KR-102963752-B1 - CELL-ENCAPSULATED MICRODROPLET SEPARATION DEVICE AND METHOD

KR102963752B1KR 102963752 B1KR102963752 B1KR 102963752B1KR-102963752-B1

Abstract

The present invention relates to a cell-carrying microdroplet separation apparatus and method, wherein the apparatus comprises: a cell inlet unit providing a cell fluid flow from a cell carrier through a first spiral channel; a bead inlet unit providing a bead fluid flow from a bead carrier through a second spiral channel; a microdroplet generating unit that generates microdroplets by combining the end of a carrier oil channel providing a carrier oil flow from a carrier oil carrier, the end of the cell inlet unit, and the end of the bead inlet unit; a microdroplet path providing a movement path for the microdroplets; and an acoustic emitting unit disposed in the microdroplet path and emitting acoustic waves to the microdroplets to separate and discharge target microdroplets and waste materials, respectively.

Inventors

박진수
김우혁
무스타크 알리

Assignees

전남대학교산학협력단

Dates

Publication Date: 20260512
Application Date: 20241031

Claims (11)

A cell inlet unit that provides cell fluid flow from a cell support through a first spiral channel; A bead inlet unit that provides bead fluid flow from a bead support through a second spiral channel; A microdroplet generating unit that generates microdroplets by combining the end of a carrier oil channel providing a flow of carrier oil from a carrier oil support, the end of a cell inlet portion, and the end of a bead inlet portion; A microdroplet path providing a movement path for the above microdroplets; and A cell-carrying microdroplet separation device comprising an acoustic emitting unit disposed in the microdroplet transport path and emitting acoustic waves to the microdroplets to separate target microdroplets and waste and discharge them separately.
In claim 1, the cell inlet portion A cell-carrying microdroplet separation device characterized by placing the cell carrier in the center of the first spiral channel and implementing the end of the first spiral channel in a curved manner.
In claim 1, the bead inlet portion A cell-supported microdroplet separation device characterized by placing the bead carrier in the center of the second spiral channel and flattening the end of the second spiral channel.
In claim 1, the microdroplet generating part A cell-supported microdroplet separation device characterized by arranging the carrier oil carriers symmetrically on both sides of the bonding site and forming the carrier oil channels that each provide a flow path for the carrier oil.
In paragraph 4, the microdroplet generating part A cell-carrying microdroplet separation device characterized by generating microdroplets by wrapping and stabilizing a fluid mixed with the cell and the bead with the carrier oil through flow focusing.
In paragraph 1, A cell-carrying microdroplet separation device characterized by further including a sheath inlet unit that provides additional carrier oil to the microdroplets moving in the microdroplet passage.
In paragraph 1, the sound emitting part A cell-carrying microdroplet separation device characterized by emitting surface acoustic waves in the surface acoustic wave transmission section of the microdroplet transport path to induce acoustic radiation force on the microdroplets.
In Clause 7, the above sound emitting part A cell-carrying microdroplet separation device characterized by emitting surface acoustic waves to deflect the microdroplets moving through the surface acoustic wave transmission section in different directions according to their material properties, thereby separating them into a collection inlet or a waste inlet.
In Clause 7, the above sound emitting part A cell-carrying microdroplet separation device characterized by controlling the separation precision of the microdroplets by adjusting the frequency or intensity of the surface acoustic waves emitted in the surface acoustic wave transmission section.
In Clause 7, the above sound emitting part A piezoelectric substrate that generates the above surface acoustic waves; and A cell-carrying microdroplet separation device characterized by comprising an IDT electrode including first and second electrode patterns arranged interlockingly with a comb-like structure having a constant spacing and thickness, and forming an acoustic pressure field within the surface acoustic wave transmission section through which the microdroplet passes.
In a method for separating cell-loaded microdroplets, A cell inlet step that provides cell fluid flow from a cell support through a first spiral channel; A bead inlet step that provides bead fluid flow from a bead support through a second spiral channel; A microdroplet generation step of generating microdroplets by combining the end of a carrier oil channel providing a flow of carrier oil from a carrier oil support, the end of a cell inlet, and the end of a bead inlet; A microdroplet movement step providing a movement path for the above microdroplet; and A cell-carrying microdroplet separation method comprising an acoustic emission step disposed in the microdroplet transport path and emitting acoustic waves to the microdroplets to separate the target microdroplets and waste and discharge them separately.

Description

Cell-encapsulated microdroplet separation device and method The present invention relates to a cell-supported microdroplet separation technology, and more specifically, to a cell-supported microdroplet separation apparatus and method capable of selectively separating a droplet in which polymer particles and cells are co-supported from an empty droplet through a microfluidic system. Recently, the in-chip manipulation of droplets containing biological samples has garnered attention as an innovative technology in the field of microfluidics, as it enables high-throughput cell analysis and precise control. This has involved encapsulating cells, nucleic acids, or proteins within individual droplets and controlling them via microfluidic systems. By encapsulating cells within microdroplets, individual cells or particles can be provided with distinct environments for analysis, offering advantages such as high-throughput screening and minimized use of biological samples. Furthermore, the generated individual microdroplets can function as miniature reactors, facilitating rapid mixing and the promotion of controlled chemical reactions. In the field of microfluidics, the co-encapsulation of cells and microparticles within microdroplets is being effectively utilized for biological analysis in numerous areas, such as single-cell genetics and drug screening, due to the ability to encapsulate single cells within microdroplets. However, conventional co-loading techniques within microdroplets are limited by the need to strike a balance between the cell/particle pairing control rate and the feasibility of loading single cells within individual microdroplets; consequently, they face the limitation that it is difficult to load single cells within individual microdroplets by matching cell/particle pairs at high speeds. This significantly restricts the generation rate of single-pair cell/particle droplets. Conventional co-loading techniques are based on random loading, so increasing the proportion of single-cell/particle loaded droplets entails an increased possibility of generating multiple-cell/particle loaded droplets, which has a negative impact on single-cell sequencing. One method to reduce the formation of multiple-cell/particle loaded droplets is to decrease the concentration of the liquid containing cells, but this has the limitation of limiting the overall droplet generation rate of single-cell loaded droplets. Furthermore, conventional microfluidic techniques for the separation of cell-embedded microdroplets utilize various detection methods, including fluorescence, absorbance, Raman spectroscopy, and image-based techniques. Generally, fluorescent labeling techniques have been widely used because they allow for the control of microdroplets through simple microscope manipulation. However, the aforementioned conventional techniques have limitations in that they are utilized only in specific, restricted areas, thereby limiting technological advancement through application in diverse fields. Factors limiting the use of the aforementioned detection methods include, first, the significant increase in the overall cost of lab-on-a-chip systems when detection elements are included in microfluidic devices, and second, the possibility of cross-contamination between embedding cells when using microdroplet labeling techniques, which consequently limits the integrity of the results. FIG. 1 is a top view of a cell-carrying microdroplet separation device according to one embodiment of the present invention. Figure 2 is a diagram illustrating the microdroplet generating section of Figure 1. Figure 3 is a diagram illustrating the sound emitting part of Figure 1. Figure 4 is a diagram explaining the principle of droplet separation through acoustic wave interaction. FIG. 5 is a flowchart illustrating a cell-carrying microdroplet separation method according to the present invention. FIG. 6 is a diagram showing a photograph of the particle separation process of a separable microfluidic chip device according to the present invention. Figure 7 is a diagram showing a photograph of an example of a microdroplet separation process according to the size of the sample contained within the microdroplet. Figure 8 is a diagram showing a photograph of an example of a microdroplet separation process according to the compression ratio of a sample contained within a microdroplet. The description of the present invention is merely an example for structural or functional explanation, and therefore the scope of the present invention should not be interpreted as being limited by the examples described in the text. That is, since the examples are subject to various modifications and may take various forms, the scope of the present invention should be understood to include equivalents capable of realizing the technical concept. Furthermore, the objectives or effects presented in the present invention do not imply that a specific example must include all of them