Title of invention: Microchip for collecting fine particles and fine particle collecting apparatus
Technical field
[0001]
　The present invention relates to a microchip for collecting fine particles and a fine particle collecting apparatus.
Background technology
[0002]
　Conventionally, various microchips for collecting fine particles for a flow cytometer have been developed.
　For example, in Patent Document 1, a microstructure for a flow cytometer, which is provided with a flow channel structure that suppresses a vortex-like flow field generated after the merging of a sample liquid laminar flow and a sheath liquid laminar flow and does not cause turbulence of the sample liquid laminar flow A chip is disclosed.
[0003]
　Specifically, the microchip is provided with a
　first introduction flow path and a
　first introduction flow path sandwiched therebetween, and a second introduction flow path that laterally joins the first introduction flow path. And
　the first and second introduction flow paths are formed, and a
　confluence flow path is formed in which fluids sent from these flow paths merge and flow, and the first introduction flow is formed in the confluence flow path.
A structure is adopted in which a taper portion is formed so that the flow passage width in the sandwiching direction of the second introduction flow passage with respect to the passage gradually increases in the fluid flow direction .
　With such a structure, the laminar flow of the sample liquid can be focused and sent to the center of the flow path.
[0004]
　Further, Patent Document 2 discloses a microparticle sorting microchip capable of rapidly and stably taking out only desired fine particles from a sheath flow flowing through a flow path.
　The microparticle sorting microchip is specifically the main channel in which a main channel through which a liquid containing fine particles flows, a sorting chamber into which the fine particles are taken, and a pressure chamber in which negative pressure is generated are arranged. And a preparative flow path communicating with the preparative flow path, wherein a vertical cross section of the preparative chamber and the pressure chamber with respect to the flow direction of the liquid is perpendicular to the flow direction of the liquid in the other part of the preparative flow path. It is formed larger than the cross section.
Prior art documents
Patent literature
[0005]
Patent Document 1: Japanese Patent Laid-Open No. 2011-179945
Patent Document 2: Japanese Patent Laid-Open No. 2017-058375
Summary of the invention
Problems to be Solved by the Invention
[0006]
　The main purpose of the present technology is to provide a microparticle sorting microchip capable of further speeding up, high purity, and high acquisition rate of microparticles as compared to the microparticle sorting microchip.
Means for solving the problem
[0007]
　In order to solve the above problems, the present technology,
　a main
　flow path through which a liquid containing fine particles flows, a capture flow path that is in coaxial communication with the main flow path, and a
　capture chamber that is in communication with the capture flow path,
　A gate flow path that intersects with the capture flow path
,
　wherein the capture flow path has an opening that intersects with the gate flow path, and
　along the direction in which the liquid flows, upstream of the opening. Provided is a
microchip for sorting fine particles , wherein the cross-sectional area is smaller than the cross-sectional area downstream of the opening
.
　A plurality of the gate flow channels may be connected to the front end of the lower capture flow channel so as to be symmetrical with respect to the center of the flow of the fine particle-containing liquid from the opening at the rear end of the upper capture flow channel. preferable.
　It is preferable that the gate flow liquid always flows at a constant flow rate.
　In the lower capture channel, the gate diversion liquid may branch and flow from the gate flow channel in the direction of the upper capture channel and the direction of the pressure chamber.
　The capture chamber is preferably a pressure chamber.
　The pressure chamber may have a diaphragm.
　The inside of the pressure chamber can be repeatedly controlled to negative pressure, positive pressure, or normal pressure.
　The main flow path may include a particle detection region.
　A connection channel may be provided between the capture channel and the capture chamber.
　The main flow path can be connected to a fine particle-containing liquid introducing section for introducing the fine particle-containing liquid.
　The main flow path can be connected to a sheath liquid introducing section for introducing a sheath liquid.
　The main flow channel may be branched upstream of the capture flow channel and may be connected to a branch flow channel in which the liquid that has not flowed into the capture flow channel flows.
　The branch channel can be connected to a sheath liquid container and/or a gate liquid container.
　A particle recovery unit for recovering the captured particles may be provided downstream of the capture chamber.
　Further, when the fine particles reach the front end of the upper capture channel, the pressure chamber becomes negative pressure from normal pressure, and the fine particle-containing liquid is drawn into the upper capture channel, After being discharged from the opening at the rear end of the capture channel to the capture channel in the lower stage, a jet (jet) is generated, and the
　fine particles pass through the capture channel in the lower stage and reach the pressure chamber, The pressure inside the chamber can be changed from negative pressure or normal pressure to positive pressure. 　It is preferable that the volume of the
　fine particle-containing liquid discharged from the opening at the rear end of the upper capture channel to the lower capture channel is
half or more of the volume of the upper capture channel.
　In addition, when the pressure of the pressure chamber is changed from normal pressure to negative pressure, the volume of the fine particle-containing liquid discharged from the opening at the rear end of the upper capture channel to the lower capture channel is:
　It is preferable that the volume is equal to or less than the volume of the liquid for gate flow flowing from the gate flow channel to the capture flow channel in the lower stage.
　Furthermore, the volume of the fine particle-containing liquid discharged from the lower capture channel toward the pressure chamber when the pressure inside the pressure chamber is changed from normal pressure to negative pressure is
　less than or equal to the volume of the lower capture channel. Is preferred.
[0008]
　Further, the present technology is
　directed to a main channel
　through which a liquid containing fine particles flows, a capture channel coaxially communicating with the main channel, a
　capture chamber communicating with the
　capture channel , and the capture channel. And a gate channel that intersects with the 　trap channel
, the
capture channel has an opening that intersects the gate channel, and
　a cross-sectional area upstream of the opening is along the direction in which the liquid flows. Characterized in that it is smaller than the cross-sectional area of ​​the downstream of the opening
, mounted microparticle sorting microchip, a microchip mounting portion,
　a light irradiation unit for irradiating light to the particle detection region contained in the main flow path,
　A detection unit that detects scattered light and/or fluorescence emitted from the fine particles, and the
　capture chamber is a pressure chamber, and a pressure chamber control unit that makes the inside of the pressure chamber negative pressure or positive pressure is
included. A fine particle sorting apparatus
is provided.
　The pressure chamber control unit may have a piezo element.
Effect of the invention
[0009]
　According to the present technology, it is possible to provide a new microchip for collecting fine particles for a flow cytometer, and it is possible to achieve high-speed, high-purity, high acquisition rate of fine particles.
　Note that the effects described here are not necessarily limited and may be any effects described in the present disclosure.
Brief description of the drawings
[0010]
FIG. 1 is a diagram showing an example of a basic structure of a microchip for collecting fine particles.
FIG. 2 is an enlarged view of a capture unit of a microchip for collecting fine particles.
FIG. 3 is a diagram schematically showing a vertical cross section of a microchip for sorting fine particles.
FIG. 4 is a diagram showing movement of fine particles by a jet generated in a microchip for sorting fine particles.
FIG. 5 is an enlarged view of a capturing unit and a pressure chamber of a microchip for sorting microparticles.
FIG. 6 is an enlarged view of a portion in which a gate flow path is connected in a trapezoidal shape to a capture flow path of a microchip for sorting microparticles.
FIG. 7 is a view schematically showing the relationship between the length of the capture channel of the microparticle sorting microchip, the volume with which the sample flow is drawn, and the fine particles.
FIG. 8 is a diagram schematically showing a drawing operation and a discharging operation of a liquid containing fine particles in an example in which a capture channel and a gate channel of the microparticle sorting microchip intersect in a cross shape.
FIG. 9 is a diagram schematically showing a drawing operation and a discharging operation of the liquid containing fine particles in the capture channel when the gate flow of the microchip for sorting fine particles is constantly flowing at a constant flow rate.
FIG. 10 is a diagram showing a two-stage capture channel of a microchip for sorting microparticles.
FIG. 11 is a diagram showing a two-stage capture channel of a microchip for sorting microparticles.
FIG. 12 is a diagram showing a particle capturing operation in a capturing channel of a microchip for sorting particles.
FIG. 13 is a diagram schematically showing a capture channel having a first-stage capture channel and a second-stage capture channel.
FIG. 14 is a diagram schematically showing the relationship between the drawing volume of the capture channel and the inflow volume of the gate flow.
FIG. 15 is a diagram schematically showing the relationship between the drawing volume of the capture channel and the inflow volume of the gate flow.
FIG. 16 is a diagram showing a state in which a gate channel is asymmetrically connected to a capture channel.
FIG. 17 is a diagram showing a state in which the gate channels are symmetrically connected to the capture channel.
FIG. 18 is a diagram showing an example in which the second-stage capture channel is a tapered channel.
FIG. 19 is a diagram showing an example in which the second-stage capture channel is a tapered channel.
FIG. 20 is a diagram showing an example in which the first-stage capture channel is a tapered channel.
FIG. 21 is a diagram showing an example in which the first capture channel 1 and the second capture channel 2 are tapered channels.
FIG. 22 is a diagram showing an example having three capture paths.
FIG. 23 is a diagram showing an example having three capture channels.
FIG. 24 is a diagram showing an example having three capture channels.
FIG. 25 is a diagram showing an example having three capture channels.
FIG. 26 is a diagram showing an example in which a connection channel is provided in a microchip for sorting fine particles.
FIG. 27 is a diagram showing a specific example of gate flow path connection symmetrical with respect to the flow path center.
FIG. 28 is a diagram showing a specific example of gate flow channel connection symmetrical with respect to the flow channel center, which is different from FIG. 27.
MODE FOR CARRYING OUT THE INVENTION
[0011]
　Hereinafter, a suitable mode for carrying out the present technology will be described.
　Note that the embodiments described below show representative embodiments of the present technology, and the scope of the present technology is not narrowly interpreted by this.
　The description will be given in the following order.
[0012]
1. Basic structure of microchip for microparticle sorting
　(1) Basic structure of microchip
　(2) Movement of microparticles in capture part
　(3) Pressure control of pressure chamber
　(4) Re-emission of
　microparticles (5) Preparative performance of microparticles Improvement
　(6) Relationship between the shape of the capture channel and the drawing volume
2. Embodiment 1
3. Embodiment 2
4. Embodiment 3
5. Embodiment 4
6. Embodiment 5
7. Embodiment 6
8. Embodiment 7
9. Embodiment 8
10. Embodiment 9
11. Embodiment 10
12. Embodiment 11
13. Particulate collection system
　(1) Configuration
　(2) Particulate collection program
[0013]
　Here, in the present technology, “fine particles” include biological fine particles such as cells, microorganisms, and liposomes, and synthetic particles such as latex particles, gel particles, and industrial particles.
　Examples of the biological microparticles include chromosomes that constitute various cells, liposomes, mitochondria, organelles (organelles), and the like.
　The cells include, for example, animal cells (hematocytes etc.) and plant cells.
　Examples of the microorganism include bacteria such as Escherichia coli, viruses such as tobacco mosaic virus, fungi such as yeast and the like.
[0014]
　The biological microparticles also include biological macromolecules such as nucleic acids, proteins and complexes thereof.
　Further, the synthetic particles include particles made of, for example, an organic or inorganic polymer material or a metal. Examples of the organic polymer material include polystyrene, styrene/divinylbenzene, and polymethylmethacrylate. Examples of the inorganic polymer material include glass, silica, and magnetic material. Examples of the metal include gold colloid and aluminum.
[0015]
　The shape of the microparticles may be generally spherical or nearly spherical, or non-spherical. The size and mass of the fine particles can be appropriately selected by those skilled in the art depending on the size of the flow path of the microchip. On the other hand, the size of the flow path of the microchip can be appropriately selected depending on the size and mass of the fine particles.
[0016]
　In the present technology, the fine particles can be labeled with a chemical or biological label, such as a fluorescent dye, if necessary. The label may make the detection of the microparticles easier. The label can be appropriately selected by those skilled in the art.
[0017]
<1. Basic Structure of Microchip for Separation of Fine Particles>
(1)
　Basic Structure of Microchip for Separation of Microparticles FIG. 1 shows an example of a basic structure of a microchip for separating microparticles.
　In the microchip of FIG. 1, the fine particle-containing liquid is introduced from the fine particle-containing liquid introducing section 101. Further, a sheath liquid introducing section 103 is provided, and the sheath liquid is introduced from here.
[0018]
　In the sheath liquid, in the sheath flow generation unit 112, the fine particle-containing liquid and the fine particle-containing liquid merge (for example, merge with the fine particle-containing liquid from both sides of the fine particle-containing liquid), and the sheath liquid surrounds the fine particle-containing liquid. The enclosed laminar flow or the particulate-containing liquid forms a laminar flow in which the sheath liquid is sandwiched, that is, a so-called sheath flow. The laminar flow flows toward the detection unit 105.
[0019]
　In the detection unit 105, for example, the fine particles in the fine particle-containing liquid are irradiated with light to detect the fine particles. Based on the fluorescence and/or the scattered light generated by the irradiation of the light, it is determined whether the fine particles should be acquired. The detection unit 105 may be one spot or a plurality of spots. Further, according to the present technology, it is possible to irradiate one spot with a plurality of laser lights.
[0020]
　The capturing unit 113 is branched into a plurality of two such as two and three, and in the case where the capturing unit 113 is branched into three, for example, the branch channel in the middle is the capturing channel and leads to the capturing chamber. Further, when branched into two, these may be a waste flow channel and a capture flow channel. It is desirable that the capture channel is located at a position where the fine particle-containing liquid in the sheath flow reaches, and has a sectional shape in which all the fine particle-containing liquid enters.
　The fine particles determined to be acquired by the detection unit 105 flow to the three branched portions of the capture unit 113, and when reaching, flow into the capture chamber for containing the target fine particles. In the present technology, the capture chamber is not particularly limited as long as it can accommodate the target fine particles, but is preferably a pressure chamber. In this specification, the case where the capture chamber is a pressure chamber is assumed and described. In this case, the pressure chamber 114 is controlled to have a negative pressure, and the fine particles are drawn into the pressure chamber 114 and flow into the acquisition channel 111.
　The upper and lower branch flow channels are connected to the waste liquid flow channel 110, and the liquid flowing in the waste liquid flow channel 110 can be discharged to the outside of the microchip.
[0021]
　It should be noted that in the capture channel, a channel referred to as a “gate channel” in the present specification is provided so as to be connected to one or more capture channels or to intersect, for example, vertically. 27 and 28 are diagrams showing specific examples of the gate flow channel connection symmetrical with respect to the flow channel center. In these figures, the gate flow channel is symmetrical with respect to the flow channel center in the orifice of the second stage. An example of arrangement is shown.
　FIG. 27 shows an embodiment in which a gate flow path is formed in a direction perpendicular to the main flow path surface (chip plane) and symmetrically connected to the second stage of the orifice. In this case, the gate flow introduction channel can be formed on the surface opposite to the surface on which the main channel is formed, and layers (not shown) to be the lid of the channel can be provided on the upper and lower surfaces of the chip. Further, in this case, a sufficient space around the orifice can be secured, the thickness of the flow channel wall can be maintained because the flow channels are not adjacent, and the bonding area of ​​the chip bonding surface can be reduced. It can be increased, which is advantageous in terms of mechanical strength.
　FIG. 28 shows an embodiment in which the gate flow path is formed in the same plane as the main flow path surface and symmetrically connected to the second stage of the orifice. In this case, since the introduction flow path of the gate flow is formed on the same surface as the surface on which the main flow path is formed, it is possible to manufacture with a two-layer structure.
　The liquid that flows in the gate channel serves as the main solvent for the fine particles that are recovered after fractionation, so various liquids can be selected according to the application. For example, a liquid medium used for the fine particle-containing liquid, a sheath liquid, or a buffer liquid in which a pH containing a surfactant is adjusted when the fine particles are proteins can be flowed at a constant flow rate according to the fine particles.
　In particular, when the fine particles are cells, a cell culture solution, a cell preservation solution, etc. can be used. When a cell culture medium is used, it is suitable for performing the next step of applying to the cells recovered after fractionation, for example, the step of cell culture, cell activation, gene transfer and the like. When using a cell preservation solution, it is suitable for storing and transporting the recovered cells. Further, when the cells to be separated and collected are undifferentiated cells such as iPS cells, a differentiation inducing liquid can be used, and the next work can be efficiently advanced.
　A solution having a blocking effect can also be used as the liquid that flows in the gate channel. This makes it possible to suppress non-specific adsorption of the collected cells to the collection container or bag. Examples of the blocking agent include a solution containing a protein such as albumin, a solution containing an amino acid such as glycine, and a solution containing a nonionic surfactant such as Pluronic F68.
　Furthermore, a solution having a cell lysing action can be used as the liquid to be flown in the gate channel. This makes it possible to extract the intracellular substance as it is after collecting the target cell population. Examples of the cell lysate include a solution containing a surfactant.
　As the sheath liquid, various liquids can be similarly selected. In the present specification, the flow formed by the liquid (liquid for gate flow) flowing in the gate flow path is referred to as “gate flow”.
[0022]
　The upstream side of the gate flow channel can be introduced independently from a gate flow channel introduction unit (not shown) and flowed at an appropriate flow rate. In the present technology, the flow rate of the liquid introduced into the gate channel is smaller than the flow rate of the liquid introduced into the sheath channel. It is economical to use.
　Further, the gate flow can be generated by branching from the sheath liquid flow.
　The sheath flow passage after the sheath liquid introducing portion 103 of FIG. 1 is connected to the upstream end of the gate flow passage 10 of FIG. 2 or 3 so that the sheath liquid flow is branched and also flows into the gate flow passage. It can also be a gate style. In that case, it is necessary to appropriately design the flow path resistance of the gate flow path so that the gate flow rate becomes an appropriate flow rate.
[0023]
　At the intersection of the gate flow passage and the capture flow passage, a gate flow that tries to go straight through the gate flow passage and a gate flow that goes toward the detection unit side and the pressure chamber side are generated. The gate flow toward the detection unit side and the pressure chamber can prevent fine particles that should not be acquired from entering the pressure chamber side of the capture channel. The gate flow that has flowed through the gate flow path flows out into the capture flow path, and is branched into a gate flow toward the detection section side and the pressure chamber side of the capture flow path. The gate flow toward the detection section side of the capture channel can prevent fine particles that should not be acquired from entering the pressure chamber side of the capture channel.
[0024]
(2) Movement of Fine Particles in Capture Portion
　FIG. 2 shows an enlarged view of the capture portion 113 portion in FIG.
　In the capture part, the upper and lower branch flow paths and the narrow central flow path connected to the pressure chamber are branched into three. The circle in the central channel is the connection with the gate channel.
　The gate channel 10 intersects the central channel vertically. A constant flow rate of the gate diversion liquid always flows through the gate flow path 10.
　The gate diversion liquid forms a branched flow (arrows 22 and 23) at the intersection of the gate flow channel 10 and the capture flow channel. The flow of arrow 22 prevents the retention of particulates at the entrance of the capture channel. The flow indicated by the arrow 23 transfers the particles captured in the pressure chamber to the acquisition channel 111 of FIG.
[0025]
(3) Pressure Control of Pressure Chamber
　FIG. 3 schematically shows a vertical cross section of the microchip for collecting fine particles.
　The detection channel 301 is coaxially connected to the capture channel 303. One or more gate flow paths 10 are connected to the capture flow path 303 in, for example, a T-shape or intersect in a cross shape.
　Furthermore, the capture channel 303 is coaxially connected to the pressure chamber 114. The pressure chamber 114 includes a vibrating plate 305, and a piezo element 306 is installed on the vibrating plate 305 via a spacer. The deformation of the piezo element 306 causes the vibration plate 305 to vibrate, and the pressure chamber 114 is repeatedly controlled from normal pressure to negative pressure, or from negative pressure or normal pressure to positive pressure.
　Due to the negative pressure and the positive pressure or the normal pressure of the pressure chamber 114, a jet is generated in the flow from the capture channel 303 to the pressure chamber 114.
[0026]
　FIG. 4 shows the movement of fine particles due to the jet generated in the pressure chamber 114.
　The lower part of FIG. 4 shows the drive waveform of the piezo element, and the upper part shows the movement of the fine particles corresponding to the deformation of the piezo element.
　The particles (left in FIG. 4) that have arrived before the capture channel immediately before the deformation of the piezo element are drawn into the pressure chamber when the piezo element is deformed and the pressure chamber becomes negative pressure (center of FIG. 4 ). Then, when the piezo element is deformed and the pressure in the pressure chamber returns, the pulling force is released and the force for pushing out the fine particles works (right in FIG. 4).
[0027]
(4) Re-emission of fine particles
　FIG. 5 shows an enlarged view of the capture unit 113 and the pressure chamber 114 in FIG.
　As shown in the upper left of FIG. 5, in the flow in the pressure chamber, a main flow (dotted line) and a swirling flow induced by the jet at the time of capturing the particles (solid line) occur. The captured fine particles may ride on the swirling flow and swirl in the pressure chamber. The upper part of the vertical triad in the lower left of FIG. 5 shows the fine particles on the swirling flow.
[0028]
　If the particles carried on the swirl flow happen to be in the vicinity of the capture flow path during the subsequent discharge operation for acquiring the particles (FIG. 5, the middle row of the vertical three-row diagram in the lower left), the particles are discharged outside the pressure chamber (vertical direction). (Bottom row of triplet diagram). Therefore, the acquisition rate of fine particles decreases.
　In particular, at a high event rate, that is, when there are more trapped particles than particles that can be carried by advection (the vertical three-row diagram in FIG. 5), the swirling flow also becomes a steady flow, and the particle density in the pressure chamber becomes uniform and high. The probability of re-release to the outside of the pressure chamber increases.
　Therefore, a study was conducted to further improve the fine particle capturing performance.
[0029]
(5) Improvement of
　fine particle capture performance In order to improve fine particle capture performance, it is conceivable to shorten the length of the capture flow path and reduce the flow path resistance to speed up the fractionation. Be done. In addition to shortening the length of the capture channel, it is conceivable that the diameter of the capture channel is reduced to reduce the volume drawn into the pressure chamber, thereby achieving higher purification and higher acquisition rate of capture. Furthermore, by reducing the volume drawn in the direction of the pressure chamber, the concentration of cells diluted from the sheath fluid can be recovered (concentrated) to some extent.
[0030]
　FIG. 6 shows an enlarged view of the portion where the gate channel is connected in a trapezoidal shape to the capture channel.
　In order to speed up the capturing operation, it is necessary to reduce the channel resistance of the capturing channel. Therefore, it is desired to shorten the capture channel length 603, but there is a limit to shortening because of the structure in which the gate channel is connected to the capture channel as shown in FIG.
[0031]
　Further, as shown by the streamline on the right side of FIG. 6, when the gate flow reaches the capture flow channel, it is divided into the detection flow channel side and the pressure chamber side, and flow separation occurs. That is, the flow is separated at the connection portion and branched into the detection flow channel side and the pressure chamber side. As a result, the flow in the vicinity of the gate flow path connecting portion of the capture flow path tends to generate a flow closer to one side of the flow path.
[0032]
　Considering the above, in order to exert the particle retention effect at the capture channel inlet, the capture channel length 603 needs to be twice or more the capture channel diameter 602 from the connecting portion with the gate channel 10. It is thought that there is (604). Considering the mechanical strength, it is considered that the capture channel length 603 needs to be 2 to 3 times or more the capture channel diameter 602.
[0033]
　In the example of FIG. 6, assuming that the capture channel diameter 602 is 60 μm, the length of the capture channel inlet side 604 is 120 μm, the gate channel diameter is 60 μm, and the capture channel outlet side 605 is 50 μm. It is considered that the capture channel length 603 needs to be about 230 μm.
[0034]
　Next, considering the pulling-in operation at the time of capturing fine particles, in the pulling-in operation at the time of capturing fine particles, it is necessary to pull in a volume such that the liquid flow containing the fine particles containing the fine particles to be acquired passes through the capture flow path.
　FIG. 7 schematically shows the relationship between the length of the capture channel, the volume of the liquid flow containing fine particles, and the fine particles.
　When the fine particles to be acquired and the unnecessary particles are close to each other, as shown in the upper part of FIG. 7, if the capture channel is long, the volume drawn into the capture channel becomes large. Are drawn together into the capture channel.
　On the other hand, as shown in the lower part of FIG. 7, if the capture channel is short, the volume drawn into the capture channel becomes small, so that the fine particles to be acquired and unnecessary particles are not drawn into the capture channel together. Absent. Therefore, it is possible to distinguish the fine particles to be acquired from the unnecessary fine particles. As a result, the purity of the fine particles to be acquired can be increased and the acquisition rate can be increased.
[0035]
(6) Relationship between shape of trap channel and drawing volume
　FIG. 8 schematically shows the drawing operation and the pushing operation of the fine particle-containing liquid in the catch channel.
　The gate channel 10 intersects with the capture channel. For example, the capture channel diameter can be 60 μm, and the total capture channel length can be 250 μm. Assuming that the volume of the capture channel (the part surrounded by the white line) is 0.9 nl (0 μsec), the fine particle-containing liquid of the order of 0.9 nl, which is close to the volume of the capture channel, is drawn from the capture channel to the pressure chamber side (50 μsec). .. Next, the fine particle-containing liquid is discharged in an amount close to the volume of the capture channel (100 μsec, leftward direction of the arrow), but when discharged, the fine particle-containing liquid in the pressure chamber also flows into the capture channel.
[0036]
　FIG. 9 schematically shows the drawing operation and the pushing operation of the fine particle-containing liquid in the capture channel when the gate flow is constantly flowing at a constant flow rate.
　As in the above, the fine particle-containing liquid is drawn from the capture channel to the pressure chamber side and extruded.
　The gate flow is flowing in during the pulling and pushing operations. For example, assuming that the drawing operation to the pressure chamber side and the pushing operation from the pressure chamber side are performed for 100 μsec and the gate flow rate to the pressure chamber side is 120 μl/min (left in FIG. 9), part of the gate flow is divided. Starts to flow into the capture channels on both sides (center of FIG. 9), and after 100 μsec, 0.2 nl also flows into the pressure chamber side (on the right in FIG. 9, the part surrounded by the white line).
　Since the order of 0.9 nl and 0.2 nl is close to each other, it is assumed that the drawing and extruding volume or the trap channel volume is reduced and the gate flow volume that flows in during the particulate capturing operation is from the trap channel to the pressure chamber side. The idea was that if a structure that discharges into the chamber could be created, the volume of extrusion from the pressure chamber could be reduced and re-emission could be suppressed.
[0037]
<2. Embodiment 1> In
　Embodiment 1, the acquired fine particles that have been once captured to the pressure chamber side are not released again. Therefore, as an example, a structure in which the capture channel is formed in two stages is adopted.
　That is, the microchip for sorting microparticles of the present technology includes
　a main
　flow path through which a liquid containing fine particles flows, a capture flow path that communicates with the main flow path coaxially, and a capture flow path that communicates coaxially with the
　capture flow path. a pressure chamber which
comprises,
　the capture channel includes a capture passage of the first and second stages of the trap passage,
　the capture flow path of the first stage includes a first ejection portion,
　said The capture passage of the second
　stage includes a second discharge part, the capture passage of the
　first stage has an opening that functions as an orifice at the rear end, and the opening area of ​​the capture passage of the first stage is The same or smaller than the minimum cross-sectional area of ​​the second-stage capture channel, the
　second-stage capture channel is connected to one or more gate channels at its front end, for example, in the shape of a letter, or in a cross shape. So that the gate flow paths intersect each other.
　The opening at the rear end of the first-stage capture channel is connected at the position of the gate channel that is connected or intersects at the front end of the second-stage capture channel.
[0038]
　FIG. 10 shows a capture channel having two stages.
　The first stage of the capture channel includes the first discharge part 1 having a channel diameter of 30 μm and a length of 50 μm. At the rear end of the first stage of the capture channel, there is an opening 7 that functions as an orifice (upper stage of FIG. 10).
　In the first stage of the capture channel, the particulate-containing liquid stream forms a jet when drawn. In the first stage, the flow path length is shortened and/or the diameter is reduced to reduce the volume ((1) in the lower stage of FIG. 10).
[0039]
　The second stage of the capture channel has a larger diameter (channel diameter 60 μm) than the first stage channel diameter to reduce the resistance. Further, a gate flow is connected to the second stage (gate flow path diameter 60 μm). The volume of the second stage is set to a size that can hold the inflowing gate flow volume or a size that can hold the inflowing and extruding volume without separating the inflowing gate flow ((2) in the lower stage of FIG. 10).
[0040]
　The structure of FIG. 10 is shown three-dimensionally in FIG.
　A gate flow is formed in the gate channel 10 arranged so as to intersect between the first-stage capture channel 1 and the second-stage capture channel 2, and an opening of the first-stage capture channel 1 is formed. A jet is generated from 7 toward the gate flow. The gate flow of the jet flows to the capture channel of the second stage having a cross-sectional area larger than that of the opening of the first stage and heads toward the pressure chamber side. On the other hand, a gate flow that branches into the detection unit side and the pressure chamber side also occurs. The gate flow branched to the detection unit side can prevent particles that should not be acquired from entering the pressure chamber side.
[0041]
　FIG. 12 shows a particle capturing operation in the capture channel in this embodiment.
　In the upper left diagram, the gate flow is always introduced, for example, 120 μl/min from the up and down arrows. At this time (0 μsec), the pressure in the pressure chamber is normal (normal pressure). When the particles to be acquired (white circles in FIG. 12) come to the entrance of the capture channel, the negative pressure control of the pressure chamber is started.
[0042]
　In the figure at the center of the upper stage (25 μsec), the pressure chamber becomes negative pressure, and the flow at the rear end of the first stage of the capture channel separates to generate a jet.
[0043]
　In the upper right diagram (50 μsec), the pressure chamber returns from negative pressure to normal pressure.
　Next, in the lower left diagram (75 μsec), even if the pressure chamber has a positive pressure and the extrusion operation is started, the fine particles on the jet are not immediately discharged. In addition, the gate flow continues to flow in during the extrusion operation, and the liquid flow containing fine particles is replaced with the gate flow from the vicinity of the rear end of the first stage of the capture channel.
[0044]
　In the lower center diagram (100 μsec), the pressure chamber returns from positive pressure to normal pressure.
　The jet wake and the gate flow are discharged in the pushing operation, and the discharge from the pressure chamber is small. That is, re-emission of fine particles is suppressed.
[0045]
　In the lower right diagram (125 μsec), even if fine particles (black circles in FIG. 12) in the pressure chamber enter the trapping channel of the second stage during the pushing operation, the pressure chamber is in a gate flow until the next trapping operation. Pushed back to.
[0046]
　FIG. 13 schematically shows a capture channel including the first and second stage capture channels.
　The capture channel of the first stage has a circular cross section. Then, the capture channel of the first stage, that is, the rear end of the first discharge part 1 here is connected to the gate channel 10, and a constant flow rate of the gate diversion liquid flows through the gate channel 10.
　The pressure inside the pressure chamber is repeatedly controlled by a piezo element from negative pressure to positive pressure.
　When the pressure in the pressure chamber is negative, the fine particle-containing liquid flow containing the fine particles to be acquired is drawn into the pressure chamber side from the first-stage capture channel.
[0047]
　To simply accelerate and decelerate the first discharge part 1 in a sine wave shape with a circular cross section, in order for the fine particles at the center of the front end of the first discharge part 1 to reach from the front end to the rear end (the circle (fine particles in FIG. 13) It is possible to approximate that it is necessary to draw in a volume which is half (1/2) of the volume of the first discharge part 1. Therefore, it is preferable that the flow of the fine particle-containing liquid flow from the first discharge part 1 be at least half the volume of the first discharge part 1 (the first capture channel).
[0048]
　FIG. 14 schematically shows the relationship between the volume drawn in from the first discharge part 1 and the inflow volume of the gate flow.
　When the pressure in the pressure chamber is set to a negative pressure, the fine particle-containing liquid flow is drawn from the first discharge part 1. Then, the gate flow flows into the first discharge unit 1 while the pressure control in the pressure chamber is performed from the negative pressure to the positive pressure or the normal pressure.
　The inflow volume of the gate flow at this time is preferably the same as or larger than the inflow volume of the fine particle-containing liquid flow.
[0049]
　This is because, if the gate flow inflow volume during the trapping operation is larger than the fine particle-containing liquid flow intake volume, almost the same volume as the intake area near the rear end of the first discharge part 1 is discharged, and therefore the volume discharged from the pressure chamber. Because it can be reduced.
[0050]
　FIG. 15 schematically shows the relationship between the discharge volume into the second capture channel 2 and the inflow volume of the gate flow.
　The rear end of the first discharge part 1 of the first-stage capture channel has an opening that functions as an orifice, and the gate channel 10 is arranged on the pressure chamber side thereof. A constant amount of the liquid for gate flow always flows through the gate channel to form a gate flow. A part of the gate flow is branched from the second capture channel into the detection channel side direction and the pressure chamber side direction (second discharge portion direction).
[0051]
　When the fine particles first reach the front end of the first-stage capture channel, the pressure inside the pressure chamber is set to a negative pressure, and the fine-particle-containing liquid flow is drawn into the first-stage capture channel. A jet is generated from the rear end (opening) of the first capture channel, and the particles on the jet pass through the second capture channel and reach the pressure chamber. Turn to. Then, the fine particle-containing liquid flow and the gate flow that have been once drawn into the pressure chamber side are pushed out toward the first capture channel side.
[0052]
　At this time, the volume (dotted line portion) of the second-stage capture channel in FIG. 15 is the same as the volume pushed out in the direction of the capture channel 1 when the pressure in the pressure chamber is positive pressure or normal pressure. It is preferably a volume or a volume larger than that.
　If the volume of the second-stage capture channel is smaller than the volume extruded in the extruding operation as shown in FIG. 15, even if a large amount of the gate flow is introduced, it cannot be held and is scattered into the pressure chamber. This is because the liquid in the pressure chamber is extruded during the extruding operation.
　Next, the pressure in the pressure chamber is set to a negative pressure so that the liquid is pushed out of the second-stage capture channel into the pressure chamber, and the volume of the liquid is equal to or less than the volume of the second-stage capture channel. Is preferred.
[0053]
<3. Embodiment 2>
　The microchip for sorting microparticles of the present technology has a plurality of gate channels, and captures the second stage so as to be symmetric with respect to the center of the flow of the liquid containing fine particles from the opening that functions as an orifice. It is preferable to connect to the front end of the flow channel.
　If the gate flow path is connected asymmetrically with respect to the center of the capture flow path, the flow in the capture flow path will also become asymmetric, an unintended secondary flow will occur, and the particles will enter the capture flow path from the pressure chamber. Because there are things.
[0054]
　FIG. 16 shows an example in which the gate channel is asymmetrically connected to the capture channel.
　If the gate flow channel is connected to the capture flow channel on only one side, flow separation occurs at the connection when the gate flow rate is high. Then, it sticks to the opposite surface of the connection part that has flowed in the capture channel of the second stage, and an unintended secondary flow occurs.
[0055]
　Therefore, the gate channel is arranged symmetrically with respect to the center of the capture channel, that is, the center of the flow of the fine particle-containing liquid from the opening that functions as an orifice.
　By arranging the gate flow paths symmetrically, the flow rate from each gate flow path can be reduced, the flow velocity can be reduced at the portion where the flow channels are connected, and flow separation can be reduced.
　Further, since the flow in the capture channel is also symmetrical, an unintended secondary flow is unlikely to occur.
　The number of gate flow paths can be one as shown in FIG. 1 or two or more as shown in FIGS. 27 and 28, but it is preferable that it is two or more (plural). Further, the gate flow paths may be not only even numbers such as two and four, but also odd numbers such as one, three and five.
[0056]
　FIG. 17 shows an example in which the gate channels are symmetrically connected to the capture channel.
　If, for example, two gate channels are provided from the top and the bottom, and a flow rate of ½ is introduced, separation of the flow at the connection with the capture channel becomes small and symmetrical, and an unintended secondary flow Is less likely to occur.
[0057]
<4. Embodiment 3>
　The shape of the cross section of the capture channel is, for example, a square, a rectangle, a circle, or an ellipse, and is not particularly limited. Further, the capture channel may be a parallel channel as well as a tapered channel.
　FIG. 18 shows an example in which the taper starts immediately after the gate flow channel connecting part, and FIG. 19 shows an example in which the taper starts from the front end of the second capture channel.
　In FIG. 18, when the second discharge part 2 of the second capture channel is a tapered flow channel, the volume of the second capture channel near the first capture channel decreases, and the pressure chamber side It is possible to efficiently store the gate flow that flows out during the discharge operation from, and reduce the resistance to the jet during the drawing operation. The volume of the second capture channel near the first capture channel is reduced, and the resistance to the jet during the drawing operation can be reduced.
[0058]
<5. Embodiment 4>
　Alternatively, the capture channel in the first stage may be a tapered channel.
　FIG. 20 shows an example in which the first-stage capture channel is a tapered channel.
　If such a structure is adopted, the jet resistance can be efficiently generated at the rear end without increasing the flow passage resistance of the capture passage of the first stage.
[0059]
<6. Fifth Embodiment>
　Furthermore, the first capture channel and the second capture channel may be tapered channels.
　FIG. 21 shows an example in which the first-stage capture channel and the second-stage capture channel are tapered channels.
Also in this case, the minimum cross-sectional area of ​​the capture channel 2 of the second stage is made larger than the minimum cross-sectional area of ​​the capture channel 1 of the first stage.
　If such a structure is adopted, the force of the jet from the opening at the rear end of the first-stage capture channel can be increased, and the volume near the first-stage capture channel in the second-stage capture channel can be increased. And the resistance to the jet during the retracting operation can be reduced. Further, it is possible to efficiently generate the jet at the rear end without increasing the flow resistance of the capture flow path of the first stage.
[0060]
<7. Embodiment 6> The number of
　capture channels is not limited to two and may be three or more.
　In FIG. 22, the first capture channel (including the first discharge part 1), the second capture channel (including the second discharge part 2), and the third capture channel (third discharge part). (Including 3).
　The rear ends of the first, second, and third capture channels may each have an opening that functions as an orifice. Each opening area is the same or smaller than the minimum cross-sectional area of ​​the capture channel immediately after.
　The gate channel 10 may intersect the front end of the second-stage capture channel and the front end of the third-stage capture channel.
[0061]
<8. Embodiment 7> When the number of
　capture channels is three, there may be a mode in which the gate channel 10 intersects with the front end of the capture channel of the second stage and the gate channel is not provided in the capture channel of the third stage. ..
　FIG. 23 shows an example in which the gate channel intersects with the front end of the capture channel of the second stage and the gate channel does not exist in the capture channel of the third stage.
[0062]
<9. Embodiment 8> In
　the case where the number of capture channels is three, there may be a mode in which the gate channel 10 intersects with the front end of the capture channel in the third stage.
　FIG. 24 shows an example in which there is no gate flow path at the front end of the second-stage capture flow path, and the gate flow paths are connected to intersect at the front end of the third-stage capture flow path. At this time, the cross-sectional area of ​​the second-stage capture channel and the cross-sectional area of ​​the third-stage capture channel may be the same.
[0063]
<10. Embodiment 9> In
　FIG. 25, there are three capture channels, the second and third capture channels are tapered, and the gate channel is provided at the front end of the second capture channel. An example is shown in which 10 intersects with each other and there is no gate channel in the third capture channel.
[0064]
　The shape of the capture channel in the present technology is not limited to the example of the above-mentioned embodiment, and various shapes of the capture channel can be adopted as long as the effects of the present invention are not lost.
[0065]
<11. Embodiment 10>
　The microchip for sorting microparticles according to an embodiment of the present technology may be provided with a connection channel between the capture channel and the pressure chamber.
　FIG. 26 shows an example in which a connection channel is provided.
　The cross-sectional area of ​​the connection channel may be larger than the cross-sectional area of ​​the capture channel. A particle detection region or the like can be arranged in the connection channel.
[0066]
　When the connection channel is not provided, for example, when a piezo element is connected to the pressure chamber, even if it is desired to install components other than the piezo element, the space for that is not sufficient. However, by adopting the above structure, a wider space can be secured around the pressure chamber, and the degree of freedom in design increases.
[0067]
<12. Eleventh Embodiment>
　The microchip for collecting fine particles of the present technology may include a fine particle-containing liquid introducing section, a sheath liquid introducing section, a fine particle collecting section, and the like, and a structure in which the respective sections are appropriately connected can also be adopted.
　As described above with reference to FIG. 1, the fine particle-containing liquid is introduced from the fine particle-containing liquid introducing section 101. The sheath liquid is introduced from the sheath liquid introducing unit 103. The particle-containing liquid and the sheath liquid form a laminar flow in the sheath flow generation unit 112 and flow in the main flow path.
　Immediately before the capture channel through which the main channel communicates, the main channel branches into a coaxial capture channel and a plurality of branch channels. The gate channel intersects the capture channel.
[0068]
　The upstream side of the gate flow channel can be introduced independently from a gate flow channel introduction unit (not shown) and flowed at an appropriate flow rate. Further, the gate flow can be generated by branching from the sheath liquid flow. As shown in FIG. 1, it is also possible to connect the sheath flow path after the sheath liquid introduction part and the upstream end of the gate flow path so that the sheath liquid flow is branched and also flows into the gate flow path to form a gate flow. In that case, it is necessary to properly design the flow path resistance of the gate flow path so that the gate flow rate becomes an appropriate flow rate.
[0069]
　In addition, the connection channel can be connected to the sheath liquid container and the gate liquid container.If necessary, a pump can be installed to control the flow of the sheath liquid, or a filter can be installed to pass the sheath liquid through the filter. May be. With such a structure, the sheath liquid can be reused.
[0070]
　Further, a fine particle recovery unit for recovering the captured fine particles may be provided downstream of the pressure chamber. The collected fine particles are used for measurement and analysis, and when the fine particles are cells, they are also used for proliferation and the like.
[0071]
<12. Fine Particle Sorting Apparatus>
(1) Configuration
　The fine particle sorting apparatus of the present technology irradiates light on the
microchip mounting portion on which the above-described fine particle sorting microchip can be mounted and the
fine particle detecting area of ​​the fine particle sorting microchip. The light irradiation unit includes a light irradiation unit,
a detection unit that detects scattered light and/or fluorescence emitted from the fine particles by light irradiation ,
and a pressure chamber control unit that sets the pressure chamber to a negative pressure or a positive pressure
.
[0072]
　In the microchip mounting portion of the particle sorting apparatus, the particle sorting microchip may be easily attached and detached. In this case, the microchip for collecting fine particles may be disposable, or may be one that can be reused after washing.
[0073]
　The light irradiation unit of the particle sorting device irradiates the particles flowing through the main flow path or the communication passage with light such as excitation light. The light irradiator may include a light source that emits light and an objective lens that focuses the light on the fine particles that flow through the main channel or the communication passage.
　The light source can be appropriately selected from a laser diode, an SHG laser, a solid-state laser, a gas laser, a high-intensity LED, and the like, depending on the purpose of determining the fine particles to be acquired.
　In addition to the light source and the objective lens, the light irradiation section may include other optical elements as needed.
[0074]
　The detection unit of the particle sorting apparatus can detect scattered light and/or fluorescence emitted from the particles due to the irradiation of light by the light irradiation unit. The detection unit may include a condenser lens that collects fluorescence and/or scattered light generated from the particles and a detector.
　Specific detectors include, but are not limited to, PMTs, photodiodes, CCDs, CMOSs, and the like.
　In addition to the condenser lens and the detector, the detection unit may include other optical elements as needed.
[0075]
　The fluorescent light is, for example, fluorescent light generated from the fine particles themselves, and fluorescent light generated from a substance labeled on the fine particles, such as a fluorescent substance, but is not limited thereto.
　The scattered light is, for example, geometrical optical scattering, Rayleigh scattering, and/or Mie scattering, and is generally detected as forward scattered light or side scattered light, but is not limited thereto.
[0076]
　The pressure chamber control unit of the particle sorting apparatus, based on the data detected by the detection unit, advances the particles flowing through the main channel or the communication channel of the particle sorting microchip to the branch channel or the particle sorting flow. Control whether to inhale in the street.
[0077]
　The fluorescence and scattered light detected by the detector can be converted into an electric signal.
　In this case, the particulate collection device is provided with an electric signal converter. The electric signal conversion unit may be included in the pressure chamber control unit or may not be included in the pressure chamber control unit.
　The pressure chamber control unit receives the electric signal and can determine the optical characteristics of the particles based on the electric signal.
[0078]
　Based on the determination result of the optical characteristics of the microparticles, the pressure chamber control unit, if the microparticles are to be acquired, flows through the flow path so that the microparticles pass through the opening functioning as an orifice to the capture flow path. Control the pressure in the pressure chamber to change. In order to draw the fine particles into the capture channel, the pressure inside the pressure chamber may be made negative.
　The flow in the flow passage can be changed by, for example, repeatedly controlling the pressure chamber from negative pressure to positive pressure or normal pressure, or from positive pressure or normal pressure to negative pressure. When the pressure chamber is set to a positive pressure or a normal pressure, the flow in the flow path can be changed again. Such specific pressure control of the pressure chamber can be performed by deforming the piezo element using an actuator, for example, a piezo element. The pressure chamber control unit can have the same configuration as that of the drive unit described in JP-A-2014-036604, for example.
[0079]
(2) Fine Particle Sorting Program
　The pressure chamber control unit of the fine particle sorting apparatus can store a fine particle sorting program for executing the above operation. Alternatively, it is possible to connect the pressure chamber control unit to a computer equipped with a particle sorting program.
[0080]
　The program is stored/held in the hard disk, read into the memory under the control of the CPU and the OS, and executes the above-mentioned capture operation.
　The program can be recorded in a computer-readable recording medium. The recording medium is not particularly limited as long as it is a computer-readable recording medium, but specifically, for example, a disk-shaped recording medium such as a flexible disk or a CD-ROM is used. Alternatively, a tape type recording medium such as a magnetic tape may be used. Further, a part of the processing is configured by hardware such as DSP (Digital Signal Processor), ASIC (Application Specific Integrated Circuit), PLD (Programming Logic Device), and FPGA (Field-Programmable Gate Array), and the above software program and A configuration in which high-speed processing is performed in cooperation with each other can also be adopted.
[0081]
　Note that the present technology can also take the following configurations.
[1] a main channel that fine particle-containing liquid is flowing,
　said a main channel and a capture passage communicates coaxially,
　a pressure chamber in communication with the capture passage coaxially
include,
　the capture The flow channel includes an upper capture channel and a lower capture channel, the
　upper capture channel has an opening functioning as an orifice at a rear end thereof, and
　an opening area of ​​a rear end of the upper capture channel. Is equal to or
smaller than the minimum cross-sectional area of ​​the
　lower capture channel, the lower capture channel is provided with one or more connecting gate channels or intersecting gate channels at the front end thereof, and the
　upper capture channel is
A microchip for sorting microparticles , wherein the opening at the rear end of the flow channel and the gate flow channel at the front end of the lower capture channel are connected .
[2] The plurality of gate channels are connected to the front end of the lower capture channel so as to be symmetrical with respect to the center of the flow of the liquid containing fine particles from the opening at the rear end of the upper capture channel. The microchip for collecting fine particles according to [1],
[3] The microchip for fractionation of fine particles according to [1] or [2], in which the gate flow liquid always flows at a constant flow rate through the gate flow path.
[4] The fine particle component according to claim 3, wherein the gate flow liquid branches and flows in the lower capture channel from the gate flow channel in the direction of the upper capture channel and in the direction of the pressure chamber. Microchip for taking.
[5] The microchip for fractionating fine particles according to any one of [1] to [4], wherein the pressure chamber has a vibrating plate.
[6] The microchip for fractionating fine particles according to any one of [1] to [5], wherein the pressure chamber is repeatedly controlled to a negative pressure, a positive pressure, and a normal pressure.
[7] The microchip for sorting fine particles according to any one of [1] to [6], wherein the main flow path includes a fine particle detection region.
[8] The microchip for fractionating fine particles according to any one of [1] to [7], including a connection channel between the capture channel and the pressure chamber.
[9] The microchip for fractionation of fine particles according to any one of [1] to [8], wherein the main flow path is connected to a fine particle-containing liquid introducing section for introducing the fine particle-containing liquid.
[10] The microchip for collecting fine particles according to any one of [1] to [9], wherein the main flow path is connected to a sheath liquid introducing section for introducing a sheath liquid.
[11] The main flow path is branched upstream of the capture flow path, and is connected to a branch flow path in which a liquid that did not flow into the capture flow path flows, and is connected to any one of [1] to [10]. Microchip for collecting fine particles.
[12] The microchip for collecting fine particles according to [11], wherein the branched flow path is connected to a sheath liquid container and/or a gate liquid container.
[13] The microchip for collecting fine particles according to any one of [1] to [12], further including a fine particle collecting unit for collecting the captured fine particles downstream of the pressure chamber.
[14] When the fine particles reach the front end of the upper capture channel, the pressure chamber is changed from normal pressure to negative pressure, and the fine particle-containing liquid is drawn into the upper capture channel. Is discharged from the opening at the rear end of the capture channel to the lower capture channel to generate a jet flow, and the
　fine particles pass through the lower capture channel to reach the pressure chamber, and then the inside of the pressure chamber The
microchip for fractionating fine particles according to any one of [1] to [13], wherein the negative pressure or the normal pressure is changed to the positive pressure .
[15] The volume of the fine particle-containing liquid from the opening of the upper capture channel rear discharged to the capture passage of the second stage,
　at least half of the volume of the upper capture channel,
[1 ] A microchip for fractionating fine particles according to any one of [14].
[16] When the pressure of the pressure chamber is changed from normal pressure to negative pressure, the volume of the fine particle-containing liquid discharged from the opening at the rear end of the upper capture channel to the subsequent capture channel is the
　gate. The microchip for collecting fine particles according to any one of [1] to [15], which has a volume of a liquid for gate flow flowing from a channel to the capture channel of the latter stage or less.
[17] When the pressure in the pressure chamber is changed from normal pressure to negative pressure, the volume of the fine particle-containing liquid discharged from the lower capture channel toward the pressure chamber is smaller than that of the
　lower capture channel. Less than or equal to the volume,
The microchip for collecting fine particles according to any one of [1] to [16].
[18] a main channel that fine particle-containing liquid is flowing,
said a main channel and a capture passage communicates coaxially,
a pressure chamber in communication with the capture passage coaxially
include,
the capture The flow channel includes an upper capture channel and a lower capture channel, the
upper capture channel has an opening functioning as an orifice at a rear end thereof, and
an opening area of ​​a rear end of the upper capture channel. Is equal to or smaller than the minimum cross-sectional area of ​​the
lower capture channel, the lower capture channel is provided with one or more connecting gate channels or intersecting gate channels at its front end, and the
upper capture channel is
A microchip mounting portion for mounting a microchip for collecting fine particles , in which an opening at the rear end of the flow passage and the gate flow passage at the position at the front end of the lower capture passage are connected, and fine particles
　contained in the main flow passage A light irradiation unit that irradiates the detection region with light,
　a detection unit that detects scattered light and/or fluorescence emitted from the fine particles ,
　and a pressure chamber control unit that makes the inside of the pressure chamber negative pressure or positive pressure,
A fine particle sorting apparatus including:
[19] The fine particle sorting apparatus according to [18], wherein the pressure chamber control unit has a piezo element.
Explanation of symbols
[0082]
1 1st discharge part
2 2nd discharge part
3 3rd discharge part
7 Opening
10 Gate flow path
101 Fine particle containing liquid introduction part
103 Sheath liquid introduction part
105 Detection part
107 Branch part
110 Waste liquid flow path
111 Acquisition flow path
112 Sheath flow generation Part
113 Capture part
114, 213 Pressure chamber
211, 303 Capture channel
212 Connection channel
301 Detection channel
305 Vibration plate
306 Piezo element
602 Capture channel diameter
603 Capture channel length
claims
[Claim 1]
　A main channel
　through which the fine particle-containing liquid flows, a capture channel that communicates with the main channel coaxially,
　a capture chamber that communicates with the
　capture channel, and a gate channel that intersects the capture channel. When,
wherein the
　said trapping passage has an opening which intersects with the gate passage,
　the liquid is along a direction of flowing, cross-sectional area of the upstream of the opening downstream of the cross-sectional of the opening
A microchip for collecting fine particles characterized by having a smaller area .
[Claim 2]
　The plurality of gate channels are connected to the front end of the lower capture channel so as to be symmetric with respect to the center of the flow of the fine particle-containing liquid from the opening at the rear end of the upper capture channel. The microchip for collecting fine particles according to claim 1.
[Claim 3]
　The microchip for fractionation of fine particles according to claim 1, wherein the gate flow liquid always flows through the gate channel at a constant flow rate.
[Claim 4]
　The microparticle fractionation micro according to claim 3, wherein the gate flow liquid branches and flows in the lower capture channel from the gate flow channel in the direction of the upper capture channel and the direction of the pressure chamber. Chips.
[Claim 5]
　The microchip for sorting microparticles according to claim 1, wherein the capture chamber is a pressure chamber.
[Claim 6]
　The microchip for sorting fine particles according to claim 5, wherein the pressure chamber has a vibration plate.
[Claim 7]
　The microchip for sorting microparticles according to claim 5, wherein the pressure chamber is repeatedly controlled to have negative pressure, positive pressure, and normal pressure.
[Claim 8]
　The microchip for collecting fine particles according to claim 1, wherein the main flow path includes a fine particle detection region.
[Claim 9]
　The microchip for sorting microparticles according to claim 1, further comprising a connection channel between the capture channel and the capture chamber.
[Claim 10]
　The microchip for collecting microparticles according to claim 1, wherein the main channel is connected to a microparticle-containing liquid introducing section for introducing the microparticle-containing liquid.
[Claim 11]
　The microchip for sorting microparticles according to claim 1, wherein the main flow path is connected to a sheath liquid introducing section for introducing a sheath liquid.
[Claim 12]
　The microchip for microparticle fractionation according to claim 1, wherein the main flow channel branches upstream of the capture flow channel and is connected to a branch flow channel in which a liquid that has not flowed into the capture flow channel flows.
[Claim 13]
　The microchip for sorting microparticles according to claim 11, wherein the branch channel is connected to the sheath liquid container and/or the gate liquid container flow channel.
[Claim 14]
　The microchip for collecting microparticles according to claim 1, further comprising a microparticle recovery unit that recovers the captured microparticles, downstream of the capture chamber.
[Claim 15]
　When the fine particles reach the front end of the upper capture channel, the pressure chamber is changed from normal pressure to negative pressure, the fine particle-containing liquid is drawn into the upper capture channel, and the upper capture flow is obtained. After being discharged from the opening at the rear end of the passage to the lower capture channel, a jet flow is generated, and after the
　fine particles pass through the lower capture channel and reach the pressure chamber, a negative pressure in the pressure chamber or The
microchip for collecting fine particles according to claim 5, wherein the microchip is changed from normal pressure to positive pressure .
[Claim 16]
16. The fine particles according to claim 15 　,
　wherein the volume of the fine particle-containing liquid discharged from the opening at the rear end of the upper capture channel to the lower capture channel is at least half the volume of the upper capture channel.
Preparative microchip.
[Claim 17]
　When the pressure inside the pressure chamber is changed from normal pressure to negative pressure, the volume of the fine particle-containing liquid discharged from the opening at the rear end of the upper capture channel to the lower capture
　channel is from the gate channel. 16. The microchip for sorting microparticles according to claim 15, wherein the volume is equal to or less than the volume of the liquid for gate flow flowing into the lower capture channel.
[Claim 18]
　When the pressure of the pressure chamber is changed from normal pressure to negative pressure, the volume of the fine particle-containing liquid discharged from the lower capture channel in the direction of the pressure chamber is
　equal to or less than the volume of the lower capture channel. The
microchip for fractionation of fine particles according to claim 15, wherein.
[Claim 19]
　A main channel
　through which the fine particle-containing liquid flows, a capture channel that communicates with the main channel coaxially,
　a capture chamber that communicates with the
　capture channel, and a gate channel that intersects the capture channel. When,
wherein the
　said trapping passage has an opening which intersects with the gate passage,
　the liquid is along a direction of flowing, cross-sectional area of the upstream of the opening downstream of the cross-sectional of the opening
A microchip mounting portion for mounting a microparticle sorting microchip , characterized by being smaller than the
　area, a light irradiation portion for irradiating light to a fine particle detection region included in the main flow path, and a light
　emitted from the fine particles. A fine particle sorting apparatus comprising: a detection unit that detects scattered light and/or fluorescence; and the
　capture chamber is a pressure chamber, and a pressure chamber control unit that sets the pressure chamber to a negative pressure or a positive pressure
.
[Claim 20]
　The fine particle sorting apparatus according to claim 19, wherein the pressure chamber control unit has a piezo element.