KR102077643B1 - Apparatus for Separating Particles of Sample with Meniscuss and Method thereof - Google Patents

Abstract

The present invention discloses a device for separating particles by moving a sample having a meniscus curved surface on a substrate, particularly as a particle separation device. The particle separation device of the present invention comprises a base portion; And a slot unit formed on one surface of the base unit and provided with a first group of slots for storing at least some particles separated from a sample containing particles to be separated. It includes.

Description

Apparatus for Separating Particles of Sample with Meniscuss and Method thereof

The present invention relates to a particle separation device and method. More particularly, it relates to a device for separating desired particles in a sample.

Molecules are the smallest particles with the properties of a substance, and in biological fluids, exosomes are small particles of a membrane structure that are secreted from various types of cells. Exosomes (exosomes) are small vesicles of a membrane structure, and are reported to be approximately 30-100 nm in diameter.

Exosomes originate from specific compartments in cells called polycysts and are released and secreted out of the cells, and when fusion of the polycysts and the plasma membrane occurs, such vesicles are released into the extracellular environment, which are called exosomes. It is known that exosomes are released separately from a number of different cell types under both conditions, normal and pathological. Exosomes contain various RNAs, and studies on how to diagnose the disease by detecting the presence or absence and presence of the RNA are active. However, in order to make an accurate diagnosis using exosomes, it is important to know the exact amount of exosomes present in a human body having a disease.

Conventional techniques for isolating and purifying exosomes include methods such as exosome isolation reagent, exomir using two filters of pore size, exoquick and ultracentrifugation, but the current technique is a considerable time and cost. And there are limitations that require specialized research facilities.

In addition, the conventional methods for measuring exosomes include quantitative methods using specific immune responses of antibodies and antigens, but are cumbersome and have limitations that can be applied only to microvesicles and exosomes.

Therefore, there is a need to develop a technology that solves the problem of a complex exosome quantification method and improves the accuracy of a diagnostic method by separating a specific exosome.

Korean Registered Patent No. 10-1738117 (Public)

The present invention has been devised to solve the above problems, and discloses a particle separation device. In particular, a device for separating particles in a sample having a meniscus curved surface is disclosed.

The present invention has been devised to achieve the above object, the particle separation device of the present invention comprises a base portion; And a slot unit formed on one surface of the base unit and provided with a first group of slots for storing at least some particles separated from a sample containing particles to be separated. It includes.

In the present invention, the slot unit may further include a second group of slots having a different depth or width from the slots of the first group.

In the present invention, the particle separation device is at least partially from the sample through dragging at a predetermined angle in a state where the sample is positioned in a predetermined shape between the first group of slots and the surface on which the second group of slots is formed. Dragging unit for separating the particles of the; It may further include.

In the present invention, the depth or width of the slots of the first group and the slots of the second group is preset according to the size of the particles to be separated, and the slots of the first group and the slots of the second group are Particles having the predetermined size may be classified and stored.

In the present invention, the slot part may include different mask patterns including at least a first photoresist pattern and a second photoresist pattern provided in consideration of the depth or width of the first group of slots and the second group of slots. It may be formed by applying on the base portion and etching the base portion using the applied mask pattern as an etch mask.

In the present invention, the alignment aid for aligning the particles included in the sample according to size and holding particles aligned according to the size in the sample; Further comprising, the dragging unit may separate the particles by dragging the sample containing the particles held in an aligned state.

In the present invention, the particle separation device includes an extraction unit for extracting the particles stored in the first group of slots and the second group of slots; Further comprising, the extraction unit may be extracted by separating the particles stored in the slots of the first group and the slots of the second group from the sample.

In the present invention, the alignment auxiliary unit is formed on the other surface of the dragging unit where the sample is located, and a power supply unit that forms an electric field in the sample through at least two or more electrodes fastened to the dragging unit; Further comprising, it is possible to arrange the size of the particles according to the direction of the formed electric field.

In the present invention, the alignment auxiliary unit is an ion selective permeable membrane connected to one end of the electrode selectively transmitting the particles according to different charge characteristics of the particles included in the sample; Further comprising, the transmitted particles may be polarized according to the electric field formed in the sample through the electrode.

In the present invention, the particles are exosomes as vesicles containing at least one protein and RNA having different sizes, and the sample may be prepared as an electrolytic sample including at least one ionized particle.

In the present invention, the sample includes a first tail formed adjacent to the dragging portion and a second tail formed adjacent to the slot portion on the opposite side of the direction in which the dragging portion is dragged, and the first tail and the second The surface of the sample between the tails may be formed by a meniscus curved surface that is concavely bent in the direction in which the dragging portion is dragged.

In the present invention, the meniscus curved surface, the length of the first tail and the second tail, the thickness of the first tail and the second tail, and the volume of the sample included in the first tail and the second tail are the base portion , It may be formed differently according to the hydrophilicity of the slot portion and the dragging portion, the predetermined angle and the attraction force between the particles contained in the sample.

In the present invention, the slots of the first group and the slots of the second group are formed for each group in at least a portion of the slot part, and the particles in the sample included in the first tail and the second tail are the first tail And as the distance from the second tail increases, the size may gradually increase.

In addition, in order to achieve the above object, the particle separation method of the present invention is formed on one surface of the base portion, the first group of slots for storing at least some of the particles separated from the sample containing the particles to be separated and the agent Positioning the sample on at least one side of a slot portion in which a second group of slots having different depths or widths from one group of slots is provided; Aligning the dragging unit for separating at least some of the particles from the sample through dragging at a predetermined angle in a state where the sample is positioned in a predetermined shape; And dragging the aligned dragging portion on the slot portion to separate and store particles included in the sample in the slots of the first group and the slots of the second group. It includes.

In the present invention, the method for separating particles comprises sorting the particles included in the sample according to size and holding particles aligned according to the size in the sample; Further comprising, the dragging unit may separate the particles by dragging the sample containing the particles held in an aligned state.

In the present invention, the holding step includes forming an electric field in the sample through at least two or more electrodes formed on the other surface of the dragging portion where the sample is located and fastened to the dragging portion; And selectively transmitting the particles according to different charge characteristics of the particles included in the sample through an ion-selective permeable membrane connected to one end of the electrode. Further comprising, it may be arranged according to the size of the particles by polarizing the transmitted particles according to the electric field formed in the sample through the electrode.

In the present invention, the sample includes a first tail formed adjacent to the dragging portion and a second tail formed adjacent to the slot portion on the opposite side of the direction in which the dragging portion is dragged, and the first tail and the second The surface of the sample between the tails may be formed by a meniscus curved surface that is concavely bent in the direction in which the dragging portion is dragged.

In the present invention, the meniscus curved surface, the length of the first tail and the second tail, the thickness of the first tail and the second tail, and the volume of the sample included in the first tail and the second tail are the base portion , The hydrophilicity of the slot portion and the dragging portion, the predetermined angle and formed differently according to the attraction force between the particles contained in the sample, the particles in the sample included in the first tail and the second tail are the first The size may be gradually increased as the distance from the tail and the second tail increases.

In the present invention, the depth or width of the slots of the first group and the slots of the second group is preset according to the size of the particles to be separated, and the slots of the first group and the slots of the second group are The particles having the predetermined size are separately stored, and the slots of the first group and the slots of the second group may be formed for each group in at least a portion of the slot portion.

In the present invention, the slot unit may include different mask patterns including at least a first photoresist pattern and a second photoresist pattern provided in consideration of the depth or width of the first group of slots and the second group of slots. It may be formed by applying on the base portion and etching the base portion using the applied mask pattern as an etch mask.

According to the present invention, target particles can be separated from the sample.

In particular, particles in a sample having a meniscus curved surface can be easily separated by dragging between the substrates.

1 is a block diagram of a particle separation device according to an embodiment of the present invention.
Figure 2 shows the operation preparation state of the particle separation device according to an embodiment of the present invention.
Figure 3 shows a detailed configuration of the particle separation device according to an embodiment of the present invention.
4 shows a configuration of an alignment assistant according to an embodiment of the present invention.
Figure 5 shows the state of the particles separated using the particle separation device of the present invention.
6 shows a configuration of an alignment assistant according to another embodiment of the present invention.
7 is a flowchart of a particle separation method according to an embodiment of the present invention.
8 is an enlarged flow chart of the holding step in the embodiment of FIG. 7.

Hereinafter, an embodiment of the present invention will be described in detail with reference to the accompanying drawings.

In the description with reference to the accompanying drawings, the same or corresponding components are assigned the same reference numbers and redundant description thereof will be omitted.

In addition, in describing the present invention, when it is determined that detailed descriptions of related well-known configurations or functions may obscure the subject matter of the present invention, detailed descriptions thereof may be omitted.

Terms used in the present application are only used to describe specific embodiments, and are not intended to limit the terms. Singular expressions include plural expressions unless the context clearly indicates otherwise. Each of the steps described below may be provided with one or several software modules or may be implemented with hardware in charge of each function, or a combination of software and hardware. The specific meaning and examples of each term will be described below in the order of each drawing. Hereinafter, the configuration of the particle separation device 10 according to the embodiment of the present invention will be described in detail with reference to the associated drawings.

1 is a block diagram of a particle separation device 10 according to an embodiment of the present invention. The configuration of the particle separation device 10 will be described with reference to FIG. 2.

The particle separation device 10 of the present invention includes a base portion 100, a dragging portion 200, an alignment auxiliary portion 300, and an extraction portion 400. For example, the particle separation device 10 of the present invention is a particle to be separated by moving a sample containing particles located between two substrates (plates) on the base portion 100 using the dragging portion 200 You can separate them. The particles to be separated by the particle separation device 10 of the present invention are particles having a size, and the smallest particle having the property of a substance (Molecule) and RNA and various proteins in a biological fluid are exosomes (Exosome) ). The particle separation device 10 of the present invention can extract only the desired exosomes by separating and separating approximately 30 to 100 nm in diameter exosomes by size, which are known to be secreted specifically for the purpose of signal transmission between cells. This will be described with reference to FIG. 3.

The base portion 100 includes a slot portion 110. For example, the base part 100 forms a slot part 110 on at least one surface so that at least the first group of slots and the second group of slots storing particles to be separated are disposed on the upper surface. The base unit 100 may be disposed on the lower surface of the slot unit 110 to support the slot unit 110. The base portion 100 of the present invention may be made of a glass wafer (Glass Wafer) or a silicon wafer (Silicon Wafer), silicon germanium or silicon-germanium material.

In addition, the base portion 100 of the present invention may be provided as a bulk wafer or an epitaxial layer, and may be formed of an SOI substrate, a gallium-arsenide substrate, or the like. A cleaning process may be performed prior to providing storage members including the first group of slots 120 and the second group of slots 140. The base portion 100 is a wet etchant used for fine processing, or is formed through a process for etching a thin film of silicon dioxide or silicon nitride using Buffered Oxide Etch (BOE), or by using a piranha etching technique It can be produced by adding an additional process to remove the organic residue on the base portion with a mixture of sulfuric acid and hydrogen peroxide. Since the base portion 100 and the slot portion 120 of the present invention are cleaned through the Piranha etching process, the surface of the base portion may be hydroxylated to have higher hydrophilicity. As will be described later, the hydrophilicity of the base portion 100 and the dragging portion 200 of the present invention determines the shape of the meniscus curve of the sample located between the base portion 100 and the dragging portion 200, and It is an important component that affects separation.

The slot unit 110 is at least one storage member for storing particles to be separated. The first group of slots 120 and the second group of slots 140 are formed on a surface to store particles. For example, the storage members of the slot unit 110 include at least a first group of slots 120 and a second group of slots 140 according to depth or width, and the first group of slots ( 120) and the second group of slots 140 may be formed for each group in the slot portion 110 formed on the upper surface of the base portion. For example, the depth or width of the first group of slots 120 and the second group of slots 140 is preset according to the size of particles to be separated, and the first group of slots 120 And the second group of slots 140 may be divided and stored particles having a predetermined size. Of course, the slot unit 110 of the present invention may further include slots for separating particles having different specifications in addition to the aforementioned first group of slots 120 and second group of slots 140. .

Also, the first group of slots 120 and the second group of slots 140 formed on at least a portion of the slot unit 110 of the present invention may be formed through a photolithography process. For example, the first group of slots 120 and the second group of slots 140 may take into account the depth or width of the first group of slots 120 and the second group of slots 140. Different mask patterns including at least the first photoresist patterns 162, 164, and 168 and the second photoresist patterns 172, 174, and 178 are provided on the base portion 100, and the applied mask The base portion 100 may be etched using a pattern as an etching mask. In the present invention, the etch mask may be made of various materials according to a desired pattern application, and may be made of a metal such as tungsten or aluminum, a semiconductor such as polysilicon, and a conductive metal nitride.

First photoresist patterns 162, 164 and 168 and second photoresist patterns 172, 174 and 178 for forming the first group of slots 120 and the second group of slots 140 of the present invention ) May be formed through a photolithography process, for example, Immersion Lithography or EUV (Extreme Ultraviolet) lithography technology may be used. Immersion lithography technology is a technique that improves resolution by increasing the numerical aperture by filling a high refractive index fluid between the lens and the object to be exposed. The first group of slots 120 and the second group of slots 140 of the present invention etch the first photoresist pattern 162, 164 and 168 and the second photoresist pattern 172, 174 and 178 The base portion 100 may be formed by etching as a mask, and the etching process includes dry etching, wet etching, or reactive ion etching (RIE). For example, when the etching mask is made of a carbon-containing film, a plasma etching process using a mixed gas of oxygen and argon may be performed. In forming the first group of slots 120 and the second group of slots 140, the first photoresist pattern 162, 164 and 168 and the second photoresist pattern 172, 174 and 178 are etched It can be removed after the process. When using the reactive ion etching method (RIE) when forming the first group of slots 120 and the second group of slots 140 formed in the slot unit 110 of the present invention, the first group of slots Different levels of etching techniques may be used according to the depth and width of the fields 120 and the second group of slots 140.

The dragging unit 200 is dragged at a predetermined angle in a state where the sample is positioned in a predetermined shape between the first group of slots 120 and the second group of slots 140 are formed. At least some of the particles are separated from the sample. For example, the dragging unit 200 stands at an angle on the slot unit 110 and at least one end contacts the slot unit 110 or approaches at a predetermined interval, and the slot unit 110 and the dragging unit 200 By placing the sample including the particles (542, 544) in the open space of the, it is possible to move the positioned sample on the slot portion 100. The dragging part 200 of the present invention may further include a fastening part through which an electrode penetrates and is fastened as described later.

In the present invention, the sample located between the slot part 110 and the dragging part 200 includes an electrolytic sample including at least one ionized particles, and the particles included in the sample have at least one protein having different sizes. And exosomes as vesicles containing RNA.

For example, the sample located on the upper surface of the slot part 110 and below the dragging part 200 is formed first adjacent to the dragging part 200 on the opposite side of the direction in which the dragging part 200 moves. A tail 520 and a second tail 540 formed adjacent to the slot portion 110, and between the first tail 520 and the second tail 540, the surface of the sample is the dragging portion It may be formed of a meniscus curved surface 560 that is concavely bent in the direction in which the 200 moves. For example, in the present invention, since the dragging part 200 is dragged at a predetermined angle from the slot part 110, the particle separation device 10 is in contact with the top surface of the dragging part 200 and the slot part 110. It may include a space provided at an acute angle or an obtuse angle between.

In the present invention, the sample containing the particles to be separated is located in a space formed narrowly at an acute angle between the dragging part 200 and the slot part 110 (the upper surface of the slot part and the lower surface of the dragging part), and the dragging part 200 is obtuse. The particles may be separated as they are dragged in the spatial direction formed by, but the dragging unit (located in the space formed broadly at an obtuse angle between the dragging unit 200 and the slot unit 110 (the upper surface of the slot unit and the upper surface of the dragging unit) As 200) is dragged in a narrow space direction formed at an acute angle, particles may be separated. That is, as described later, the direction in which the dragging unit 200 is dragged on the upper surface of the base unit 100 may vary according to the direction in which the sample of the present invention is located between the dragging unit 200 and the base unit 100, In addition, the meniscus curved surface 560 formed between the first tail 520 and the second tail 540 of the sample may also be formed differently.

In the present invention, the meniscus curved surface, the length of the first tail and the second tail, the thickness of the first tail and the second tail, and the volume of the sample included in the first tail and the second tail are the base portion And the hydrophilicity of the dragging part, the predetermined angle, and the attraction force between particles contained in the sample. In addition, the meniscus curved surface 560 of the present invention, the hydrophilicity of the base portion 100 and the dragging portion 200, the height of the contact portion of the base portion 100 and the dragging portion 200, the dragging portion 200 A may be formed differently depending on the angle and dragging angle located on the upper surface of the base portion 100, the volume of the sample located between the base portion 100 and the dragging portion 200. For example, in the sample located between the slot portion 110 and the dragging portion 200 of the present invention, the first tail 520 and the second tail 540 are connected to the slot portion 110 and the dragging portion 220. As a part formed by a narrow area of an adjacent narrow height, only small particles 5440 in the sample may be located.

For example, the meniscus curved shape formed between the slot part 110 and the dragging part 200 in the sample of the present invention includes the attractive force of particles in the sample, the surface tension of the solvent, the base part 100, and the slot part ( 110) and the dragging part 200 may be formed differently depending on the hydrophilicity (Hydrophilicity) of the surface. The particle separation device 10 of the present invention can effectively separate particles by maintaining the shape of the sample as a meniscus curved surface between the slot portion 110 and the dragging portion 200. Of course, the meniscus curved surface may be formed of different meniscus curved surfaces according to the heights of the contact portions of the base portion 100 and the dragging portion 200.

In addition, the particle separation device 10 of the present invention can adjust the degree of hydrophilicity of the base portion 100, the slot portion 110 and the dragging portion 200 in order to adjust the meniscus shape. For example, the base part 100, the slot part 110, and the dragging part 200 of the present invention may be formed with functional groups on the surface using etching means including ionized plasma gas or heat at high temperature, Through this, the surface can be hydroxylated (-oh) to improve hydrophilicity. In addition, the base portion 100, the slot portion 110, and the dragging portion 200 of the present invention can control the hydrophilicity by smoothly or artificially bending the surface through a micro / nano process in terms of structure. Through the phosphorus method, the hydrophilicity of the surfaces of the base portion 100, the slot portion 110, and the dragging portion 200 may be improved. This will be described with reference to FIG. 4.

The alignment aid 300 aligns the particles included in the sample according to the size, and holds the particles aligned according to the size in the sample. For example, the alignment auxiliary unit 300 may include the dragging unit 200 through the electrophoresis, ion concentration polarization, and surface acoustic wave, the slot portion of the upper surface of the base unit 100 ( 110) It is possible to align and hold the particles in the sample before moving on. The alignment auxiliary unit 300 may align and hold the particles in the sample before the dragging unit 200 moves, so that the sample is sequentially separated according to the size of the particles when the sample moves as the dragging unit 200 moves. .

Alignment auxiliary unit 300 of the present invention is formed on the other side of the dragging portion where the sample is located, further comprising a power supply unit 320 for forming an electric field in the sample through at least two electrodes fastened to the dragging unit, the power supply unit ( 320) the size of the particles can be aligned according to the direction of the electric field formed by the. For example, the alignment auxiliary unit 300 is a dragging unit to connect an electrode in a sample located between the slot 110 and the dragging unit 200 on the other surface of the dragging unit 200 adjacent to the slot unit 110. An electric field may be formed in the sample through the fastening part included in the 200. The alignment auxiliary unit 300 of the present invention may form an electric field in the sample, and use the properties of particles in the sample having different electrical affinity to maintain the particles in the sample in an aligned state before the sample moves.

In addition, the alignment assistant 300 further includes an ion-selective permeable membrane 340 connected to one end of the electrode selectively transmitting the particles according to different charge characteristics of the particles included in the sample in addition to the power source 320. And, the transmitted particles can be polarized according to the electric field formed in the sample through the electrode. The ion selective permeable membrane 340 of the present invention may be made of Nafion. For example, the ion-selective permeable membrane of the present invention includes at least one nano-permeable membrane, and discharges only ionized particles that are charged by forming an ion depletion region near the nano-permeable membrane in consideration of different charge characteristics of the particles. can do.

More specifically, the ion-selective permeable membrane 340 behaves predominantly with respect to cations that do not coincide with ion conductivity in the electrolytic sample, so that an ion concentration gradient is generated on both sides of the ion-selective permeable membrane. When the ion concentration polarization (ICP) by the ion-selective permeable membrane 340 is induced near the ion-selective permeable membrane, both the cation and anion concentrations decrease at the anode side of the junction surface and increase at the cathode side, resulting in ion deficiency or excess. A phenomenon appears, and through this phenomenon, an ion depletion region can be formed. The excessive or depleted ionized region formed by the ion-selective permeable membrane 340 may also be applied to charged particles, cells, and other small colloids to form an excessive region of ionized particles and a region lacking ionized particles. It can also be applied to the particle separation device 10 for separating particles by moving a sample containing ionized particles of the present invention. Particles in a sample located between the slot part 110 and the dragging part 200 of the present invention may be self-assembled in the sample and crystallized. In addition, the particles in the sample included in the first tail and the second tail may be provided to gradually increase in size as they move away from the first tail and the second tail.

The extraction unit 400 extracts the particles stored in the first group of slots 120 and the second group of slots 140. For example, the extraction unit 400 may use negative pressure or other known methods to separate and extract the separated particles stored in the storage members from the sample. In addition, the extraction unit 400, the first group of slots 120 and the second group of slots 120 and the second group of the particles are stored in order to extract the particles stored in the slots 140 and the second group of groups 120 A PDMS film may be formed on the slots 140 of the particles, and particles may be extracted based on the formed PDMS film.

2 shows an alignment state of the particle separation device 10 according to an embodiment of the present invention.

Before moving the sample by dragging the dragging unit 200 on the slot unit 110 of the present invention, it is necessary to align the slot unit 110 and the dragging unit 200. For example, the shape of a sample having a meniscus curved surface positioned between the slot portion 110 and the dragging portion 200 may be adjusted in consideration of a contact angle on the slot portion 110 of the dragging portion 200. . In the present invention, the sample is made of a meniscus curved shape including a first tail 520 and a second tail 540, and the shape of the meniscus curved surface is a group of the slot part 110 and the dragging part 200. It can be set differently according to the set angle, the hydrophilicity of the slot portion 110 and the dragging portion 200, the surface tension of the sample, and the attractive force of particles in the sample. In addition, the meniscus curved surface 560 of the sample is located on the top surface of the slot part 110 and below the dragging part 200 or on the top surface of the slot part 110 and the top surface of the dragging part 200. It can be formed differently depending on the case as described above.

Figure 3 shows a detailed configuration of the particle separation device according to an embodiment of the present invention.

The base unit 100 includes a slot unit 110 including at least one slot for storing particles on an upper surface. The slot unit 110 includes at least one slot for storing particles to be separated. For example, in the present invention, the slot unit 110 includes at least a portion of the first group of slots 120 and the second group of slots 140. Depth 122, width 124, first slot spacing 126 of the first group of slots 120 and depth 142, width 144, second of the second group of slots 140 The slot spacing 146 and the third slot spacing 150 between the first group of slots and the second group of slots are provided differently depending on the size and type of particles to be separated and the moving speed of the dragging unit 200 Can be.

In the present invention, to form the first group of slots 120 and the second group of slots 140, the base portion is utilized by using predetermined photoresist patterns 162, 164, 168, 172, 174, and 178 The specific method of forming (100) by etching is the same as described above, and thus is omitted.

The sample located between the slot portion 100 and the dragging portion 200 of the present invention may be positioned in a meniscus shape before being moved on the base portion 100. The sample positioned in the meniscus shape of the present invention includes a first tail 520 and a second tail 540, and particles having a small size may be located in the first tail 520 and the second tail 540. You can. In the present invention, the meniscus shape of the sample may be determined according to the hydrophilicity of the solvent in the sample, the interparticle attraction of the particles in the sample, the degree of hydrophilicity of the base part 100, the slot part 110, and the dragging part 200. .

In addition, the particle separation device 10 of the present invention can separate particles using a coffee stain effect (coffee ring effect). For example, the sample located in the slot portion 110 and the dragging portion 200 may be formed in a hat shape due to the attraction of the internal particles and the solvent, the contact angle of the liquid in contact with the solid, etc., wherein the particles in the sample They may be provided in a spinn-ring state. In order for the liquid to be rounded in the shape of a hat, a predetermined threshold must be satisfied, and the threshold is determined only by the interface energy of the liquid, the substrate and air, and the contact angle of the liquid droplet. When the liquid is positioned round, the amount of evaporation at the center and edge is greatly changed, and the amount of evaporation is generated because the contact area of the edge is large, so that the flow of the internal liquid supplies the liquid to the edge, and the particles inside the evaporation amount It can show the phenomenon of flowing into this large part.

For example, particles in a sample located in a meniscus curved shape between the slot part 110 and the dragging part 200 may be mainly located in the first tail 520 and the second tail 540 according to the coffee stain effect. have. At this time, depending on the size of the particles, the mass and the surface tension of the liquid, small particles may be mainly located in the narrow portions of the first tail 520 and the second tail 540. Therefore, large-sized particles are mainly located on the side of the direction of movement of the dragging unit 200. When the sample including the particles is aligned on the slot unit 110, the large-sized particles are slots of the second group. It passes through the field 140 is stored in the first group of slots 120, the small particles 544 are stored in the second group of slots (140).

The particle separation device 10 of the present invention may further include an alignment auxiliary unit 300 to further improve the alignment state according to the size of particles in the sample, and the alignment auxiliary unit 300 may hold and align particles in the sample Since the specific method is as described above, it is omitted.

Figure 4 shows the configuration of the alignment aid 300 according to an embodiment of the present invention.

The alignment assistant 300 of the present invention sorts the particles included in the sample according to the size, and holds the particles aligned according to the size in the sample. For example, the alignment auxiliary unit 300 further includes a power source unit 320 and an ion-selective permeable membrane 340, and the particles 542 and 544 included in the sample are aligned and held in different sizes. For example, in the present invention, the ion-selective permeable membrane 340 may be provided as Nafion, as described above, and an electrode for supplying power of the power supply unit 320 into the sample is fastened in the dragging unit 200. It is as described above that the fastening portion for becoming may be further included in the dragging portion 200.

Figure 5 shows the state of the particles separated using the particle separation device of the present invention.

The particle separation device 10 of the present invention moves the sample including particles 543 and 544 to be separated between the slot portion 110 and the dragging portion 200 on the slot portion 110, The particles can be separated. For example, the particle separation device 10 of the present invention drags the dragging unit 200 and performs a cleaning process to process particles not stored in the first slots 120 and the second slots 140. It may further include.

6 shows a configuration of the alignment aid 300 according to another embodiment of the present invention.

The alignment auxiliary unit 300 of the present invention sorts the particles included in the sample according to size, and generates an acoustic wave to generate a surface acoustic wave in order to hold the particles arranged according to the size in the sample. 360 may be further included, and the particles included in the sample may be sorted according to the size using the surface acoustic wave generated by the seismic generator 360. For example, the alignment auxiliary unit 300 of the present invention is one surface of the dragging unit 200 positioned in the direction of the sample in order to transmit the surface acoustic wave generated by the acoustic wave generating unit 360 in the sample Further included, it is possible to transmit a surface acoustic wave (Surface Acoustic Wave) generated by the seismic generator through the seismic connection portion in the sample.

7 is a flowchart of a particle separation method according to an embodiment of the present invention.

The particle separation method of the present invention includes the following steps that the particle separation device 10 performs in time series.

In S100, the particle separation device 10 is formed on one surface of the base portion, the first group of slots 120 and the first group for storing at least some of the particles separated from the sample containing the particles to be separated The sample is placed on at least one side of the slot unit 110 in which the second group of slots 140 having different depths or widths from the slots of the are provided. Although the particle separation device 10 uses the dragging unit 200 to place the sample to be moved on the slot unit 100, there is no particular limitation, but the first group of slots 120 and the second group of slots It may be located on at least one surface of the slot portion 110 where 140 is not located.

In the present invention, the sample is a first tail 520 formed adjacent to the dragging portion 200 and a second tail formed adjacent to the slot portion 110 on the opposite side of the direction in which the dragging portion 200 moves ( 540), the surface of the sample between the first tail and the second tail may be formed as a meniscus curved surface that is concavely bent in a direction in which the dragging part moves, so it is omitted. .

In addition, the first group of slots 120 and the second group of slots 140 may depend on the depth or width of the first group of slots 120 and the second group of slots 140. Grouped by group in at least a portion of the slot portion 110 may be formed. At least a first photo provided in consideration of the depth or width of the first group of slots 120 and the second group of slots 140 formed in the slot unit 110 located on the upper surface of the base unit 100 As described above, different mask patterns including a resist pattern and a second photoresist pattern may be formed by coating the base part and etching the base part 100 using the applied mask pattern as an etching mask. same.

In S200, the particle separation device 10 aligns the dragging unit 200 for separating at least some particles from the sample through dragging at a predetermined angle in a state where the sample is positioned in a predetermined shape. For example, the particle separation device 10 may include a menis that includes a first tail 520 and a second tail 540 suitable for separating particles between samples between the slot 110 and the dragging unit 200. The dragging part 200 may be aligned on the slot part 110 so as to have a curse shape. At this time, the pre-set angle at which the dragging unit 200 is located is the size of the particles, the hydrophilicity of the sample, the attraction of the particles in the sample, the hydrophilicity of the base unit 100, the slot unit 110 and the dragging unit 200, the first It may be set in consideration of the length of the tail 520 and the second tail 540 and the curvature of the meniscus surface.

In the present invention, the meniscus curved surface 560, the length of the first tail 520 and the second tail 540, the thickness of the first tail 520 and the second tail 540 and the first tail ( 520) and the volume of the sample included in the second tail 540 is between the hydrophilicity of the base portion 100, the slot portion 110 and the dragging portion 200, the predetermined angle and the particles contained in the sample It is formed differently according to the attraction of the, the particles in the sample included in the first tail 520 and the second tail 540 is further away from the first tail 520 and the second tail 540 The size can be gradually increased as described above.

In S300, the alignment assistant 300 aligns the particles included in the sample according to the size, and holds the particles aligned according to the size in the sample. For example, the alignment aid 300 forms an electric field to size the particles contained in the sample, including electrophoresis to align the ionized particles, ion concentration polarization to separate particles by ion concentration polarization, and surface acoustic wave. It can be held in alignment.

In S400, the particle separation device 10 drags the aligned dragging portion on the slot portion to store the particles included in the sample in the first group of slots 120 and the second group of slots 140 To separate. For example, the particle separation device 10 drags the aligned dragging part 200 on the slot part 100 to move the sample over the first group of slots 120 and the second group of slots 140 and At this time, particles of a predetermined size may be stored separately in slots of the corresponding group.

7 is an enlarged flow diagram of the holding step in the embodiment of FIG. 6.

In S320, the power supply unit 320 included in the alignment auxiliary unit 300 is formed on the other side of the dragging unit 200 in which the sample is located, and the electric field in the sample through at least two or more electrodes fastened to the dragging unit 200 To form. As described above, in the present invention, an electrode fastened to a fastening part formed in the dragging part 200 may be coupled to supply power for forming an electric field in a sample.

In S340, the ion selective permeable membrane 340 selectively permeates the particles according to different charge characteristics of the particles included in the sample through the ion selective permeable membrane connected to one end of the electrode. For example, the alignment auxiliary unit 300 of the present invention may polarize the transmitted particles according to the electric field formed in the sample through the electrode and sort them according to the size of the particles.

All or part of the method of one embodiment of the present invention described above may be embodied in the form of a computer-executable recording medium (or computer program product), such as a program module executed by a computer. Here, the computer-readable medium may include a computer storage medium (eg, memory, hard disk, magnetic / optical medium, solid-state drive (SSD), or the like). Computer readable media can be any available media that can be accessed by a computer and includes both volatile and nonvolatile media, removable and non-removable media.

In addition, all or part of the method according to an embodiment of the present invention includes instructions executable by a computer, and the computer program includes programmable machine instructions processed by a processor, and is a high-level programming language. Language), object-oriented programming language, assembly language, or machine language.

Means or modules in this specification may mean hardware capable of performing functions and operations according to each name described in the specification, and computer program code capable of performing specific functions and operations It may mean or an electronic recording medium on which computer program code capable of performing a specific function and operation is mounted, for example, a processor or a microprocessor. In other words, a means or module may mean a functional and / or structural combination of hardware for performing the technical idea of the present invention and / or software for driving the hardware.

Accordingly, a method according to an embodiment of the present invention may be implemented by executing a computer program as described above by a computing device. The computing device may include at least a portion of a processor, a memory, a storage device, a high-speed interface connected to the memory and a high-speed expansion port, and a low-speed interface connected to the low-speed bus and the storage device. Each of these components is connected to each other using various buses, and may be mounted on a common motherboard or mounted in other suitable ways.

The above description is merely illustrative of the technical idea of the present invention, and those of ordinary skill in the art to which the present invention pertains may make various modifications, changes, and substitutions without departing from the essential characteristics of the present invention. will be. Therefore, the embodiments disclosed in the present invention and the accompanying drawings are not intended to limit the technical spirit of the present invention, but to explain the scope of the technical spirit of the present invention. . The scope of protection of the present invention should be interpreted by the claims below, and all technical spirits within the scope equivalent thereto should be interpreted as being included in the scope of the present invention.

Claims (20)

Base portion;
The first group of slots for storing at least some of the particles formed on one surface of the base portion and separated from the sample containing the particles to be separated, and the second group of different depths or widths from the slots of the first group A slot portion provided with slots; And
For separating at least some of the particles from the sample through dragging at a predetermined angle in a state where the sample is positioned in a predetermined shape between the slots of the first group and the slots of the second group. It includes a dragging part,
The sample includes a first tail formed adjacent to the dragging portion and a second tail formed adjacent to the slot portion on the opposite side of the direction in which the dragging portion is dragged, and between the first tail and the second tail. Particle separation apparatus characterized in that the surface of the sample is formed of a meniscus curved surface that is concavely bent in the direction in which the dragging portion is dragged. delete delete According to claim 1,
The depth or width of the slots of the first group and the slots of the second group is preset according to the size of the particles to be separated, and the slots of the first group and the slots of the second group are preset Particle separation device characterized in that to store the particles having a size. According to claim 4, The slot portion
Different mask patterns including at least a first photoresist pattern and a second photoresist pattern provided in consideration of the depth or width of the slots of the first group and the slots of the second group are coated on the base portion,
Particle separation apparatus characterized by being formed by etching the base portion using the applied mask pattern as an etching mask. According to claim 1,
An alignment aid for aligning the particles included in the sample according to a size and holding the particles aligned according to the size in the sample; Further comprising,
The dragging unit is a particle separation device, characterized in that by dragging the sample containing the particles held in an aligned state to separate the particles. According to claim 1,
An extraction unit for extracting the particles stored in the first group of slots and the second group of slots; Further comprising,
The extracting unit extracts the particles stored in the slots of the first group and the slots of the second group separately from the sample. According to claim 6, The alignment aid
A power supply unit formed on the other surface of the dragging unit where the sample is located, and forming an electric field in the sample through at least two or more electrodes fastened to the dragging unit; Further comprising,
Particle separation device characterized in that to align the size of the particles according to the direction of the formed electric field. According to claim 8, The alignment aid
An ion selective permeable membrane connected to one end of the electrode selectively transmitting the particles according to different charge characteristics of the particles included in the sample; Further comprising,
Particle separation device characterized in that the transmitted particles are polarized according to the electric field formed in the sample through the electrode. The method of claim 6,
The particles are exosomes as vesicles containing at least one protein and RNA having different sizes,
The sample is a particle separation device, characterized in that provided as an electrolytic sample comprising at least one ionized particles. delete According to claim 1,
The meniscus curved surface, the length of the first tail and the second tail, the thickness of the first tail and the second tail, and the volume of the sample included in the first tail and the second tail are
Particle separation device characterized in that the base portion, the slot portion and the dragging portion is formed differently according to the attraction between the hydrophilicity, the predetermined angle and the particles contained in the sample. According to claim 1,
The slots of the first group and the slots of the second group are formed for each group in at least a portion of the slot part,
Particle separation apparatus characterized in that the particles in the sample included in the first tail and the second tail gradually increase in size as they move away from the first tail and the second tail. A first group of slots for storing at least a part of particles separated from a sample containing particles to be separated, formed on one surface of the base part, and a second group of slots having a different depth or width from the slots of the first group Placing the sample on at least one side of a slot portion in which the particles are provided;
Aligning the dragging unit for separating at least some of the particles from the sample through dragging at a predetermined angle in a state where the sample is positioned in a predetermined shape; And
Dragging the aligned dragging portion on the slot portion to separate and store particles included in the sample in the first group of slots and the second group of slots; It includes,
The sample includes a first tail formed adjacent to the dragging portion and a second tail formed adjacent to the slot portion on the opposite side of the direction in which the dragging portion is dragged, and between the first tail and the second tail. Particle separation method characterized in that the surface of the sample is formed by a meniscus curved surface that is concavely bent in the direction in which the dragging portion is dragged. The method of claim 14,
Aligning the particles included in the sample according to size, and holding particles aligned according to the size in the sample; Further comprising,
The dragging unit is a particle separation method characterized in that by dragging the sample containing the particles held in an aligned state to separate the particles. The method of claim 15, wherein the holding step
Forming an electric field in the sample through at least two or more electrodes formed on the other surface of the dragging portion where the sample is located, and fastened to the dragging portion; And
Selectively transmitting the particles according to different charge characteristics of the particles included in the sample through an ion-selective permeable membrane connected to one end of the electrode; Further comprising,
Particle separation method characterized in that the transmitted particles are polarized according to the electric field formed in the sample through the electrode to be aligned according to the size of the particles. delete The method of claim 14,
The meniscus curved surface, the length of the first tail and the second tail, the thickness of the first tail and the second tail, and the volume of the sample included in the first tail and the second tail are the base portion, the slot It is formed differently according to the hydrophilicity of the part and the dragging part, the predetermined angle and the attraction force between particles contained in the sample,
Particle separation method characterized in that the particles in the sample included in the first tail and the second tail gradually increase in size as they move away from the first tail and the second tail. The method of claim 14,
The depth or width of the slots of the first group and the slots of the second group is preset according to the size of the particles to be separated, and the slots of the first group and the slots of the second group are preset The particles with size are stored separately.
The method of claim 1, wherein the first group of slots and the second group of slots are formed for each group in at least a portion of the slot. The method of claim 19, wherein the slot portion
Different mask patterns including at least a first photoresist pattern and a second photoresist pattern provided in consideration of the depth or width of the slots of the first group and the slots of the second group are coated on the base portion,
Particle separation method characterized in that it is formed by etching the base portion using the applied mask pattern as an etch mask.

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