Centrifugal device, sample separation method, controller, and medium

Abstract

The invention provides a centrifugal device, a sample separation method, a controller and a medium. The centrifugal device comprises a cup body with a centrifugal space; the cup body comprises a bottom plate located at the bottom of the centrifugal space and an annular side plate arranged around the bottom plate, and the annular side plate comprises a straight plate section and an inclined section connected between the straight plate section and the bottom plate; the cup body can rotate around the rotating axis; the distance between the inclined section and the rotating axis is gradually reduced from top to bottom along the rotating axis; a pipetting channel is arranged on the bottom plate; two ends of the pipetting channel are respectively provided with a first inlet and outlet and a second inlet and outlet, the pipetting channel is communicated with the centrifugal space through the first inlet and outlet, and the inclined surface of the inclined section close to the rotating axis is overlapped and connected with at least one part of the edge of the first inlet and outlet. According to the invention, liquid residue is avoided, and the pipetting efficiency is improved.

Description

Technical Field

[0001] The present invention belongs to the field of centrifugal technology, and in particular relates to a centrifugal device, a sample separation method, a controller and a medium. Background Art

[0002] At present, automated centrifugal devices generally have channels for entering and exiting liquids, so that all liquids in the centrifugal device can be discharged through the channels after centrifugation is completed, or part of the liquid can be selectively discharged through the channels during the centrifugation process (for example, when the cell fluid in the centrifugal device is concentrated, it is necessary to extract useless liquid from the centrifugal device, and then the liquid remaining in the centrifugal device is the concentrated product, and the volume of the product is called the minimum concentrated volume). In the prior art, when the centrifugal device empties all liquids through the channel, there is often residual liquid. At this time, the centrifugal device needs to be opened manually or other means are used to discharge the residual liquid, which has problems such as slow processing speed and long processing time; on the other hand, when selectively discharging part of the liquid in the centrifugal device through the channel in the prior art, the accuracy of the discharge of the part of the liquid is often unable to be guaranteed. Summary of the invention

[0003] The present invention aims at the technical problems in the prior art such as residual liquid when a centrifugal device is discharging liquid, and provides a centrifugal device, a sample separation method, a controller and a medium.

[0004] In view of the above technical problems, an embodiment of the present invention provides a centrifugal device, comprising a cup body provided with a centrifugal space; the cup body comprises a bottom plate located at the bottom of the centrifugal space, and an annular side plate arranged around the bottom plate, the annular side plate comprises a straight plate segment and an inclined segment connected between the straight plate segment and the bottom plate; the cup body can rotate around a rotation axis; along the direction from top to bottom of the rotation axis, the distance between the inclined segment and the rotation axis gradually decreases;

[0005] A pipetting channel is provided on the bottom plate; a first inlet and a second inlet are respectively provided at both ends of the pipetting channel, the pipetting channel is connected to the centrifugal space through the first inlet and the inclined surface of the inclined section close to the rotation axis overlaps and is connected with at least a portion of an edge of the first inlet and the outlet.

[0006] A sample separation method, the sample separation method is performed by the centrifugal device, and the sample separation method comprises:

[0007] Controlling a first preset dose of separation liquid to flow into the centrifugal space in the cup body of the centrifugal device;

[0008] After controlling the cup body to rotate around the rotation axis at a first preset speed, controlling a second preset dose of sample liquid to flow into the centrifugal space at a preset flow rate;

[0009] Controlling the cup body to rotate at a second preset speed for a first preset time to centrifuge the liquid in the centrifugal space in the cup body, so that the liquid adheres to the straight plate section and the inclined section to form a liquid ring; the top surface of the liquid ring adheres to the top wall of the centrifugal space, and the bottom surface of the liquid ring covers the first inlet and outlet;

[0010] The liquid in the liquid ring covering the first inlet and outlet is discharged from the centrifugal space through the pipetting channel.

[0011] A controller comprises a processor and a memory, wherein the memory stores an executable program, and the processor is used to execute the executable program to implement the sample separation method.

[0012] A computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the sample separation method is implemented.

[0013] The centrifugal device provided by the present invention comprises a cup body provided with a centrifugal space; the cup body comprises a bottom plate located at the bottom of the centrifugal space, and an annular side plate arranged around the bottom plate, the annular side plate comprises a straight plate section and an inclined section connected between the straight plate section and the bottom plate; the cup body can rotate around a rotation axis; along the direction from top to bottom of the rotation axis, the distance between the inclined section and the rotation axis gradually decreases; a liquid transfer channel is provided on the bottom plate; a first inlet and a second inlet and a second inlet are respectively provided at both ends of the liquid transfer channel, the liquid transfer channel communicates with the centrifugal space through the first inlet and a second inlet and a second inlet are overlapped and connected with the inclined surface of the inclined section close to the rotation axis and at least a part of the edge of the first inlet and a second inlet. The present invention arranges an inclined section connected to the bottom plate on the annular side plate, and overlaps and connects the inclined surface of the inclined section close to the rotation axis and at least a part of the edge of the first inlet and a second inlet, so that the liquid can be guided through the inclined surface and the overlapping connection position between the bottom surface of the inclined surface and the edge of the first inlet and a second inlet, so that the liquid in the centrifugal space can be quickly introduced into the first inlet and a second inlet from the overlapping connection position through the inclined surface, and the residual liquid in the centrifugal space can be discharged through the liquid transfer channel as much as possible, thereby avoiding liquid residue and improving the liquid transfer efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The present invention is further described below in conjunction with the accompanying drawings and embodiments.

[0015] Figure 1 It is a schematic structural diagram of a centrifugal device provided in one embodiment of the present invention.

[0016] Figure 2 It is a flow chart of a sample separation method provided by an embodiment of the present invention.

[0017] The reference numerals in the specification are as follows:

[0018] 100, cup body; 110, centrifugal space; 120, bottom plate; 121, pipetting channel; 1211, first entrance and exit; 1212, second entrance and exit; 1213, bending section; 1214, pipetting section; 122, pipetting groove; 123, blind hole; 130, annular side plate; 131, straight plate section; 132, inclined section; 1321, inclined surface; 140, cup cover; 141, entrance and exit channel; 200, mounting portion; 300, guide portion. DETAILED DESCRIPTION

[0019] In order to make the technical problems, technical solutions and beneficial effects solved by the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0020] It should be understood that the directions or positional relationships indicated by terms such as "upper", "lower", "left", "right", "front", "back", and "middle" are based on the directions or positional relationships shown in the accompanying drawings and are only for the convenience of describing the present invention and simplifying the description. They do not indicate or imply that the device or element referred to must have a specific direction, be constructed and operated in a specific direction, and therefore should not be understood as a limitation of the present invention.

[0021] like Figure 1 As shown, an embodiment of the present invention provides a centrifugal device, comprising a cup body 100 provided with a centrifugal space 110; the cup body 100 comprises a bottom plate 120 located at the bottom of the centrifugal space 110, and an annular side plate 130 arranged around the bottom plate 120, the annular side plate 130 comprises a straight plate section 131 and an inclined section 132 connected between the straight plate section 131 and the bottom plate 120; the cup body 100 can rotate around a rotation axis; along the direction from top to bottom of the rotation axis, the distance between the inclined section 132 and the rotation axis gradually decreases; a pipetting channel 121 is provided on the bottom plate 120; a first inlet and outlet 1211 and a second inlet and outlet 1212 are respectively provided at both ends of the pipetting channel 121, the pipetting channel 121 is connected to the centrifugal space 110 through the first inlet and outlet 1211, and an inclined surface 1321 of the inclined section 132 close to the rotation axis overlaps and is connected with at least a portion of an edge of the first inlet and outlet 1211.

[0022] It can be understood that liquid (for example, fluids such as cell fluid, separation fluid or gas) can be input into or output from the centrifugal space 110 of the cup body 100 through the pipetting channel 121. The cup body 100 is in a state of rotating around the rotation axis during the centrifugation process. Therefore, when the centrifugal device rotates around the rotation axis at a certain speed (for example, the second preset speed mentioned later), if the centrifugal space 110 contains liquid (for example, cell fluid including target cells), it adheres to the straight plate section 131 and the inclined section 132 of the annular side plate 130 under the action of centrifugal force to form a liquid ring (that is, the liquid adheres to the side wall of the centrifugal space 110, thereby forming a cavity in the middle of the liquid). When the centrifugal device rotates and causes the liquid in the centrifugal space 110 to form a liquid ring, all the liquid in the liquid ring covering the first port and outlet 1211 can be discharged from the centrifugal space 110 through the first port and outlet 1211 through the first port and outlet 1211 (after the liquid covering the first port and outlet 1211 is extracted, the liquid located on the side of the first port and outlet 1211 close to the rotation axis flows outward under the action of centrifugal force and then re-covers the first port and outlet 1211, so that it can continue to be extracted through the first port and outlet 1211 until no liquid is covered on the first port and outlet 1211) through the pipetting channel 121, and the liquid ring remaining in the centrifugal space 110 will be extracted to the position corresponding to the first port and outlet 1211 and can no longer be extracted.

[0023] If it is necessary to drain the liquid in the centrifugal space 110 after the centrifugal operation is completed, at this time, the liquid in the centrifugal space 110 can flow downward along the inclined section 132 under the action of gravity. If the preset centrifugal force is 1G (Gravity), since the inclined surface 1321 of the inclined section 132 close to the rotation axis overlaps and connects with at least a portion of the edge of the first inlet and outlet 1211, the liquid in the centrifugal space 110 is guided to the overlapping connection position by the inclined section 132, and enters the first inlet and outlet 1211 after passing through the overlapping connection position, and then the liquid is completely discharged through the pipetting channel 121, and the liquid residue will not be accumulated at the bottom of the centrifugal space. Specifically, the angle between the inclined surface 1321 of the inclined section 132 close to the rotation axis and the straight plate section 131 can be set as needed. In one embodiment, the angle between the inclined surface 1321 of the inclined section 132 close to the rotation axis and the straight plate section 131 is 100° to 160°. Furthermore, the angle between the inclined surface 1321 of the inclined section 132 close to the rotation axis and the straight section 131 is 135°.

[0024] The present invention provides an inclined section 132 connected to the bottom plate 120 on the annular side plate 130, and overlaps and connects the inclined surface 1321 of the inclined section 132 close to the rotation axis with at least a portion of the edge of the first inlet and outlet 1211 of the pipetting channel 121, thereby guiding the liquid through the inclined surface 1321 and the overlapping connection position between the bottom surface of the inclined surface 1321 and the edge of the first inlet and outlet 1211, so as to quickly introduce the liquid in the centrifugal space 110 from the overlapping connection position into the first inlet and outlet 1211 through the inclined surface 1321, and then discharge all the residual liquid in the centrifugal space 110 through the pipetting channel 121 as much as possible, thereby avoiding liquid residue and improving the pipetting efficiency. Compared with the solution without the inclined surface 1321 or the overlapping connection position, the present invention has a faster speed for the liquid to flow to the first inlet and outlet 1211, and less residual liquid.

[0025] In one embodiment, the inclined section 132 is an annular inclined section; the first inlet and outlet 1211 is arranged between the inclined surface 1321 and the rotation axis. It can be understood that the inclined section 132 is an annular inclined section, that is, the inclined section 132 completely covers the bottom edge of the bottom plate 120, and the projection of the inclined section on the bottom plate 120 is annular. The bottom edge of the inclined surface 1321 of the inclined section 132 and the edge of the first inlet and outlet 1211 can be connected by points or lines. The first inlet and outlet 1211 is located at the lowest position of the inclined section 132, and the residual liquid can be gathered at the first inlet and outlet 1211 under the action of gravity, and then discharged through the pipetting channel 121.

[0026] In one embodiment, when the bottom edge of the inclined surface 1321 is connected to the edge of the first inlet and outlet 1211 by a line, the first inlet and outlet 1211 may be annular, that is, the projection of the first inlet and outlet 1211 on the bottom plate 120 is annular, and the outer ring of the annular first inlet and outlet 1211 is fitted and connected to the annular bottom edge of the inclined surface of the annular inclined section, that is, the overlapping connection position of the annular first inlet and outlet 1211 and the annular inclined section is also a closed annular connecting line, so that the liquid left at any position of the inclined surface 1321 can directly flow into the annular first inlet and outlet 1211 through the annular connecting line, and then be discharged through the pipetting channel 121, further avoiding liquid residue in the centrifugal space 110.

[0027] Understandably, when the inclined section 132 is an annular inclined section with uniform width (the width of the projection ring of the inclined section 132 on the bottom plate 120 is consistent), since the inclined surface 1321 of the inclined section 132 close to the rotation axis overlaps and connects with at least a portion of the edge of the first inlet and outlet 1211, when the centrifugal device rotates and causes the liquid in the centrifugal space 110 to form a liquid ring, after all the liquid in the liquid ring covering the first inlet and outlet 1211 is discharged from the centrifugal space 110 through the first inlet and outlet 1211 through the pipetting channel 121, the maximum thickness of the liquid ring remaining in the centrifugal space 110 will be equal to the width of the projection ring of the inclined section 132 on the bottom plate 120. The width of the projection ring of the inclined section 132 on the bottom plate 120 can be set as needed, so that the width of the liquid ring remaining in the centrifugal space 110 can be controlled, and the volume of the liquid ring remaining in the centrifugal space 110 can be controlled by the size parameters of the centrifugal space 110. In addition, since the annular inclined section is arranged at the edge of the bottom plate 120, when the centrifugal device rotates and causes the cell fluid containing the target cells in the centrifugal space 110 to form a liquid ring, the target cells will be directly attached to the straight plate section 131 and the inclined section 132. At this time, since there is a certain distance between the bottom edge of the inclined section 132 and the straight plate section 131, when the liquid is discharged through the first inlet and outlet 1211, the inclined section 132 will form a certain blocking force, so that the target cells attached to the straight plate section 131 and the inclined section 132 will basically not be discharged with the liquid through the first inlet and outlet 1211, thereby improving the purity of the target cells in the cell fluid remaining in the centrifugal space 110 when a concentration operation is required.

[0028] like Figure 1 As shown, in one embodiment, the inclined section 132 is a C-shaped inclined section with an opening; the first inlet and outlet 1211 is located in the opening. It can be understood that the inclined section 132 is a C-shaped inclined section, that is, the inclined section 132 is a ring-shaped inclined section with a notch (that is, an opening), and at this time, the projection of the inclined section 132 on the bottom plate 120 is C-shaped. The first inlet and outlet 1211 is located in the opening, and the inclined surface 1321 of the inclined section 132 close to the rotation axis overlaps and connects with at least a part of the edge of the first inlet and outlet 1211, that is, the first inlet and outlet 1211 is located at the lowest position of the inclined section 132 and is connected to the inclined surface 1321 by point or line. The liquid in the centrifugal space can be gathered at the first inlet and outlet 1211 under the action of gravity, and then discharged through the pipetting channel 121.

[0029] In one embodiment, the centrifugal device further includes a liquid pushing plate (not shown) connected to the bottom plate 120 and located in the centrifugal space 110; when the cup body 100 rotates around the rotation axis, the liquid pushing plate rotates synchronously with the cup body 100, thereby pushing the liquid to rotate synchronously in the same rotation direction as the cup body 100. It can be understood that when the cup body 100 rotates, the liquid pushing plate rotates synchronously with the cup body 100, thereby reducing the relative movement between the outer layer liquid near the annular side plate 130 and the inclined section 132 and the inner layer liquid near the rotation axis. The power source for the rotation of the inner layer liquid is no longer the viscous resistance between the outer layer liquid and the inner layer liquid, but the thrust of the liquid pushing plate, thereby avoiding the shearing effect on the target cells in the liquid when the viscous resistance acts on the target cells, and avoiding the shear damage to the target cells during the centrifugation process. Further, the specific number of liquid pushing plates can be 1, 2, 3 or 4, etc., which is not specifically limited here.

[0030] like Figure 1 As shown, in some embodiments, the centrifugal device further includes a cup cover 140 installed on the side of the annular side plate 130 away from the bottom plate 120; the cup cover 140 is provided with an access channel 141; the access channel 141 is connected to the pipetting channel 121 through the second access port 1212. It can be understood that the cup cover 140 and the annular side plate 130 can be detachably connected (the detachable method can include but is not limited to a snap connection, etc.) or can be an integral connection structure. The specific structural design of the access channel 141 is not limited to Figure 1 As shown in, the flow channel can be specifically designed according to the needs. For example, different control valves can be designed on the inlet and outlet channels 141 to control the opening and closing of the inlet and outlet channels 141, and pistons can be set at the position where the inlet and outlet channels 141 are connected to the cup body 100 or other components to cooperate with sealing or flow channel direction change. The end of the inlet and outlet channel 141 away from the second outlet is connected to the external structure, so that the fluid (for example, fluid such as cell fluid, separation liquid or gas) can be input or output from the centrifugal space 110 of the cup body 100 through the inlet and outlet channel 141. The cup body 100 is in a state of rotating around the rotation axis during the centrifugation process. Therefore, the second inlet and outlet 1212 is generally arranged at the rotation axis (not easy to be displaced when the cup body 100 rotates), and the inlet and outlet channel 141 is generally also arranged at the rotation axis (will not be displaced when the cup body 100 rotates) and is connected to the second inlet and outlet 1212 through a connection method such as a rotating seal.

[0031] Furthermore, the cup cover 140 is provided with an air hole (not shown) connected to the centrifugal space 110. There is at least one air hole, and a breathable membrane is arranged in the air hole. Specifically, when the centrifugal device is used for cell culture, the breathable membrane can prevent external bacteria from entering the centrifugal space 110. At the same time, the breathable membrane can realize gas exchange between the centrifugal space 110 and the outside world, ensuring the normal respiration of the cells in the centrifugal space 110. Furthermore, the breathable membrane is preferably a hydrophobic breathable membrane.

[0032] like Figure 1 As shown, in one embodiment, the pipetting channel 121 includes a bending section 1213 and a pipetting section 1214 connected to the bending section 1213; the first inlet and outlet 1211 is arranged at one end of the bending section 1213 away from the pipetting section 1214; the second inlet and outlet 1212 is arranged at one end of the pipetting section 1214 away from the bending section 1213; the minimum value of the distance between each inflection point on the bending section 1213 and the rotation axis is greater than the maximum value of the distance between any point on the first inlet and outlet 1211 and the rotation axis. It can be understood that the bending section 1213 can be one or more, and the inflection point refers to the point where each bending section 1213 turns, and the inflection point includes an outer inflection point and an inner inflection point. For example, in Figure 1 In the embodiment shown, the inflection point includes an outer inflection point located at the leftmost side of the bending section 1213, and also includes an inner inflection point located at a position corresponding to the outer inflection point (i.e. Figure 1 In this embodiment, the minimum value of the distance between each inflection point on the bending section 1213 and the rotation axis is greater than the maximum value of the distance between any point on the first entrance 1211 and the rotation axis, so that the bending section 1213 can always be located outside the entire first entrance 1211 away from the rotation axis (for example, Figure 1 Only one bending section 1213 is provided in the centrifugal space 110, and the bending section 1213 is located on the outer side of the first inlet and outlet 1211 away from the rotation axis). The bending section 1213 is located on the side of the inclined surface 1321 away from the rotation axis, that is, the bending section 1213 is bent in a direction away from the rotation axis; thus, when the liquid in the centrifugal space 110 is extracted through the pipetting channel 121, the liquid will enter the bending section 1213 from the first inlet and outlet 1211. Since the bending section 1213 is bent in a direction away from the rotation axis, the flow direction of the fluid entering the bending section 1213 from the first inlet and outlet 1211 will be from the rotation axis toward the annular side plate 130.

[0033] In a specific embodiment, when the centrifugal device rotates and forms a liquid ring in the centrifugal space 110, the cup body 100 is in a state of rotating around the rotation axis during the centrifugation process, and the direction of the centrifugal force is also from the rotation axis toward the annular side plate 130. At this time, if the liquid is extracted through the first inlet and outlet 1211, the extracted liquid will never generate bubbles during the extraction process until the liquid is extracted to the outer edge of the first inlet and outlet 1211 away from the rotation axis, so that the pipeline connected to the pipetting channel 121 can detect the liquid flow state through a sensor (such as a bubble sensor, etc.) (for example: when the gas is detected by the sensor, it is confirmed that the extraction is completed. If there are bubbles in the pipeline, it may cause misjudgment). Specifically, when the bending section 1213 is not provided, after the liquid is extracted to the inner edge close to the rotation axis, in the process of the liquid being extracted from the inner edge to the outer edge of the first inlet and outlet 1211 away from the rotation axis, the extracted liquid will always contain certain bubbles, so that the bubbles in the extracted liquid affect the next use of the liquid. For example, when the bubble sensor is used to determine whether the liquid in the centrifugal space has been exhausted, the bubbles in the extracted liquid will affect the measurement result of the bubble sensor, and the liquid extraction may be terminated prematurely according to the erroneous measurement result, thereby bringing adverse effects to the subsequent steps; for example, if the next step calculation is required based on the volume of the extracted liquid, at this time, the gas contained in the extracted liquid will affect the measurement result of the volume, etc.; in this embodiment, after the bending section 1213 is provided, in the process of the liquid being extracted from the inner edge to the outer edge of the first inlet and outlet 1211 away from the rotation axis, the liquid will always cover the entire cross-section of the tube section corresponding to the inflection point of the bending section, so that the bubbles will not be extracted from the pipetting channel 121 with the liquid, thereby ensuring the measurement or extraction accuracy.

[0034] like Figure 1As shown, in some embodiments, the bottom plate 120 is provided with a pipetting groove 122 and a blind hole 123 connected to the pipetting groove 122; the centrifugal device further comprises a mounting portion 200 and a guide portion 300 connected to the mounting portion 200 on a side away from the rotation axis; the mounting portion 200 is inserted into the pipetting groove 122 and forms the pipetting channel 121 with the groove wall of the pipetting groove 122; the guide portion 300 is inserted into the blind hole 123 and forms the bending section 1213 between the blind hole 123. It can be understood that the pipetting section 1214 and / or the bending section 1213 can be generated by directly processing the bottom plate 120, or can be formed by enclosing multiple parts. In this embodiment, in order to reduce the difficulty of processing, the pipetting section 1214 is inserted into the pipetting groove 122 by the mounting portion 200 and is surrounded by the groove wall of the pipetting groove 122, and the bending section 1213 is inserted into the blind hole 123 by the guide portion 300 and is surrounded by the blind hole 123.

[0035] In one embodiment, the bending section 1213 includes U-shaped bending sections and / or J-shaped bending sections that are interconnected. Figure 1 In the centrifugal device shown in FIG. 1 , only one bending section 1213 is provided. Since the lengths of the first inlet and outlet 1211 and the pipetting section 1214 can be set according to requirements, Figure 1 The bending section 1213 can be U-shaped or J-shaped according to the different length settings of the first inlet and outlet 1211 and the pipetting section 1214; the bending section 1213 can also be set as required to be composed of multiple U-shaped bending sections and / or J-shaped bending sections connected to each other, which will not be repeated here.

[0036] In one embodiment, the mounting portion 200 is connected to the liquid pushing plate, and the liquid pushing plate is connected to the bottom plate 120 through the mounting portion 200. It can be understood that the liquid pushing plate is connected to the side of the mounting portion 200 away from the bottom plate 120, and the liquid pushing plate and the mounting portion 200 can be detachably connected or integrally connected. In this way, the liquid pushing plate is fully utilized, and components are reduced while reducing the difficulty of processing.

[0037] In some embodiments, a liquid-passing gap is provided between the liquid-pushing plate and the annular side plate 130 of the cup body 100; a preset distance is provided between the side of the liquid-pushing plate away from the liquid-passing gap and the rotation axis, and the preset distance is greater than the liquid-passing gap. In this embodiment, during the centrifugation of the cup body 100, the target cells in the sample liquid gradually approach the annular side plate 130 of the cup body 100, pass through the liquid-passing gap, and finally adhere to the annular side plate 130 of the cup body 100, thereby avoiding the target cells from being concentrated and accumulated on the liquid-pushing plate. In addition, the sample liquid can freely pass through the liquid-passing gap, thereby ensuring that the liquid level heights on both sides of the liquid-pushing plate remain consistent. Setting the preset distance between the liquid-pushing plate and the rotation axis requires providing a certain operating space for the installation and removal of the liquid-pushing plate. Understandably, since the liquid-passing gap should not be set too large, the liquid-pushing plate cannot achieve a good liquid-pushing effect when the liquid-passing gap is set too large, and the sample liquid close to the rotation axis has a smaller linear velocity during the centrifugation process, the interval setting of the preset distance has little effect on the centrifugal effect of the sample liquid (and it needs to provide a larger operating space), therefore, the preset distance needs to be greater than the liquid-passing gap.

[0038] like Figure 2 As shown, an embodiment of the present invention further provides a sample separation method, which is performed by the centrifugal device, and the sample separation method includes:

[0039] S10, control the first preset dose of separation liquid to flow into the centrifugal space 110 in the cup body 100 of the centrifugal device. In one embodiment, by controlling the peristaltic pump to rotate (clockwise or counterclockwise, or other liquid pumps) to make the separation liquid flow into the centrifugal space 110, the first preset dose of separation liquid can be accurately controlled to flow into the centrifugal space 110 in the cup body 100 of the centrifugal device. The first preset dose of separation liquid can flow into the centrifugal space 110 in the cup body 100 of the centrifugal device through the pipetting channel 121. The first preset dose can be set according to actual conditions, as long as it plays a buffering role when the sample liquid enters the centrifugal space 110 in the cup body 100, for example, considering the length of the pipeline or the capacity of the centrifugal space 110 in the cup body 100. In one embodiment, the first preset dose is 120mL to 150mL, so as to ensure that after the separation liquid enters the centrifugal space 110 in the cup body 100 through the pipeline, it plays a buffering role for the sample liquid entering the centrifugal space 110 in the cup body 100 in step S20. It can be understood. In step S10, the centrifugal device can also inject the separation liquid after starting the cup body 100 to rotate (the specific rotation speed can be set according to demand, for example, set to be the same as the first preset speed. In this case, the first preset speed can be maintained in step S20 without being turned on again).

[0040] S20, after controlling the cup body 100 to rotate around the rotation axis at a first preset speed, controlling the second preset dose of sample liquid to flow into the centrifugal space 110 at a preset flow rate. It can be understood that controlling the cup body 100 to rotate around the rotation axis at the first preset speed is to control the cup body 100 to start the centrifugal operation. It can be understood that controlling the cup body 100 to rotate around the rotation axis at the first preset speed can be before step S10, that is, controlling the cup body 100 to rotate around the rotation axis at the first preset speed, and then controlling the first preset dose of separation liquid to flow into the centrifugal space 110 in the cup body 100 of the centrifugal device; controlling the cup body 100 to rotate around the rotation axis at the first preset speed can also be after step S10, or simultaneously with step S10, thereby saving processing time. The range of the preset flow rate can be set to 0 to 300 mL / min, and the realization of the preset flow rate can be achieved by rotating a non-contact peristaltic pump to squeeze the pipeline. In this embodiment, the cup body 100 is first controlled to rotate around the rotation axis at a first preset speed, and then the sample liquid of the second preset dose is controlled to flow into the centrifugal space 110 at a preset flow rate, so that the sample liquid can be centrifuged at the same time as entering the centrifugal space 110, thereby saving processing time and improving processing efficiency. The first preset speed can be set according to actual conditions, so as to achieve the best centrifugal effect according to different types of sample liquid. In one embodiment, the first preset speed is in the range of 2000rpm~2500rpm (Revolutions Per minute, revolutions per minute), so as to achieve the best centrifugal effect for the sample liquid containing PBMC cells (peripheral blood mononuclear cell, peripheral blood mononuclear cells) (the PBMC cells in the sample liquid are the target cells). The second preset dose can be set according to actual conditions, generally considering factors such as the capacity of the sample liquid bag or the size of the centrifugal space 110. In one embodiment, the second preset dose is 150mL~200mL.

[0041] S30, control the cup body 100 to rotate at a second preset speed for a first preset time to centrifuge the liquid in the centrifugal space 110 in the cup body 100, so that the liquid adheres to the straight plate section 131 and the inclined section 132 to form a liquid ring; the top surface of the liquid ring adheres to the top wall of the centrifugal space 110, and the bottom surface of the liquid ring covers the first inlet and outlet 1211. It can be understood that after the first preset dose of separation liquid and the second preset dose of sample liquid are injected into the centrifugal space 110, the liquid in the centrifugal space 110 can be continuously centrifuged. The first preset time can be set according to actual conditions, so as to achieve the best centrifugal effect according to different sample liquid types or doses, such as making the liquid stratified into multiple liquid layers, and then in the subsequent steps, the liquid in the liquid layer located on the inner side of the liquid ring is discharged through the first inlet and outlet 1211, and only the liquid layer located on the outer side of the first inlet and outlet 1211 is retained in the centrifugal space 110, so as to achieve accurate discharge of the liquid. In one embodiment, the first preset time is 150 seconds, so that the best centrifugation effect can be achieved for a sample solution of preferably 150 mL. The second preset speed can be set according to actual conditions (the second preset speed can be the same as or different from the first preset speed), so as to achieve the best centrifugation effect according to different sample liquid types. In one embodiment, the second preset speed is preferably 2000 rpm, so that the best centrifugation effect can be achieved for a sample solution containing PBMC cells (target cells).

[0042] S40, the liquid in the liquid ring covering the first inlet and outlet 1211 is discharged from the centrifugal space 110 through the pipetting channel 121. It can be understood that when the cup body 100 continues to rotate at the first preset speed or the second preset speed, the step S40 can discharge the liquid in the liquid ring covering the first inlet and outlet 1211 from the centrifugal space 110 through the pipetting channel 121, so that a preset concentrated volume of sample liquid (for example,) can be obtained. Specifically, when the sample liquid is cell liquid, after the liquid in the centrifugal space 110 in the cup body 100 is centrifuged, the cup body 100 continues to rotate so that the sample liquid forms a liquid ring, and the liquid is stratified into multiple liquid layers. The target cells in the sample liquid (the target cells are generally relatively heavy, such as PBMC cells) will be located in the liquid layer close to the annular side plate 130 of the cup body 100. When the liquid in the centrifugal space 110 is extracted through the pipetting channel 121, the liquid in the liquid layer located on the inner side of the liquid ring will be discharged through the first inlet and outlet 1211, and the liquid layer in the centrifugal space 110 located on the outer side of the first inlet and outlet 1211 will be retained, and the retained liquid layer is the sample liquid with the minimum concentrated volume containing the target cells. The above method improves the cell centrifugation efficiency. When the cup body 100 is not rotating, since the first inlet and outlet 1211 is located on the bottom plate 120 and at least a portion of the edge of the first inlet and outlet 1211 overlaps and is connected with the inclined surface 1321 of the inclined section 132 close to the rotation axis, the liquid in the centrifugal space 110 can flow downward along the inclined section 132 under the action of gravity to the overlapping connection position between the first inlet and outlet 1211 and the inclined section 132. At this time, after step S30, the liquid in the centrifugal space 110 can also be completely discharged through the overlapping connection position through the first inlet and outlet 121 and the pipetting channel 121 to avoid liquid residue.

[0043] In some embodiments, the pipetting channel 121 includes a bending section 1213 and a pipetting section 1214 connected to the bending section 1213; the first inlet and outlet 1211 is arranged at one end of the bending section 1213 away from the pipetting section 1214; the second inlet and outlet 1212 is arranged at one end of the pipetting section 1214 away from the bending section 1213; the minimum value of the distance between each inflection point on the bending section 1213 and the rotation axis is greater than the maximum value of the distance between any point on the first inlet and outlet 1211 and the rotation axis; further, the step S40, the liquid in the liquid ring covering the first inlet and outlet 1211 is discharged from the centrifugal space 110 through the pipetting channel 121, includes:

[0044] S41, the liquid in the liquid ring covering the first inlet and outlet 1211 is discharged from the centrifugal space 110 through the first inlet and outlet 1211, the bent section 1213 and the transfer section 1214 of the transfer channel 121 in sequence. It can be understood that when the cup body 100 continues to rotate at the first preset speed or the second preset speed, the sample liquid forms a liquid ring; the bent section 1213 can always be located outside the entire first inlet and outlet 1211 away from the rotation axis, so the flow direction of the fluid entering the bent section 1213 from the first inlet and outlet 1211 will be from the rotation axis toward the annular side plate 130, so the liquid covering the first inlet and outlet 1211 can be discharged from the centrifugal space 110 through the first inlet and outlet 1211, the bent section 1213 and the transfer section 1214 of the transfer channel 121 in sequence, and no bubbles will be generated during the discharge process. The specific process can refer to the specific embodiment of the above-mentioned centrifugal device, which will not be repeated here.

[0045] It is understandable that the above sample separation method is not limited to that described in the above embodiment. The specific configuration of the sample separation method of the present application corresponds to the above centrifugal device one by one, and will not be described in detail here.

[0046] An embodiment of the present invention further provides a controller, the controller comprising a processor and a memory, the memory storing an executable program, and the processor being configured to execute the executable program to implement the sample separation method.

[0047] The specific settings of the controller of the present application correspond to the above-mentioned sample separation method one by one, and will not be repeated here. Each module in the above-mentioned controller can be implemented in whole or in part by software, hardware and a combination thereof. The above-mentioned modules can be embedded in or independent of the controller in the form of hardware, or can be stored in the controller in the form of software, so that the controller can call and execute the operations corresponding to the above modules.

[0048] An embodiment of the present invention further provides a computer-readable storage medium, wherein the computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the above-mentioned sample separation method is implemented.

[0049] Those of ordinary skill in the art can understand that all or part of the processes in the above-mentioned embodiment methods can be implemented by instructing related hardware through computer-readable instructions, and the computer-readable instructions can be stored in a non-volatile readable storage medium or a volatile readable storage medium. When the computer-readable instructions are executed, they may include the processes of the embodiments of the above-mentioned methods. Among them, any reference to memory, storage, database or other media used in the embodiments provided in this application may include non-volatile and / or volatile memory. Non-volatile memory may include read-only memory (ROM), programmable ROM (PROM), electrically programmable ROM (EPROM), electrically erasable programmable ROM (EEPROM) or flash memory. Volatile memory may include random access memory (RAM) or external cache memory. By way of illustration and not limitation, RAM is available in many forms, such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), double data rate SDRAM (DDRSDRAM), enhanced SDRAM (ESDRAM), synchronous link (Synchlink) DRAM (SLDRAM), memory bus (Rambus) direct RAM (RDRAM), direct memory bus dynamic RAM (DRDRAM), and memory bus dynamic RAM (RDRAM), etc.

[0050] Those skilled in the art will clearly understand that for the sake of convenience and brevity of description, only the division of the above-mentioned functional units and modules is used as an example. In actual applications, the above-mentioned functions can be distributed and completed by different functional units and modules as needed, that is, the internal structure of the system can be divided into different functional units or modules to complete all or part of the functions described above.

[0051] The above are merely embodiments of the centrifugal device, sample separation method, controller and medium of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A centrifugal device, characterized in that: The invention comprises a cup body provided with a centrifugal space; the cup body comprises a bottom plate located at the bottom of the centrifugal space, and an annular side plate arranged around the bottom plate, the annular side plate comprises a straight plate section and an inclined section connected between the straight plate section and the bottom plate; the cup body can rotate around a rotation axis; along the direction from top to bottom of the rotation axis, the distance between the inclined section and the rotation axis gradually decreases; A pipetting channel is provided on the bottom plate; a first inlet and a second inlet are respectively provided at both ends of the pipetting channel, the pipetting channel is connected to the centrifugal space through the first inlet and the inclined surface of the inclined section close to the rotation axis overlaps and is connected with at least a portion of an edge of the first inlet and the outlet.

2. The centrifugal device according to claim 1, characterized in that The angle between the inclined surface of the inclined section close to the rotation axis and the straight plate section is 100° to 160°.

3. The centrifugal device according to claim 1, characterized in that The inclined section is an annular inclined section; the first inlet and outlet are arranged between the inclined surface and the rotation axis; or The inclined section is a C-shaped inclined section with an opening; the first entrance and exit is located in the opening.

4. The centrifugal device according to claim 1, characterized in that The centrifugal device also includes a liquid pushing plate connected to the bottom plate and located in the centrifugal space; When the cup body rotates around the rotation axis, the liquid pushing plate rotates synchronously with the cup body, thereby pushing the liquid to rotate synchronously in the same rotation direction as the cup body.

5. The centrifugal device according to claim 1, characterized in that The centrifugal device also includes a cup cover installed on the side of the annular side plate away from the bottom plate; the cup cover is provided with an inlet and outlet channel; the inlet and outlet channel is connected to the transfer channel through the second inlet and outlet.

6. The centrifugal device according to any one of claims 1 to 5, characterized in that The pipetting channel includes a bending section and a pipetting section connected to the bending section; the first inlet and outlet are arranged at one end of the bending section away from the pipetting section; the second inlet and outlet are arranged at one end of the pipetting section away from the bending section; The minimum value of the distance between each inflection point on the bending section and the rotation axis is greater than the maximum value of the distance between any point on the first entrance and exit and the rotation axis.

7. The centrifugal device according to claim 6, characterized in that The bending section includes a U-shaped bending section and / or a J-shaped bending section that are interconnected.

8. The centrifugal device according to claim 6, characterized in that The bottom plate is provided with a pipetting groove and a blind hole connected to the pipetting groove; the centrifugal device also includes a mounting portion and a guide portion connected to the mounting portion on a side away from the rotation axis; the mounting portion is inserted into the pipetting groove and forms the pipetting channel with the groove wall of the pipetting groove; the guide portion is inserted into the blind hole and forms the bending section between the blind holes.

9. A sample separation method, characterized in that: The sample separation method is performed by a centrifugal device according to any one of claims 1 to 5, and the sample separation method comprises: Controlling a first preset dose of separation liquid to flow into the centrifugal space in the cup body of the centrifugal device; After controlling the cup body to rotate around the rotation axis at a first preset speed, controlling a second preset dose of sample liquid to flow into the centrifugal space at a preset flow rate; Controlling the cup body to rotate at a second preset speed for a first preset time to centrifuge the liquid in the centrifugal space in the cup body, so that the liquid adheres to the straight plate section and the inclined section to form a liquid ring; the top surface of the liquid ring adheres to the top wall of the centrifugal space, and the bottom surface of the liquid ring covers the first inlet and outlet; The liquid in the liquid ring covering the first inlet and outlet is discharged from the centrifugal space through the pipetting channel.

10. The sample separation method according to claim 9, characterized in that: The pipetting channel includes a bending section and a pipetting section connected to the bending section; the first inlet and outlet are arranged at one end of the bending section away from the pipetting section; the second inlet and outlet are arranged at one end of the pipetting section away from the bending section; the minimum value of the distance between each inflection point on the bending section and the rotation axis is greater than the maximum value of the distance between any point on the first inlet and outlet and the rotation axis; The step of discharging the liquid in the liquid ring covering the first inlet and outlet from the centrifugal space through the liquid transfer channel comprises: The liquid in the liquid ring covering the first inlet and outlet is discharged from the centrifugal space sequentially through the first inlet and outlet, the bending section and the pipetting section of the pipetting channel.

11. A controller, characterized in that: The controller includes a processor and a memory, wherein the memory stores an executable program, and the processor is configured to execute the executable program to implement the sample separation method according to claim 9 or 10.

12. A computer-readable storage medium storing a computer program, characterized in that: When the computer program is executed by a processor, the sample separation method according to claim 9 or 10 is implemented.

Images