KR20190096566A - Apparatus for Separating Particles of Sample with Meniscuss and Method thereof - Google Patents

Abstract

The present invention discloses a device for separating particles and, specifically, a device for moving a sample having a meniscus curved surface on a substrate to separate particles. According to the present device, the device for separating particles comprises: a base part; and a slot part formed on one surface of the base part, and provided with slots of a first group for storing at least some particles separated from the sample including particles to be separated.

Description

Apparatus for Separating Particles of Sample with Meniscuss and Method

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

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

Exosomes originate from specific compartments within cells called polycystic bodies and are released and secreted out of cells. When fusion occurs between polycystic bodies and plasma membranes, these vesicles are released into the extracellular environment, which is called exosomes. Exosomes are known to be released separately from many different cell types under both normal and pathological conditions. Exosomes contain a variety of RNA, and there is a lot of research on how to diagnose the disease by detecting the presence and presence of it. However, for accurate diagnosis using exosomes, it is important to know the exact amount of exosomes present in the human body with the disease.

As a technique for separating and purifying conventional exosomes, there exist methods such as exosome isolation reagent, exomir, exoquick, and ultracentrifugation using two pore-sized filters, but current techniques have significant time and cost. And specialized research facilities.

In addition, the conventional exosome measurement method has a quantitative method using a specific immune response of the antibody and antigen, but it is cumbersome, there is a limit that can be applied only to microvesicles and exosomes.

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

Korea Patent Registration No. 10-1738117 (Public)

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

The present invention has been made to achieve the above object, the particle separation device of the present invention is a base portion; And a slot part formed on one surface of the base part and having a first group of slots for storing at least some particles separated from a sample including 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 than 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 while the sample is positioned in a predetermined shape between the slots of the first group and the surface on which the slots of the second group are formed. Dragging unit for separating the particles of; 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 preset 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 in consideration of a depth or a width of the slots of the first group and the slots of the second group. The substrate may be coated on the base part, and the base part may be etched using the applied mask pattern as an etching mask.

An alignment aid for aligning the particles included in the sample according to size and holding the particles aligned according to the size in the sample; Further comprising, The dragging unit may separate the particles by dragging the sample including 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 slots of the first group and the slots of the second group; The extracting unit may further extract and separate 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 aid is formed on the other surface of the dragging portion in which the sample is located, the power supply for forming an electric field in the sample through at least two electrodes fastened to the dragging portion; Further comprising, it is possible to align the size of the particles according to the direction of the formed electric field.

In the present invention, the alignment aid may include an ion selective permeable membrane connected to one end of the electrode for selectively transmitting the particles according to different charge characteristics of the particles included in the sample; Further comprising, the transmitted particles may be polarized in accordance with 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 particles.

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

In the present invention, the meniscus 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 contained in the first tail and the second tail is the base portion The hydrophilicity of the slot part and the dragging part may be formed differently according to the attraction force between the predetermined angle and the particles included in the sample.

In the present invention, the slots of the first group and the slots of the second group are formed in groups 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 a size may gradually increase as the distance from the second tail increases.

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 particles separated from the sample containing the particles to be separated and the first Positioning the sample on at least one side of a slot part in which a second group of slots having a depth or width different from that of the first group of slots is provided; Arranging a dragging unit for separating at least some particles from the sample by dragging at a predetermined angle while the sample is positioned in a predetermined shape; And dragging the aligned dragging unit on the slot part to store and separate particles included in the sample into slots of the first group and slots of the second group. It includes.

In the present invention, the particle separation method comprises the steps of aligning the particles contained in the sample according to the size, holding the particles aligned in the sample in the sample; Further comprising, The dragging unit may separate the particles by dragging the sample including the particles held in an aligned state.

In the present invention, the holding step may include forming an electric field in the sample through at least two electrodes formed on the other surface of the dragging part in which the sample is located, and fastened to the dragging part; 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, by polarizing the transmitted particles according to the electric field formed in the sample through the electrode can be aligned according to the size of the particles.

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

In the present invention, the meniscus 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 contained in the first tail and the second tail is the base portion And differently formed according to the hydrophilicity of the slot part and the dragging part, the predetermined angle, and the attraction force between the particles contained in the sample, wherein the particles in the sample included in the first tail and the second tail are formed in the first part. It may be arranged to gradually increase in size as it moves away from the tail and the second tail.

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 classified and stored, and the slots of the first group and the slots of the second group may be formed in groups in at least a portion of the slot part.

In the present invention, the slot part may include different mask patterns including at least a first photoresist pattern and a second photoresist pattern in consideration of a depth or a width of the slots of the first group and the slots of the second group. The substrate may be coated on the base part, and the base part may be etched using the applied mask pattern as an etching mask.

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

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

1 is a block diagram of a particle separation device according to an embodiment of the present invention.
2 is a view illustrating an operation preparation state of a particle separation device according to an embodiment of the present invention.
3 shows a detailed configuration of a particle separation device according to an embodiment of the present invention.
4 illustrates a configuration of an alignment aid 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 aid according to another embodiment of the present invention.
7 is a flow chart of a particle separation method according to an embodiment of the present invention.
8 is an enlarged flowchart of a 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 will be given the same reference numerals and redundant description thereof will be omitted.

In describing the present invention, when it is determined that detailed descriptions of related well-known structures or functions may obscure the gist of the present invention, the detailed description may be omitted.

The terminology used herein is for the purpose of describing particular example embodiments only and is not intended to be limiting of terms. Singular expressions include plural expressions unless the context clearly indicates otherwise. Each step described below may be provided by one or several software modules or may be implemented by hardware that is responsible for each function, or may be a combination of software and hardware. Specific meanings and examples of each term will be described below in the order of the drawings. Hereinafter, the configuration of the particle separation device 10 according to an embodiment of the present invention will be described in detail with reference to the accompanying drawings.

1 is a block diagram of a particle separation device 10 according to an embodiment of the present invention. The structure of the particle separation apparatus 10 is demonstrated with reference to FIG.

The particle separation device 10 of the present invention includes a base part 100, a dragging part 200, an alignment aid part 300, and an extraction part 400. For example, the particle separation device 10 of the present invention is to move the sample containing the particles located between the two substrates (plate) on the base portion 100 using the dragging unit 200 to separate the particles to be separated Can be separated. Particles to be separated by the particle separation device 10 of the present invention is a particle having a size, the smallest particle having the properties of the material (Molecule) and exosomes embedded with RNA and various proteins in biological fluids (Exosome) ). The particle separation device 10 of the present invention can extract only the desired exosomes by separating and separating approximately 30-100 nm exosomes with diameters known to be specifically secreted for the purpose of intercellular signal transmission. It demonstrates with reference to FIG.

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

In addition, the base part 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 included prior to preparing storage members including the first group of slots 120 and the second group of slots 140. The base part 100 is a wet etchant used for microfabrication, and is formed through etching a thin film of silicon dioxide or silicon nitride using Buffered Oxide Etch (BOE), or using a Piranha etching technique. And a further process of removing the organic residue on the base with a mixture of sulfuric acid and hydrogen peroxide. Since the base part 100 and the slot part 120 of the present invention are cleaned through the Piranha etching process, the surface of the base part 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 curved surface of the sample located between the base portion 100 and the dragging portion 200, It is an important configuration that affects separation.

The slot unit 110 is at least one storage member for storing particles to be separated, and thus the first group of slots 120 and the second group of slots 140 may be formed on a surface thereof to store the particles. For example, the storage members of the slot portion 110 may include at least the first group of slots 120 and the second group of slots 140, depending on the depth or width, and the slots of the first group ( 120 and the slots 140 of the second group may be formed for each group in the slot part 110 formed on the upper surface of the base part. For example, the depth or width of the slots 120 of the first group and the slots 140 of the second group is preset according to the size of the particles to be separated, and the slots 120 of the first group And the slots 140 of the second group may distinguish and store particles having a preset size. Slot portion 110 of the present invention may further include slots for separating particles having a different standard in addition to the slots 120 and the slots 140 of the first group described above, of course. .

In addition, the first group of slots 120 and the second group of slots 140 formed in at least a portion of the slot part 110 of the present invention may be formed through a photolithography process. For example, the slots 120 of the first group and the slots 140 of the second group may be configured in consideration of the depth or width of the slots 120 of the first group and the slots 140 of the second group. Different mask patterns including at least the first photoresist patterns 162, 164, and 168 and the second photoresist patterns 172, 174, and 178 are coated on the base part 100, and the applied masks are applied. The base part 100 may be etched using a pattern as an etching mask. In the present invention, the etch mask may be formed of various materials according to the intended pattern of use, 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. ) May be formed through a photolithography process, for example, using Immersion Lithography or Extreme Ultraviolet (EUV) lithography techniques. Immersion lithography technology improves the resolution by increasing the numerical aperture by filling a high refractive index fluid between the lens and the subject. The first group of slots 120 and the second group of slots 140 may etch the first photoresist patterns 162, 164, and 168 and the second photoresist patterns 172, 174, and 178. The base part 100 may be etched using a mask, and the etching process may include a dry etching, a wet etching, or a reactive ion etching method (RIE). For example, when the etching mask is formed of a carbon containing film, a plasma etching process using a mixed gas of oxygen and argon may be performed. In forming the slots 120 of the first group and the slots 140 of the second group, the first photoresist patterns 162, 164, and 168 and the second photoresist patterns 172, 174, and 178 are etched. It can be removed after the process. In the case of using the reactive ion etching method (RIE) in forming the first group of slots 120 and the second group of slots 140 formed in the slot portion 110 of the present invention, the first group of slots Different degrees of etching may be used depending on the depth and width of the holes 120 and the slots 140 of the second group.

The dragging unit 200 drags at a predetermined angle in a state in which the sample is positioned in a predetermined shape between the slots 120 of the first group and the surface on which the slots 140 of the second group are formed. Separating at least some of the particles from the sample through. For example, the dragging unit 200 is in an oblique state on the slot unit 110, at least one end of the dragging unit 200 contacts the slot unit 110 or approaches at a predetermined interval, and the slot unit 110 and the dragging unit 200 are separated. By placing a sample including particles 542 and 544 in the gaping space of the sample, the positioned sample may be moved on the slot part 100. Dragging unit 200 of the present invention may further include a fastening portion through which the electrode is fastened therein as will be described later.

In the present invention, the sample located between the slot portion 110 and the dragging portion 200 includes an electrolytic sample including at least one ionized particles, the particles contained in the sample at least one protein having a different size And exosomes as vesicles comprising RNA.

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

In the present invention, the sample containing the particles to be separated is located in the space (the upper surface of the slot and the lower side of the dragging portion) is formed in a narrow narrow angle between the dragging portion 200 and the slot portion 110, the dragging portion 200 is an obtuse angle Particles may be separated by being dragged in the spatial direction formed by the dragging part, which is located in the space (the upper surface of the slot part and the upper surface of the dragging part) formed at an obtuse angle between the dragging part 200 and the slot part 110. Particles may be separated as 200 is dragged in a narrow space direction formed at an acute angle. That is, according to the direction in which the sample of the present invention is located between the dragging unit 200 and the base unit 100 as described below, the direction in which the dragging unit 200 is dragged on the upper surface of the base unit 100 may vary. 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 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 contained in the first tail and the second tail is the base portion And a hydrophilicity of the dragging unit, the predetermined angle, and the attraction force between the particles included in the sample. In addition, the meniscus curved surface 560 of the present invention is 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 It may be formed differently depending on the angle located on the upper surface of the base portion 100 and the dragging angle, 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 part 110 and the dragging part 200 of the present invention, the first tail 520 and the second tail 540 are connected to the slot part 110 and the dragging part 220. Only small particles 5440 in the sample may be located as a portion formed by a narrow area of an adjacent narrow height.

For example, the meniscus curved shape formed between the slot portion 110 and the dragging portion 200 in the sample of the present invention is the attraction of the particles in the sample, the surface tension of the solvent, the base portion 100, the slot portion ( 110 and the dragging unit 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 the particles by maintaining the shape of the sample between the slot portion 110 and the dragging portion 200 to the meniscus curved surface. The meniscus curved surface may be formed of different meniscus curved surfaces according to the heights of the contact portions of the base part 100 and the dragging part 200.

In addition, the particle separation device 10 of the present invention may adjust the degree of hydrophilicity of the base portion 100, the slot portion 110 and the dragging portion 200 to adjust the meniscus shape. For example, the base portion 100, the slot portion 110 and the dragging portion 200 of the present invention may be formed with a functional group on the surface by using an etching means including a plasma gas or heat ionized at a high temperature, Through this, the surface may 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 smoothing or artificially curved surface through the micro / nano process in terms of the structure, and other chemical By using the method, the hydrophilicity of the surface of the base part 100, the slot part 110, and the dragging part 200 may be improved. It demonstrates with reference to FIG.

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 assistant 300 may move the dragging unit 200 through the slot portion of the upper surface of the base unit 100 through electrophoresis, ion concentration polarization, and surface acoustic wave. The particles in the sample may be aligned and held prior to moving on 110. The alignment assistant 300 may arrange and hold particles in the sample before the dragging unit 200 moves so that the sample may be sequentially separated according to the size of the particles when the sample moves as the dragging unit 200 moves. .

Alignment assistant 300 of the present invention is formed on the other side of the dragging portion where the sample is located, further includes a power supply unit 320 for forming an electric field in the sample through at least two or more electrodes coupled to the dragging portion, the power supply unit ( The size of the particles may be aligned according to the direction of the electric field formed by 320. For example, the alignment assistant 300 may include a dragging unit for connecting an electrode in a sample positioned between the slot unit 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 assistant 300 of the present invention may form an electric field in the sample, and maintain the particles in the sample in an aligned state before the sample moves by using the properties of the particles in the sample having different electrical affinity.

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 supply 320. In addition, the transmitted particles may be polarized according to an 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 comprises at least one nano permeable membrane and emits only ionized particles that are charged by forming an ion depletion region near the nano permeable membrane in consideration of the 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 match the ion conductivity in the electrolytic sample so that ion concentration gradients are generated on both sides of the ion selective permeable membrane. When the ion concentration polarization (ICP) caused by the ion selective permeable membrane 340 is induced near the ion selective permeable membrane, both the cation and anion concentration decrease at the anode side of the junction 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. Excessive or deficient ionized regions formed by the ion-selective permeable membrane 340 may also be applied to charged particles, cells, and other small colloids to form regions in which ionized particles are excessive and regions in which ionized particles are deficient. This may also be applied to the particle separation device 10 for separating the particles by moving a sample including the ionized particles of the present invention. Particles in a sample located between the slot portion 110 and the dragging portion 200 of the present invention may be self-assembled and crystallized in the sample. 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 extractor 400 extracts the particles stored in the slots 120 of the first group and the slots 140 of the second group. For example, the extractor 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 extractor 400 extracts the particles stored in the slots 120 of the first group and the slots 140 of the second group, and the slots 120 and the second group of the first group in which the particles are stored. A PDMS film may be formed on the slots 140, and particles may be extracted based on the formed PDMS film.

2 shows the alignment 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 110 and the dragging unit 200 may be adjusted in consideration of the contact angle on the slot 110 of the dragging unit 200. . In the present invention, the sample is made of a meniscus curved surface shape including the first tail 520 and the second tail 540, the shape of the meniscus curved surface of the slot 110 and the dragging unit 200 As described above, it may be differently set by the set angle, the hydrophilicity of the slot part 110 and the dragging part 200, the surface tension of the sample, and the attraction force of the particles in the sample. In addition, the meniscus curved surface 560 of the sample is located on the upper surface of the slot portion 110 and the dragging portion 200 or on the upper surface of the slot portion 110 and the upper surface of the dragging portion 200. As described above may be formed differently depending on the case.

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

The base portion 100 includes a slot portion 110 including at least one slot for storing particles on the top surface. The slot part 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 slots 120 of the first group and the slots 140 of the second group. Depth 122, width 124, first slot spacing 126 and depth 142, width 144, second of slots 140 of the first group of slots 120 of the first group The slot spacing 146 and the third slot spacing 150 between the slots of the first group and the slots of the second group are different 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, the base part is formed by utilizing predetermined photoresist patterns 162, 164, 168, 172, 174, and 178 to form the slots 120 of the first group and the slots 140 of the second group. The specific method of etching and forming (100) is as described above, and will be omitted.

The sample located between the slot part 100 and the dragging part 200 of the present invention may be positioned in a meniscus shape before being moved on the base part 100. The sample positioned in the meniscus shape of the present invention includes a first tail 520 and a second tail 540, and small particles may be located in the first tail 520 and the second tail 540. Can be. In the present invention, the meniscus shape of the sample may be determined according to the hydrophilicity of the solvent in the sample, the attraction between 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 the particles using the coffee stain effect (coffee ring effect). For example, the sample located in the slot 110 and the dragging unit 200 may be formed in a round cap shape due to the attraction of the internal particles and the solvent, the contact angle of the liquid in contact with the solid, and at this time, the particles in the sample They may be provided in a spin-ring state. In order for the liquid to be rounded in the shape of a cap, a predetermined threshold must be satisfied, which is determined only by the interfacial energy of the liquid, the substrate and the air, and the contact angle of the liquid drop. When the liquid is located in a round position, the evaporation amount of the center and the edge is greatly changed, and the contact area of the edge is large, so that the evaporation amount is larger, the flow of the internal liquid supplies the liquid to the edge, and the particles inside the evaporation amount This phenomenon can be shown to flow in a large portion.

For example, particles in a sample located in a meniscus curved shape between the slot portion 110 and the dragging portion 200 may be mainly located in the first tail 520 and the second tail 540 according to the coffee stain effect. have. In this case, small particles may be mainly located in the narrow portions of the first tail 520 and the second tail 540 according to the size, mass, and surface tension of the liquid. Therefore, the large particles are mainly located on the side of the dragging unit 200 in the moving direction, and when the sample including the aligned particles is moved on the slot unit 110, the large particles are slots of the second group. The particles 140 are stored in the slots 120 of the first group and the small particles 544 are stored in the slots 140 of the second group.

Particle separation apparatus 10 of the present invention may further include an alignment assistant 300 to further improve the alignment according to the size of the particles in the sample, the alignment assistant 300 is to hold the particles in the sample to align The detailed method is the same as described above, and will be omitted.

4 illustrates a configuration of the alignment assistant 300 according to an embodiment of the present invention.

The alignment assistant 300 of the present invention 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 assistant 300 may further include a power supply 320 and an ion selective permeable membrane 340, and arrange and hold the particles 542 and 544 included in the sample 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 from the power supply unit 320 into the sample is fastened in the dragging unit 200. As described above, the fastening part may be further included in the dragging part 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 a sample including particles 543 and 544 to be separated between the slot part 110 and the dragging part 200 on the slot part 110. The particles can be separated. For example, the particle separation device 10 of the present invention performs a cleaning process to drag the dragging unit 200 and to process particles that are not stored in the first slots 120 and the second slots 140. It may further include.

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

The alignment assistant 300 of the present invention arranges the particles included in the sample according to the size and generates a surface acoustic wave to hold the particles aligned according to the size in the sample. 360 may be further included, and the particles included in the sample may be aligned according to the size by using the surface acoustic waves generated by the acoustic wave generator 360. For example, the alignment aid 300 of the present invention is in one side of the dragging unit 200 located in the direction of the sample to the acoustic wave connection in order to deliver the surface acoustic wave (Surface Acoustic Wave) generated in the acoustic wave generator 360 in the sample Further, the surface acoustic wave generated in the acoustic wave generating unit through the elastic wave connection unit (Surface Acoustic Wave) can be delivered to the sample.

7 is a flow chart 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 performed by the particle separation device 10 in time series.

In S100, the particle separation device 10 is formed on one surface of the base part, so that the first group of slots 120 and the first group for storing at least some particles separated from the sample including the particles to be separated The sample is positioned on at least one side of the slot part 110 in which the slots 140 of the second group having a depth or width different from those of the slots are provided. The particle separation device 10 places the sample to be moved on the slot portion 100 by using the dragging portion 200, but there is no limitation, but the slots 120 of the first group and the slots of the second group are not limited. It may be located on at least one surface of the slot portion 110 where the 140 is not located.

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

In addition, the slots 120 of the first group and the slots 140 of the second group of the present invention may vary according to the depth or width of the slots 120 of the first group and the slots 140 of the second group. At least a portion of the slot part 110 may be formed by grouping by group. At least a first photo provided in consideration of the depth or width of the slots 120 of the first group and the slots 140 of the second group formed in the slot 110 located on the upper surface of the base part 100. It can be formed by applying different mask patterns including a resist pattern and a second photoresist pattern on the base portion, and etching the base portion 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 in which the sample is positioned in a predetermined shape. For example, the particle separation device 10 includes a menis including a first tail 520 and a second tail 540 suitable for separating a particle from a sample between the slot 110 and the dragging 200. In order to have a curse shape, the dragging part 200 may be aligned on the slot part 110. In this case, the preset angle at which the dragging unit 200 is positioned may include the size of the particles, the hydrophilicity of the sample, the attraction force of the particles in the sample, the base unit 100, the hydrophilicity of the slot unit 110 and the dragging unit 200, and the first angle. The length of the tail 520 and the second tail 540 and the curvature of the meniscus curved surface may be set.

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 Differently formed according to the attraction force of the particles, the particles in the sample included in the first tail 520 and the second tail 540 are farther away from the first tail 520 and the second tail 540. The size can be increased gradually 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 may include particles in a sample including electrophoresis to form ionized particles to align ionized particles, ion concentration polarization to separate particles by ion concentration polarization, and surface acoustic wave. It can be aligned and held accordingly.

In S400, the particle separation device 10 stores the particles included in the sample in the slots 120 and the slots 140 of the second group by dragging the aligned dragging units on the slots. Separate. For example, the particle separation device 10 drags the aligned dragging unit 200 on the slot unit 100 to move the sample over the slots 120 of the first group and the slots 140 of the second group. In this case, particles having a predetermined size may be separately stored in slots of the corresponding group.

7 is an enlarged flowchart of a holding step in the embodiment of FIG. 6.

In S320, the power supply unit 320 included in the alignment assistant 300 is formed at 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. In the present invention, as described above, the electrode coupled to the fastening part formed in the dragging part 200 may be coupled to supply power for forming an electric field in the sample.

In S340, the ion selective permeable membrane 340 selectively transmits 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 aid 300 of the present invention may be aligned according to the size of the particles by polarizing the transmitted particles according to the electric field formed in the sample through the electrode.

All or part of the method of one embodiment of the present invention described above may be implemented in the form of a computer-executable recording medium (or a 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 media or solid-state drive, etc.). 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, the computer program including programmable machine instructions processed by a processor, and a high-level programming language. Language, an object-oriented programming language, an assembly language, or a machine language.

Means or modules in the present specification may mean hardware capable of performing functions and operations according to each name described in the present specification, and computer program code capable of performing specific functions and operations. It may mean an electronic recording medium, for example, a processor or a microprocessor, on which computer program code capable of performing specific functions and operations may be implemented. 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.

Thus, 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 are connected to each other using a variety of buses and may be mounted on a common motherboard or otherwise mounted in a suitable manner.

The above description is merely illustrative of the technical idea of the present invention, and various modifications, changes, and substitutions may be made by those skilled in the art without departing from the essential characteristics of the present invention. will be. Accordingly, 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 describe the present invention, and the scope of the technical idea of the present invention is not limited by the embodiments and the accompanying drawings. . The scope of protection of the present invention should be interpreted by the following claims, and all technical ideas within the scope equivalent thereto should be construed as being included in the scope of the present invention.

Claims (20)

A base portion; And
A slot part formed on one surface of the base part and provided with a first group of slots for storing at least some particles separated from a sample including particles to be separated; Particle separation apparatus comprising a. The method of claim 1, wherein the slot portion
And a second group of slots having a different depth or width than the slots of the first group. The method of claim 2,
For separating at least some particles from the sample by dragging at a predetermined angle with the sample positioned in a predetermined shape between the slots of the first group and the slot on which the slots of the second group are formed. Dragging unit; Particle separation apparatus comprising a further. The method of claim 2,
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 for storing the particles having a size. The method of claim 4, wherein the slot portion
Applying different mask patterns including at least a first photoresist pattern and a second photoresist pattern on the base part in consideration of depth or width of the slots of the first group and the slots of the second group;
And etching the base part using the applied mask pattern as an etch mask. The method of claim 3,
An alignment aid for aligning the particles included in the sample according to size and holding the particles aligned in the sample in the sample; More,
The dragging unit is a particle separation device, characterized in that for separating the particles by dragging the sample including the particles held in an aligned state. The method of claim 2,
An extraction unit configured to extract the particles stored in the slots of the first group and the slots of the second group; More,
And the extractor extracts particles stored in the slots of the first group and the slots of the second group by separating the sample from the sample. The method of claim 6, wherein the alignment aid is
A power supply unit formed at the other surface of the dragging unit in which the sample is located and forming an electric field in the sample through at least two electrodes fastened to the dragging unit; More,
Particle separation apparatus, characterized in that for aligning the size of the particles in the direction of the formed electric field. The method of claim 8, wherein the alignment aid is
An ion selective permeable membrane connected to one end of the electrode for selectively permeating the particles according to different charge characteristics of the particles included in the sample; More,
And wherein the transmitted particles are polarized according to an 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,
And said sample is provided with an electrolytic sample comprising at least one ionized particle. The method of claim 6, wherein the sample
A first tail formed adjacent to the dragging unit and a second tail formed adjacent to the slot unit on the opposite side of the dragging unit in a direction in which the dragging unit is dragged;
And the surface of the sample is formed between the first tail and the second tail as a meniscus curved surface that is concavely curved in the direction in which the dragging portion is dragged. The method of claim 11,
The meniscus curved surface, the length of the first and second tails, the thickness of the first and second tails and the volume of the sample contained in the first and second tails are
Particle separation apparatus, characterized in that formed differently depending on the hydrophilicity of the base portion, the slot portion and the dragging portion, the predetermined angle and the attraction between the particles contained in the sample. The method of claim 11,
Slots of the first group and slots of the second group are formed in groups in at least a portion of the slot portion,
The particles in the sample contained in the first tail and the second tail is characterized in that the particle size gradually increases with distance from the first tail and the second tail. Slots of a first group for storing at least some particles separated from a sample including particles to be separated and formed on one surface of the base part, and slots of a second group having a different depth or width from those of the slots of the first group Positioning the sample on at least one side of the slot portion in which the recesses are provided;
Arranging a dragging unit for separating at least some particles from the sample by dragging at a predetermined angle while the sample is positioned in a predetermined shape; And
Dragging the aligned dragging unit on the slot unit to separate and store particles included in the sample into slots of the first group and slots of the second group; Particle separation method comprising a. The method of claim 14,
Aligning the particles included in the sample according to size and holding the particles aligned according to the size in the sample; More,
The dragging unit is a particle separation method characterized in that for separating the particles by dragging the sample including the particles held in an aligned state. The method of claim 15, wherein the holding step
Forming an electric field in the sample through the at least two electrodes formed on the other surface of the dragging part in which the sample is located; And
Selectively permeating 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; More,
And permeate the transmitted particles according to the electric field formed in the sample through the electrode to align the particles according to the size of the particles. The method of claim 14, wherein the sample
A first tail formed adjacent to the dragging unit and a second tail formed adjacent to the slot unit on the opposite side of the dragging unit in a direction in which the dragging unit is dragged;
And a surface of the sample formed between the first tail and the second tail is formed as a meniscus curved surface that is concavely curved in a direction in which the dragging portion is dragged. The method of claim 17,
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 contained in the first tail and the second tail are the base portion, the slot It is formed differently depending on the hydrophilicity between the portion and the dragging portion, the predetermined angle and the particles contained in the sample,
And 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. Classify and store particles of size,
And the slots of the first group and the slots of the second group are formed in groups in at least a portion of the slot portion. The method of claim 19, wherein the slot portion
Applying different mask patterns including at least a first photoresist pattern and a second photoresist pattern on the base part in consideration of depth or width of the slots of the first group and the slots of the second group;
And etching the base part using the applied mask pattern as an etching mask.

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