CN120112624A

CN120112624A - Sorting fluid system and related equipment, fluid sorting method and cell sorting method

Info

Publication number: CN120112624A
Application number: CN202380073403.6A
Authority: CN
Inventors: 查朋飞; 朱礼浩; 孟宗发; 罗杰; 邓兵龙; 洪艳; 沈梦哲; 黎宇翔; 陈奥; 章文蔚; 徐讯
Original assignee: BGI Shenzhen Co Ltd
Current assignee: BGI Shenzhen Co Ltd
Priority date: 2023-01-03
Filing date: 2023-01-03
Publication date: 2025-06-06
Also published as: WO2024145732A1

Abstract

The sorting fluid system comprises a centrifugal mechanism, pipeline consumable materials and a pipeline time sequence control assembly, wherein the centrifugal mechanism is used for accommodating a sample accommodating part, a rotary connecting part which is respectively communicated with the sample accommodating part and the pipeline consumable materials is arranged on the centrifugal mechanism, the centrifugal mechanism drives liquid in the sample accommodating part to realize centrifugal layering and extrude corresponding layers of liquid into the pipeline consumable materials, and the pipeline time sequence control assembly is arranged on the pipeline consumable materials and is used for controlling the liquid entering the pipeline consumable materials to flow according to a preset time sequence and a preset path. The provided sorting fluid system can directly obtain target cells, and reduce the risk of cell pollution. In addition, the sample containing piece is extruded through the centrifugal machine, all layers are extruded into the pipeline consumable, the fact that a pump is independently arranged to pump all layers in the sample containing piece is avoided, and the system structure is simplified.

Description

Sorting fluid system, related equipment, fluid sorting method and cell sorting method

Technical Field

The application relates to the technical field of cell sorting, in particular to a sorting fluid system, a cell sorting system, a sorter, a fluid sorting method and a cell sorting method.

Background

The market space of cell therapy in China is increased from 13 hundred million yuan in 2021 to 584 hundred million yuan in 2030, and the annual acceleration rate is up to 53%. The treatment of cellular genes and the like requires centrifugation of blood and the like by a centrifuge, and each component and the like after centrifugal separation needs to be pumped out by a pump and the like and transported to another device for subsequent processes such as magnetic separation, and the like, so that the target liquid in the component cannot be directly obtained, and the pollution risk is increased in the transfer process. In addition, after centrifugation, additional pumping pumps are required to pump out components and the like, and the whole system has a complex structure.

In summary, how to reduce the complexity of the system, facilitate the direct acquisition of the target liquid, and reduce the risk of contamination of the target liquid is a problem to be solved by those skilled in the art.

Content of the application

The application provides a sorting fluid system, a cell sorting system, a sorter, a fluid sorting method and a cell sorting method, which are used for reducing the complexity of the system, facilitating the direct acquisition of target liquid and reducing the pollution risk of the target liquid.

In order to achieve the above object, the present application provides the following technical solutions:

in a first aspect, the present application provides a sort fluid system comprising a centrifugal mechanism, a tubing consumable, and a tubing timing control assembly;

The centrifugal mechanism is used for accommodating a sample accommodating part, a rotary connecting part which is respectively communicated with the sample accommodating part and the pipeline consumable is arranged on the centrifugal mechanism, and the centrifugal mechanism drives the sample in the sample accommodating part to realize centrifugal layering and extrudes corresponding layer components into the pipeline consumable;

The pipeline time sequence control assembly is arranged on the pipeline consumable and used for controlling samples or components entering the pipeline consumable to flow according to a preset time sequence and a preset path.

In some possible aspects of the present application, the sorting fluidic system further includes a sample source holding member, a waste liquid holding member, and a non-waste liquid component collecting holding member, which are respectively communicated with the consumable materials of the pipeline;

The sample source containing piece is internally provided with a sample source, and the pipeline time sequence control assembly can control the on-off of the sample source containing piece and the sample containing piece so as to realize the input of a sample into the sample containing piece;

The pipeline time sequence control assembly can also control the on-off of the waste liquid containing piece and the sample containing piece so as to realize the input of waste liquid in components separated by the sample containing piece into the waste liquid containing piece;

The pipeline time sequence control assembly can also control the on-off of the non-waste liquid component collecting and containing piece and the sample containing piece so as to realize the input of the non-waste liquid component in the components separated by the sample containing piece into the non-waste liquid component collecting and containing piece.

In some possible solutions of the present application, the non-waste liquid component collecting and containing member includes a target containing member and a non-target containing member, which are respectively communicated with the consumable of the pipeline;

The target holding piece is used for holding target liquid in the non-waste liquid component separated by the sample holding piece, and the non-target holding piece is used for holding non-target liquid in the non-waste liquid component separated by the sample holding piece.

In some possible aspects of the application, the sorting fluidic system further comprises a buffer containing member in communication with the tubing consumable;

The buffer solution containing piece is used for containing buffer solution, and the pipeline time sequence control assembly can control the buffer solution containing piece to be communicated with each preset path in the pipeline consumable, so that the buffer solution is input into the pipeline consumable to positively clean each preset path of the pipeline consumable.

In some possible aspects of the application, the sorting fluid system further comprises a reverse flushing fluid holding member in communication with the tubing consumable;

The pipeline time sequence control assembly can control the conduction of the reverse flushing liquid containing piece and each preset path in the pipeline consumable material so as to reversely clean each preset path of the pipeline consumable material by inputting cleaning liquid into the pipeline consumable material.

In some possible solutions of the present application, the pipeline timing control assembly may control the reverse flushing liquid containing member to be in communication with the buffer liquid containing member, so as to convey the buffer liquid in the buffer liquid containing member into the reverse flushing liquid containing member;

The cleaning liquid in the reverse flushing liquid containing part is buffer liquid.

In some possible aspects of the application, the sorting fluidic system further comprises a back flush waste collection member in communication with the tubing consumable;

The pipeline timing control assembly can also control the reverse flushing waste liquid collecting piece to be switched to collect the waste liquid after the pipeline consumable is reversely cleaned.

In some possible aspects of the application, the sort fluid system further comprises a filter;

the filtering piece is communicated with the pipeline consumable, and the pipeline time sequence control assembly can control the filtering piece to be communicated with the sample containing piece so as to filter non-waste liquid components in the output components of the sample containing piece.

In some possible aspects of the application, the sorting fluid system further comprises a power member mounted on the tubing consumable;

the power piece is used for providing power.

In some possible aspects of the present application, the power member is at least disposed on a path of a consumable material in a pipeline through which the sample in the sample source holder enters the sample holder in the centrifuge, so as to convey the sample in the sample source holder into the sample holder.

In some possible aspects of the application, the sorting fluidic system further comprises an airtight detection element;

The airtight detection piece is installed on the pipeline consumable, and at least 1 airtight detection piece is arranged at two ends of the power piece respectively and used for detecting the air tightness of the pipeline consumable.

In some possible solutions of the present application, the airtight detection component is a pressure sensor clamped on the pipeline consumable.

In some possible solutions of the present application, the pipeline timing control assembly includes a plurality of on-off valves for controlling on-off of each position of the pipeline consumable.

In some possible solutions of the present application, the on-off valve is clamped on the pipeline consumable.

In some possible aspects of the application, the pipeline timing control assembly further comprises a detector group clamped on the pipeline consumable to detect the condition of the sample or each component flowing through the pipeline consumable.

In a second aspect, the present application provides a cell sorting system comprising a magnetic bead holder, a binding solution holder, an eluent holder, a sorting column, and a sorting fluidic system according to any of the preceding claims;

The magnetic bead containing piece, the binding solution containing piece, the eluent containing piece and the sorting column are respectively communicated with pipeline consumable materials in the sorting fluid system;

the pipeline time sequence control assembly in the sorting fluid system can control the connection between the binding solution containing piece and the sample containing piece in the sorting fluid system so as to convey the binding solution in the binding solution containing piece into the sample containing piece, and the binding solution and the sample in the sample containing piece are uniformly mixed through centrifugal motion of the centrifugal mechanism, and target cells are marked;

The pipeline time sequence control assembly can control the magnetic bead containing piece to be communicated with the sample containing piece so as to convey the magnetic bead solution in the magnetic bead containing piece into the sample containing piece, and the magnetic bead solution is uniformly mixed with a sample in the sample containing piece through centrifugal motion of the centrifugal mechanism and combined with the marked target cells;

the pipeline time sequence control assembly can control the separation column to be communicated with the sample containing piece, the sample containing piece is extruded through the centrifugal mechanism to convey the sample mixed with the binding solution and the magnetic beads into the separation column, the separation column in a magnetic field state can magnetically adsorb the magnetic beads, and non-target cells in the sample are discharged into the non-target containing piece of the separation fluid system;

the pipeline time sequence control assembly can control the separation column to be communicated with the eluent containing piece so as to inject the eluent in the eluent containing piece into the separation column in a state of being separated from a magnetic field, so as to elute the target cells into the target containing piece of the separation fluid system.

In some possible aspects of the application, the cell sorting system further comprises a pre-sorting column;

the pre-separation column is communicated with the pipeline consumable;

The pipeline time sequence control assembly can control the pre-sorting column to be respectively communicated with the sample containing piece and the sorting column, so that samples in the sample containing piece flow through the pre-sorting column and then enter the sorting column.

In some possible aspects of the application, the pipeline timing control assembly can control the pre-sorting column to be communicated with a filter element of the sorting fluid system, so that samples in the sample containing element enter the pre-sorting column after being filtered by the filter element.

In some possible aspects of the application, the cell sorting system further comprises a color sensor;

The color sensor is arranged at the position where the pipeline consumable material is communicated with the sample containing piece and is used for detecting each layer of the sample in the sample containing piece which is output after centrifugal layering.

In a third aspect, the present application provides a sorter comprising an equipment mount and a sorting fluidic system according to any one of the preceding claims or a cell sorting system according to any one of the preceding claims;

the sorting fluidic system or the cell sorting system is mounted on the equipment mounting frame.

In some possible solutions of the present application, the device mounting rack is provided with a centrifugal mounting table, a pipeline panel and a controller;

The centrifugal mounting table is used for mounting a centrifugal mechanism of the sorting fluid system;

The pipeline panel is obliquely or vertically arranged with the centrifugal mounting table and is used for mounting pipeline consumable materials, a pipeline time sequence control assembly and the controller of the sorting fluid system;

The controller is respectively in communication connection with the pipeline time sequence control assembly and the centrifugal mechanism.

In some possible aspects of the application, the sorter further comprises a loading magnetic field assembly mounted on the pipeline panel;

the loading magnetic field assembly is used for applying or disconnecting a magnetic field to a sorting column in the cell sorting system.

In some possible solutions of the present application, the loading magnetic field assembly includes a permanent magnet and a driving member, the permanent magnet is provided with a receiving slot for receiving the sorting column, and a driving end of the driving member is connected with the permanent magnet and is used for driving the permanent magnet to approach or depart from the sorting column so as to apply a magnetic field to the sorting column or break the magnetic field;

Or alternatively

The loading magnetic field assembly comprises an electromagnet and a power supply piece, wherein the electromagnet is provided with a containing groove for containing the sorting column, the power supply piece is connected with the electromagnet in a ferroelectric manner, and the power supply piece is used for electrifying or de-electrifying the electromagnet so that the sorting column can apply a magnetic field or disconnect the magnetic field.

In some possible solutions of the present application, the driving member is a rack-and-pinion structure, an electric push rod structure, a hydraulic cylinder or an air cylinder.

In some possible solutions of the present application, a hanging post is provided at the top end of the pipeline panel, and a hook is installed on the hanging post for hanging each containing piece.

In some possible aspects of the present application, the controller includes a controller main body and a touch screen in signal connection with the controller main body.

In a fourth aspect, the present application provides a method of sorting fluids comprising:

providing a sort fluidic system according to any of the preceding claims, and placing a sample holder within the centrifugation mechanism;

starting the centrifugal mechanism to rotate and driving the sample holding piece to centrifugally rotate so as to laminate the sample in the sample holding piece;

Switching the pipeline timing control assembly to conduct the sample holding member and the waste liquid holding member of the sorting fluid system;

starting the centrifugal mechanism to extrude the sample containing piece, and extruding the waste liquid in the separated components in the sample containing piece into the waste liquid containing piece;

switching the pipeline timing control assembly to turn on the sample holder and the non-waste component collection holder of the sorting fluidic system;

and starting the centrifugal mechanism to squeeze the sample containing piece, and conveying each non-waste liquid component separated by the sample containing piece into the corresponding non-waste liquid component collecting containing piece.

In a fifth aspect, the present application provides a cell sorting method comprising:

providing a cell sorting system according to any one of the preceding claims and placing a sample holder within the centrifuge mechanism;

Starting the centrifugal mechanism to centrifugally layer the sample in the sample holding piece;

Switching the pipeline time sequence control assembly to be communicated with the sample containing piece and the waste liquid containing piece, and extruding the waste liquid in the centrifugally layered sample into the waste liquid containing piece by the centrifugal mechanism;

Switching the pipeline time sequence control assembly to be communicated with the combined solution containing piece and the sample containing piece, and conveying the combined solution in the combined solution containing piece into the sample containing piece;

switching the pipeline time sequence control assembly to be communicated with the magnetic bead containing piece and the sample containing piece, and conveying the magnetic bead solution in the magnetic bead containing piece into the sample containing piece;

Switching the pipeline time sequence control assembly to the state that the sample holding part is communicated with the sorting column, applying a magnetic field to the sorting column, extruding the sample holding part into the sorting column under the magnetic field state by the centrifugal mechanism, adsorbing magnetic beads in the sorting column, and discharging non-target cells in a sample into the non-target holding part through the sorting column;

And switching the pipeline time sequence control assembly to the eluent containing piece to be communicated with the sorting column, and conveying the eluent in the eluent containing piece into the sorting column so as to elute target cells in the sorting column into the target containing piece.

According to the technical scheme, the application provides a sorting fluid system, when the sorting fluid system is used, a sample containing piece is placed in a centrifugal mechanism and is communicated with a pipeline consumable through a rotary connecting piece, the centrifugal mechanism drives the sample containing piece to rotate so as to realize centrifugal layering of samples in the sample containing piece, the centrifugal mechanism is matched with a switching pipeline time sequence control assembly, the sample containing piece is extruded through the centrifugal mechanism to input separated layers of components into the pipeline consumable, and the components flow according to a preset time sequence and a preset path so as to achieve acquisition of target fluid. The sorting fluid system provided by the application can directly obtain the target fluid, and reduce the risk of pollution of the target fluid by cells. In addition, the sample containing piece is extruded through the centrifugal machine, all layers are extruded into the pipeline consumable, the fact that a pump is independently arranged to pump all layers in the sample containing piece is avoided, and the system structure is simplified.

Drawings

In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It is apparent that the drawings in the following description are only some examples or embodiments of the present application, and it is possible for those of ordinary skill in the art to obtain other drawings from the provided drawings without inventive effort, and to apply the present application to other similar situations from the provided drawings. Unless otherwise apparent from the context of the language or otherwise specified, like reference numerals in the figures refer to like structures or operations.

FIG. 1 is a schematic diagram of a front view of a sorting fluid system according to an embodiment of the present application without a pipeline timing control assembly installed;

FIG. 2 is a schematic diagram of a front view of a sorting fluidic system according to an embodiment of the present application;

FIG.3 is a schematic diagram of a front view of a sorting fluidic system according to another embodiment of the present application;

Fig.4 is a schematic view of a part of a three-dimensional structure of a sorting fluid system according to an embodiment of the present application.

Wherein:

Sorting fluidic system 100, sample holder 200, centrifuge 101, tubing consumable 102, tubing timing control assembly 103, rotary connector 101a, magnetic sorting mechanism 104, sorting column 104a, loading magnetic field assembly 104b, permanent magnet 104b-1, holding tank 104b-1a, pre-sorting column 104c, fluid loading mechanism 105, buffer holder 105a, filter 105b, sample source holder 105c, backflush waste liquid collector 105d, backflush liquid holder 105e, power member 106, airtight detection member 107, component collection mechanism 108, waste liquid holder 108a, destination holder 108b, non-destination holder 108c, first main tube 102-1, second main tube 102-2, third main tube 102-3, loading tube 102-4, waste liquid tube 102-5 the power line 102-6, the distribution line 102-7, the destination line 102-8, the non-destination line 102-9, the fourth main line 102-10, the filter line 102-11, the preseparation line 102-12, the buffer line 102-13, the binding solution line 102-14, the bead line 102-15, the eluent line 102-16, the sample input line 102-17, the first sub-line 102-18, the second sub-line 102-19, the first sampling bag 102-20, the first drip chamber 102-21, the first filter 102-22, the second sampling bag 102-23, the second drip chamber 102-24, the first backup line 102-25, the second backup line 102-26, the adapter 102-27, the second filter 102-28, the third filter 102-29, the first switch valve 103-1, the second switch valve 103-2, the third switch valve 103-3, the fourth switch valve 103-4, the fifth switch valve 103-5, the sixth switch valve 103-6, the seventh switch valve 103-7, the eighth switch valve 103-8, the ninth switch valve 103-9, the tenth switch valve 103-10, the eleventh switch valve 103-11, the color sensor 103-12, the first bubble sensor 103-13, the second bubble sensor 103-14, the twelfth switch valve 103-15, the thirteenth switch valve 103-16, the filtration switch valve 103-17, the pre-selection switch valve 103-18, the buffer switch valve 103-19, the binding solution switch valve 103-20, the magnetic bead switch valve 103-21, the eluent switch valve 103-22, the sample switch valve 103-23, the auxiliary switch valve 103-24, the fourth bubble sensor 103-25, the third bubble sensor 103-26, the device mounting rack 109, the centrifuge mounting rack 109a, the pipeline panel 109b, the controller 109c, the hanging column 109b-1, the hanging column 109b, the cell sorting system 1000-2.

Detailed Description

The application is described in further detail below with reference to the drawings and examples. It is to be understood that the specific embodiments described herein are merely illustrative of the application and not limiting of the application. The described embodiments are only some, but not all, embodiments of the application. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.

Referring to fig. 1-4, a sorting fluid system 100 according to an embodiment of the present application is used to reduce complexity of the system, facilitate direct acquisition of a target fluid, and reduce risk of contamination of the target fluid. It should be noted that, the sorting fluid system 100 is not limited to sorting blood cells, but can be applied to amplification, enrichment, and split charging of cells, and can also be applied to component separation of other multicomponent liquids or preparation, enrichment, and the like of nano immunomagnetic beads.

Specifically, the sorting fluidic system 100 includes a centrifugal mechanism 101, a tubing consumable 102, and a tubing timing control assembly 103.

The centrifugal mechanism 101 is used for accommodating the sample holder 200. It should be noted that, the centrifugal mechanism 101 is a mechanism with the ability to centrifuge and squeeze the sample holder 200, and the squeezing may be achieved by a mechanical structure, or an effect of squeezing may be produced by a change in fluid volume, in this embodiment, taking the mechanical structure to produce squeezing as an example, specifically, the centrifugal mechanism 101 includes a frame, a centrifugal chamber, a squeezing platform, a spindle, a rotation driving assembly, and a flat pushing driving assembly, where the centrifugal chamber is rotatably mounted on the frame, and the centrifugal chamber has an inner cavity. The mandrel is connected with the centrifugal chamber in a synchronous rotation way, and one end of the mandrel extends into the centrifugal chamber and is connected with an extrusion platform arranged in the centrifugal chamber. The extrusion platform is used for bearing the sample containing piece 200, and the rotary driving assembly drives the centrifugal chamber to rotate, so that the sample containing piece to be sorted is driven to perform centrifugal motion. The flat pushing driving assembly drives the mandrel to move in the centrifugal chamber along the direction of the rotation axis of the centrifugal chamber. The flat pushing driving component is rotatably connected with the mandrel, so that the rotation of the mandrel is prevented from being influenced when the mandrel is pushed flat. Specifically, the flat pushing component can be any component capable of realizing the movement of the flat pushing mandrel, and the rotary driving component is any component capable of driving the centrifugal chamber to rotate, and the mandrel is connected with the centrifugal chamber through spline transmission.

The sample holder 200 may be a disposable plastic film product, which is inexpensive and is used for holding samples such as blood. The shape of the sample holder 200 is not limited to a circular shape, but may be an elliptical shape, a polygonal shape, or the like, and may be a piston-type centrifugal cup.

The centrifugal mechanism 101 is provided with a rotary connector 101a which is respectively communicated with the sample holding member 200 and the pipeline consumable 102, and specifically, the rotary connector 101a is a rotary joint or the like so as to realize the communication of a rotary part (the sample holding member 200) and a stationary part (the pipeline consumable 102). The centrifugal mechanism 101 drives the sample in the sample holder 200 to realize centrifugal delamination and squeeze the corresponding layer components into the tubing consumable 102.

The pipeline timing control assembly 103 is installed on the pipeline consumable 102, and is used for controlling samples or components entering the pipeline consumable 102 to flow according to a preset timing sequence and a preset path.

The pipeline timing control assembly 103 may be specifically a switch valve installed at each position of the pipeline consumable 102 and used for controlling on-off of each position, and the switching of different paths is realized by opening and closing each switch valve. The preset time sequence and the preset path are specifically set according to the types of the samples to be sorted.

To facilitate the installation of the various on-off valves, the present application discloses that the on-off valves are clamped on the tubing consumable 102.

The sorting fluid system 100 provided by the application realizes the direct acquisition of the target fluid, avoids the pollution risk in the transfer process caused by the sorting carried to different equipment, i.e. reduces the pollution risk of the target fluid. In addition, the sample containing piece 200 is extruded through the centrifugal mechanism 101, and each layer of centrifugally separated components are extruded into the pipeline consumable 102, so that each layer in the sample containing piece 200 is prevented from being drawn out by a pump, and the system structure is simplified.

In some embodiments, the sorting fluidic system 100 further comprises a fluid loading mechanism 105 and a component collection mechanism 108, wherein the fluid loading mechanism 105 is configured to load a sample or component or the like of fluid into the tubing consumable 102.

Further, the application discloses a fluid loading mechanism 105 comprising a sample source holding member 105c, wherein a sample source is stored in the sample source holding member 105c, specifically, the sample source holding member 105c is a bag-shaped plastic product, and the cost is low, so that the processing and the manufacturing are convenient. It will be appreciated that the sample source holder 105c is a bag-like plastic article and is merely one embodiment of the present application, and that in practice, the sample source holder 105c may be provided as another article.

The sample source holding member 105c is communicated with the pipeline consumable 102, and the pipeline timing control assembly 103 can control the on-off of the sample source holding member 105c and the sample holding member 200 so as to realize the input of a sample into the sample holding member 200. The setting of the sample source holding member 105c realizes the input of the sample into the sample holding member 200, avoids the opening of the centrifugal mechanism 101, and can manually inject the sample into the sample holding member 200 or manually replace the sample holding member 200, thereby saving the manpower and improving the automation. It should be noted that, the number of the sample source holding members 105c may be2 or more, and different samples may be respectively held, so as to implement sorting and use of different samples in the same sorting fluid system 100, thereby improving the versatility of the sorting fluid system 100.

Further, the application discloses that the sample source holding piece 105c is also communicated with the first sampling bag 102-20 through the pipeline consumable 102, so that samples in the sample source holding piece 105c can be sampled conveniently, and the samples can be sampled conveniently at any time for analysis.

Further, as shown in fig. 2, the present application discloses that the pipeline consumable 102 is further connected to the first drip chamber 102-21, and the sample in the sample source holding member 105c is conveyed into the sample holding member 200 after entering the first drip chamber 102-21. The first drip chamber 102-21 is provided to prevent the sample from being sucked back into the sample source holding member 105c, and to smoothly flow down the sample in the first drip chamber 102-21.

The filter 105b may be a plastic bag or other articles.

Further, the application discloses that the component collection mechanism 108 comprises a waste liquid containing member 108a and a non-waste liquid component collection containing member, wherein the waste liquid containing member 108a and the non-waste liquid component collection containing member are respectively communicated with the pipeline consumable 102.

Specifically, the waste liquid containing member 108a and the non-waste liquid component collecting containing member may be bag-shaped plastic products, which is low in cost and convenient for processing and manufacturing. It should be understood that the plastic bag-shaped waste liquid container 108a and the non-waste liquid component collecting container are only one specific embodiment of the present application, and in practical application, the waste liquid container 108a and the non-waste liquid component collecting container may be other products.

The pipeline timing control assembly 103 can also control the on-off of the waste liquid containing member 108a and the sample containing member 200 so as to input the waste liquid separated by the sample containing member 200 into the waste liquid containing member 108 a. The waste liquid includes cleaning fluid used to clean the tubing consumable 102 or other parts of the sorting fluid system 100, and also includes waste liquid after centrifugal stratification of the sample.

The pipeline timing control assembly 103 can also control the on-off of the non-waste liquid component collecting and containing piece and the sample containing piece 200 so as to input components separated by the sample containing piece 200 into the non-waste liquid component collecting and containing piece. The non-waste liquid component herein refers to a portion of the sample separated from the sample after the waste liquid is removed. Taking the sample as blood as an example, the waste liquid refers to blood plasma after blood centrifugation and delamination, and the rest is useful non-waste liquid components.

Specifically, the non-waste component collection receptacles include a destination receptacle 108b and a non-destination receptacle 108c, each in communication with the tubing consumable 102.

The destination container 108b is used for containing the destination liquid in the component separated by the sample container 200, and the non-destination container 108c is used for containing the non-destination liquid in the component separated by the sample container 200.

It should be noted that, the target accommodating element 108b and the non-target accommodating element 108c may be bag-shaped plastic products, which is low in cost and convenient for processing and manufacturing. It should be understood that the plastic bag-shaped articles for the purpose of holding the articles 108b and the non-purpose of holding the articles 108c are only one specific embodiment of the present application, and that the purpose of holding the articles 108b and the non-purpose of holding the articles 108c may be other articles in practical application.

The number of the target storage pieces 108b and the non-target storage pieces 108c is not limited to 1, and may be 2 or more.

In some embodiments, the fluid loading mechanism 105 further comprises a buffer reservoir 105a in communication with the tubing consumable 102, wherein the buffer reservoir 105a is configured to hold buffer.

The pipeline timing control assembly 103 can control the buffer containing part 105a to be communicated with each preset path in the pipeline consumable 102, so as to input buffer into the pipeline consumable 102 to clean each preset path of the pipeline consumable 102 in a forward direction. Of course, it should be noted that the buffer may also clean other components in the sort fluid system 100.

The buffer container 105a may be a plastic bag, which is low in cost and convenient to manufacture. It should be understood that the buffer container 105a is a plastic bag, which is only one embodiment of the present application, and that the buffer container 105a may be other products in practical applications.

Further, as shown in fig. 2, the application discloses that the pipeline consumable 102 is further communicated with the first filter 102-22, and the first filter 102-22 is communicated with the buffer liquid containing member 105a, so that the buffer liquid in the buffer liquid containing member 105a is filtered by the first filter 102-22 and then is conveyed to other parts to be cleaned, and impurities in the buffer liquid are prevented from entering the sorting fluid system 100.

Specifically, the first filter 102-22 is a square filter.

In some embodiments, the fluid loading mechanism 105 further includes a backflush fluid holder 105e in communication with the tubing consumable 102.

The pipeline timing control assembly 103 can control the conduction of the backflushing liquid containing part 105e and each preset path in the pipeline consumable 102 so as to input cleaning liquid into the pipeline consumable 102 to reversely clean each preset path of the pipeline consumable 102, thereby improving the cleaning effect.

The back flushing liquid containing member 105e may be a bag-shaped plastic product, which has low cost and is convenient for processing and manufacturing. It should be understood that the use of the back flush fluid container 105e as a plastic bag is only one embodiment of the present application, and that in practical applications, the back flush fluid container 105e may be provided as another plastic bag.

Further, the pipeline timing control assembly 103 can control the conduction between the reverse flushing liquid containing member 105e and the buffer liquid containing member 105a, so as to convey the buffer liquid in the buffer liquid containing member 105a into the reverse flushing liquid containing member 105e, that is, the buffer liquid in the reverse flushing liquid containing member 105e is provided by the buffer liquid containing member 105a, and the cleaning liquid in the reverse flushing liquid containing member 105e is the buffer liquid, so that forward flushing and reverse flushing can be realized only by injecting the buffer liquid into the buffer liquid containing member 105 a.

In some embodiments, the present disclosure discloses that the fluid loading mechanism 105 further includes a backflushing waste collection member 105d in communication with the tubing consumable 102.

The pipeline timing control assembly 103 can also control the switching of the back flush waste liquid collecting member 105d to collect the waste liquid after the back cleaning of the pipeline consumable 102, so as to collect the waste liquid after the back cleaning.

The back flushing waste liquid collecting member 105d may be a plastic bag, which is low in cost and convenient to manufacture. It will be appreciated that the use of the back flush waste collection member 105d as a pouch plastic article is only one embodiment of the present application, and that in practice the back flush waste collection member 105d may be provided as another article.

In some embodiments, the fluid loading mechanism 105 further comprises a filter 105b, the filter 105b being in communication with the tubing consumable 102, the tubing timing control assembly 103 being capable of controlling the communication of the filter 105b with the sample holder 200 to filter non-waste components output by the sample holder 200.

The filter 105b is provided to facilitate removal of impurities from the non-waste liquid component and to obtain a target liquid having a higher purity. Specifically, the application discloses that the filter element 105b is a filter bag, and the filter bag is also communicated with the second sampling bag 102-23 through the pipeline consumable 102, so as to facilitate sampling of samples in the filter bag.

The filter 105b may be a plastic bag or other articles.

Further, as shown in fig. 2, the present application discloses that the pipeline consumable 102 is further connected to the second dropping funnel 102-24, and the non-waste liquid component in the filter 105b enters the second dropping funnel 102-24 and then is conveyed to the subsequent required components.

The second drip chamber 102-24 is provided to prevent the non-waste liquid component from being sucked back into the filter 105b, and to smoothly flow down the sample in the second drip chamber 102-24.

In some embodiments, the sorting fluidic system 100 further includes a power member 106 mounted on the tubing consumable 102, the power member 106 for powering components or samples and the like flowing through the tubing consumable 102.

In particular, the power member 106 may be a peristaltic pump or the like, which may be clamped to the tubing consumable 102 for ease of installation.

Further, the power member 106 is disposed at least on the path of the tubing consumable 102 through which the sample in the sample source holder 105c enters the sample holder 200 in the centrifuge, to convey the sample in the sample source holder 105c into the sample holder 200.

In order to realize the air tightness detection of the pipeline consumable 102, the application discloses a sorting fluid system 100 which further comprises an air tightness detection piece 107, wherein the air tightness detection piece 107 is arranged on the pipeline consumable 102, and at least 1 air tightness detection piece 107 is respectively arranged at two ends of the power piece 106 and is used for detecting the air tightness of the pipeline consumable 102.

Specifically, the airtight detecting member 107 is a pressure sensor interposed on the piping consumable 102.

Further, the tubing timing control assembly 103 further includes a detector set interposed on the tubing consumable 102 to detect the condition of the sample or each component flowing through the tubing consumable 102.

Referring to fig. 1-3, a second aspect of the present application provides a cell sorting system 1000 for sorting cells.

Cell sorting system 1000 includes sorting column 104a and sorting fluid system 100 in any of the embodiments described above.

The fluid loading mechanism 105 further includes a magnetic bead containing member, a binding solution containing member, and an eluent containing member, and the magnetic bead containing member, the binding solution containing member, the eluent containing member, and the sorting column 104a are respectively in communication with the tubing consumable 102 in the sorting fluid system 100.

The pipeline timing control assembly 103 in the sorting fluidic system 100 can control the connection between the binding solution containing member and the sample containing member 200 in the sorting fluidic system 100, so as to convey the binding solution in the binding solution containing member into the sample containing member 200, and mix the binding solution with the sample in the sample containing member 200 uniformly through centrifugal motion of the centrifugal mechanism 101, and mark the target cells. In this example, the binding solution is exemplified as an IgG solution.

The pipeline time sequence control assembly 103 can control the magnetic bead containing piece to be communicated with the sample containing piece 200 so as to convey the magnetic bead solution in the magnetic bead containing piece into the sample containing piece 200, and the magnetic bead solution is uniformly mixed with the sample in the sample containing piece 200 through centrifugal motion of the centrifugal mechanism 101 and combined with the marked target cells.

The pipeline timing control assembly 103 can control the separation column 104a to be communicated with the sample containing piece 200, the sample containing piece 200 is extruded by the centrifugal mechanism 101 to convey the sample mixed with the binding solution and the magnetic beads into the separation column 104a, the separation column 104a in a magnetic field state can magnetically adsorb the magnetic beads, and other components in the sample are discharged into the non-target containing piece 108c of the separation fluid system 100.

The pipeline timing control assembly 103 can control the sorting column 104a to be communicated with the eluent containing piece so as to inject the eluent in the eluent containing piece into the sorting column 104a in a state of being separated from the magnetic field, so as to elute the target cells into the target containing piece 108b of the sorting fluid system 100, thereby being beneficial to the preservation of the target cells.

In the application, when the cell sorting system 1000 is used, a sample to be sorted is injected into a sample containing piece 200, a centrifugal mechanism 101 is started to centrifugally layer the sample in the sample containing piece 200, then a pipeline time sequence control assembly 103 is switched to conduct the sample containing piece 200 and a waste liquid containing piece 108a, the centrifugal mechanism 101 extrudes the centrifugally layered waste liquid into the waste liquid containing piece 108a, then the pipeline time sequence control assembly 103 is switched to conduct the combined solution containing piece and the sample containing piece 200, the combined solution in the combined solution containing piece is conveyed into the sample containing piece 200, then the pipeline time sequence control assembly 103 is switched to conduct the magnetic bead containing piece and the sample containing piece 200, the magnetic bead solution in the magnetic bead containing piece is conveyed into the sample containing piece 200, then the pipeline time sequence control assembly 103 is switched to conduct the sample containing piece 200 and a sorting column 104a, a magnetic field is applied to the sorting column 104a, the centrifugal mechanism 101 extrudes the sample containing piece 200 into the sorting column 104a in the magnetic field state, the magnetic bead is adsorbed into the sorting column 104a, the combined solution in the cell containing piece is conveyed into the sorting column 104a, the magnetic bead containing piece is conveyed into the non-column 104 b, and finally the eluting component is conveyed into the sorting column 104 b, and the eluting component is conveyed into the sorting column 104 b. Namely, in the application, the cell sorting is realized in the same cell sorting system 1000, so that the risk of cell pollution is reduced.

In some embodiments, the cell sorting system 1000 further includes a pre-sorting column 104c, the pre-sorting column 104c being in communication with the tubing consumable 102.

The pipeline timing control assembly 103 can control the pre-sorting column 104c to be respectively communicated with the sample containing member 200 and the sorting column 104a, so that the sample (at this time, the sample in the sample containing member 200 contains target cells, non-target cells, magnetic beads, binding liquid and the like) after the waste liquid is removed in the sample containing member 200 flows through the pre-sorting column 104c and then enters the sorting column 104a, and the filtration of the sample entering the sorting column 104a is realized.

In some embodiments, the tubing timing control assembly 103 can control the pre-sort column 104c to communicate with the filter 105b of the sort fluidic system 100 such that the sample within the sample holder 200 enters the pre-sort column 104c after being filtered by the filter 105 b.

In some embodiments, the cell sorting system 1000 further includes a color sensor 103-12, where the color sensor 103-12 is disposed at a position where the tubing consumable 102 communicates with the sample holder 200 for detecting each layer of the sample output after centrifugal delamination within the sample holder 200.

Taking a sample as blood as an example, after centrifugation, the blood is divided into a red blood cell layer, a white membrane layer and a plasma layer, the centrifugal mechanism 101 extrudes the sample containing piece 200 so as to discharge plasma into the waste liquid containing piece 108a, the emitting end of the color sensor 103-12 emits infrared rays with certain intensity, and after the infrared rays detect the white membrane in a pipeline, the infrared ray receiving end of the color sensor 103-12 stops rising of the centrifugal mechanism 101 due to light intensity change signal feedback.

As shown in FIG. 1, the tubing consumable 102 includes a first main tubing 102-1, a second main tubing 102-2, a third main tubing 102-3, a loading tubing 102-4, a waste tubing 102-5, a power tubing 102-6, a distribution tubing 102-7, a destination tubing 102-8, and a non-destination tubing 102-9.

One end of the first main line 102-1 communicates with the sample holding member 200 through the rotary connector 101a, one end of the second main line 102-2 communicates with the first position of the first main line 102-1, and the other end of the second main line 102-2 communicates with the second position of the waste line 102-5.

The power piece 106 is mounted on the power pipeline 102-6, one end of the power pipeline 102-6 is communicated with the first position of the second main pipeline 102-2, the other end of the power pipeline 102-6 is divided into a first communication pipeline, a second communication pipeline and a third communication pipeline, the first communication pipeline is communicated with the other end of the first main pipeline 102-1, the second communication pipeline is communicated with one end of the third main pipeline 102-3, and the third communication pipeline is communicated with one end of the waste liquid pipeline 102-5, which is far away from the waste liquid containing piece 108 a.

The sorting column 104a is connected to the third main line 102-3, and the other end of the third main line 102-3 is connected to the first position of the distribution line 102-7, one end of the distribution line 102-7 is connected to the waste line 102-5, the other end of the distribution line 102-7 is connected to the destination line 102-8, the non-destination line 102-9 is connected to the second position of the distribution line 102-7, and the second position of the distribution line 102-7 is located between the first position of the distribution line 102-7 and the end of the connected waste line 102-5.

One end of the loading line 102-4 communicates with the second location of the first main line 102-1 and the other end of the loading line 102-4 communicates with the fluid loading mechanism 105. Specifically, the magnetic bead holder, the binding solution holder, the eluent holder, and the sample source holder 105c are all in communication with the loading line 102-4.

As shown in fig. 2, the switching valve group includes a first switching valve 103-1, a second switching valve 103-2, a third switching valve 103-3, a fourth switching valve 103-4, a fifth switching valve 103-5, a sixth switching valve 103-6, a seventh switching valve 103-7, an eighth switching valve 103-8, a ninth switching valve 103-9, a tenth switching valve 103-10, and an eleventh switching valve 103-11.

The first switch valve 103-1, the second switch valve 103-2 and the third switch valve 103-3 are all installed on the first main pipeline 102-1, the first switch valve 103-1 is located between the rotary joint 101a and the second main pipeline 102-2, the second switch valve 103-2 is located between the second main pipeline 102-2 and the loading pipeline 102-4, and the third switch valve 103-3 is located between the loading pipeline 102-4 and the first connecting pipeline.

The fourth switch valve 103-4 and the fifth switch valve 103-5 are both installed on the second main line 102-2, and the fourth switch valve 103-4 is located between the first main line 102-1 and the power line 102-6, and the fifth switch valve 103-5 is located between the power line 102-6 and the waste line 102-5.

The sixth on-off valve 103-6 is installed on the third main line 102-3, and the sixth on-off valve 103-6 is located between the second communication line and the sorting column 104 a.

The seventh switching valve 103-7 and the eighth switching valve 103-8 are both installed on the waste liquid line 102-5, and the seventh switching valve 103-7 is located between the third communication line and the second main line 102-2, and the eighth switching valve 103-8 is located between the distribution line 102-7 and the end of the waste liquid line 102-5 remote from the distribution line 102-7.

The ninth switching valve 103-9 and the tenth switching valve 103-10 are both installed on the distribution pipe 102-7, and the ninth switching valve 103-9 is located between the third main pipe 102-3 and the non-destination pipe 102-9, and the tenth switching valve 103-10 is located between the third main pipe 102-3 and the destination pipe 102-8.

The eleventh on-off valve 103-11 is mounted on the non-destination line 102-9.

When the waste liquid in the sample-holding member 200 needs to be output, the first switch valve 103-1, the fourth switch valve 103-4, the fifth switch valve 103-5 and the eighth switch valve 103-8 are opened, and the other switch valves are closed.

When it is desired to load non-waste liquid components, buffer, bead solution, binding solution or eluent in the filter member into the sample-holding member 200, the third switch valve 103-3, fourth switch valve 103-4 and first switch valve 103-1 are opened, the other switch valves are closed, and the power member 106 starts to operate.

When it is necessary to output the non-target cells, the first switching valve 103-1, the fourth switching valve 103-4, the fifth switching valve 103-5, the seventh switching valve 103-7, the sixth switching valve 103-6, the ninth switching valve 103-9 and the eleventh switching valve 103-11 are opened, and the other switching valves are closed.

When the target cell is to be output, the first switching valve 103-1, the fourth switching valve 103-4, the fifth switching valve 103-5, the seventh switching valve 103-7, the sixth switching valve 103-6 and the tenth switching valve 103-10 are opened, and the other switching valves are closed.

The detector group includes a first bubble sensor 103-13 and a second bubble sensor 103-14, the first bubble sensor 103-13 is mounted on the first main line 102-1, the first bubble sensor 103-13 is used for controlling the centrifugal mechanism 101 to stop pressing the sample holder 200 when detecting bubbles, and the second bubble sensor 103-14 is mounted on the loading line 102-4 for stopping loading when detecting bubbles.

In some embodiments, the tubing consumable 102 further comprises a fourth main tubing 102-10, a filter tubing 102-11, and a pre-sorting tubing 102-12, one end of the fourth main tubing 102-10 is in communication with the third location of the first main tubing 102-1, the other end of the fourth main tubing 102-10 is in communication with the top end of the filter 105b of the fluid loading mechanism 105, one end of the filter tubing 102-11 is in communication with the bottom end of the filter 105b of the fluid loading mechanism 105, the other end of the filter tubing 102-11 is in communication with the loading tubing 102-4, one end of the pre-sorting tubing 102-12 is in communication with the first communication tubing and one end of the first main tubing 102-1, the pre-sorting column 104c of the magnetic sorting mechanism 104 is mounted on the pre-sorting tubing 102-12, the other end of the pre-sorting tubing 102-12 is in communication with the third main tubing 102-3, and is located between the sixth switching valve 103-6 and the sorting column 104 a.

The switch valve block further includes a twelfth switch valve 103-15, a thirteenth switch valve 103-16, a filter switch valve 103-17, and a pre-sorting switch valve 103-18, the twelfth switch valve 103-15 being mounted on the fourth main line 102-10, the thirteenth switch valve 103-16 being mounted on the first main line 102-1 and between the fourth main line 102-10 and the loading line 102-4, the filter switch valve 103-17 being mounted on the filter line 102-11, the pre-sorting switch valve 103-18 being mounted on the pre-sorting line 102-12 and between the first main line 102-1 and the pre-sorting column 104 c.

When it is desired to filter the sample outputted from the sample holding member 200 and deliver the filtered sample to the sorting column 104a, the first, second and twelfth switching valves 103-1, 103-2 and 103-15 are opened, the other switching valves are closed, the centrifugal mechanism 101 presses the sample holding member 200 to press the sample into the filter member 105b, all of the first, second and twelfth switching valves 103-1, 103-2 and 103-15 are closed, the filter switching valves 103-17, thirteenth switching valve 103-16, 103-2, 103-4 and 103-18 are opened to deliver the sample to the sorting column 104a, and when the non-target cells in the sorting column 104a are sorted, the ninth and eleventh switching valves 103-9 and 103-11 are simultaneously opened, and when the target cells in the sorting column 104a are sorted, the tenth switching valve 103-10 is simultaneously opened.

The detector set further includes a third bubble sensor 103-26, the third bubble sensor 103-26 being mounted on the first main line 102-1 and located between the second switch valve 103-2 and the thirteenth switch valve 103-16.

In some embodiments, the tubing consumable 102 further comprises a buffer tubing 102-13, a binding solution tubing 102-14, a magnetic bead tubing 102-15, an eluent tubing 102-16, and a sample input tubing 102-17; buffer solution line 102-13, combined solution line 102-14, magnetic bead line 102-15, eluent line 102-16 and sample input line 102-17 are sequentially connected to loading line 102-4 at intervals from one end of loading line 102-4 connected to filtering line 102-11 to one end connected to first main line 102-1, buffer solution line 102-13 is used for conveying buffer solution for cleaning each part of sorting fluid system 100, sample input line 102-17 is used for injecting preset amount of sample into sample containing piece 200 after air tightness detection and cleaning of consumable 102, combined solution line 102-14 is used for conveying combined solution to sample containing piece 200 after waste liquid is output from sample containing piece 200 and washed by buffer solution, magnetic bead line 102-15 is used for conveying magnetic bead solution to sample containing piece 200 after sample in sample containing piece 200 is uniformly mixed with combined solution, eluent line 102-16 is used for conveying sample mixed with magnetic beads in sample containing piece 200 to filtering piece 105b for filtering in centrifugal mechanism 101, and injecting non-sorting column 104a for non-sorting column 104a, and eluting cell column 104a is completed.

In order to facilitate adding other reagents or solutions to the cell sorting system 1000, the application discloses that the pipeline consumable 102 is further reserved with a standby pipeline for communicating standby holding parts for holding other reagents or solutions, and it should be noted that the number of standby pipelines is not limited.

The adapter 102-27 and the second filter 102-28 are respectively arranged on the solution pipeline 102-14, the magnetic bead pipeline 102-15 and the first standby pipeline 102-25, the adapter 102-27 is arranged to be convenient for communicating with a corresponding containing piece or standby containing piece and the like, and the second filter 102-28 is arranged to realize the filtration of solution or reagent and the like entering the pipeline consumable 102. Specifically, the second filter 102-28 is a circular filter.

The eluent line 102-16 and the second backup line 102-26 are respectively provided with a third filter 102-29 and a puncture outfit 102-30. The puncture outfit 102-30 is arranged to facilitate the puncture of the eluent containing piece, the reagent bag or the solution bag, and the third filter 102-29 filters the eluent in the pipeline consumable 102 or the reagent or the solution in the standby containing piece.

The switch valve block further includes a buffer switch valve 103-19 mounted on the buffer line 102-13, a binding solution switch valve 103-20 mounted on the binding solution line 102-14, a magnetic bead switch valve 103-21 mounted on the magnetic bead line 102-15, an eluent switch valve 103-22 mounted on the eluent line 102-16, and a sample switch valve 103-23 mounted on the sample input line 102-17.

In some embodiments, the tubing consumable 102 further includes a first secondary tubing 102-18 and a second secondary tubing 102-19 in communication with the fourth primary tubing 102-10, respectively, the first secondary tubing 102-18 for outputting non-target fluid cells within the tubing consumable 102 and the second secondary tubing 102-19 for inputting or outputting buffer.

The switch valve block further includes a secondary switch valve 103-24 mounted on the second secondary line 102-19.

The detector group further includes a fourth bubble sensor 103-25 mounted on the second sub-piping 102-19, and when a bubble is detected, the sub-switching valve 103-24 is closed.

Referring to fig. 1-4, a third aspect of the present application provides a sorter comprising an equipment mount 109 and a sorting fluid system 100 as in any of the embodiments described above or a cell sorting system 1000 as in any of the embodiments described above.

Either the sorting fluid system 100 or the cell sorting system 1000 is mounted on the equipment mount 109. The equipment mount 109 may be a frame structure, a plate welded structure, or the like, and is specifically set as needed.

In some embodiments, a centrifuge mounting block 109a, a conduit panel 109b, and a controller 109c are provided on the equipment mount 109, the centrifuge mounting block 109a being used to mount the centrifuge mechanism 101 of the sorting fluidic system 100.

The conduit panel 109b is disposed obliquely or vertically to the centrifugal mounting stage 109a for mounting the conduit consumable 102, the conduit timing control assembly 103, and the controller 109c of the sorting fluid system 100. Specifically, the pipe panel 109b is provided with a notch 109b-2, and the controller 109c is mounted at the position of the notch 109 b-2.

The controller 109c is communicatively connected to the pipeline timing control assembly 103 and the centrifugal mechanism 101, respectively, to control the operation of the pipeline timing control assembly 103 and the centrifugal mechanism 101.

The top end of the pipeline panel 109b is provided with a hanging column 109b-1, and the hanging column 109b-1 is provided with a hanging hook 109b-1a for hanging each containing piece.

In some embodiments, the sorter also includes a loading magnetic field assembly 104b mounted on the pipeline panel 109b, the loading magnetic field assembly 104b being used to apply or break a magnetic field to the sorting column 104a in the cell sorting system 1000.

Specifically, the loading magnetic field assembly 104b includes a permanent magnet 104b-1 and a driving member, the permanent magnet 104b-1 is provided with a receiving slot 104b-1a for receiving the sorting column 104a, and a driving end of the driving member is connected to the permanent magnet 104b-1 for driving the permanent magnet 104b-1 to approach or separate from the sorting column 104a so as to apply or break a magnetic field to the sorting column 104 a. The driving piece can be a gear rack structure, an electric push rod structure, a hydraulic cylinder or an air cylinder and other structures.

It should be noted that, the structure of the loading magnetic field assembly 104b disclosed above is only one specific embodiment of the present application, and in practical applications, the loading magnetic field assembly 104b may be provided with other structures, for example, the loading magnetic field assembly 104b includes an electromagnet and a power supply member, the electromagnet is provided with a receiving slot 104b-1a for receiving the sorting column 104a, the power supply member is connected with the electromagnet in a ferroelectric manner, and the power supply member energizes or de-energizes the electromagnet so as to enable the sorting column 104a to apply or break the magnetic field.

In some embodiments, the controller 109c includes a main body of the controller 109c and a touch screen in signal connection with the main body of the controller 109c, where the main body of the controller 109c controls operation of each part of the sorter, and the touch screen can display operation states of each part of the sorter in time, and can also facilitate interaction with a person, etc.

A fourth aspect of the application provides a method of sorting a fluid, comprising:

providing a sort fluidic system 100 according to any of the embodiments described above, and placing a sample holder 200 within a centrifuge 101;

starting the centrifugal mechanism 101 to rotate and driving the sample holding piece 200 to centrifugally rotate so as to layer the sample in the sample holding piece 200;

switching the line timing control assembly 103 to turn on the sample holder 200 and the waste holder 108a of the sorting fluidic system 100;

Starting the centrifugal mechanism 101 to squeeze the sample holder 200 and squeeze the waste liquid in the sample holder 200 into the waste liquid holder 108 a;

Switching the line timing control assembly 103 to turn on the sample holder 200 and the non-waste component collection holder of the sorting fluidic system 100;

The centrifugal mechanism 101 is started to squeeze the sample holder 200, and each non-waste liquid component separated by the sample holder 200 is conveyed into the corresponding non-waste liquid component collecting holder.

The application realizes the direct acquisition of the target liquid and reduces the risk of pollution of the target liquid.

Providing a cell sorting system 1000 according to any one of the above, and placing the sample holder 200 in the centrifugation mechanism 101;

starting the centrifugal mechanism 101 to centrifugally layer the sample in the sample holding piece 200;

Switching the pipeline time sequence control assembly 103 to the sample holding part 200 to be communicated with the waste liquid holding part 108a, and extruding the waste liquid subjected to centrifugal layering into the waste liquid holding part 108a by the centrifugal mechanism 101;

Switching the pipeline timing control assembly 103 to the connection state between the binding solution containing member and the sample containing member 200, and conveying the binding solution in the binding solution containing member into the sample containing member 200;

Switching the pipeline time sequence control assembly 103 to the magnetic bead containing piece to be communicated with the sample containing piece 200, and conveying the magnetic bead solution in the magnetic bead containing piece into the sample containing piece 200;

Switching the pipeline timing control assembly 103 to the sample containing piece 200 to be communicated with the sorting column 104a, applying a magnetic field to the sorting column 104a, extruding the sample of the sample containing piece 200 into the sorting column 104a under the magnetic field state by the centrifugal mechanism 101, adsorbing magnetic beads in the sorting column 104a, and discharging non-target cells in the sample into the non-target containing piece 108c through the sorting column 104 a;

the pipeline timing control assembly 103 is switched to the eluent containing piece to be communicated with the sorting column 104a, and the eluent in the eluent containing piece is conveyed into the sorting column 104a so as to elute the target cells in the sorting column 104a into the target containing piece 108 b.

Taking a sample as a blood sample and a binding solution as an IgG as an example, the cell sorting method comprises the steps of switching a pipeline time sequence control group 103 to detect the tightness of a pipeline consumable 102 so as to ensure the smooth proceeding of cell sorting; then, switching the pipeline timing control assembly 103 to connect the sample containing member 105c with the sample containing member 200, injecting blood sample into the sample containing member 200, then, starting the centrifugal mechanism 101 to centrifugally layer the blood sample in the sample containing member 200, separating the blood sample in the sample containing member 200 into red blood cell layers, white membrane layers and plasma layers along the radial direction of the sample containing member 200 under the action of centrifugal force, then, switching the pipeline timing control assembly 103 to connect the sample containing member 200 with the waste liquid containing member 108a, extruding the blood plasma of the plasma layers in the centrifugally layered sample containing member 200 into the pipeline consumable member 102 and conveying the blood sample into the waste liquid containing member 108a, then, switching the pipeline timing control assembly 103 to inject IgG solution into the sample containing member 200, driving the centrifugal mechanism 101 to positively reverse the sample containing member 200 so as to uniformly mix the sample in the sample containing member 200, thereby realizing labeling of target cells, then, switching the pipeline timing control assembly 103 to inject the red blood sample into the sample containing member 200, extruding the pre-arranged magnetic bead layer into the sample containing member 200, extruding the sample containing member 104 b to sequentially, extruding the sample containing member 104 b to the sample containing member 105b to sequentially, extruding the sample containing member 104 b to the sample containing member, and conveying the sample containing member 104 b to the sample containing member, and sequentially extruding the sample containing member 105b to the sample containing member, and filtering the sample containing member 104 b to sequentially, the magnetic beads combined with the target cells are adsorbed in the separation column 104a, the non-target cells in the sample are discharged into the non-target holder 108c through the separation column 104a, and then the magnetic field applied to the separation column 104a is turned off, and the line timing control assembly 103 is switched to inject the eluent into the separation column 104a to elute the target cells in the separation column 104a into the target holder 108 b.

In order to facilitate the transportation of the blood sample into the sample containing member 200, the method further comprises the steps of injecting buffer solution and eluent into the pipeline consumable 102 through the buffer solution containing member 105a and the eluent containing member to realize the rinsing of the pipeline consumable 102, and injecting a certain amount of buffer solution into the back flushing liquid collecting member 105d and the back flushing liquid containing member 105e respectively, so that the buffer solution in the back flushing liquid containing member 105e can be used to realize the thorough cleaning during the subsequent back flushing.

In order to ensure that the blood sample can be completely loaded into the sample holder 200, after the blood sample is injected into the sample holder 200, the method further comprises the steps of inputting buffer solution into the sample source holder 105c for rinsing, switching to the connection between the sample source holder 105c and the sample holder 200, mixing the rinsed buffer solution and the blood sample in the sample source holder 105c, and inputting the mixture into the sample holder 200. It should be noted that the above steps may be performed only 1 time, or may be repeated a plurality of times until the blood sample in the sample source holder 105c is thoroughly rinsed into the sample holder 200. It will be appreciated that when the total volume of blood sample within sample source holder 105c is greater than the maximum volume allowed for sample holder 200 (as controlled by the input sample volume parameters), the sample may be loaded and volume adjusted multiple times, with the sample source holder 105c being rinsed only the last time.

In order to realize thorough discharge of the blood plasma in the sample holder 200, after discharging the blood plasma to the waste liquid holder, the method further comprises injecting buffer solution into the sample holder 200 and performing centrifugal delamination, and outputting the mixed solution of the blood plasma and the buffer solution to the waste liquid holder 108a. The above steps may be performed only 1 time, or may be repeated a plurality of times until the plasma is completely discharged into the waste liquid container 108a.

In order to realize the discharge of the magnetic beads which are not combined with the target cells in the sample containing piece 200, after injecting the magnetic bead solution into the sample containing piece 200 and incubating for a preset time, the method further comprises the steps of injecting buffer solution into the sample containing piece 200 to clean the magnetic beads, rotating the sample containing piece 200 through the centrifugal mechanism 101 to drive the sample in the sample containing piece 200 to centrifugally layer, and extruding the sample containing piece 200 by the centrifugal mechanism 101 to discharge the magnetic beads which are not combined with the target cells in the sample containing piece 200 and the buffer solution into the waste liquid containing piece 108 a. The above steps may be performed only 1 time, or may be repeated a plurality of times until the magnetic beads, buffer, and the like, which are not bound to the target cells, are completely discharged into the waste liquid container 108 a.

In order to thoroughly squeeze the sample such as target cells after being combined with the magnetic beads in the sample holder 200 into the filter 105b, after squeezing the sample holder 200 to squeeze the sample into the filter 105b, the method further comprises injecting buffer solution into the sample holder 200 to rinse the sample holder 200, and squeezing the sample holder 200 by the centrifugal mechanism 101 to squeeze the sample after washing the sample holder 200 into the filter 105 b. The above steps may be performed only 1 time, or may be repeated a plurality of times until the sample labeled with the magnetic beads is completely discharged into the filter 105 b.

In order to thoroughly discharge the sample in the filter 105b, the non-target cells in the sample are discharged into the non-target holding part 108c through the sorting column 104a, and then buffer solution is input into the filter 105b for cleaning, and the cleaned sample and the buffer solution are mixed and conveyed into the pre-sorting column 104c and the sorting column 104a for sorting again. The above steps may be performed only 1 time, or may be repeated a plurality of times until the sample in the filter element 105b is completely discharged and sorted. In this step, the reverse flushing liquid containing member 105e communicates with the filter member 105b to input the buffer liquid into the filter member 105b for washing, and the buffer liquid flows through the pre-sorting column 104c in the reverse direction (the reverse direction is opposite to the direction of flowing through the pre-sorting column 104c when the sample is sorted) during the process of inputting the buffer liquid into the filter member 105b, thereby washing the pre-sorting column 104 c.

In order to facilitate the removal of non-target cells and the like from the separation column 104a, the filter 105b is washed and then a buffer solution is injected into the separation column 104a to wash the separation column 104a. The above steps may be performed only 1 time, or may be repeated a plurality of times until the column 104a is thoroughly washed.

In summary, each time a sample, components, or reagent solutions (including an eluent, a buffer, an IgG solution, and a magnetic bead solution) and the like flow through the tube consumable 102, the tube consumable 102 and the respective holding members (including the fluid loading mechanism 105 and the component collection mechanism 108) need to be cleaned, so as to improve the purity of the collected target cells.

In order to avoid that waste liquid in the tubing consumable 102 is injected into the sample holder 200 along with the IgG solution, the path for loading the IgG solution is pre-filled with the IgG solution before the IgG solution is injected into the sample holder 200. Specifically, the combined solution switching valve 103-20, the third switching valve 103-3, the fifth switching valve 103-5 and the eighth switching valve 103-8 are opened, the other switching valves are closed, and the combined solution switching valve 103-20, the third switching valve 103-3, the fourth switching valve 103-4 and the first switching valve 103-1 are opened after the loading is completed, and the other switching valves are closed, so that the IgG solution is injected into the sample holder 200.

It should be noted that, for convenience of description, only a portion related to the related application is shown in the drawings. Embodiments of the application and features of the embodiments may be combined with each other without conflict.

It is to be understood that the terms "system," "apparatus," "unit," and/or "module" as used herein are one means for distinguishing between different components, elements, parts, portions, or assemblies at different levels. However, if other words can achieve the same purpose, the word can be replaced by other expressions.

As used in the specification and in the claims, the terms "a," "an," "the," and/or "the" are not specific to a singular, but may include a plurality, unless the context clearly dictates otherwise. In general, the terms "comprises" and "comprising" merely indicate that the steps and elements are explicitly identified, and they do not constitute an exclusive list, as other steps or elements may be included in a method or apparatus. The inclusion of an element as defined by the phrase "comprising one does not exclude the presence of additional like elements in a process, method, article, or apparatus that comprises an element.

In the description of the embodiment of the present application, unless otherwise indicated, "/" means or, for example, a/B may represent a or B, and "and/or" herein is merely an association relationship describing an association object, which means that three relationships may exist, for example, a and/or B, and that three cases, i.e., a alone, a and B together, and B alone, exist. In addition, in the description of the embodiments of the present application, "plurality" means two or more than two.

The terms "first" and "second" are used below for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature.

A flowchart is used in the present application to describe the operations performed by a system according to embodiments of the present application. It should be appreciated that the preceding or following operations are not necessarily performed in order precisely. Rather, the steps may be processed in reverse order or simultaneously. Also, other operations may be added to or removed from these processes.

The above description is only illustrative of the preferred embodiments of the present application and the technical principles applied, and is not intended to limit the present application. Various modifications and variations of the present application will be apparent to those skilled in the art. The scope of the present application is not limited to the specific combination of the above technical features, but also includes other technical features formed by any combination of the above technical features or their equivalents without departing from the spirit of the present application. Such as the above-mentioned features and the technical features disclosed in the present application (but not limited to) having similar functions are replaced with each other.

Claims

A sorting fluidic system (100) characterized by comprising a centrifugal mechanism (101), a pipeline consumable (102) and a pipeline timing control assembly (103);

The centrifugal mechanism (101) is used for accommodating a sample accommodating part (200), a rotary connecting part (101 a) which is respectively communicated with the sample accommodating part (200) and the pipeline consumable (102) is arranged on the centrifugal mechanism (101), and the centrifugal mechanism (101) drives a sample in the sample accommodating part (200) to realize centrifugal layering and extrude corresponding layer components into the pipeline consumable (102);

The pipeline time sequence control assembly (103) is arranged on the pipeline consumable (102) and is used for controlling samples or components entering the pipeline consumable (102) to flow according to a preset time sequence and a preset path.
The sorting fluidic system (100) of claim 1, further comprising a sample source holder (105 c), a waste liquid holder (108 a), and a non-waste liquid component collection holder in communication with the tubing consumable (102), respectively;

The sample source holding part (105 c) is internally provided with a sample source, and the pipeline time sequence control assembly (103) can control the on-off of the sample source holding part (105 c) and the sample holding part (200) so as to realize the input of a sample into the sample holding part (200);

The pipeline time sequence control assembly (103) can also control the on-off of the waste liquid containing piece (108 a) and the sample containing piece (200) so as to realize the input of waste liquid in components separated by the sample containing piece (200) into the waste liquid containing piece (108 a);

the pipeline time sequence control assembly (103) can also control the on-off of the non-waste liquid component collection containing piece and the sample containing piece (200) so as to realize the input of the non-waste liquid component collection containing piece into the sample containing piece (200) to separate out each non-waste liquid component in the components.
The sortation fluidic system (100) of claim 2, wherein said non-waste component collection receptacles include a destination receptacle (108 b) and a non-destination receptacle (108 c) in communication with said piping consumable (102), respectively;

The target holding piece (108 b) is used for holding target liquid in the non-waste liquid component separated by the sample holding piece (200), and the non-target holding piece (108 c) is used for holding non-target liquid in the non-waste liquid component separated by the sample holding piece (200).
The sorting fluidic system (100) of claim 1, further comprising a buffer reservoir (105 a) in communication with the tubing consumable (102);

The buffer solution containing piece (105 a) is used for containing buffer solution, the pipeline time sequence control assembly (103) can control the buffer solution containing piece (105 a) to be conducted with each preset path in the pipeline consumable (102), so that the buffer solution is input into the pipeline consumable (102) to positively clean each preset path of the pipeline consumable (102).
The sorting fluidic system (100) of claim 4, further comprising a backflushing fluid holder (105 e) in communication with the tubing consumable (102);

The pipeline time sequence control assembly (103) can control the conduction of the reverse flushing liquid containing piece (105 e) and each preset path in the pipeline consumable (102) so as to input cleaning liquid into the pipeline consumable (102) to reversely clean each preset path of the pipeline consumable (102).
The sorting fluidic system (100) of claim 5, wherein the pipeline timing control assembly (103) is capable of controlling the communication of the backflushing fluid holder (105 e) with the buffer holder (105 a) to deliver buffer within the buffer holder (105 a) into the backflushing fluid holder (105 e);

the cleaning liquid in the reverse flushing liquid containing piece (105 e) is buffer liquid.
The sorting fluidic system (100) of claim 5, further comprising a back flush waste collection (105 d) in communication with the tubing consumable (102);

The pipeline timing control assembly (103) can also control the switching of the back flushing waste liquid collecting piece (105 d) to collect the waste liquid after the reverse cleaning of the pipeline consumable (102).
The sortation fluidic system (100) as recited in claim 1, further comprising a filter (105 b);

The filtering piece (105 b) is communicated with the pipeline consumable (102), and the pipeline time sequence control assembly (103) can control the filtering piece (105 b) to be communicated with the sample containing piece (200) so as to filter non-waste liquid components output by the sample containing piece (200).
The sortation fluidic system (100) as recited in claim 2, further comprising a power component (106) mounted on said piping consumable (102);

the power piece (106) is used for providing power.
The sorting fluidic system (100) of claim 9, wherein the power member (106) is disposed at least in the path of a tubing consumable (102) through which a sample in the sample source holder (105 c) enters the sample holder (200) in the centrifuge to transport the sample in the sample source holder (105 c) into the sample holder (200).
The sorting fluidic system (100) of claim 9, further comprising an airtight detection element (107);

The airtight detection piece (107) is installed on the pipeline consumable (102), and at least 1 airtight detection piece (107) is arranged at two ends of the power piece (106) respectively and is used for detecting the air tightness of the pipeline consumable (102).
The sorting fluidic system (100) of claim 11, wherein the airtight detection means (107) is a pressure sensor clamped on the tubing consumable (102).
The sorting fluidic system (100) of any of claims 1-12, wherein the tubing timing control assembly (103) includes a plurality of on-off valves that control the on-off of the respective positions of the tubing consumable (102).
The sortation fluidic system (100) as recited in claim 13, wherein said on-off valve is sandwiched on said piping consumable (102).
The sorting fluidic system (100) of claim 13, wherein the tubing timing control assembly (103) further comprises a detector set clamped to the tubing consumable (102) to detect the condition of a sample or components flowing through the tubing consumable (102).
A cell sorting system (1000) comprising a magnetic bead holder, a binding solution holder, an eluent holder, a sorting column (104 a), and a sorting fluidic system (100) according to any of claims 1-15;

the magnetic bead containing piece, the binding solution containing piece, the eluent containing piece and the sorting column (104 a) are respectively communicated with a pipeline consumable (102) in the sorting fluid system (100);

The pipeline time sequence control assembly (103) in the sorting fluid system (100) can control the connection between the binding solution containing piece and the sample containing piece (200) in the sorting fluid system so as to convey the binding solution in the binding solution containing piece into the sample containing piece (200), and the binding solution and the sample in the sample containing piece (200) are uniformly mixed through centrifugal motion of the centrifugal mechanism (101) and mark target cells;

The pipeline time sequence control assembly (103) can control the magnetic bead containing piece to be communicated with the sample containing piece (200) so as to convey the magnetic bead solution in the magnetic bead containing piece into the sample containing piece (200), and the magnetic bead solution is uniformly mixed with a sample in the sample containing piece (200) through centrifugal motion of the centrifugal mechanism (101) and combined with marked target cells;

The pipeline time sequence control assembly (103) can control the separation column (104 a) to be communicated with the sample containing piece (200), the centrifugal mechanism (101) is used for extruding the sample containing piece (200) to convey a sample mixed with a binding solution and magnetic beads into the separation column (104 a), the separation column (104 a) in a magnetic field state can magnetically adsorb the magnetic beads, and non-target cells in the sample are discharged into a non-target containing piece (108 c) of the separation fluid system (100);

The pipeline timing control assembly (103) can control the sorting column (104 a) to be communicated with the eluent containing piece so as to inject the eluent in the eluent containing piece into the sorting column (104 a) in a state of being separated from a magnetic field, and elute the target cells into a target containing piece (108 b) of the sorting fluid system (100).
The cell sorting system (1000) of claim 16, further comprising a pre-sorting column (104 c);

the pre-separation column (104 c) is communicated with the pipeline consumable (102);

The pipeline timing control assembly (103) can control the pre-sorting column (104 c) to be respectively communicated with the sample containing piece (200) and the sorting column (104 a), so that samples in the sample containing piece (200) flow through the pre-sorting column (104 c) and then enter the sorting column (104 a).
The cell sorting system (1000) of claim 17, wherein the pipeline timing control assembly (103) is capable of controlling the pre-sorting column (104 c) to communicate with a filter (105 b) of the sorting fluidic system (100) such that samples within the sample holder (200) enter the pre-sorting column (104 c) after being filtered by the filter (105 b).
The cell sorting system (1000) according to any of claims 16-18, further comprising a color sensor (103-12);

The color sensor (103-12) is arranged at the position where the pipeline consumable (102) is communicated with the sample containing piece (200) and is used for detecting each layer of the sample containing piece (200) which is output after centrifugal layering of the sample.
A sorter comprising an equipment mount (109) and a sorting fluidic system (100) according to any one of claims 1 to 15 or a cell sorting system (1000) according to any one of claims 16 to 19;

The sorting fluidic system (100) or the cell sorting system (1000) is mounted on the equipment mounting frame (109).
The sorter of claim 20, wherein the equipment mount (109) is provided with a centrifugal mounting table (109 a), a piping panel (109 b), and a controller (109 c);

The centrifugal mounting table (109 a) is used for mounting a centrifugal mechanism (101) of the sorting fluid system (100);

the pipeline panel (109 b) is obliquely or vertically arranged with the centrifugal mounting table (109 a) and is used for mounting pipeline consumable (102), a pipeline time sequence control assembly (103) and the controller of the sorting fluid system (100);

The controller (109 c) is respectively in communication connection with the pipeline time sequence control assembly (103) and the centrifugal mechanism (101).
The sorter (100) of claim 21 further comprising a loading magnetic field assembly (104 b) mounted on the conduit panel (109 b);

the loading magnetic field assembly (104 b) is used to apply or break a magnetic field to a sorting column (104 a) in the cell sorting system (1000).
The sorter (100) of claim 22 wherein the loading magnetic field assembly (104 b) comprises a permanent magnet (104 b-1) and a driving member, the permanent magnet (104 b-1) having a receiving slot (104 b-1 a) for receiving the sorting column (104 a), the driving end of the driving member being coupled to the permanent magnet (104 b-1) for driving the permanent magnet (104 b-1) toward or away from the sorting column (104 a) to apply or break a magnetic field to the sorting column (104 a);

Or alternatively

The loading magnetic field assembly (104 b) comprises an electromagnet and a power supply piece, wherein the electromagnet is provided with a containing groove for containing the sorting column (104 a), the power supply piece is connected with the electromagnet in a ferroelectric manner, and the power supply piece is used for electrifying or de-electrifying the electromagnet so that the sorting column (104 a) can apply a magnetic field or break the magnetic field.
The sorter (100) of claim 23 wherein the drive is a rack and pinion structure, an electric push rod structure, a hydraulic cylinder, or an air cylinder.
The sorter (100) of claim 21 wherein a hanging post (109 b-1) is provided at the top end of the conduit panel (109 b), and a hanger (109 b-1 a) is mounted on the hanging post (109 b-1) for hanging each of the receptacles.
The sorter (100) of any of claims 21-25 wherein the controller includes a controller body and a touch screen (109 c) in signal communication with the controller body.
A method of sorting a fluid, comprising:

Providing a sorting fluidic system (100) according to any of claims 1-15, and placing a sample holder (200) within the centrifugation mechanism (101);

starting the centrifugal mechanism (101) to rotate and driving the sample holding piece (200) to centrifugally rotate so as to laminate the sample in the sample holding piece (200);

Switching the pipeline timing control assembly (103) to conduct the sample holder (200) and the waste holder (108 a) of the sorting fluidic system (100);

Starting the centrifugal mechanism (101) to extrude the sample containing piece (200), and extruding the waste liquid in the separated components in the sample containing piece (200) into the waste liquid containing piece (108 a);

Switching the pipeline timing control assembly (103) to communicate the sample holder (200) with a non-waste component collection holder of the sorting fluidic system (100);

And starting the centrifugal mechanism (101) to squeeze the sample containing piece (200), and conveying each non-waste liquid component separated by the sample containing piece (200) into the corresponding non-waste liquid component collecting containing piece.
A method of cell sorting comprising:

Providing a cell sorting system (1000) according to any of claims 16-19, and placing a sample holder (200) within the centrifugation mechanism (101);

Starting the centrifugal mechanism (101) to centrifugally layer the sample in the sample holding piece (200);

Switching the pipeline time sequence control assembly (103) to be communicated with the sample containing piece (200) and the waste liquid containing piece (108 a), and extruding the waste liquid in the centrifugally layered sample into the waste liquid containing piece (108 a) by the centrifugal mechanism (101);

Switching the pipeline timing control assembly (103) to be communicated with the binding solution containing piece and the sample containing piece (200), and conveying the binding solution in the binding solution containing piece into the sample containing piece (200);

Switching the pipeline time sequence control assembly (103) to be communicated with the magnetic bead containing piece and the sample containing piece (200) to convey the magnetic bead solution in the magnetic bead containing piece into the sample containing piece (200);

Switching the pipeline time sequence control assembly (103) to the state that the sample containing piece (200) is communicated with the sorting column (104 a) and applying a magnetic field to the sorting column (104 a), wherein the centrifugal mechanism (101) extrudes the sample in the sample containing piece (200) into the sorting column (104 a) under the magnetic field state, magnetic beads are adsorbed in the sorting column (104 a), and non-target cells in the sample are discharged into a non-target containing piece (108 c) through the sorting column (104 a);

Switching the pipeline time sequence control assembly (103) to the eluent containing piece to be communicated with the sorting column (104 a), and conveying the eluent in the eluent containing piece into the sorting column (104 a) so as to elute target cells in the sorting column (104 a) into a target containing piece (108 b).