Liquid drop fusion system
By using microfluidic technology to amplify picolinate droplets into larger droplets, and utilizing the detection and electrode control of droplet sorting and fusion modules, the problem of difficult picolinate droplet expansion culture was solved, and the formation of stable monoclonal cell populations and efficient expansion culture were achieved.
Patent Information
- Application Number
- CN202422093831.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-08-28
- Publication Date
- 2025-11-07
- Estimated Expiration
- 2034-08-28
AI Technical Summary
In existing technologies, the number of droplets after picolinate sorting is large, and they are easily affected by surfactants during scale-up culture. Furthermore, traditional methods are difficult to scale up on 96-well plates or plates.
Microfluidic technology is used to amplify picolinol droplets into larger droplets. Controllable flow, cultivation, and sorting of droplets are achieved through droplet sorting and fusion modules. Droplet fusion is controlled using detection devices and electrodes, and gas injection is used to increase the droplet spacing.
This technology enables further cell expansion and culture, forming a large and stable monoclonal population, overcoming the difficulties of traditional expansion culture, and improving operational efficiency and cell growth stability.
Smart Images
Figure CN223522561U_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The utility model relates to micro -fluidic field, concretely is a kind of droplet fusion system. BACKGROUND
[0002] Existing high-flux pico-liter droplet, after sorting, droplet is often placed on 96 well plate or flat plate culture, but the droplet after sorting is still as high as thousands, and the further expansion culture to well plate or flat plate brings huge pressure.And the cell quantity in pico-liter droplet is less, and is inhibited in growth in the process of expansion culture by surfactant.
[0003] Microliter-level droplet has good microorganism culture performance and higher flux, and is easy to operate, can provide sufficient nutrient substance for cell growth, so that cell grows at high speed and metabolizes, obtains a large number of stable monoclonal population, and can be directly placed into shake flask, well plate, flat plate and expanded culture. UTILITY MODEL CONTENT
[0004] In order to solve the above problems, the utility model enlarges small droplet to large droplet through micro-fluidic technology, so that it can flow, culture and sort in pipeline, and therefore the utility model provides a kind of droplet fusion system.
[0005] A kind of droplet fusion system, including droplet sorting module and droplet fusion module, the droplet sorting module includes generation part and sorting part, the generation part is formed by the intersection of several pipelines, pipeline is used to input oil phase and water phase, generates the droplet of " water in oil", the sorting part outside is equipped with detection device and sorting electrode, the detection device is used to detect positive droplet in pipeline, the sorting part downstream is equipped with positive droplet pipeline and negative droplet pipeline;The droplet fusion module includes fusion part, the fusion part outside is equipped with fusion electrode, at least 2 communication parts are included in the upstream of the fusion electrode, the communication part is formed by the intersection of pipeline.
[0006] Preferably, the droplet fusion module further includes spacing part, the spacing part is connected with gas pipeline, and gas is injected into the space between droplets to generate bubbles and increase the spacing between droplets.
[0007] Preferably, the fusion module includes two fusion parts, which are a first fusion part and a second fusion part.
[0008] Preferably, the first fusion part upstream includes a first communication part and a second communication part, and the second fusion part upstream includes a third communication part and a fourth communication part.
[0009] Preferably, the first fusion part upstream includes a first communication part and a second communication part, and the second fusion part upstream includes a third communication part and a fourth communication part.
[0010] Preferably, the third communication part is formed by the intersection of the second oil phase pipeline and the second water phase pipeline, and a second droplet pipeline is arranged downstream of the third communication part.
[0011] Preferably, the droplet transfer module comprises a collection container and a power source, the collection container is connected to the sorting part for collecting the sorted positive droplets, and the power source inputs the positive droplets in the collection container into the droplet fusion module.
[0012] The utility model discloses to solve the problem of the expansion and subculture of pico-liter droplets, overcome the problem of traditional direct sorting to 96-hole plate or flat plate for expansion and culture, combine the characteristics of micro-liter droplets, fuse and amplify pico-liter droplets and micro-liter droplets, realize the further expansion and culture of cells, and form a large number of stable monoclonal populations. BRIEF DESCRIPTION OF DRAWINGS
[0013] The above and / or additional aspects and advantages of the utility model will become apparent and more readily appreciated from the following description of the embodiments, with reference to the following drawings, in which:
[0014] Figure 1 is the structural schematic diagram of the embodiment of the utility model;
[0015] Figure 2 is the structural schematic diagram of the generating part of the embodiment of the utility model;
[0016] Figure 3 is the structural schematic diagram of the sorting part of the embodiment of the utility model;
[0017] Figure 4 is the structural schematic diagram of the droplet fusion module of the embodiment of the utility model. DETAILED DESCRIPTION
[0018] The embodiments of the utility model will be described in detail below, and the examples of the embodiments are shown in the drawings, wherein the same or similar reference signs represent the same or similar elements or elements with the same or similar functions throughout. The embodiments described below with reference to the drawings are exemplary and are only used to explain the utility model, and cannot be understood as a limitation on the utility model.
[0019] In the description of the utility model, it needs to be understood that the orientation or positional relationship indicated by the terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise" and the like is the orientation or positional relationship shown based on the drawings, and is only for the convenience of describing the utility model and simplifying the description, and therefore cannot be understood as a limitation on the utility model that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the utility model.
[0020] In the description of the utility model, it needs to be understood that the orientation or positional relationship indicated by the terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise" and the like is the orientation or positional relationship shown based on the drawings, and is only for the convenience of describing the utility model and simplifying the description, and therefore cannot be understood as a limitation on the utility model that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the utility model.
[0021] The following disclosure provides many different embodiments or examples for implementing different structures of the utility model. In order to simplify the disclosure of the utility model, the components and settings of a specific example are described below. Of course, they are only examples, and the purpose is not to limit the utility model.
[0022] As shown in the figure, the utility model embodiment relates to a kind of droplet fusion systems, including droplet sorting module 1 and droplet fusion module 2, droplet sorting module 1 includes generating part 11 and sorting part 12, generating part 11 is formed by the intersection of several pipelines, respectively by pipeline injection oil phase and water phase, when passing through generating part 11, oil phase cuts water phase, forms the droplet of the interval of oil phase, water phase.
[0023] In the embodiment, generating part 11 is "X" type structure, and in some embodiments, it can also be Y type, T type structure, coaxial structure and the like.
[0024] Further, detection device 14 and sorting electrode 13 are provided outside sorting part 12, positive droplet pipeline 121 and negative droplet pipeline 122 are provided downstream sorting part 12, detection device 14 is at least one of fluorescence detection, optical density detection, Raman detection, infrared detection, scattered light detection, impedance detection or image recognition device, and the detected positive droplet enters positive droplet pipeline 121 under the action of sorting electrode 13, and negative droplet enters negative droplet pipeline 122.
[0025] Further, the system comprises a droplet transfer module 3, the droplet transfer module 3 comprises a collection container 32 and a power source 31, the collection container 32 collects the sorted positive droplets, and the power source 31 inputs the positive droplets in the collection container 32 into the droplet fusion module 2 through a droplet sample pipe 211.
[0026] The droplet fusion module 2 comprises a fusion section, and a fusion electrode 212 is arranged outside the fusion section, the fusion electrode 212 changes the surface tension of the droplets through the electrode, so that adjacent droplets in the pipe can be fused, and the fusion electrode 212 upstream comprises at least two communication sections, and the communication sections are formed by the intersection of the pipes.
[0027] The droplet fusion module 2 further comprises a spacing section 203, and the spacing section 203 is connected with a gas pipe 216, and the gas pipe 216 injects gas into the space between the droplets to generate bubbles, so as to increase the spacing between the droplets and facilitate subsequent operation.
[0028] In some embodiments, the fusion section comprises two fusion sections, i.e. a first fusion section 201 and a second fusion section 202, in the microfluidic system, the picoliter microdroplets are difficult to be accurately controlled due to their small volume, and the embodiment is intended to expand the volume of the microdroplets through two times of fusion, so that the microdroplets are easy to be detected and controlled.
[0029] Specifically, the picoliter droplets are expanded by one time of fusion through the first fusion section 201, and the volume of the droplets is expanded by two times of fusion through the second fusion section 202, and the expansion multiple of each time is, for example, 1 times, 2 times, 5 times, 10 times, etc. It should be noted that the embodiment only illustrates the principle of expanding the volume of the microdroplets by two times of fusion, and does not limit the size of the initial microdroplets and the expansion multiple of each fusion, and the initial size of the microdroplets and the expansion multiple should be determined according to actual needs, which can be adjusted by changing the pipe size, solution flow rate and other factors.
[0030] Further, the first fusion section 201 upstream comprises a first communication section 204 and a second communication section 205, and the second fusion section 202 upstream comprises a third communication section 206 and a fourth communication section 207.
[0031] Specifically, the first communication section 204 is formed by the intersection of a first oil phase pipe 208 and a first water phase pipe 209, and the oil phase and the water phase form an intermediate droplet with an oil phase and a water phase spacing after passing through the first communication section 204, the second communication section 205 is formed by the intersection of a first droplet pipe 210 and the droplet sample pipe 211, the positive droplets enter the first droplet pipe 210 from the second communication section 205, and the intermediate droplets and the positive droplets are arranged adjacent to each other after passing through the second communication section 205, and flow through the first fusion section 201, and under the action of the fusion electrode 212, one adjacent intermediate droplet and the positive droplet will fuse with each other; the intermediate droplet fused with the positive droplet continues to move downstream along the first droplet pipe 210.
[0032] Specifically, the second oil phase pipeline 213, the second water phase pipeline 214, and the third communication part 206 are formed by the intersection of the second oil phase pipeline 213 and the second water phase pipeline 214, and the oil phase and the water phase form large liquid drops after passing through the third communication part 206, and the downstream of the third communication part 206 is configured as the second liquid drop pipeline 215, and the first liquid drop pipeline 210 and the second liquid drop pipeline 215 intersect at the fourth communication part 207.
[0033] The medium liquid drops and the large liquid drops meet at the fourth communication part 207, and are arranged in an adjacent and spaced manner by controlling the flow rate, and flow through the second fusion part 202 and are fused under the action of the fusion electrode 212, and the size of the liquid drops is further enlarged.
[0034] It should be emphasized that, in the embodiments of the utility model, the liquid drops at each stage in the same experiment are named as positive liquid drops, medium liquid drops, large liquid drops, and the description of the fusion and enlargement multiple of the liquid drops is for better explanation and help for the understanding of the technical scheme by the relevant technical personnel. In the microfluidic system, the volume of the liquid drops is usually small, and can be several picoliters of small liquid drops to microliters of liquid drops, the volume of the medium liquid drops can be several times to tens of thousands of times of the positive liquid drops, and the volume of the large liquid drops can be several times to tens of thousands of times of the medium liquid drops, the description of the volume and multiple of the liquid drops in the embodiments of the utility model is only for more intuitive explanation of the principle and process of the step-by-step enlargement of the micro liquid drops, and the size of the liquid drops at each stage is not limited.
[0035] Although the embodiments of the utility model have been shown and described, those skilled in the art can understand that various changes, modifications, replacements and variations can be made to the embodiments without departing from the principles and purposes of the utility model, and the scope of the utility model is defined by the claims and the equivalents thereof.
Claims
1. A droplet fusion system, characterized by, The application relates to a liquid droplet sorting and fusing module, which comprises a liquid droplet sorting module and a liquid droplet fusing module.
2. A droplet fusion system according to claim 1, wherein, The liquid droplet fusing module further comprises a spacing part connected with a gas pipeline, which injects gas into the liquid droplets to generate bubbles and increase the spacing between the liquid droplets.
3. The droplet fusion system of claim 1, wherein, The liquid droplet fusing module further comprises a spacing part connected with a gas pipeline, which injects gas into the liquid droplets to generate bubbles and increase the spacing between the liquid droplets.
4. A droplet fusion system according to claim 3, wherein, The first fusing part upstream comprises a first connecting part and a second connecting part, and the second fusing part upstream comprises a third connecting part and a fourth connecting part.
5. A droplet fusion system according to claim 4, wherein, The first fusing part upstream comprises a first connecting part and a second connecting part, and the second fusing part upstream comprises a third connecting part and a fourth connecting part.
6. A droplet fusion system according to claim 5, wherein, The first fusing part upstream comprises a first connecting part and a second connecting part, and the second fusing part upstream comprises a third connecting part and a fourth connecting part.
7. The droplet fusion system of claim 1, wherein, The first fusing part upstream comprises a first connecting part and a second connecting part, and the second fusing part upstream comprises a third connecting part and a fourth connecting part. The liquid droplet fusing module further comprises a spacing part connected with a gas pipeline, which injects gas into the liquid droplets to generate bubbles and increase the spacing between the liquid droplets.