A centrifugal device and a separation method for a centrifugal device

Abstract

The application provides a centrifugal device and a separation method for the centrifugal device, the centrifugal device comprising a centrifugal cup, a first fluid channel, a second fluid channel and a third fluid channel; the center port of the first fluid channel, the inner port of the second fluid channel and the outer port of the third fluid channel are all in fluid communication with the centrifugal cup; the outer port is close to the outer wall of the centrifugal cup, and the center port is close to the center of the centrifugal cup; the inner port is between the outer port and the center port; after liquid centrifugal separation is carried out in the centrifugal cup, the liquid is divided into multiple layers from outside to inside in the centrifugal cup, and the target object is extracted through the combination of the outer port, the inner port and the center port. The separation method for the centrifugal device provided by the application can realize the rapid separation and extraction of multi-component sample components flexibly through the three-port liquid inlet and outlet scheme of the center port, the inner port and the outer port, is convenient to operate and has higher efficiency.

Description

Technical Field

[0001] This invention belongs to the field of biological cell processing, specifically relating to a centrifuge device and a separation method for the centrifuge device. Background Technology

[0002] In the field of biological cell processing, centrifuges are mainly used for cell separation, concentration of cell slurry after target cell culture, and washing. After cell separation, the target cell slurry is obtained. This slurry is then concentrated to remove excess culture medium and waste liquid, followed by washing and other subsequent operations. The separation technology used in cell separation and concentration primarily relies on the high-speed rotation of the centrifuge cup. Within the centrifuge cup, the cell sample experiences centripetal force due to the high-speed rotation. Because of the mass differences among the components within the cells, the centripetal forces acting on each component are also different, resulting in varying radii of rotation for each component within the centrifuge cup, thus achieving cell separation.

[0003] Existing centrifuge cups are typically single-port or two-port designs. For single-port designs, the single port is used for both sample injection (usually liquid) and sample effluent. While the single-port design is structurally simple, it cannot achieve continuous flow operation; sample must be injected first, then centrifuged, and finally effluent using the same port, with the effluent sequence from the outside in. For two-port designs, there is an inlet (or outlet) near the center of rotation, and an outlet (or inlet) outside the center of rotation. The two-port design allows for more flexible inlet and outlet operations, making it suitable for two-component extraction. However, for multi-component extraction, more complex process control is required. Therefore, there is an urgent need to develop a novel separation method to address these technical problems. Summary of the Invention

[0004] To at least solve one of the above-mentioned technical problems, the present invention provides a centrifuge device and a separation method for the centrifuge device, which adopts a three-port liquid inlet and outlet scheme, and can flexibly realize the rapid separation and extraction of multi-component sample components.

[0005] To achieve at least one of the above objectives, the technical solution adopted by the present invention is as follows:

[0006] This invention provides a separation method for a centrifuge apparatus, the centrifuge apparatus comprising a centrifuge cup, a first fluid channel, a second fluid channel, and a third fluid channel; the central port of the first fluid channel, the inner port of the second fluid channel, and the outer port of the third fluid channel are all in fluid communication with the centrifuge cup; the outer port is close to the outer wall of the centrifuge cup, and the central port is close to the center of the centrifuge cup; the inner port is located between the outer port and the central port; after liquid centrifugation separation in the centrifuge cup, the liquid is divided into multiple layers from the outside to the inside of the centrifuge cup, and the target analyte is extracted through the combination of the outer port, the inner port, and the central port.

[0007] Furthermore, the liquid inside the centrifuge cup is divided into a first layer, a second layer, a third layer, and a fourth layer from the outside to the inside; the first layer corresponds to the position of the outer opening; and the second layer corresponds to the position of the inner opening.

[0008] Furthermore, the material of the first layer is extracted through the external port; the remaining liquid in the centrifuge cup moves towards the outer wall of the centrifuge cup under the action of centrifugal force.

[0009] Furthermore, the complete removal of all materials in the first layer can be estimated by measuring the sample size, or by visually monitoring the boundary between the first and second layers to determine if all materials in the first layer have been removed.

[0010] Further, a portion of the material from the second layer is extracted through the outer opening until the third layer moves to the area between the inner and outer openings.

[0011] Furthermore, the material of the fourth layer is extracted through the inner opening; the remaining material in the centrifuge cup is the target substance.

[0012] Furthermore, most of the material in the second layer and all the material in the third layer are extracted through the inner opening and stored in a temporary container.

[0013] Further, the remaining material in the centrifuge cup is extracted until the centrifuge cup is emptied.

[0014] Furthermore, most of the material in the second layer and all of the material in the third layer of the temporary container are discharged into the centrifuge cup through the central opening.

[0015] Furthermore, the liquid is leukocyte fluid, the first layer is a red blood cell layer, the second layer is a Ficoll fluid layer, the third layer is a white blood cell membrane layer, and the fourth layer is a plasma layer.

[0016] The present invention also provides a centrifugation device, including a centrifuge cup body, a centrifuge cup lid, and an inlet / outlet connection seat; the centrifuge cup lid is fixedly disposed on the centrifuge cup body; the inlet / outlet connection seat is rotatably supported on the centrifuge cup lid; a plurality of fluid channels are provided between the inlet / outlet connection seat and the centrifuge cup lid, the plurality of fluid channels connecting the outside to the inside of the centrifuge cup body; the inlet / outlet positions of the plurality of fluid channels are radially distanced from the center position of the centrifuge cup body.

[0017] Furthermore, the plurality of fluid channels includes a first fluid channel, a second fluid channel, and a third fluid channel; portions of the first fluid channel, the second fluid channel, and the third fluid channel on the centrifuge cup lid are evenly distributed along the circumference of the centrifuge cup lid.

[0018] Furthermore, the central opening of the first fluid channel, the inner opening of the second fluid channel, and the outer opening of the third fluid channel are all in fluid communication with the centrifuge cup; the outer opening is close to the outer wall of the centrifuge cup, and the central opening is close to the center of the centrifuge cup; the inner opening is located between the outer opening and the central opening.

[0019] Furthermore, the centrifuge cup lid includes a lid body and a rotating shaft; the rotating shaft is provided with a first flow channel, a second flow channel and a third flow channel; the first flow channel, the second flow channel and the third flow channel are all arranged radially along the rotating shaft, and are staggered in both the axial and circumferential directions of the rotating shaft.

[0020] Furthermore, the first flow channel is connected to the first shaft hole inside the rotating shaft; the first shaft hole is connected to the first conduit disposed on the cover; the second flow channel is connected to the second shaft hole inside the rotating shaft; the second shaft hole is connected to the second conduit disposed on the cover; the third flow channel is connected to the third shaft hole inside the rotating shaft, and the third shaft hole is connected to the third conduit disposed on the cover.

[0021] Furthermore, the end of the first conduit is close to the center of the rotation axis of the centrifuge cup, the end of the third conduit is close to the outer wall of the centrifuge cup, and the end of the second conduit is located between the center of the rotation axis and the outer wall.

[0022] Furthermore, the ends of the first catheter and / or the second catheter and / or the third catheter extend to the bottom wall of the centrifuge cup.

[0023] Furthermore, the inlet / outlet liquid connection seat is provided with a shaft groove, and the open end of the shaft groove is provided with a bearing groove, in which a bearing is installed; the rotating shaft is rotatably supported in the shaft groove by the bearing.

[0024] Furthermore, a first annular groove, a second annular groove, and a third annular groove are sequentially spaced at the other end of the shaft groove; there is an annular gap between the first annular groove, the second annular groove, and the third annular groove and the rotating shaft; a sealing ring is provided between the first annular groove, the second annular groove, and the third annular groove, and the sealing ring is rotatably and sealingly connected to the rotating shaft, and the sealing ring achieves sealing isolation between the first annular groove, the second annular groove, and the third annular groove.

[0025] Furthermore, the inlet / outlet liquid connection seat is provided with a first connector that is in fluid communication with the first annular groove, the first flow channel is correspondingly provided with the first annular groove, and forms a first fluid channel through the first shaft hole and the first conduit in sequence.

[0026] Furthermore, the inlet / outlet liquid connection seat is provided with a second connector that is in fluid communication with the second annular groove, the second flow channel is correspondingly provided with the second annular groove, and forms a second fluid channel through the second shaft hole and the second conduit in sequence.

[0027] Furthermore, the inlet / outlet liquid connection seat is provided with a third connector that is in fluid communication with the third annular groove, the third flow channel is correspondingly provided with the third annular groove, and forms a third fluid channel through the third shaft hole and the third conduit in sequence.

[0028] This invention provides a centrifuge apparatus and a separation method for the centrifuge apparatus. Compared with the prior art, the advantages of this invention are as follows:

[0029] The design of the shaft holes on the centrifuge cup lid, connecting the rotating shaft to the inlet / outlet connectors, creates a first fluid channel from the first connector to the first conduit, a second fluid channel from the second connector to the second conduit, and a third fluid channel from the third connector to the third conduit. These three fluid channels are distributed radially within the centrifuge cup, allowing for fluid entry and exit from the outermost, middle, and innermost parts of the cup, respectively, near the center of rotation. This allows for rapid separation and extraction of sample components as needed, unlike single-port centrifuge cups where the sample exit sequence must follow an outside-to-inside order. Furthermore, the combination of these three fluid channels enables PBMC separation, concentration washing, or centrifugal displacement operations, eliminating the need for complex process control required for multi-component extraction in dual-port centrifuge cups.

[0030] It should be understood that the above general description and the following detailed description are exemplary and explanatory only, and do not limit the scope of protection of this application. Attached Figure Description

[0031] Figure 1 This is a schematic diagram of the centrifuge device in the embodiments of this application;

[0032] Figure 2 This is an explosion diagram of the centrifuge device in the embodiments of this application;

[0033] Figure 3 This is a schematic diagram of the internal structure of the centrifuge cup lid in an embodiment of this application;

[0034] Figure 4 This is a schematic diagram of the internal structure of the centrifuge cup lid from another perspective in an embodiment of this application.

[0035] Figure 5 This is a schematic diagram of the internal structure of the centrifuge cup lid from another perspective in an embodiment of this application;

[0036] Figure 6 This is a schematic diagram of the internal structure of the inlet / outlet connection seat in the embodiments of this application;

[0037] Figure 7 This is a schematic diagram of the internal structure of the inlet / outlet liquid connection seat from another perspective in an embodiment of this application.

[0038] Figure 8 This is another perspective view of the internal structure of the inlet / outlet liquid connection seat in the embodiments of this application;

[0039] Figure 9 This is a schematic diagram of the leukocyte fluid after centrifugation in an embodiment of this application;

[0040] Figure 10 This is a schematic diagram of red blood cell extraction through an external opening in an embodiment of this application;

[0041] Figure 11 This is a schematic diagram showing the red blood cells after extraction in an embodiment of this application;

[0042] Figure 12 This is a schematic diagram illustrating the extraction of a portion of Ficoll liquid through an external port in an embodiment of this application.

[0043] Figure 13 This is a schematic diagram of the Ficoll solution extracted in one of the embodiments of this application;

[0044] Figure 14 This is a schematic diagram illustrating the extraction of plasma through the internal opening in an embodiment of this application;

[0045] Figure 15 This is a schematic diagram showing the plasma extraction process in an embodiment of this application.

[0046] Figure 16 This is a schematic diagram of extracting the target object through the inner opening in another embodiment of this application;

[0047] Figure 17 This is a schematic diagram of the target object after extraction in another embodiment of this application;

[0048] Figure 18 This is a schematic diagram of extracting the remaining material through an external opening in another embodiment of this application;

[0049] Figure 19 This is a schematic diagram of the centrifuge cup being emptied in another embodiment of this application;

[0050] Figure 20 This is a schematic diagram of the target object being discharged through the central opening in another embodiment of this application;

[0051] Among them, 1 centrifuge cup body, 2 centrifuge cup lid, 2-1 lid body, 2-2 liquid guiding part, 2-3 rotating shaft, 2-311 first flow channel, 2-312 first shaft hole, 2-321 second flow channel, 2-322 second shaft hole, 2-331 third flow channel, 2-332 third shaft hole, 2-4 first guide tube, 2-5 second guide tube, 2-6 third guide tube, 3 inlet / outlet connection seat, 3-1 shaft groove, 3-2 first annular groove, 3-3 second annular groove, 3-4 third annular groove, 4 connector, 4-1 first connector, 4-2 second connector, 4-3 third connector, 5 bearing, 6 sealing ring, 7 red blood cell layer, 8 Ficoll liquid layer, 9 white film layer, 10 plasma layer. Detailed Implementation

[0052] To enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be further described in detail below with reference to specific embodiments. Please note that the embodiments described below are exemplary and are only used to explain the present invention, and should not be construed as limiting the present invention. Where specific techniques or conditions are not specified in the embodiments, they shall be performed in accordance with the techniques or conditions described in the literature in the art or in accordance with the product manual.

[0053] In the description of this invention, it should be understood that the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this invention, "a plurality of" means two or more, unless otherwise explicitly specified.

[0054] In this invention, unless otherwise explicitly specified and limited, the terms "installation," "connection," "linking," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0055] The separation method for a centrifuge provided by the present invention will be described in detail below through specific embodiments:

[0056] like Figure 1-8 As shown, this embodiment provides a centrifuge device, which mainly includes components such as a centrifuge cup body 1, a centrifuge cup lid 2, an inlet / outlet liquid connection seat 3, and a connector 4.

[0057] The centrifuge cup lid 2 is fixedly mounted on the centrifuge cup body 1, and an inlet / outlet liquid connection seat 3 is rotatably supported on the centrifuge cup lid 2; three connectors 4 are also fixedly mounted on the inlet / outlet liquid connection seat 3. The centrifuge cup lid 2 includes a lid body 2-1, a liquid guiding part 2-2, and a rotating shaft 2-3. The three liquid guiding parts 2-2 are mounted on the lid body 2-1 and connected to the rotating shaft 2-3; the three liquid guiding parts 2-2 are arranged separately along the circumference of the lid body 2-1.

[0058] The rotating shaft 2-3 is located at the center of the cover 2-1; the rotating shaft 2-3 is provided with a first flow channel 2-311, a second flow channel 2-321 and a third flow channel 2-331. The first flow channel 2-311, the second flow channel 2-321 and the third flow channel 2-331 are all arranged radially along the rotating shaft 2-3, and are staggered in both the axial and circumferential directions of the rotating shaft 2-3.

[0059] The first flow channel 2-311 communicates with the first shaft hole 2-312 inside the rotating shaft 2-3; the first shaft hole 2-312 communicates with the first conduit 2-4 disposed in a liquid guiding section 2-2. The second flow channel 2-321 communicates with the second shaft hole 2-322 inside the rotating shaft 2-3; the second shaft hole 2-322 communicates with the second conduit 2-5 disposed in another liquid guiding section 2-2. The third flow channel 2-331 communicates with the third shaft hole 2-332 inside the rotating shaft 2-3; the third shaft hole 2-332 communicates with the third conduit 2-6 disposed in another liquid guiding section 2-2.

[0060] The ends of the first catheter 2-4, the second catheter 2-5, and the third catheter 2-6 are all connected to the internal cavity of the centrifuge cup 1, and the distances from the ends of the first catheter 2-4, the second catheter 2-5, and the third catheter 2-6 to the center of the centrifuge cup lid increase sequentially. That is, the end of the first catheter 2-4 is closest to the center of the rotation axis, the end of the third catheter 2-6 is closest to the outer wall of the centrifuge cup 1, and the end of the second catheter 2-5 is in between.

[0061] The inlet / outlet connection seat 3 is provided with a shaft groove 3-1, and a bearing groove is provided at the open end of the shaft groove 3-1, in which a bearing 5 is installed. At the other end of the shaft groove 3-1, a plurality of annular grooves are provided at intervals, and a sealing ring 6 is installed between each annular groove. The rotating shaft 2-3 of the centrifuge cup lid 2 is rotatably supported in the shaft groove 3-1 by the bearing 5.

[0062] The annular grooves include a first annular groove 3-2, a second annular groove 3-3, and a third annular groove 3-4. Each annular groove has an annular gap with the rotating shaft 2-3; each sealing ring 6 is rotatably sealed to the rotating shaft 2-3, thereby achieving sealing isolation between the multiple annular gaps through the sealing rings.

[0063] The inlet / outlet liquid connection seat 3 is provided with a first connector 4-1 that is in fluid communication with the first annular groove 3-2. The first annular groove 3-3 is correspondingly provided with the first flow channel 2-311 of the rotating shaft 2-3, and is connected to the sealed cavity in sequence through the first shaft hole 2-312 and the first conduit 2-4 to form a first fluid channel from the first connector 4-1 to the first conduit 2-4.

[0064] The inlet / outlet liquid connection seat 3 is also provided with a second connector 4-2 that is in fluid communication with the second annular groove 3-3. The second annular groove 3-4 is correspondingly provided with the second flow channel 2-321 of the rotating shaft 2-3, and is connected to the sealed cavity in sequence through the second shaft hole 2-322 and the second conduit 2-5 to form a second fluid channel from the second connector 4-2 to the second conduit 2-5.

[0065] The inlet / outlet liquid connection seat 3 is also provided with a third connector 4-3 that is in fluid communication with the third annular groove 3-4. The third annular groove 3-5 is correspondingly provided with the third flow channel 2-331 of the rotating shaft 2-3, and is connected to the sealed cavity in sequence through the third shaft hole 2-332 and the third conduit 2-6, forming a third fluid channel from the third connector 4-3 to the third conduit 2-6.

[0066] Because the ends of the three fluid channels are at different distances from the rotation center of the centrifuge cup, the liquid can enter and exit directly from the outermost part of the centrifuge cup 1 through the third fluid channel; the liquid can enter and exit from the middle position through the second fluid channel; and the liquid can enter and exit from the center of the centrifuge cup 1 through the first fluid channel.

[0067] This embodiment provides a separation method using the centrifuge device provided in this application, which is described in detail below:

[0068] like Figure 9 The embodiment shown takes the separation of white blood cells from blood as an example. After high-speed centrifugation in the centrifuge device, the sample blood and the cell separation solution Ficoll solution are separated into 4 layers, which are, from the outside to the inside of the centrifuge cup 1, the layers are: red blood cell layer 7, Ficoll solution layer 8, white membrane layer 9 (the layer where the target cells are located, which is less numerous) and plasma layer 10. At this time, the red blood cell layer 7 corresponds to the outer opening of the end of the third fluid channel, and the Ficoll solution layer corresponds to the inner opening of the end of the second fluid channel.

[0069] Next, as follows Figure 10As shown, an external power device extracts red blood cells from the red blood cell layer 7 through the outer opening of the third fluid channel of the centrifuge device. During the drainage process, the centrifuge cup continuously rotates at high speed, and the liquid in the cup is driven by outward centrifugal force to move towards the outer wall of the cup. The number of red blood cells is estimated by the sample size, or the boundary between the red blood cell layer and the Ficoll liquid layer is monitored by a visual camera to determine whether all red blood cells have been drained.

[0070] like Figure 11 As shown, after the red blood cells have been drained from the external opening, the remaining layers in the centrifuge cup, from the outside to the inside, are Ficoll liquid layer 8, white membrane layer 9, and plasma layer 10. At this time, Ficoll liquid layer 8 corresponds to both the external opening at the end of the third fluid channel and the internal opening at the end of the second fluid channel.

[0071] Then as Figure 12 As shown, an external power unit draws a portion of the Ficoll liquid through the outer port of the third fluid channel of the centrifuge. This continues until the white film layer 9 is moved to the area between the inner and outer ports, as shown. Figure 13 As shown. At this time, the Ficoll liquid layer 8 corresponds to the outer opening of the third fluid channel, and the plasma layer 10 corresponds to the inner opening of the second fluid channel.

[0072] Then as Figure 14 As shown, the external power equipment stops after drawing all the plasma from the plasma layer 10 through the inner port of the second fluid channel of the centrifuge. Figure 15 As shown, the remaining layers in the centrifuge container at this point, from the outside to the inside, are Ficoll liquid layer 8 and white film layer 9, which are the target substances. If an even smaller retention volume is needed, under visual control, the external device's third fluid channel continues to draw some Ficoll liquid, further reducing the retention volume until the requirement is met.

[0073] In summary, the separation method using centrifugation devices is used to separate and obtain the target substance, which can then be used for subsequent preparation processes.

[0074] Another embodiment of this application also provides a separation method using the centrifuge device provided in this application, which is described in detail below:

[0075] This embodiment also uses the separation of white blood cells from blood as an example. After high-speed centrifugation in the centrifuge device, the sample blood and the cell separation fluid Ficoll are separated into four layers, which are, from the outside to the inside of the centrifuge cup 1, as follows: red blood cell layer 7, Ficoll fluid layer 8, white blood cell layer 9 (the layer containing the target cells, which is relatively few in number), and plasma layer 10. At this time, the red blood cell layer 7 corresponds to the outer opening of the end of the third fluid channel, and the Ficoll fluid layer corresponds to the inner opening of the end of the second fluid channel, as detailed below. Figure 9 As shown.

[0076] Then as Figure 16 As shown, the external power equipment draws Ficoll liquid through the inner port of the second fluid channel of the centrifuge device. Because the centrifuge cup is continuously rotating at high speed during the drainage process, the liquid in the centrifuge cup is driven by outward centrifugal force to move towards the outer wall of the centrifuge cup. Therefore, a small portion of the Ficoll liquid near the red blood cell layer 7 cannot be drawn from outside the inner port.

[0077] like Figure 17 The number of red blood cells is estimated by sample size or monitored and controlled by a visual camera until most of the Ficoll solution and white film layer 9, i.e., the target material, are extracted and stored in an external temporary container. At this point, the remaining layers in the centrifuge cup, from the outside to the inside, are red blood cell layer 7, Ficoll solution layer 8, and plasma layer 10, all of which are non-target materials.

[0078] Then as Figure 18 As shown, the external power equipment extracts all remaining material through the outer opening of the third fluid channel of the centrifuge and discharges it into the waste liquid bag. After a predetermined time, until the centrifuge cup is empty, as... Figure 19 As shown.

[0079] Then as Figure 20 As shown, the external power equipment discharges the Ficoll liquid and white film layer stored in the external temporary container into the centrifuge cup through the central port of the first fluid channel of the centrifuge device.

[0080] In summary, the separation method using centrifugation devices is used to separate and obtain the target substance, which can then be used for subsequent preparation processes.

[0081] In this invention, unless otherwise explicitly specified and limited, "above" or "below" the second feature can mean that the first feature is in direct contact with the second feature, or that the first feature is in indirect contact with the second feature through an intermediate medium. Furthermore, "above," "over," and "on top" of the second feature can mean that the first feature is directly above or diagonally above the second feature, or simply that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature can mean that the first feature is directly below or diagonally below the second feature, or simply that the first feature is at a lower horizontal level than the second feature.

[0082] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. Moreover, without contradiction, those skilled in the art can combine and integrate the different embodiments or examples described in this specification, as well as the features of different embodiments or examples.

Claims

1. A centrifuge device, characterized in that, The centrifuge container includes a centrifuge cup body, a centrifuge cup lid, and an inlet / outlet connection seat. The centrifuge cup lid is fixedly mounted on the centrifuge cup body. The inlet / outlet connection seat is rotatably supported on the centrifuge cup lid. Multiple fluid channels are provided between the inlet / outlet connection seat and the centrifuge cup lid, connecting the outside to the interior of the centrifuge cup body. The inlet / outlet positions of the multiple fluid channels are radially distanced from the center of the centrifuge cup body. The multiple fluid channels include a first fluid channel, a second fluid channel, and a third fluid channel. The center opening of the first fluid channel, the inner opening of the second fluid channel, and the outer opening of the third fluid channel are all in fluid communication with the centrifuge cup body. The outer opening is close to the outer wall of the centrifuge cup body, and the center opening is close to... The centrifuge cup body is located at its center; the inner opening is located between the outer opening and the center opening; the centrifuge cup lid includes a lid body and a rotating shaft; the rotating shaft is provided with a first flow channel, a second flow channel, and a third flow channel; the first flow channel, the second flow channel, and the third flow channel are all arranged radially along the rotating shaft and are staggered in both the axial and circumferential directions of the rotating shaft; the first flow channel communicates with a first shaft hole inside the rotating shaft; the first shaft hole communicates with a first conduit provided on the lid body; the second flow channel communicates with a second shaft hole inside the rotating shaft; the second shaft hole communicates with a second conduit provided on the lid body; the third flow channel communicates with a third shaft hole inside the rotating shaft, and the third shaft hole communicates with a third conduit provided on the lid body; The end of the first conduit is close to the center of the rotation axis of the centrifuge cup, the end of the third conduit is close to the outer wall of the centrifuge cup, and the end of the second conduit is located between the center of the rotation axis and the outer wall. A central groove is provided on the inlet / outlet connection seat, and a bearing groove is provided at the open end of the central groove, in which a bearing is installed. The rotating shaft is rotatably supported in the central groove by the bearing. A first annular groove, a second annular groove, and a third annular groove are sequentially spaced at the other end of the central groove. Each of the first, second, and third annular grooves has an annular gap with the rotating shaft. A sealing ring is provided between the first, second, and third annular grooves, and the sealing ring is rotatably and sealingly connected to the rotating shaft. The first, second, and third annular grooves are sealed and isolated from each other. The inlet / outlet connection seat is provided with a first connector that is in fluid communication with the first annular groove. The first flow channel is correspondingly arranged with the first annular groove and forms a first fluid channel through the first shaft hole and the first conduit in sequence. The inlet / outlet connection seat is provided with a second connector that is in fluid communication with the second annular groove. The second flow channel is correspondingly arranged with the second annular groove and forms a second fluid channel through the second shaft hole and the second conduit in sequence. The inlet / outlet connection seat is provided with a third connector that is in fluid communication with the third annular groove. The third flow channel is correspondingly arranged with the third annular groove and forms a third fluid channel through the third shaft hole and the third conduit in sequence.

2. A separation method for a centrifuge apparatus, comprising the centrifuge apparatus of claim 1, characterized in that, The separation method is as follows: After the liquid is centrifuged in the centrifuge cup, the liquid is divided into multiple layers from the outside to the inside of the centrifuge cup. The target substance is extracted through the combination of the outer port, inner port and center port. The ends of the three fluid channels are at different distances from the rotation center of the centrifuge cup. The liquid can enter and exit directly from the outermost part of the centrifuge cup through the third fluid channel; the liquid can enter and exit from the middle position through the second fluid channel; and the liquid can enter and exit from the center of the centrifuge cup through the first fluid channel.

3. The separation method for a centrifuge according to claim 2, characterized in that, The liquid inside the centrifuge cup is divided into four layers from the outside to the inside: a first layer, a second layer, a third layer, and a fourth layer. The first layer corresponds to the position of the outer opening, and the second layer corresponds to the position of the inner opening.

4. The separation method for a centrifuge according to claim 3, characterized in that, The first layer of material is extracted through the external port; the remaining liquid in the centrifuge cup moves towards the outer wall of the centrifuge cup under the action of centrifugal force.

5. The separation method for a centrifuge according to claim 4, characterized in that, The complete removal of the first layer of material can be determined by estimating the sample size or by visually monitoring the boundary between the first and second layers.

6. The separation method for a centrifuge according to claim 5, characterized in that, A portion of the material from the second layer is extracted through the outer opening until the third layer moves to the area between the inner and outer openings.

7. The separation method for a centrifuge according to claim 6, characterized in that, The material from the fourth layer is extracted through the inner opening; the remaining material in the centrifuge cup is the target substance.

8. The separation method for a centrifuge according to claim 3, characterized in that, Most of the material in the second layer and all of the material in the third layer are extracted through the inner opening and stored in a temporary container.

9. The separation method for a centrifuge according to claim 8, characterized in that, Extract the remaining material from the centrifuge container until the centrifuge container is empty.

10. The separation method for a centrifuge according to claim 8, characterized in that, Most of the material in the second layer and all of the material in the third layer of the temporary container are discharged into the centrifuge cup through the central opening.

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