Particle sorting apparatus and particle sorting method

Abstract

An object of the present invention is to provide a technique that can form stable droplets. A particle sorting apparatus is provided, which includes a microchip including: a main channel through which a liquid including particles flows; a sheath liquid channel that communicates with the main channel and through which a sheath liquid flows; and a sheath liquid introduction portion that introduces the sheath liquid. The particle sorting apparatus vibrates the sheath liquid flowing through the sheath liquid introduction portion. Further, a particle sorting method is provided, in which, in the method, in a microchip including: at least one main channel through which a liquid including particles flows; a sheath liquid channel that communicates with the main channel and supplies a sheath liquid; and a sheath liquid introduction portion that introduces the sheath liquid, the sheath liquid flowing through the sheath liquid introduction portion is vibrated.

Description

Technical Field

[0001] This technology relates to particle sorting devices and particle sorting methods. Background Technology

[0002] Various devices for sorting microparticles have been developed to date, and specifically, devices for sorting cells are called "cell sorters." In a cell sorter, vibration is typically applied to flowing cells or microchips via a vibrating element or the like, causing fluid exiting the flow path to form droplets. After applying a positive (+) or negative (-) charge to the droplets separated from the fluid, the direction of droplet movement is changed by a deflector plate or the like, and the droplets are collected in a predetermined container or the like.

[0003] For example, Patent Document 1 discloses a method for forming droplets by applying vibration to the orifice of a microchip during extraction using a microchip.

[0004] Existing technical documents

[0005] Patent documents

[0006] Patent Document 1: Japanese Patent Application Publication No. 2017-219521 Summary of the Invention

[0007] The problem to be solved by the present invention

[0008] Control techniques for the stable formation of droplets in flow cytometers are one of the important factors for improving sorting accuracy. Here, it is known that when droplet formation is unstable, for example when the break point (BOP) of the fluid exiting the outlet of the flow channel is unstable, the time it takes for the droplet to be charged also becomes unstable, and therefore, the sorting of particles also becomes unstable.

[0009] On the other hand, because the microchip has a large number of high-order eigenvalues ​​and corresponding mode shapes at frequencies near the excitation frequency used by flow cytometry for droplet formation (approximately several 10 kHz to 100 kHz), the microchip can have multiple frequencies affected by these eigenvalues. Therefore, when vibration is applied to the orifice of the microchip, the vibration intensity changes in a complex manner according to the frequency, which can lead to instability at the breakpoint.

[0010] Therefore, the main objective of this technology is to provide a technique capable of forming stable droplets.

[0011] Solution to the problem

[0012] This technology first provides a particle sorting device, which includes: a microchip, the microchip including: a main flow path through which liquid containing particles flows; a sheath fluid flow path connected to the main flow path through which sheath fluid flows; and a sheath fluid inlet for introducing sheath fluid, wherein the sheath fluid flowing through the sheath fluid inlet vibrates.

[0013] This technology may further include a connecting member that can be attached to a microchip and has a sheath fluid inlet coupling portion coupled to a sheath fluid inlet portion.

[0014] In this technology, the vibrating element can be attached to the connecting member.

[0015] In this technology, the driving frequency of the vibrating element may be different from the resonant frequency of the flow path in the microchip.

[0016] In this technology, the driving frequency of the vibrating element can be within ±10% of the resonant frequency of the flow path in the microchip.

[0017] In this technology, the sheath fluid introduction coupling portion may include a sheath fluid convergence portion, the width of which gradually or partially narrows from one side of the vibrating element toward the side of the sheath fluid introduction portion.

[0018] In this technology, the height of the sheath fluid convergence section can be gradually or partially reduced from one side of the vibrating element toward the side of the sheath fluid inlet section.

[0019] The technology may further include a connecting portion having a tubular portion that communicates with the distal end of the sheath fluid converging portion between the sheath fluid converging portion and the sheath fluid inlet portion.

[0020] In this technology, a tubular member can be inserted into the interior of a tubular portion.

[0021] In this technology, at least a portion of the tubular portion and / or tubular member may be formed from at least one selected from the group consisting of elastomers, resins, and metals.

[0022] In this technology, the sheath fluid convergence portion can be a generally conical shape, a generally pyramidal shape, or a body of revolution of an exponential function or a parabola.

[0023] In this technology, the sheath fluid confluence portion can be formed of resin, metal, or a transparent component.

[0024] In this technology, the electrode can be inserted into the sheath fluid convergence zone.

[0025] In this technology, eddies that cause the sheath fluid to swirl can be generated in the sheath fluid convergence zone.

[0026] In this technology, the sheath fluid inlet for introducing sheath fluid into the sheath fluid confluence section can be located at a position far from the center of the sheath fluid confluence section.

[0027] Furthermore, this technology provides a particle sorting device, comprising: a microchip, the microchip including: a main flow path through which a liquid containing particles flows; a sheath fluid flow path connected to the main flow path and supplying sheath fluid; a sheath fluid inlet for introducing sheath fluid; a light irradiation unit for irradiating particles with light; a light detection unit for detecting light from particles; and a processing unit for processing signals obtained from the light detection unit, wherein the sheath fluid flowing through the sheath fluid inlet vibrates.

[0028] In addition, this technology also provides a particle sorting method, which includes in a microchip: at least a main flow path through which a liquid containing particles flows; a sheath fluid flow path connected to the main flow path through which sheath fluid flows; and a sheath fluid inlet for introducing the sheath fluid, through which the sheath fluid vibrates and flows. Attached Figure Description

[0029] Figure 1 This is a view showing an example configuration of a particle sorting apparatus according to a first embodiment.

[0030] Figure 2 This is a view where A and B show examples of microchip configurations.

[0031] Figure 3 This is a view where A through C shows an example configuration of the apertures of a microchip.

[0032] Figure 4 This is a view showing the standing waves generated in the flow path.

[0033] Figure 5 This is a graph showing the relationship between the fluctuating pressure in the nozzle and the driving frequency.

[0034] Figure 6 This is a view showing examples of applicable configurations for a piezoelectric element unit.

[0035] Figure 7 This is a view showing an example configuration using a bent piezoelectric element.

[0036] Figure 8 This is a view showing the electrodes positioned near the sheath fluid convergence point.

[0037] Figure 9 This is a view showing an example configuration of the particle sorting apparatus according to the second embodiment.

[0038] Figure 10 This is a diagram showing an example configuration of a particle sorting apparatus according to a third embodiment.

[0039] Figure 11It is a graph showing the results of measuring the frequency characteristics of the fracture point under conditions of vibration applied using existing technology and under conditions of vibration applied using this technology.

[0040] Figure 12 This is a view schematically illustrating pattern example 1 of the connection part.

[0041] Figure 13 This is a graph showing the relationship between the drive voltage and the BOP length from 27 kHz to 31 kHz.

[0042] Figure 14 This is a schematic view illustrating pattern example 2 of the connection part.

[0043] Figure 15 This is a schematic view illustrating pattern example 3 of the connection section.

[0044] Figure 16 This is a view schematically illustrating pattern example 4 of the connection part. Detailed Implementation

[0045] In the following description, preferred embodiments for carrying out the present technology will be described with reference to the accompanying drawings. It should be noted that the embodiments described below are examples of representative embodiments of the present technology, and the scope of the present technology is not to be interpreted narrowly by them. Note that the descriptions will proceed in the following order.

[0046] 1. First Embodiment (Particle Sorting Device 100)

[0047] (1) Microchip M

[0048] (2) Connecting component C

[0049] 2. Second Embodiment (Particle Sorting Device 100)

[0050] 3. Third Embodiment (Particle Sorting Device 100)

[0051] [Example 1 of the connection pattern]

[0052] [Example 2 of the connection pattern]

[0053] [Example 3 of the connection pattern]

[0054] [Example 4 of the connection pattern]

[0055] 4. Fourth Embodiment (Particle Sorting Device 100)

[0056] (1) Light irradiation unit 103

[0057] (2) Optical detection unit 104

[0058] (3) Processing unit 105

[0059] (4) Sorting unit 106 (including charging unit 106c)

[0060] (5) Storage unit 107

[0061] (6) Display unit 108

[0062] (7) Input Unit 109

[0063] (8) Control Unit 110

[0064] 5. Fifth Implementation Method (Particle Sorting Method)

[0065] 1. First Embodiment (Particle Sorting Device 100)

[0066] Figure 1 This is a view showing an example configuration of the particle sorting apparatus 100 according to the first embodiment.

[0067] The particle sorting apparatus 100 according to this embodiment includes a microchip M, which includes: a main flow path M2 through which a liquid containing particles flows; a sheath fluid flow path M41 connected to the main flow path and providing sheath fluid; and a sheath fluid inlet M4 for introducing sheath fluid, wherein the sheath fluid flowing through the sheath fluid inlet M4 vibrates. Each unit will be described in detail below.

[0068] (1) Microchip M

[0069] Figure 2 This is a view showing an example configuration of microchip M. Figure 3 This is a view showing an example configuration of the aperture M1 of the microchip M. Figure 2 A is a schematic top view, and Figure 2 B is a schematic cross-sectional view corresponding to section PP in A. Furthermore, Figure 3 A is the top view. Figure 3 B is a cross-sectional view. Figure 3 C is the front view.

[0070] like Figure 2As shown in Figure A, the microchip M includes a sheath fluid flow path M41 connected to the main flow path M2 and through which sheath fluid flows, a sheath fluid inlet M4 for introducing sheath fluid, a sample fluid flow path M31 connected to the main flow path M2 and through which sample fluid containing microparticles flows, a sample fluid inlet M3 for introducing sample fluid, and a merging section where the sample flow is introduced and merged with the sheath fluid. The sheath fluid introduced from the sheath fluid inlet M4 is fed in two directions respectively, and then the sample fluid is combined with the sample fluid introduced from the sample fluid inlet M3 by clamping the sample fluid from both directions at the merging section. Therefore, a three-dimensional laminar flow with the sample fluid laminar flow located at the center of the sheath fluid laminar flow is formed at the merging section.

[0071] Figure 2 M51, shown in section A, represents the suction flow path, used to apply negative pressure to the interior of the main flow path M2 when blockage or air bubbles occur in the main flow path M2, thereby temporarily reversing the flow and eliminating the blockage or air bubbles. A suction port M5, which connects to a negative pressure source such as a vacuum pump, is formed at one end of the suction flow path M51. Furthermore, the other end of the suction flow path M51 is connected to the main flow path M2 via a connecting port M52.

[0072] The laminar flow width in three-dimensional laminar flow is at the narrowing point M61 (see...) Figure 2 A) and M62 (see A) and M62 Figure 3 The flow path narrows at points A and B, with the narrowing sections M61 and M62 formed such that the area of ​​the cross-section perpendicular to the flow direction gradually decreases from upstream to downstream in the flow direction. Subsequently, the three-dimensional laminar flow is discharged as a fluid flow from the orifice M1 located at one end of the flow path.

[0073] Here, in the prior art, the fluid flow ejected from the orifice M1 is converted into droplets by applying vibration to the vibrating element of the orifice M1, but in this art, the fluid flow ejected from the orifice M1 is converted into droplets by applying vibration to the sheath fluid flowing through the sheath fluid inlet M4, as described later.

[0074] An aperture M1 opens in the direction of the end faces of substrate layers Ma and Mb, and a cutout M11 is disposed between the opening of aperture M1 and the end face of the substrate layers. The cutout M11 is formed by removing substrate layers Ma and Mb between the opening of aperture M1 and the end face of the substrate, such that the diameter L1 of the cutout M1 is larger than the opening diameter L2 of aperture M1 (see [reference]). Figure 3 (C). The diameter L1 of the cut portion M11 is preferably formed to be more than twice the opening diameter L2 of the orifice M1 so as not to impede the movement of droplets discharged from the orifice M1.

[0075] In this technology, "micro" means that at least a portion of the flow path contained in the microchip M has a size on the order of μm, specifically, a cross-sectional size on the order of μm. That is, in this technology, a "microchip" refers to a chip that includes flow paths on the order of μm, specifically, a chip that includes flow paths with a cross-sectional size on the order of μm. For example, a chip that includes a particle sorting unit configured with flow paths having a cross-sectional size on the order of μm can be referred to as a microchip according to this technology.

[0076] Microchip M can be manufactured by methods known in the art. For example, microchip M is formed by bonding substrate layers Ma and Mb, on which a main flow path M2 is formed. The main flow path M2 can be formed in substrate layers Ma and Mb, for example, by injection molding a thermoplastic resin using a mold. For example, the flow path can be formed in all or more substrates, or it can be formed in only some of the two or more substrates. Furthermore, microchip M can be formed from three or more substrates by further bonding substrates from an upper direction, a lower direction, or both directions relative to the plane of the substrate on which the respective flow paths are formed.

[0077] Materials known in the art can be used as materials for forming the microchip M. Examples include, but are not limited to, polycarbonate (PC), cyclic olefin polymers, polypropylene, polydimethylsiloxane (PDMS), polymethyl methacrylate (PMMA), polyethylene, polystyrene, glass, silicon, etc. Among these, polymer materials such as polycarbonate, cyclic olefin polymers, and polypropylene are particularly preferred because they have excellent processability and can be used to manufacture microchips inexpensively using molding equipment.

[0078] The microchip M is preferably transparent. For example, in the microchip M, at least a portion through which light (laser and scattered light) passes can be transparent, and the entire microchip M can be transparent.

[0079] In this technique, the "sample" contained in the sample liquid specifically refers to microparticles, and these microparticles can be of a size capable of flowing within the flow path of the microchip M. In this technique, the microparticles can be appropriately selected by those skilled in the art. In this technique, the microparticles can include, for example, biological microparticles (such as cells, cell aggregates, microorganisms, and ribosomes) and synthetic microparticles (such as gel particles, beads, latex particles, polymer particles, and industrial particles).

[0080] Bioparticles (also known as "biological particles") can include chromosomes, ribosomes, mitochondria, organelles (small organelles), etc., that make up various cells. Cells can include animal cells (e.g., blood cells, etc.) and plant cells. These cells can specifically be blood-derived cells or tissue-derived cells. Blood cells can be, for example, suspension cells such as T cells and B cells. Tissue-derived cells can be, for example, adherent cells isolated from adherent cultured cells or tissues, etc. Cell aggregates can include, for example, spheroids, organoids, etc. Microorganisms can include bacteria such as Escherichia coli, viruses such as tobacco mosaic virus, fungi such as yeast, etc. In addition, bioparticles can also include biopolymers, such as nucleic acids, proteins, and their complexes. These biopolymers can be, for example, those extracted from cells, or those contained in blood samples or other liquid samples.

[0081] Synthetic microparticles can be, for example, particles comprising organic or inorganic polymer materials, metals, etc. Organic polymer materials can include polystyrene, styrene-divinylbenzene, polymethyl methacrylate, etc. Inorganic polymer materials can include glass, silica, magnetic materials, etc. Metals can include gold colloids, aluminum, etc. These synthetic microparticles can be, for example, gel particles, beads, or the like, and can specifically be gel particles or beads, selected from one or more combinations of oligonucleotides, peptides, proteins, or enzymes bound thereto.

[0082] The particles can be spherical or substantially spherical, or they can be non-spherical. Those skilled in the art can appropriately select the size and mass of the particles based on the flow path dimensions of the microchip M. Conversely, the flow path dimensions of the microchip M can also be appropriately selected based on the size and mass of the particles. In this technology, chemical or biological labels (e.g., fluorescent dyes, fluorescent proteins, etc.) can be attached to the particles as needed. This labeling can facilitate the detection of the particles. Those skilled in the art can appropriately select the label to be attached. Molecules that specifically react with the particles (e.g., antibodies, aptamers, DNA, RNA, etc.) can bind to the label.

[0083] In this technology, the microparticles are preferably biological particles, and in particular, they can be cells.

[0084] (2) Connecting component C

[0085] The particle sorting apparatus 100 according to this embodiment further includes a connecting member that can be attached to the microchip M and has a sheath fluid introduction coupling portion C2 coupled to the sheath fluid introduction portion M4.

[0086] Figure 1 The connecting member C shown includes at least a sample liquid introduction coupling part C1 coupled to the sample liquid introduction part M3 and a sheath liquid introduction coupling part C2 coupled to the sheath liquid introduction part M4.

[0087] By using a removable connecting component C from the microchip M, a portion of the components constituting the device can be separated when continuously sorting many different particles using a single apparatus. Therefore, even if particles contained in previously sorted fluid streams remain in the component, the entire component can be removed, reducing the risk of contamination. Furthermore, by making the microchip M and connecting component C disposable for each sample, the hassle of cleaning operations performed when changing samples is eliminated, and the operator's workload is reduced.

[0088] The sheath fluid inlet connection C2 may include a delivery tube capable of supplying liquid from the sheath fluid delivery unit 101. Alternatively, the delivery tube may have an inter-tube coupling portion that is directly connected to the sheath fluid delivery unit 101. In this case, the inter-tube coupling portion is preferably configured to prevent the liquid inside the delivery tube from contacting external air. Therefore, the cleanliness of the sheath fluid can be ensured.

[0089] The sample liquid inlet coupling C1 may have a tube fixing portion for securing the liquid feed tube that can feed liquid from the sample liquid feed unit 102. Therefore, it saves labor in installing and fixing the tubes, prevents operational complexity during measurement, and reduces the operator's burden. Furthermore, by making these components disposable for each sample, contamination can also be prevented.

[0090] The liquid delivery tube can be integrally formed with the connecting member C, or it can be formed separately. For example, the liquid delivery tube and the tube fixing part that can supply liquid from the sample liquid delivery unit 102 are formed to be detachable from the connecting member C, and can be easily connected to the sample liquid delivery unit 102 located in a different position than the sheath liquid delivery unit 101.

[0091] In this embodiment, the vibrating element C3 is attached to the connecting member C. Therefore, vibration can be propagated to the sheath fluid flowing through the sheath fluid inlet M4 of the microchip M, and droplet formation can be induced after it is ejected from the nozzle.

[0092] The driving frequency of the vibrating element C3 is preferably different from the resonant frequency of the flow path in the microchip M. The reason will be described in detail below.

[0093] like Figure 4 As shown, a standing wave is generated between the vibrating element C3 and the nozzle, but the undulating pressure near the nozzle varies depending on the shape, size, material, and driving frequency of the flow path. The state in which the undulating pressure is maximized is the so-called resonance (see [reference]). Figure 5Furthermore, for example, the resonant frequency can be reduced by increasing the length of the flow path, and the resonant frequency can be adjusted by combining multiple such characteristics. Then, the detailed frequency characteristics of the fluctuating pressure can be estimated using commercially available acoustic analysis software, etc. If the resonance occurs at the desired driving frequency, the droplet can form with small vibrational energy, but the change in fluctuating pressure reverses before and after the resonant frequency of the flow path in the microchip M, and the fluctuation is also abrupt. Therefore, droplet formation at the resonant frequency is less stable. Therefore, as... Figure 5 As shown, by slightly shifting the driving frequency from the resonant frequency, both efficiency and stability can be improved through resonance.

[0094] In this technique, whether the driving frequency is shifted to a higher or lower side or to the resonant frequency is irrelevant. Typically, the driving frequency is set to a bandwidth with a specific width for adjustment. This shift is not particularly limited, as long as the driving bandwidth does not intersect with the resonant frequency even in the presence of disturbances such as temperature changes, and it only needs to be set considering the balance between the desired efficiency improvement and the stability within the bandwidth. In this technique, for example, the driving frequency of the vibrating element can be within ±10% of the resonant frequency of the flow path in the microchip M.

[0095] In this embodiment, the sheath fluid introduction coupling portion C2 preferably includes a sheath fluid convergence portion C21, the width of which gradually narrows or partially narrows from the side of the attached vibrating element C3 toward the side of the sheath fluid introduction portion M4. Therefore, the thickness of the flow path in the sheath fluid introduction coupling portion C2 gradually decreases from the thickness around the vibrating element C3 to the thickness around the sheath fluid introduction portion M4. The scale of the dimension around the vibrating element C3 is connected to the scale of the dimension around the flow path. The vibration energy of the vibrating element C3 is concentrated near the sheath fluid introduction portion M4, and the vibration energy can be effectively transmitted to the flow path in the microchip M using a small driving voltage. The following will describe in detail how the vibration propagates to the sheath fluid near the sheath fluid introduction portion M4 of the microchip M.

[0096] Sheath fluid is supplied from sheath fluid feeding unit 101 to sheath fluid convergence section C21, and the sheath fluid is vibrated by a vibrating element C3 arranged upstream of convergence section C21. Vibrating element C3 includes, for example, a piezoelectric element unit and a piston section, and each part of the piezoelectric element unit and the piston section is firmly coupled using an adhesive or the like. The structure of the piezoelectric element unit is not limited, as long as the ultimately extracted vibration can vibrate in the X direction at the target vibration frequency and the necessary amplitude (see...). Figure 1 B) is sufficient. For example, one can think of stacked structures, square plate structures, disk structures, tube structures, etc. (see...) Figure 6(A to C). Furthermore, magnetic force, such as that of a permanent magnet or a solenoid, can be used as the vibrating element C3. Additionally, instead of the structure where a piston, which is attached to the piezoelectric element, is inserted into the converging portion C21, a structure such as... Figure 7 The structure shown depicts a curved piezoelectric element attached to the top surface of the converging portion C21. Sheath fluid is supplied to the microchip M from the sheath fluid inlet M4, and the vibration of the vibrating element C3 propagates through the sheath fluid to induce droplet formation after being ejected from the nozzle.

[0097] As the vibrating element C3, for example, a piezoelectric element such as a piezoelectric element can be used; however, as mentioned above, a vibrating element C3 that converts electrical energy into vibration through magnetic force such as a permanent magnet and a solenoid can be used. Furthermore, the frequency is not limited to the ultrasonic region above 20 kHz and can be appropriately set according to the size of the droplets to be formed.

[0098] Preferably, the height of the sheath fluid convergence portion C21 gradually or partially decreases from the vibrating element C3 side toward the sheath fluid inlet portion M4 side. Specifically, for example, the shape can be a generally conical shape, a generally pyramidal shape, or a body of revolution of an exponential function or parabola. By designing these shapes, vibrational energy can be effectively concentrated. The invention is not limited to these shapes, provided that the vibrating element C3 can generate sufficient vibrational energy and there are no problems with assemblability. Rather, a form with a gradually narrowing flow path generally has greater advantages (e.g., since the amplitude of the vibrating element C3 may be small, droplets can be formed using a low driving voltage of the piezoelectric element, and the freedom of selection and design of the piezoelectric element is increased).

[0099] When a roughly pyramidal shape is chosen as the shape of the sheath fluid convergence portion C21, a flat portion can be provided. Therefore, specifically, when the sheath fluid convergence portion C21 is formed of a transparent component, there is an advantage that the interior of the convergence portion C21 can be easily seen and bubbles can be easily identified. Furthermore, roughly pyramidal shapes with small interior angles, such as roughly triangular pyramids, are not preferred because bubbles are easily trapped at the corners, making bubble removal difficult. For example, roughly pyramidal shapes with large interior angles, such as roughly hexagonal or larger pyramidal shapes, are preferred. Additionally, to improve the effect without suppressing the vortex described later, a pyramidal shape with large interior angles is preferred. Furthermore, from the viewpoint of bubble removal or from the viewpoint of not obstructing the vortex, it is desirable not to sharpen each corner and to appropriately apply radius R. Appropriate application of R includes, for example, applying an R that is approximately 1 / 10 or greater of the radius of the inscribed circle of the polygon.

[0100] Note that it is preferable to determine the detailed shape and dimensions of the sheath fluid convergence section C21 while examining its acoustic properties. This can further improve the efficiency of vibration energy utilization.

[0101] Materials known in the art can be used as the material for forming the sheath fluid convergence portion C21, but in this art, resin, metal, or a transparent component is preferred. For example, polyetheretherketone (PEEK) can be used as a resin. Furthermore, for example, polymethyl methacrylate (PMMA), polycarbonate (PC), etc., can be used as a transparent component. By forming the sheath fluid convergence portion C21 with a transparent component, the interior of the sheath fluid convergence portion C21 can be observed. For example, stainless steel, aluminum alloy, titanium alloy, etc., can be used as a metal. By forming the sheath fluid convergence portion C21 with a metal, electrodes for droplet charging can be omitted.

[0102] Figure 8 This diagram shows electrode C4 positioned near the sheath fluid convergence point C21. (See diagram below.) Figure 8 As shown, when a charge is applied to the formed droplet, if the sheath fluid convergence portion C21 is formed of an insulator such as resin, an electrode C4 is inserted into the sheath fluid convergence portion C21, allowing the droplet to be charged through the sheath fluid. The aim is to make the distance between the droplet splitting point and the electrode C4 as close as possible, and to charge the droplet at a time closer to the ideal moment.

[0103] In this configuration, tiny air bubbles may become trapped at the boundary between the sheath fluid convergence point C21 and the electrode C4, and are particularly difficult to remove when the flow stagnates. Therefore, in this technology, it is preferable to generate a vortex that causes the sheath fluid to swirl within the sheath fluid convergence point C21. This vortex eliminates trapped portions and allows for easy removal of air bubbles.

[0104] Furthermore, the sheath fluid convergence point C21 encounters a section where the flow path thickens throughout the flow path system, and bubbles may be retained within this section. When bubbles are retained, vibrational energy is absorbed by the bubbles, and the breakage point is affected. Therefore, it is conceivable that the sheath fluid inlet C20, used to introduce sheath fluid into the sheath fluid convergence point C21, is located away from the center of the sheath fluid convergence point C21. Specifically, for example, the position of the sheath fluid inlet C20 is offset from the center of the sheath fluid convergence point C21 to the upper part, and further arranged in the upstream part. Therefore, eddies are generated in the sheath fluid convergence point C21, thereby preventing stagnant sections, thus preventing bubble retention, and because the specific gravity of bubbles is less than that of sheath fluid, bubbles easily accumulate at the center of the flow path and flow downwards.

[0105] Note that the offset does not have to be in the upward direction of the sheath fluid convergence section C21, but rather in the downward direction, as long as the intensity of the generated eddy current is sufficient to cause the bubble to flow downwards. Furthermore, there is also a configuration in which the electrode C4 is inserted into this section to charge the droplet (see...). Figure 8This allows for the capture of fine bubbles at the boundary between the pores formed in the sheath fluid convergence part C21 and the electrode C4, thus also enabling the removal of bubbles adhering to the periphery of the electrode C4 through eddy current stripping.

[0106] In this embodiment, the microchip M and the connecting member C can be appropriately disassembled as needed and can be disposable. Furthermore, the vibrating element C3 attached to the connecting member C can also be cyclically attached to the connecting member C beforehand, and in this case, the vibrating element C3 can be disposable.

[0107] 2. Second Embodiment (Particle Sorting Device 100)

[0108] Figure 9 A and B are views illustrating an example configuration of a particle sorting apparatus according to a second embodiment. Figure 9 A is a schematic top view, and Figure 9 B is a schematic sectional view.

[0109] In the first embodiment described above, a confluence portion is formed in the microchip M to introduce the sample flow and merge it with the sheath fluid, and the sample fluid is introduced into the sheath fluid in the microchip M. However, this embodiment is configured such that the flow path is in the form of introducing the sample fluid into the sheath fluid convergence portion C21 and focusing and converging the core flow containing the sample fluid through hydrodynamics. Note that, except for the different method of introducing the sample fluid into the sheath fluid, this embodiment is similar to the first embodiment described above.

[0110] In this embodiment, such as Figure 9 As shown, the vortex component of the flow is removed by a current plate formed inside the sheath fluid convergence section C21, upstream of the sample fluid inlet used to introduce the sample fluid into the sheath fluid convergence section C21, before the sample fluid is introduced. This prevents the core flow from being disturbed by vortices. The current plate can be formed from, for example, a thin plate. Here, it is preferable that the rectifier plate is not positioned at the thickest part of the flow path to avoid disturbing the vortices.

[0111] 3. Third Embodiment (Particle Sorting Device 100)

[0112] Figure 12 and Figures 14 to 16 This is a diagram showing a schematic example of the connecting portion C22 in the particle sorting apparatus 100 according to the third embodiment. It should be noted that... Figures 14 to 16 In this paper, the vibrating element C3 is omitted.

[0113] In this embodiment, the sheath fluid inlet connector C2 has a sheath fluid converging portion C21, in which the width and height gradually or partially decrease from the side connected to the vibrating element C3 toward the sheath fluid inlet portion M4, and includes a connector C22 having a tubular portion C221, which communicates with the distal end C211 of the sheath fluid converging portion C21 between the sheath fluid converging portion C21 and the sheath fluid inlet portion M4. Furthermore, in this embodiment, except for the connector C22, it is the same as in the first embodiment described above.

[0114] In this embodiment, by providing the connecting part C22, the scale of the dimensions around the vibrating element C3 and the scale of the dimensions around the flow path can be connected more effectively.

[0115] Examples of materials forming the connecting part C22 include elastomers, resins, metals, or combinations of two or more of them.

[0116] Examples of elastomers include thermosetting elastomers and thermoplastic elastomers.

[0117] Examples of thermosetting elastomers include vulcanized rubbers such as natural rubber and synthetic rubber, as well as resin elastomers such as silicone rubber and fluororubber.

[0118] Examples of thermoplastic elastomers include polystyrene-based thermoplastic elastomers, olefin / olefin-based thermoplastic elastomers, polyvinyl chloride-based thermoplastic elastomers, polyurethane-based thermoplastic elastomers, polyester-based thermoplastic elastomers, and polyamide-based thermoplastic elastomers.

[0119] Examples of resins include thermoplastic resins, thermosetting resins, etc.

[0120] Examples of thermoplastic resins include: polyolefins such as polystyrene, polyethylene, and polypropylene; polyvinyl chloride; acrylic resins; ABS resins; AS resins; engineering plastics such as polyamides, polycarbonates, polyacetals, and polyethylene terephthalate; super engineering plastics such as polyethersulfone, polyetheretherketone (PEEK), and thermoplastic polyimides; and so on.

[0121] Examples of thermosetting resins include phenolic resins, melamine resins, polyurethanes, unsaturated polyester resins, and epoxy resins.

[0122] Examples of metals include aluminum alloys, titanium alloys, and stainless steel.

[0123] The following text will describe in detail a specific example of the connecting part C22.

[0124] [Example 1 of the connection section C22]

[0125] Figure 12This is a schematic view illustrating a schematic example 1 of the connecting portion C22. In schematic example 1, the tubular member C222 is inserted into the tubular portion C221 formed of a thermosetting elastomer. Thus, upon contact with the sheath fluid, it can separate from the acoustic impedance of the sheath fluid (when the acoustic impedance is equal, vibration is diffused and vibration energy is dissipated). Therefore, it is possible to prevent vibration generated from the vibrating element C3 from diffusing to the connecting portion C22, and to efficiently transfer vibration energy to the flow path in the microchip M.

[0126] Furthermore, when the driving voltage applied to the vibrating element C3 is the same as the driving voltage when the tubular member C222 is not inserted, such as Figure 13 As shown, a shorter BOP length can be obtained, or the same BOP length can be obtained at a lower drive voltage.

[0127] Examples of materials forming the tubular member C222 include the aforementioned elastomers, resins, metals, or combinations thereof. In Example 1, polystyrene, acrylic resin, aluminum alloy, titanium alloy, or stainless steel are particularly preferred, aluminum alloy, titanium alloy, or stainless steel are more preferred, and stainless steel is even more preferred.

[0128] Furthermore, in Example 1, it is preferred that at least a portion of the tubular member C222 is formed of the aforementioned material, but more preferably, the entire tubular member C222 is formed of the aforementioned material. Additionally, in Example 1, the tubular portion C221 may be formed of a thermosetting elastomer for use as a seal.

[0129] In this embodiment, the length of the tubular portion C221 in the longitudinal direction can be the same as the length of the tubular member C222 in the longitudinal direction; however, it can be slightly shorter considering the compression for sealing. Furthermore, there is no particular limitation on the size of the inner and outer diameters of the tubular member C222, but the outer diameter of the tubular member C222 can, for example, be the same as or slightly larger than the flow path diameter of the tubular portion C221.

[0130] [Example 2 of the connection section C22]

[0131] Figure 14This is a schematic view illustrating a second example of the connecting portion C22. In this second example, the connecting portion C22, including the tubular portion 221, is formed by two-color molding of resin C22a and thermosetting elastomer C22b. Specifically, the thermosetting elastomer C22b for sealing is fused to the resin C22a by two-color molding. By forming at least a portion of the tubular portion C221 in the connecting portion C22 with resin C22a in this way, compared to the case where the entire tubular portion 221 is formed of thermosetting elastomer, it is possible to separate the acoustic impedance from the sheath fluid upon contact with the sheath fluid and to suppress the diffusion of vibration to the outside of the flow path of the tubular portion C221. Here, examples of resin C22a include those described above, and in this second example, polystyrene or acrylic resin is particularly preferred.

[0132] [Example 3 of the connection section C22]

[0133] Figure 15 This is a schematic view illustrating a schematic example 3 of the connecting portion C22. In Example 3, the connecting portion C22, including the cylindrical portion 221, is entirely formed of metal, and O-rings C22c for sealing are disposed at both ends of the connecting portion C22. By forming the entire connecting portion 22 with metal in this way, vibrations generated from the vibrating element C3 can be prevented from diffusing to the connecting portion C22, and vibrational energy can be efficiently transferred to the flow path in the microchip M. Here, examples of metals include those described above, and in this schematic example 3, aluminum alloy, titanium alloy, or stainless steel is particularly preferred, and stainless steel is more preferred. Furthermore, materials known in the art can be used as the material for forming the O-rings C22c. Examples include the elastomers described above, resins, or combinations of two or more thereof.

[0134] [Example 4 of the connection section C22]

[0135] Figure 16 This is a schematic view illustrating a fourth example of the connecting portion C22. In this example, the sheath fluid confluence portion C21 and a portion of the connecting portion C22, including the tubular portion 221, are formed of metal, and an O-ring C22c and a thermosetting elastomer C22b are provided for sealing at both ends. By forming the sheath fluid confluence portion C21 with metal in this manner, the diffusion of vibrations generated by the vibrating element C3 can be further suppressed. Examples of the metals forming the sheath fluid confluence portion C21 and a portion of the connecting portion C22 include those described above, but in this example, aluminum alloy, titanium alloy, or stainless steel is particularly preferred, with stainless steel being more preferred. Furthermore, the materials described above can be listed as materials for forming the O-ring C22c.

[0136] 4. Fourth Embodiment (Particle Sorting Device 100)

[0137] Figure 10 This is a diagram showing an example configuration of a particle sorting apparatus according to the fourth embodiment.

[0138] The particle sorting device 100 according to this embodiment includes a microchip, which includes: a main flow path M2 through which liquid containing particles flows; a sheath fluid flow path M41 connected to the main flow path M2 and supplying sheath fluid; a sheath fluid inlet M4 for introducing sheath fluid; a light irradiation unit 103 for irradiating particles; a light detection unit 104 for detecting light from particles; and a processing unit 105 for processing the signal obtained from the light detection unit 104, wherein the sheath fluid flowing through the sheath fluid inlet vibrates. Furthermore, a sorting unit 106, a storage unit 107, a display unit 108, an input unit 109, a control unit 110, etc., can be provided as needed.

[0139] Since the microchip M is similar to the one described above, its description is omitted here. Furthermore, the method of vibrating the sheath fluid through the sheath fluid inlet M4 of the microchip M is also similar to the one described above, so its description is omitted here.

[0140] (1) Light irradiation unit 103

[0141] The light illumination unit 103 illuminates the particles to be sorted with light (e.g., excitation light). The light illumination unit 103 may include a light source that emits light and an objective lens that focuses the excitation light onto particles flowing in a detection area. The light source can be appropriately selected by those skilled in the art according to the sorting purpose; for example, it may be a laser diode, an SHG laser, a solid-state laser, a gas laser, a high-brightness LED, or a combination of two or more of these. In addition to the light source and objective lens, the light illumination unit 103 may include other optical elements as needed.

[0142] (2) Optical detection unit 104

[0143] The light detection unit 104 detects light (scattered light and / or fluorescence) generated from the particles by illumination through the light illumination unit 103. The light detection unit 104 may include a condenser lens for collecting the fluorescence and / or scattered light generated from the particles, and a photodetector. PMTs, photodiodes, CCDs, CMOS, etc., can be used as photodetectors, but this technology is not limited to these. In addition to the condenser lens and photodetector, the light detection unit 104 may include other optical elements as needed. The light detection unit 104 may further include, for example, a beam splitter. Examples of optical components constituting a beam splitter may include gratings, prisms, and filters. The beam splitter can detect, for example, light having the wavelength to be detected and light having another wavelength.

[0144] The fluorescence detected by the light detection unit 104 can be fluorescence generated from the particle itself or fluorescence generated from a substance labeled in the particle (e.g., a fluorescent substance, etc.), but is not limited thereto in this art. The scattered light detected by the light detection unit 104 can be forward scattered light, side scattered light, Rayleigh scattering, Mie scattering, or a combination thereof.

[0145] (3) Processing unit 105

[0146] The processing unit 105 is connected to the light detection unit 104 and processes the signal obtained from the light detection unit 104. For example, the detection value of the light received from the light detection unit 104 can be corrected to calculate the characteristic quantity of each particle. More specifically, characteristic quantities representing the size, shape, internal structure, etc. of the particles are calculated from the received fluorescence, forward scattered light, and backscattered light detection values. Furthermore, a sorting control signal can be generated by performing sorting determination based on the calculated characteristic quantities and sorting conditions received in advance from the input unit.

[0147] The processing unit 105 can also analyze the state of particles using external analysis equipment or the like based on the detection value of the light detected by the light detection unit 104. For example, the processing unit 105 can be implemented by a personal computer or CPU, and can also be stored as a program in hardware resources including recording media (non-volatile memory (USB memory, etc.), HDD, CD, etc.) and run by a personal computer or CPU. Furthermore, the processing unit 105 can be connected to each unit of the particle sorting device 100 via a network.

[0148] (4) Sorting unit 106 (including charging unit 106c)

[0149] The sorting unit 106 includes at least a deflector plate 106a for changing the direction of charged droplets and a collection container 106b for collecting droplets. Although in Figure 4 As defined separately, the charging unit 106c is part of the sorting unit 106 and performs charging based on the sorting control signal generated by the processing unit 105.

[0150] exist Figure 10 In the particle sorting device 100 shown, the vibrating element C3 attached to the connecting member C forms droplets by propagating vibration to the sheath fluid as described above. The charging unit 106c is connected to the electrode C4 inserted into the sheath fluid convergence section C21, and based on the sorting control signal generated by the processing unit 105, the droplets discharged from the aperture M1 of the microchip M are positively or negatively charged. Then, the path of the charged droplets is changed in the desired direction by the deflection plate (counter electrode) 106a through which a voltage is applied, and the charged droplets are sorted.

[0151] (5) Storage unit 107

[0152] Storage unit 107 stores all items related to measurement, such as the values ​​detected by light detection unit 103, the feature quantities calculated by processing unit 105, sorting control signals, and sorting conditions input by input unit.

[0153] In the particle sorting apparatus 100, the storage unit 107 is not necessary and can be connected to an external storage device. For example, a hard disk or the like can be used as the storage unit 107. Furthermore, the recording unit 107 can be connected to each unit of the particle sorting apparatus 100 via a network.

[0154] (6) Display unit 108

[0155] Display unit 108 can display all items related to the measurement, such as the values ​​detected by light detection unit 103 and the characteristic quantities calculated by processing unit 105. Preferably, display unit 108 displays the characteristic quantities of each particle calculated by processing unit 105 as a scattering map.

[0156] In the particle sorting apparatus 100, the display unit 108 is not necessary and can be connected to an external display device. For example, a monitor, printer, etc., can be used as the display unit 110. Furthermore, the display unit 108 can be connected to each unit of the particle sorting apparatus 100 via a network.

[0157] (7) Input Unit 109

[0158] The input unit 109 is a component operated by a user, such as an operator. The user can access the control unit 110, described later, through the input unit 109 to control each unit of the particle sorting apparatus 100. The input unit 109 preferably sets the region of interest on a scattering map displayed in the display unit 108 to determine the sorting conditions.

[0159] In the particle sorting device 100, the input unit 109 is not necessary and can be connected to an external operating device. For example, a mouse, keyboard, etc. can be used as the input unit 109. In addition, the input unit 109 can be connected to each unit of the particle sorting device 100 via a network.

[0160] (8) Control Unit 110

[0161] The control unit 110 is capable of controlling each of the light irradiation unit 103, light detection unit 104, analysis unit 105, sorting unit 106, charging unit 106c, recording unit 107, display unit 108, and input unit 109. The control unit 110 can be arranged individually for each unit of the particle sorting device 100, or it can be located externally to the particle sorting device 100. For example, the control unit 110 can be implemented by a personal computer or CPU, and furthermore, it can be stored as a program in hardware resources including recording media (non-volatile memory (USB memory, etc.), HDD, CD, etc.) and executed by a personal computer or CPU. Furthermore, the control unit 110 can be connected to each unit of the particle sorting device 100 via a network.

[0162] 5. Fifth Implementation Method (Particle Sorting Method)

[0163] The particle sorting method according to the present technology includes: a microchip, the microchip including at least: a main flow path through which a liquid containing particles flows; a sheath fluid flow path connected to the main flow path through which sheath fluid flows; and a sheath fluid inlet for introducing the sheath fluid, the sheath fluid of which vibrates and flows through the sheath fluid inlet.

[0164] Since the particle sorting method according to this technology is the same as the method performed by the particle sorting apparatus according to this technology described above, its description will be omitted here.

[0165] Example

[0166] The invention will be described in more detail below, based on examples.

[0167] Furthermore, the examples described below are representative embodiments of the present invention, and the scope of the present invention is not to be interpreted narrowly.

[0168] In existing technologies, microchips form droplets by vibrating a vibrating element. Figure 11 It is a graph showing the results of measuring the frequency characteristics of the fracture point when vibration is applied using the prior art and when vibration is applied using the present art.

[0169] like Figure 11 As shown, in the prior art, the BOP changes drastically with frequency. This is believed to be because, as mentioned above, phenomena occur such as the vibration frequency matching the characteristic value of the microchip and causing large vibrations of the microchip (resonance, shortened BOP), or conversely, they cancel each other out and cause small vibrations of the microchip (anti-resonance, extended BOP). On the other hand, in this technology, since vibration is not applied through the microchip and is not affected by its characteristic value, it can be seen that the change in BOP with frequency becomes more gradual and the generation of droplets can be stabilized.

[0170] It should be noted that this technology may also have the following configurations.

[0171] [1] A particle sorting device, comprising:

[0172] The microchip includes: a main flow path through which a liquid containing microparticles flows; a sheath fluid flow path connected to the main flow path through which sheath fluid flows; and a sheath fluid inlet for introducing the sheath fluid, wherein...

[0173] Vibrate the sheath fluid flowing in the sheath fluid inlet.

[0174] [2] The particle sorting device according to [1] further includes a connecting member that can be attached to the microchip and has a sheath fluid inlet coupling portion coupled to the sheath fluid inlet portion.

[0175] [3] According to the particle sorting device of [2], the vibrating element is attached to the connector.

[0176] [4] According to the particle sorting device described in [3], the driving frequency of the vibrating element is different from the resonant frequency of the flow path in the microchip.

[0177] [5] According to the particle sorting device of [4], the driving frequency of the vibrating element is within ±10% of the resonant frequency of the flow path in the microchip.

[0178] [6] The particle sorting apparatus according to any one of [3] to [5], wherein the sheath fluid inlet connection includes a sheath fluid converging portion, the width of which gradually or partially narrows from one side of the vibrating element toward one side of the sheath fluid inlet.

[0179] [7] According to the particle sorting device of [6], the height of the sheath fluid convergence portion gradually or partially decreases from one side of the vibrating element toward the side of the sheath fluid inlet portion.

[0180] [8] The particle sorting apparatus according to [7] further includes a connecting portion having a tubular portion communicating with the distal end of the sheath fluid gathering portion between the sheath fluid gathering portion and the sheath fluid introduction portion.

[0181] [9] According to the particle sorting device of [8], the tubular member is inserted inside the tubular portion.

[0182]

[10] According to the particle sorting apparatus of [9], wherein at least a portion of the tubular portion and / or the tubular member is formed of at least one selected from the group consisting of elastomers, resins and metals.

[0183]

[11] The particle sorting apparatus according to any one of [6] to

[10] , wherein the sheath fluid converging portion is a generally conical shape, a generally pyramidal shape, or a body of revolution of an exponential function or a parabola.

[0184]

[12] The particle sorting apparatus according to any one of [6] to

[11] , wherein the sheath fluid converging portion is formed of a transparent member or metal.

[0185]

[13] The particle sorting apparatus according to any one of [6] to

[12] , wherein the electrode is inserted into the sheath fluid converging portion.

[0186]

[14] The particle sorting apparatus according to any one of [6] to

[13] , wherein a vortex that causes the sheath fluid to swirl is generated in the sheath fluid converging portion.

[0187]

[15] The particle sorting apparatus according to any one of [6] to

[14] , wherein the sheath fluid inlet for introducing the sheath fluid into the sheath fluid converging portion is located at a position away from the center of the sheath fluid converging portion.

[0188]

[16] A particle sorting device, comprising:

[0189] The microchip includes: a main flow path through which a liquid containing microparticles flows; a sheath fluid flow path connected to the main flow path and supplying sheath fluid; and a sheath fluid inlet for introducing the sheath fluid.

[0190] A light irradiation unit that irradiates the particles with light;

[0191] A light detection unit detects light from the particles; and

[0192] The processing unit processes the signal obtained from the optical detection unit, wherein...

[0193] Vibrate the sheath fluid flowing in the sheath fluid inlet.

[0194]

[17] A particle sorting method, comprising in a microchip: at least a main flow path through which a liquid containing particles flows; a sheath fluid flow path connected to the main flow path through which sheath fluid flows; and a sheath fluid inlet for introducing the sheath fluid.

[0195] The sheath fluid vibrates and flows through the sheath fluid inlet.

[0196] Reference Symbol List

[0197] 100-particle sorting device

[0198] 101 Sheath Fluid Feeding Unit

[0199] 102 Sample Liquid Feeding Unit

[0200] 103 light irradiation units

[0201] 104 optical detection units

[0202] 105 processing unit

[0203] 106 sorting units

[0204] 107 memory cells

[0205] 108 display units

[0206] 109 input units

[0207] 110 control unit

[0208] M Microchip

[0209] Ma, Mb substrate layer

[0210] M1 hole

[0211] M11 incision

[0212] M2 Main Road

[0213] M3 Sample Liquid Inlet Section

[0214] M31 Sample Liquid Flow Path

[0215] M4 sheath fluid inlet section

[0216] M41 sheath fluid flow path

[0217] M5 intake port

[0218] M51 Inhalation Flow Path

[0219] M52 connector

[0220] M61, 62 narrowing sections

[0221] M7 Straight Section

[0222] Diameter of L1 cut M11

[0223] The opening diameter of L2 orifice M1

[0224] C-connecting component

[0225] C1 sample solution is introduced into the coupling section.

[0226] C2 sheath fluid inlet coupling section

[0227] C20 sheath fluid inlet

[0228] C21 Sheath fluid convergence section

[0229] The front end of the C211 sheath fluid convergence section

[0230] C22 connection part

[0231] C22a resin

[0232] C22b thermosetting elastomer

[0233] C22c O-ring

[0234] C221 tubular portion

[0235] C222 tubular components

[0236] C3 Vibration Element

[0237] C4 electrode.

Claims

1. A particle sorting apparatus comprising: a microchip including: a main flow path through which a liquid containing particles flows; a sheath liquid flow path communicating with the main flow path and through which a sheath liquid flows; and a sheath liquid introduction portion that introduces the sheath liquid; a connection member attachable to the microchip and having a sheath liquid introduction coupling portion coupled to the sheath liquid introduction portion, and a vibration element attached to the connection member, wherein the sheath liquid flowing through the sheath liquid introduction portion is vibrated, and wherein the sheath liquid introduction coupling portion includes a sheath liquid converging portion that gradually narrows or partially narrows in width from one side of the vibration element toward one side of the sheath liquid introduction portion.

2. The particle sorting device of claim 1, wherein, The driving frequency of the vibration element is different from the resonance frequency of the flow path in the microchip.

3. Apparatus for sorting particles as claimed in claim 2 wherein, The driving frequency of the vibration element is within ±10% of the resonance frequency of the flow path in the microchip.

4. The particle sorting device of claim 1, wherein, The height of the sheath liquid converging portion gradually decreases or partially decreases from one side of the vibration element toward one side of the sheath liquid introduction portion.

5. The particle sorting device of claim 4, further comprising: A connection portion having a tubular portion communicating with a distal end of the sheath liquid converging portion between the sheath liquid converging portion and the sheath liquid introduction portion.

6. The particle sorting device of claim 5, wherein, A tubular member is inserted inside the tubular portion.

7. The particle sorting device of claim 6, wherein, At least a portion of the tubular portion and / or the tubular member is formed of at least one selected from an elastomer, a resin, and a metal.

8. The particle sorting device of claim 1, wherein, The sheath liquid converging portion is a conical shape, a polygonal pyramid shape, or a rotation body of an exponential function or a parabola.

9. The particle sorting device of claim 1, wherein, The sheath liquid converging portion is formed of a resin, a metal, or a transparent member.

10. The particle sorting device of claim 1, wherein, An electrode is inserted in the sheath liquid converging portion.

11. The particle sorting device of claim 1, wherein, In the sheath liquid converging portion, an eddy current that makes the sheath liquid swirl is generated.

12. The particle sorting device of claim 1, wherein, A sheath liquid introduction port for introducing the sheath liquid into the sheath liquid converging portion is located away from the center of the sheath liquid converging portion.

13. A particle sorting apparatus comprising: a microchip including: a main flow path through which a liquid containing particles flows; a sheath liquid flow path communicating with the main flow path and supplying a sheath liquid; and a sheath liquid introduction portion that introduces the sheath liquid; a light irradiation unit that irradiates the particles with light; a light detection unit that detects light from the particles; and a processing unit that processes a signal obtained from the light detection unit; a connection member attachable to the microchip and having a sheath liquid introduction coupling portion coupled to the sheath liquid introduction portion, and a vibration element attached to the connection member, wherein the sheath liquid flowing through the sheath liquid introduction portion is vibrated, and wherein the sheath liquid introduction coupling portion includes a sheath liquid converging portion that gradually narrows or partially narrows in width from one side of the vibration element toward one side of the sheath liquid introduction portion.

14. A method of sorting microparticles, the method of sorting microparticles comprising: In a microchip, the microchip includes at least: a main flow path through which a liquid containing microparticles flows; a sheath liquid flow path that communicates with the main flow path and through which a sheath liquid flows; and a sheath liquid introduction portion that introduces the sheath liquid; a connection member that is attachable to the microchip and has a sheath liquid introduction coupling portion that is coupled to the sheath liquid introduction portion, and a vibration element that is attached to the connection member, vibrating the sheath liquid flowing through the sheath liquid introduction portion, wherein the sheath liquid introduction coupling portion includes a sheath liquid converging portion that is gradually narrowed or partially narrowed in width from one side of the vibration element toward one side of the sheath liquid introduction portion.

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