CN219136736U

CN219136736U - Liquid drop single cell sorting system

Info

Publication number: CN219136736U
Application number: CN202223603171.2U
Authority: CN
Inventors: 王立言; 郭肖杰; 袁英歌
Original assignee: Luoyang Tmaxtree Biotechnology Co ltd
Current assignee: Luoyang Tmaxtree Biotechnology Co ltd
Priority date: 2022-12-30
Filing date: 2022-12-30
Publication date: 2023-06-06
Anticipated expiration: 2032-12-30

Abstract

The utility model belongs to the technical field of cell detection and separation, in particular to a liquid drop single-cell separation system, wherein a liquid drop generation and culture module comprises a power assembly, a culture cabin, a culture pipeline and a temperature controller, and the culture pipeline is connected with the power assembly; the detection and sorting module comprises an optical detection assembly and a sorting platform; the control module is used for controlling the running state of the power assembly, reading and recording the measured data value of the optical detection assembly in real time, and analyzing and processing the data. According to the utility model, micro-droplets with bubble intervals are generated by a microfluidic technology, the droplets are arranged in a micro-pipeline in a queue mode, the air-permeable micro-pipeline is used, the bubbles prevent the droplets from fusing, meanwhile, the oxygen dissolving performance of the droplets is enhanced, the culture pipeline has high permeability to various gases, evaporation is effectively reduced, and oxygen dissolving is improved.

Description

Liquid drop single cell sorting system

Technical Field

The utility model relates to the technical field of cell detection and separation, in particular to a liquid drop single cell separation system.

Background

The cell sample culture apparatus is a mechanism for culturing and detecting cells, which is usually configured such that a culture medium tank is filled with a variety of cells containing a target to be cultured, and the proliferation rate of the cells is detected at a set temperature, oxygen content, illuminance, and other set external conditions for a set period of time; in order to make the cultivation more accurate and controllable; a conduit type culture device is developed, and a culture solution is prepared into oil-water two phases to form liquid drops, and is input into a long transparent or semitransparent conduit, so that cells are propagated and grown in the conduit; at present, droplets formed by oil-water two phases slide mutually in the movement process to cause contact fusion, and a surfactant can only maintain stability of nano-scale droplets and droplets below, but can not prevent the mutual fusion of micro-scale droplets, and can not meet the oxygen dissolving requirement when being cultured in a closed container; the evaporation rate is too high when culturing in an open container; the bubbles cause serious interference to the spectrum detection and the droplet sorting of the droplets; the identification and elimination of traditional bubbles requires complex photoelectric identification devices and complex power driving logic.

Disclosure of Invention

(one) solving the technical problems

In order to overcome the defects in the prior art, the utility model provides a liquid drop single-cell sorting system, which solves the problems in the background art.

(II) technical scheme

The utility model adopts the following technical scheme for realizing the purposes:

the liquid drop single-cell sorting system comprises a liquid drop generation culture module, a liquid drop generation culture module and a liquid drop sorting module, wherein the liquid drop generation culture module comprises a power assembly, a culture cabin, a culture pipeline and a temperature controller, and the culture pipeline is connected with the power assembly; the detection and sorting module comprises an optical detection assembly and a sorting platform; the control module is used for controlling the running state of the power assembly, reading and recording the measured data value of the optical detection assembly in real time, and analyzing and processing the data.

Further, the culture cabin is internally coiled with a culture pipeline, and the temperature controller is positioned in the culture cabin.

Further, the culture pipeline is a gas-permeable pipeline.

Further, the sample injection position of the culture pipeline is set to be of a four-way structure.

Further, the detection and sorting module comprises a bubble collecting device.

Further, the detection sorting module comprises a sorting platform, wherein an X-axis moving part, a Y-axis moving part, an A-axis rotating part and a B-axis lifting part are arranged on the sorting platform, a bearing bracket is arranged on the X-axis moving part, and a liquid drop collecting container is placed on the bearing bracket.

Further, a first working carrying platform and a second working carrying platform are fixedly arranged on the sorting platform, the lifting part of the shaft B is connected with the rotating part of the shaft A to drive the rotating part of the shaft A to lift, and the bottom of the front side of the rotating part of the shaft A is provided with a clamping mechanism.

Further, a cantilever is arranged on the sorting platform, the cantilever is arranged above the X-axis moving part in parallel, the culture pipeline is connected with the cantilever, and the optical detection assembly is fixed at the tail end of the cantilever and is used for detecting liquid drops in the culture pipeline passing through the cantilever.

Further, the cantilever below parallel arrangement waste liquid collection mechanism, when detecting as the target liquid drop through optical detection subassembly, waste liquid collection mechanism staggers with the culture pipeline export, and the liquid drop is dripped in the liquid drop collection container under the action of gravity, and when detecting as non-target liquid drop through optical detection subassembly, waste liquid collection mechanism is relative with the culture pipeline export, and the liquid drop is dripped in the waste liquid collection mechanism under the action of gravity.

Further, the control module is connected with the liquid drop generation and culture module and the detection and separation module and is used for adjusting the working state of each module, recording detection data and analyzing and processing.

(III) beneficial effects

Compared with the prior art, the utility model provides a liquid drop single cell sorting system, which comprises the following components

The beneficial effects are that:

according to the utility model, micro-droplets with bubble intervals are generated by a water-gas-oil three-phase microfluidic technology, the droplets are arranged in a micro-pipeline in a queue mode, the air permeability micro-pipeline is used for preventing the droplets from fusing, meanwhile, the oxygen dissolving performance of the droplets is enhanced, the culture pipeline has high permeability to various gases, evaporation is effectively reduced (less than 2%), and dissolved oxygen (Kla approximately equal to 90/hr) is improved; the power assembly is used for controlling the stable output of the gas-liquid-oil three-phase, the generation speed of micro liquid drops is 3-5 drops/s, the volume of the liquid drops is 1-5uL, and the sorting quantity can reach 10000 drops/batch; the air is removed by using the air bubbles in the collecting pipeline of the air collecting bottle, so that the method is simple, efficient and stable; the mechanical structure with high degree of freedom is used for automatically switching and storing the sample collection containers, supports 10 sample collection containers per batch, and realizes batch classified storage and detection.

Drawings

FIG. 1 is a schematic block diagram of the workflow principle of the present utility model;

FIG. 2 is a schematic view of the structure of the culture bin of the utility model;

FIG. 3 is a schematic diagram of a detection and sorting module according to the present utility model;

in the figure: 1. a droplet generation culture module; 2. a detection and sorting module; 3. a power assembly; 4. a control module; 5. an optical detection assembly; 6. a sorting platform; 7. a culture chamber; 8. a culture pipeline; 9. an X-axis moving part; 10. a Y-axis moving part; 11. an A-axis rotating part; 12. a B-axis lifting part; 13. a clamping mechanism; 14. a droplet collection container; 15. a support bracket; 16. a first work stage; 17. a second work stage; 18. a cantilever; 19. a waste liquid collection mechanism; 21. a bubble collection device; 22. an oil phase; 23 aqueous phase; 24. a gas phase; 25. a cell; 26. a gas source; 27. an oil bottle; 28. a solution bottle; 29. and a temperature controller.

Detailed Description

Embodiments of the present utility model are described in detail below, examples of which are illustrated in the accompanying drawings, wherein the same or similar reference numerals refer to the same or similar elements or elements having the same or similar functions throughout. The embodiments described below by referring to the drawings are exemplary only for explaining the present utility model and are not to be construed as limiting the present utility model.

In the description of the present utility model, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings are merely for convenience in describing the present utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present utility model. Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a 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 of the described features. In the description of the present utility model, the meaning of "a plurality" is two or more, unless explicitly defined otherwise.

In the description of the present utility model, it should be noted that, unless explicitly specified and limited otherwise, the terms "mounted," "connected," and "connected" are to be construed broadly, and may be either fixedly connected, detachably connected, or integrally connected, for example; can be mechanically connected, electrically connected or can be communicated with each other; can be directly connected or indirectly connected through an intermediate medium, and can be communicated with the inside of two elements or the interaction relationship of the two elements. The specific meaning of the above terms in the present utility model can be understood by those of ordinary skill in the art according to the specific circumstances.

In the description of the present utility model, unless explicitly stated and limited otherwise, a first feature "above" or "below" a second feature may include both the first and second features being in direct contact, or may include both the first and second features not being in direct contact but being in contact by another feature therebetween. Moreover, a first feature being "above," "over" and "on" a second feature includes the first feature being directly above and obliquely above the second feature, or simply indicating that the first feature is higher in level than the second feature. The first feature being "under", "below" and "beneath" the second feature includes the first feature being directly under and obliquely below the second feature, or simply means that the first feature is less level than the second feature.

The following disclosure provides many different embodiments, or examples, for implementing different features of the utility model. In order to simplify the present disclosure, components and arrangements of specific examples are described below. They are, of course, merely examples and are not intended to limit the utility model. Furthermore, the present utility model may repeat reference numerals and/or letters in the various examples, which are for the purpose of brevity and clarity, and which do not themselves indicate the relationship between the various embodiments and/or arrangements discussed. In addition, the present utility model provides examples of various specific processes and materials, but one of ordinary skill in the art will recognize the application of other processes and/or the use of other materials.

As shown in fig. 1, 2 and 3, the droplet single-cell sorting system provided by the embodiment of the utility model comprises a droplet generation and culture module 1, a detection and sorting module 2 and a control module 4, wherein the droplet generation and culture module 1 comprises a power assembly 3, a culture cabin 7, a culture pipeline 8 and a temperature controller 29; the detection and sorting module 2 comprises an optical detection assembly 5 and a sorting deck 6; the control module 4 is used for controlling the operation state of the power assembly 3, reading and recording the measured data of the optical detection assembly 5 in real time, and analyzing and processing the data.

The working principle of the device is as follows: the control module 4 controls the power assembly 3, the power assembly 3 is a combination of an air pump and a liquid pump, the power assembly 3 respectively conveys bacterial liquid, air and oil from the solution bottle 28, the air source 26 and the oil bottle 27 at set flow rates, and the bacterial liquid, the air and the oil pass through a four-way structure at the end part of the culture pipeline 8 to form a three-phase state in which an oil phase 22, an aqueous phase 23 and a gas phase 24 are orderly arranged in the culture pipeline 8. The term "oil" as used herein refers to a reagent prepared by a professional and insoluble in water or bacterial liquid. The aqueous phase 23 is a 1-5. Mu.L volume of droplets, each of which contains one cell 25; the bubbles can prevent the liquid drops from sliding, so that the oxygen dissolving capacity of the liquid drops is enhanced; under the continuous pushing of the power component, the liquid drops are continuously generated, and after the set quantity is reached, the liquid drops are subjected to in-situ culture in the culture pipeline 8, and the culture time and the culture temperature can be set through the control module 4; after the culture is finished, the power assembly 3 continues to push the liquid drops to move, the bubble collecting device 21 eliminates the gas phase 24, so that the influence of the gas on the detection result is avoided, and the optical detection assembly 5 detects optical signals of cells in the water phase 23, including OD, fluorescence, chemiluminescence and the like; the detection and separation module 2 recovers target droplets into a droplet collecting container 14, wherein the droplet collecting container is a common cell culture plate in biological experiments such as 96-well plates, 48-well plates, 24-well plates and the like, and non-target droplets are recovered by a waste liquid collecting mechanism.

As shown in FIG. 2, in some embodiments, a culture pipeline 8 is coiled in a culture cabin 7, the culture pipeline 8 is a gas-permeable micro pipeline, has high permeability to various gases such as carbon dioxide, hydrogen, nitrogen, oxygen and the like, effectively reduces evaporation (< 2%), and improves dissolved oxygen (Kla approximately 90/hr).

A temperature controller 29 is located inside the culture chamber 7, and the temperature controller 29 can adjust the temperature inside the culture chamber 7 so that cells in the pipeline grow at a proper temperature.

As shown in fig. 3, in some embodiments, the detection and sorting module includes a sorting platform 6, an X-axis moving part 9, a Y-axis moving part 10, an a-axis rotating part 11, and a B-axis lifting part 12 are disposed on the sorting platform 6, a support bracket 15 is disposed on the X-axis moving part 9, and a droplet collecting container 14 is disposed on the support bracket 15; the sorting platform 6 is fixedly provided with a first workbench 16 and a second workbench 17, the B-axis lifting part 12 is connected with the A-axis rotating part 11 and drives the A-axis rotating part to lift, and the bottom of the front side of the A-axis rotating part 11 is provided with a clamping mechanism 13; the sorting platform 6 is provided with a cantilever 18, the cantilever 18 is arranged above the X-axis moving part 9 in parallel, the culture pipeline 8 is connected with the cantilever 18, and the optical detection assembly 5 is fixed at the tail end of the cantilever 18 and detects liquid drops in the culture pipeline passing through the cantilever.

Before sorting, the liquid drop collecting containers 14 are stacked and placed on the second workbench 17, the clamping mechanism 13 clamps one liquid drop collecting container 14, and the liquid drop collecting container 14 is placed on the bearing bracket 15 under the cooperative work of the A-axis rotating part 11 and the B-axis lifting part 12; the liquid waste collecting mechanism 19 is arranged below the cantilever 18 in parallel, when the liquid waste is detected as target liquid drops by the optical detection assembly 5, the liquid waste collecting mechanism 19 is staggered with the outlet of the culture pipeline 8, and the liquid drops drop into the liquid drop collecting container 14 under the action of gravity; when the optical detection component 5 detects non-target liquid drops, the liquid drops drop by gravity into the liquid drop collecting mechanism 19, and the liquid drop collecting mechanism 19 is opposite to the outlet of the culture pipeline 8.

After the collection of 1 droplet recovery container 14 is completed, the container is clamped and transferred by a clamping mechanism and placed on the first workbench 16, the clamping mechanism 13 clamps 1 new droplet recovery container 14 from the second workbench 17 and places the container on the bearing bracket 15, and the above operation is repeated until the sorting is completed.

In some embodiments the device supports 10 drop collection containers per batch, which are midway replaceable, with a spatial positioning error of <0.2mm, a rotation angle error of <0.5 °.

In some embodiments, the control module 4 is connected with the droplet generation culture module 1 and the detection sorting module 2, and is used for adjusting the working states of the modules, including adjusting the flow rate of the power assembly 3, controlling the culture temperature, the culture time, controlling the sorting action, recording detection data and performing analysis and processing.

Finally, it should be noted that: the above is only a preferred embodiment of the present utility model, and the present utility model is not limited thereto, but it is to be understood that the present utility model is described in detail with reference to the foregoing embodiments, and modifications and equivalents of some of the technical features described in the foregoing embodiments may be made by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present utility model should be included in the protection scope of the present utility model.

Claims

1. The utility model provides a liquid drop single cell sorting system, includes liquid drop generation culture module, detects sorting module and control module, its characterized in that: the liquid drop generation and culture module comprises a power assembly, a culture cabin, a culture pipeline and a temperature controller, wherein the culture pipeline is connected with the power assembly; the detection and separation module comprises an optical detection assembly and a separation platform; the control module is used for controlling the running state of the power assembly, reading and recording the measured data value of the optical detection assembly in real time, and analyzing and processing the data.

2. A droplet single cell sorting system according to claim 1, characterized in that: the culture cabin is internally coiled with a culture pipeline, and the temperature controller is positioned in the culture cabin.

3. A droplet single cell sorting system according to claim 1, characterized in that: the culture pipeline is a breathable pipeline.

4. A droplet single cell sorting system according to claim 1, characterized in that: the sample injection position of the culture pipeline is set to be of a four-way structure.

5. A droplet single cell sorting system according to claim 1, characterized in that: the detection and separation module comprises a bubble collecting device.

6. A droplet single cell sorting system according to claim 1, characterized in that: the detection sorting module comprises a sorting platform, wherein an X-axis moving part, a Y-axis moving part, an A-axis rotating part and a B-axis lifting part are arranged on the sorting platform, a bearing bracket is arranged on the X-axis moving part, and a liquid drop collecting container is placed on the bearing bracket.

7. The drop single cell sorting system of claim 6, wherein: the sorting platform is fixedly provided with a first working carrying platform and a second working carrying platform, the B-axis lifting part is connected with the A-axis rotating part to drive the A-axis rotating part to lift, and the bottom of the front side of the A-axis rotating part is provided with a clamping mechanism.

8. The drop single cell sorting system of claim 6, wherein: the sorting platform is provided with a cantilever, the cantilever is arranged above the X-axis moving part in parallel, the culture pipeline is connected with the cantilever, and the optical detection assembly is fixed at the tail end of the cantilever and used for detecting liquid drops in the culture pipeline passing through the cantilever.

9. A droplet single cell sorting system according to claim 8, wherein: and when the cantilever is detected as a non-target liquid drop by the optical detection assembly, the liquid drop collecting mechanism is opposite to the outlet of the culture pipeline, and the liquid drop drops into the liquid drop collecting mechanism under the action of gravity.

10. A droplet single cell sorting system according to claim 1, characterized in that: the control module is connected with the liquid drop generation and culture module and the detection and separation module and is used for adjusting the working state of each module, recording detection data and analyzing and processing.