liquid inlet flow channel, the second end of the ball outlet flow channel extends to the top end of the chip body and is used for being connected with a ball outlet connecting pipe, and the second end of the continuous phase liquid inlet flow channel is connected with the second pushing/absorbing mechanism which is used for being connected with a liquid storage container;
the switching mechanism is arranged at the top end of the chip body and is provided with a connecting channel, the switching mechanism can rotate between a first position and a second position relative to the chip body, when the switching mechanism is positioned at the first position, the first end of the connecting channel is communicated with the second end of the liquid storage flow channel, the second end of the connecting channel is communicated with the second end of the liquid inlet flow channel, and when the switching mechanism is positioned at the second position, the first end of the connecting channel is communicated with the second end of the liquid storage flow channel, and the second end of the connecting channel is communicated with the second end of the liquid outlet flow channel.
2. The organoid spheroid preparation device of claim 1, wherein the chip body is provided with a dispersed phase feed channel, a first liquid storage channel and a continuous phase feed channel, wherein a first end of the dispersed phase feed channel is in communication with a first end of the feed channel, a second end of the dispersed phase feed channel extends to a top end of the chip body, a first end of the first liquid storage channel is in communication with the first end of the liquid storage channel, a second end of the first liquid storage channel extends to a top end of the chip body, a first end of the continuous phase feed channel is in communication with a second end of the continuous phase feed channel, and a second end of the continuous phase feed channel extends to a top end of the chip body.
3. The organoid sphere preparation apparatus according to claim 2, wherein a dispersed phase liquid inlet connector, a liquid storage connector, a continuous phase liquid inlet connector and a ball outlet connector are provided at the top end of the chip body, wherein the dispersed phase liquid inlet connector is connected with the second end of the dispersed phase liquid inlet channel, the dispersed phase liquid inlet connector is connected with the first push/liquid suction mechanism through a first connecting pipe, the first push/liquid suction mechanism is connected with the sample injection pipe through a second connecting pipe, the liquid storage connector is connected with the second end of the first liquid storage channel, the liquid storage connector is connected with the first push/liquid suction mechanism through a third connecting pipe, the continuous phase liquid inlet connector is connected with the second end of the continuous phase liquid inlet channel, the continuous phase liquid inlet connector is connected with the second push/liquid suction mechanism through a fourth connecting pipe, the second push/liquid suction mechanism is connected with the liquid storage container through a fifth connecting pipe, the ball outlet connector is connected with the second end of the ball outlet channel, and the ball outlet connector is connected with the ball outlet connector.
4. The organoid spheroid preparation device of claim 1, wherein the chip body is provided with a dispersed phase liquid outlet channel, a second liquid outlet channel and a liquid outlet connecting channel, wherein a first end of the dispersed phase liquid outlet channel is communicated with a second end of the liquid inlet channel, the second end of the dispersed phase liquid outlet channel extends to the top end of the chip body, a first end of the second liquid outlet channel is communicated with the second end of the liquid outlet channel, the second end of the second liquid outlet channel extends to the top end of the chip body, the first end of the liquid outlet connecting channel is communicated with the second end of the liquid outlet channel, and the second end of the liquid outlet connecting channel extends to the top end of the chip body.
5. The organoid spheroid preparation device according to claim 4, wherein the switching mechanism comprises a knob and a switching piece, a ring portion is formed at the top end of the chip body, the second end of the dispersion phase liquid outlet channel, the second end of the second liquid storage channel and the second end of the liquid outlet connecting channel are all located inside the ring portion, the bottom end of the knob is contained in the ring portion and can rotate between a first position and a second position relative to the chip body, a mounting cavity is formed at the bottom end of the knob, the switching piece is arranged in the mounting cavity, the bottom end of the switching piece is in close contact with the top end of the chip body and is provided with the connecting channel, the rotation of the knob can drive the switching piece to rotate between the first position and the second position relative to the chip body, when the knob and the switching piece are located at the first position, the first end of the connecting channel is communicated with the second end of the second liquid storage channel, and when the knob and the switching piece are located at the first position, the second end of the connecting channel is communicated with the second end of the second channel.
6. The organoid sphere preparation device of claim 5, further comprising a drive mechanism disposed at a top end of said platform, said top end of said knob being coupled to said drive mechanism, said drive mechanism for driving said knob to rotate between a first position and a second position.
7. The organoid sphere preparation device of claim 1, wherein said fluid storage channel is a serpentine channel.
8. The organoid sphere preparation device of claim 1, wherein said sphere outlet flow channel has a size greater than a size of said continuous phase liquid inlet flow channel.
9. The organoid sphere preparation device of claim 1, wherein said first push/pull mechanism and said second push/pull mechanism are each syringe pumps.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202321538568.XU CN219951074U (en) | 2023-06-15 | 2023-06-15 | Organoid spheroid preparation facilities |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202321538568.XU CN219951074U (en) | 2023-06-15 | 2023-06-15 | Organoid spheroid preparation facilities |
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| CN219951074U true CN219951074U (en) | 2023-11-03 |
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| CN202321538568.XU Active CN219951074U (en) | 2023-06-15 | 2023-06-15 | Organoid spheroid preparation facilities |
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| CN (1) | CN219951074U (en) |
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