WO2020036115A1 - Dispositif fluidique - Google Patents

Dispositif fluidique Download PDF

Info

Publication number
WO2020036115A1
WO2020036115A1 PCT/JP2019/031309 JP2019031309W WO2020036115A1 WO 2020036115 A1 WO2020036115 A1 WO 2020036115A1 JP 2019031309 W JP2019031309 W JP 2019031309W WO 2020036115 A1 WO2020036115 A1 WO 2020036115A1
Authority
WO
WIPO (PCT)
Prior art keywords
syringe
conduit
outlet
cells
microcircuit
Prior art date
Application number
PCT/JP2019/031309
Other languages
English (en)
Japanese (ja)
Inventor
知大 久保
まどか 綾野
Original Assignee
株式会社 ＴＬ Ｇｅｎｏｍｉｃｓ
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 株式会社 ＴＬ Ｇｅｎｏｍｉｃｓ filed Critical 株式会社 ＴＬ Ｇｅｎｏｍｉｃｓ
Publication of WO2020036115A1 publication Critical patent/WO2020036115A1/fr

Links

Images

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03B—SEPARATING SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS
    • B03B5/00—Washing granular, powdered or lumpy materials; Wet separating
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/26—Inoculator or sampler

Definitions

  • the present invention relates to a classification device, and more particularly to a fluid device suitable for hydrodynamic classification of suspended cells such as blood cells.
  • Patent Document 1 discloses an apparatus for separating cells by filtration.
  • Patent Documents 2 and 3 disclose a method for hydrodynamically classifying fine particles.
  • suspension of suspended cells is treated as fluid. It is necessary to give a sufficiently high flow rate to the suspension to obtain a hydrodynamic effect.
  • the flow rate of simple filtration is not particularly limited, and it is preferable that the flow rate be low unless the processing time is excessively long.
  • An object of the present invention is to provide a fluid device suitable for hydrodynamic classification of floating cells.
  • ⁇ Aspect 1> An inlet for sucking a mixture containing floating cells having a particle size distribution, Syringe A, A fine circuit for hydrodynamic classification of the floating cells, An outlet for discharging a cell suspension containing nucleated cells classified by the fine circuit, A branch communicating between the intake, the syringe A and the three-way circuit comprising the microcircuit, With Taking the mixed solution into the syringe A from the inlet via the branch, Sending the mixture from the syringe A to the microcircuit via the branch, While continuously classifying the floating cells in the mixture by continuously flowing the mixture toward the inside of the microcircuit by the pressure generated by the syringe A, the cell suspension is continuously Flush from the outlet, Fluid device.
  • the floating cells include at least annucleated red blood cells and nucleated cells, The fluid device according to aspect 1.
  • ⁇ Aspect 3> In the microcircuit, against the flow of the mixture, pushing the flow of the mixture to the opposite side by contacting the flow of a clear solution containing no nucleated red blood cells and nucleated cells from the side, The nucleated red blood cells in the mixture are hydrodynamically classified from the side not in contact with the flow of the clarified liquid, and further downstream in the mixed liquid from the side not in contact with the flow of the clarified liquid. Obtaining the cell suspension by hydrodynamically classifying nucleated cells, The fluid device according to aspect 2.
  • ⁇ Aspect 4> Further comprising a syringe B for sending the clarified liquid to the fine circuit, The fluid device according to aspect 3.
  • ⁇ Aspect 5> A first conduit having said inlet and connecting to said branch on the opposite side; A second conduit located between the syringe A and the branch; A third conduit located between the branch and the microcircuit; A fourth conduit located between the syringe B and the microcircuit; Further comprising When sending the mixture from the syringe A to the microcircuit via at least the second conduit and the third conduit, the second conduit, the branch, and the third conduit are vertically aligned and the Forming a path for connecting the syringe A and the microcircuit; Throughout this path, the nucleated red blood cells and nucleated cells in the mixture can fall toward microcircuits by gravity.
  • a fifth conduit connected to the microcircuit and having the outlet on the opposite side; The nucleated cells in the cell suspension throughout the fifth conduit can fall toward the outlet by gravity.
  • a third outlet as an outlet different from the outlet, The remainder of the mixture and the clarified liquid after the classification of the nucleated red blood cells and nucleated cells are continuously flushed from the third outlet, A drain reservoir connected to and sealed to the third outlet; A drainage absorbent enclosed in the drainage reservoir, An air vent with a filter connected to the drainage reservoir, Further comprising The fine circuit, the third outlet and the drain reservoir are embedded in the case, The fluid device according to aspect 7.
  • a driving device for driving the fluid device according to aspect 7, comprising:
  • the fluid device can be attached to and detached from the driving device as a cartridge,
  • a reciprocating mechanism for pulling the piston A of the syringe A the liquid mixture is taken into the syringe A from the inlet via the branch,
  • the reciprocating mechanism pushes the piston A to cause the mixed liquid to flow into the microcircuit by pressure generated in the syringe A.
  • ⁇ Aspect 12> The mixed solution is taken into the syringe A, and when the mixed solution is further sent from the syringe A to the microcircuit, the mixed solution in the syringe A is previously arranged in the syringe A in order to stir the mixed solution. Move the stirrer with magnetic force, The driving device according to aspect 10.
  • ⁇ Aspect 13> A method for obtaining concentrated nucleated cells by classifying whole blood using the fluid device according to aspect 1.
  • a stir bar is further arranged in the syringe A in advance, and the stir bar has a size that does not fit into the discharge port of the syringe A.
  • the mixed solution is taken into the syringe A, and further, when the mixed solution is sent from the syringe A to the microcircuit, the mixed solution is stirred by the stirrer in the syringe A. 15. The method according to aspect 14.
  • a fluid device suitable for hydrodynamic classification of floating cells can be provided.
  • FIG. 3 is a circuit diagram of a fluid device.
  • FIG. 3 is a plan view of a fine circuit.
  • FIG. 3 is a partial plan view of a fine circuit.
  • FIG. 2 is a detailed partial perspective view of a fine circuit.
  • FIG. 2 is a perspective view of a fluid device. The perspective view of a case. The front view of a blood collection tube holder.
  • FIG. Flowchart of classification work.
  • the front view of a fluid device The front view of a fluid device.
  • FIG. 2 is a front view of the fluid device and the driving device.
  • classification refers to dividing floating cells into a group of large particles and a group of small particles when the cells are regarded as particles having a particle size distribution. Alternatively, it means that a suspension of floating cells is fractionated for each size of floating cells.
  • Blood includes diluted blood and undiluted blood as collected. Suspended cells contained in blood include blood cells and cells other than blood cells.
  • Hydrodynamic classification refers to hydrodynamic classification of particles using a water-based liquid as a medium.
  • water-based liquid includes plasma and water-based buffers. Plasma and buffers have the osmotic pressure and pH necessary to preserve suspended cells such as blood cells.
  • the classification that simply separates the suspension cells smaller than the mesh from the suspension cells larger than the mesh based on the size of the mesh is the hydrodynamic classification in the present invention. different.
  • the combination of hydraulic classification and simple filtration is not excluded from the present invention.
  • FCM flow cytometry
  • nucleated cells particularly refer to nucleated cells in the blood and floating cells.
  • Nucleated cells include, but are not limited to, leukocytes, immature red blood cells, so-called nucleated red blood cells, and circulating tumor cells (CTCs).
  • CTCs are an example of cells that are nucleated but not blood cells.
  • Anucleated red blood cells are blood cells but are an example of anucleated cells.
  • FIG. 1 shows a circuit diagram of the fluid device 10.
  • Fluid device 10 includes an inlet 50.
  • the fluid device 10 sucks the mixed liquid MX from the inlet 50.
  • the mixture MX is a suspension of floating cells containing a population of floating cells to be classified.
  • the population of floating cells is composed of nucleated red blood cells and nucleated cells.
  • Mixture MX contains nucleated red blood cells and nucleated cells.
  • the nucleated red blood cells and nucleated cells and other blood components are mixed with each other. Anucleated red blood cells are thought to be smaller than nucleated cells because the nuclei have been removed.
  • the fluid device 10 further includes a microcircuit 20.
  • the microcircuit 20 is a fluid circuit suitable for performing a hydrodynamic classification on a population of floating cells. Also called a microchannel.
  • the fine circuit 20 generates a cell suspension containing nucleated cells by classifying the floating cells in the mixed solution MX. Nucleated cells are included in nucleated cells (Nucleated Cell, NC).
  • a cell suspension containing nucleated cells obtained by classification is referred to as a cell suspension CS.
  • fine means that the diameter of the flow path arranged in the fine circuit is a size suitable for performing hydrodynamic classification of blood cells or other floating cells.
  • the diameter of the channel arranged in the microcircuit is in the range of 1 ⁇ m to 1000 ⁇ m.
  • the diameter of the channel is the diameter of the inscribed circle in the cross section of the channel.
  • the diameter of the flow path may be 1 ⁇ m to 100 ⁇ m.
  • the fluid device 10 further includes a syringe 30.
  • the syringe 30 functions as a syringe A.
  • the syringe 30 generates a suction pressure for sucking the mixed liquid MX into the fluid device 10.
  • the syringe 30 generates an exhaust pressure for sending the mixed liquid MX to the fine circuit 20.
  • the fluid device 10 further includes an outlet 40.
  • the fluid device 10 discharges the cell suspension CS.
  • the cell suspension CS is discharged by the discharge pressure of the syringe 30.
  • the fluid device 10 further includes a branch 15.
  • the branch 15 communicates between three sides including the inlet 50, the syringe 30, and the microcircuit 20.
  • the fluid device 10 takes the mixed liquid MX into the syringe 30 from the intake port 50 via the branch 15.
  • the fluid device 10 stops taking the mixed liquid MX.
  • the fluid device 10 sends the mixed liquid MX taken into the microcircuit 20 from the syringe 30 via the branch 15.
  • the fluid device 10 continuously flows the mixed liquid MX into the fine circuit 20 by the pressure generated by the syringe 30. Thereby, the fluid device 10 continuously classifies the floating cells in the mixed liquid MX.
  • the fluid device 10 continuously flushes the cell suspension CS from the outlet 40 while performing classification.
  • the fluid device 10 may receive power for driving the syringe 30 from outside.
  • the syringe 30 does not generate pulsation unlike a peristaltic pump or the like.
  • the syringe 30 can send out the mixed liquid MX at a constant speed and a strong constant pressure. Therefore, it is advantageous to perform the classification in the fine circuit 20 stably.
  • FIG. 2 shows the fluid device 10 as viewed from the front.
  • Fluid device 10 includes several conduits.
  • the first conduit 11a has an inlet 50.
  • the first conduit 11 a connects to the branch 15 on the opposite side of the inlet 50.
  • a blood collection tube 51 storing whole blood as a mixed liquid MX is connected to the intake port 50.
  • At the center of the first conduit 11a there is a portion which is closed and opened by the intake valve 71. How to use the intake valve 71 for the fluid device 10 will be described later.
  • the second conduit 11b is located between the syringe 30 and the branch 15 as shown in FIG.
  • the syringe 30 has a discharge port 31.
  • One end of the second conduit 11b is connected to the discharge port 31.
  • the other end of the second conduit 11b connects to the branch 15.
  • the third conduit 11c is located between the branch 15 and the microcircuit 20.
  • One end of the third conduit 11c is connected to the branch 15.
  • the other end of the third conduit 11c is connected to an inlet 21a of the microcircuit 20.
  • a pipe fitting (tube fitting) may be provided in advance at the inlet 21a.
  • the microcircuit 20 and the third conduit 11c may be connected by a pipe joint.
  • At the center of the third conduit 11c is a portion which is closed and opened by the start valve 76. The use of the start valve 76 for the fluidic device 10 will be described later.
  • the fluid device 10 includes a syringe 35 as the syringe B.
  • the syringe 35 sends the clarified liquid CL to the fine circuit 20.
  • the syringe 35 has no pulsation, so that the clear liquid CL can be sent out at a constant and low speed. Therefore, it is advantageous for classification in the fine circuit 20.
  • the fourth conduit 12 is located between the syringe 35 and the microcircuit 20 as shown in FIG.
  • One end of the fourth conduit 12 is connected to a discharge port 36 of the syringe 35.
  • the other end of the fourth conduit 12 is connected to an inlet 21b of the microcircuit 20.
  • a pipe joint may be provided in advance at the inlet 21b.
  • the microcircuit 20 and the fourth conduit 12 may be connected by a pipe joint.
  • the outlet 40 in the fluid device 10 functions as a first outlet.
  • the fluid device 10 further has an outlet different from the outlet 40.
  • the fluid device 10 has a discharge port 45a as a second discharge port.
  • the suspension containing the non-nucleated red blood cells is continuously flushed from the outlet 45a.
  • the fluid device 10 has an outlet 45b as a third outlet. From the outlet 45b, the remaining part of the mixed liquid after the classification of the floating cells and the clarified liquid, that is, the flow through, are continuously flushed.
  • the fluid device 10 shown in FIG. 2 further includes a fifth conduit 16.
  • the fifth conduit 16 is connected to an outlet 22b of the microcircuit 20.
  • a pipe joint may be provided in advance at the outlet 22b.
  • the microcircuit 20 and the fifth conduit 16 may be connected by a pipe joint.
  • the fifth conduit 16 further has an outlet 40 on the opposite side.
  • the fluid device 10 shown in FIG. 2 further includes a sixth conduit 17a.
  • the sixth conduit 17a is connected to an outlet 22a of the microcircuit 20.
  • a pipe joint may be provided in advance at the outlet 22a.
  • the microcircuit 20 and the sixth conduit 17a may be connected by a pipe joint.
  • the sixth conduit 17a further has an outlet 45a on the opposite side.
  • the fluid device 10 shown in FIG. 2 further includes a seventh conduit 17b.
  • the seventh conduit 17b is connected to an outlet 22c of the microcircuit 20.
  • a pipe joint may be provided in advance at the outlet 22c.
  • the microcircuit 20 and the seventh conduit 17b may be connected by a pipe joint.
  • the seventh conduit 17b further has an outlet 45b on the opposite side.
  • the fluid device 10 has a drainage reservoir 46.
  • the drain reservoir 46 is connected to the outlet 45a and the outlet 45b.
  • the drainage reservoir 46 has a drainage container 47a and a drainage container 47b.
  • the fluid device 10 continuously flushes the classified cell suspension CS from the outlet 40.
  • the cell suspension CS is received from the outlet 40 by the collection container 41 connected to the outlet 40.
  • the collection container 41 is a container for collecting nucleated cells in the cell suspension CS. When the collection of the predetermined amount of the cell suspension CS is completed, the collection container 41 can be removed from the fluid device 10.
  • the fluid device 10 continuously flushes the suspension containing the classified non-nucleated red blood cells from the outlet 45a.
  • the suspension containing the non-nucleated red blood cells is stored in the drainage container 47a.
  • the fluid device 10 continuously flushes the remaining liquid MX after the classification of the floating cells and the clarified liquid CL from the outlet 45b.
  • the remainder of the mixed liquid MX and the used clarified liquid CL are stored in the drainage container 47b.
  • the drainage reservoir 46 stores these liquids as drainage.
  • FIG. 3 shows the fine circuit 20 in a plan view.
  • the microcircuit 20 is a flow path chip for separating floating cells such as blood cells.
  • the fine circuit 20 has a main flow path 23. One end of the main flow path 23 is an inlet 21a. The other end of the main flow path 23 is an outlet 22c.
  • the microcircuit 20 further has a sub flow path 24.
  • the end of the sub flow path 24 is an inlet 21b.
  • the end of the sub flow path 24 is connected to the main flow path 23 at a junction 28.
  • the main flow path 23 has flow path parts 25a to 25d in order from the inlet 21a to the outlet 22c.
  • the flow path parts 25a to 25d are connected continuously from the inlet 21a to the outlet 22c.
  • Outlets 22a and 22b are further provided at one ends of the branch passages 26a and 26b connected by the passage part 25c.
  • the inlet 21a shown in FIG. 3 is connected to the syringe 30 containing the mixed liquid MX. From the syringe 30, the mixed liquid MX is sent to the inlet 21a at a predetermined flow rate. The mixed liquid MX enters the flow path part 25a via the inlet 21a.
  • the mixed liquid MX shown in FIG. 3 be diluted before being introduced into the fine circuit 20.
  • the liquid used for diluting the mixed liquid MX is referred to as a diluent DL.
  • An agent for preserving the diluent for example, sodium azide may be added to the DL.
  • the diluent DL may be injected into the syringe 30 in advance.
  • the syringe 30 may be a pre-filled syringe in which a diluent DL is previously filled in a syringe.
  • the dilution ratio can be 2 to 500 times. In this embodiment, the dilution ratio may be 50 times.
  • the flow rate of the diluted mixed solution MX per unit time can be 1 to 1000 ⁇ l / min. For example, it may be 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 ⁇ l / min.
  • the processing time may be 1 minute to 300 minutes. For example, any of 5, 10, 20, 30, 40, 50, 60, 90, 120, 150, 180, 210, 240, and 270 minutes may be used.
  • a liquid obtained by diluting, for example, 15 ml of the mixed liquid MX may be treated in one classification.
  • the sub flow path 24 is connected to the syringe 35.
  • a clear liquid CL is put in the syringe 35.
  • the clarified liquid CL may be injected into the syringe 35 in advance.
  • the syringe 35 may be a prefilled syringe in which a clarified liquid CL is previously filled in a syringe.
  • the clarified liquid CL is a liquid containing no floating cells.
  • the clarified liquid CL is a liquid that does not damage blood cells and other cells.
  • the clarified liquid CL is a buffer.
  • the clarified liquid CL may be PBS.
  • Branch channels 26 a and 26 b shown in FIG. 3 are both channels that branch off from the main channel 23.
  • the branch flow path 26a and the branch flow path 26b are branched from the main flow path 23 in this order from the upstream side.
  • branch flow paths 26a and 26b are arranged on the side facing the sub flow path 24.
  • Each of the branch passages 26a and 26b shown in FIG. 3 has a plurality of narrow passages branched from the main passage 23.
  • the narrow channels are arranged from the upstream to the downstream of the main channel 23.
  • Branch channels 26a and 26b reach outlets 22a and 22b, respectively.
  • the narrow passages in the branch passages 26a and 26b join before the outlets 22a and 22b, respectively.
  • the channel part 25d reaches the outlet 22c.
  • FIG. 4 shows the fine circuit 20 in a plan view.
  • the figure schematically shows the process of sorting floating cells by the fine circuit 20.
  • the branch channel 26a shows ten narrow channels
  • the branch channel 26b shows three narrow channels.
  • FIGS. 4 and 5 are cited from Japanese Patent Application Laid-Open No. 2007-175684 of Patent Document 3 and are partially modified so as to be suitable for the description of the present embodiment.
  • the classification mechanism is described in detail in Japanese Patent Application Laid-Open No. 2007-175684 of Patent Document 3.
  • the mixed liquid MX continuously flows from further upstream of the flow path part 25b.
  • the mixture MX contains a large amount of cells.
  • the flow of the clarified liquid CL is continuously brought into contact with the flow of the mixed liquid MX from the side. Thereby, the cells flowing through the main flow path 23 are continuously pushed in from the side of the main flow path 23. As a result, the flow of the mixed liquid MX is continuously pushed to the opposite side to the flow of the clarified liquid CL.
  • the floating cells are continuously pushed toward the branch flow channels 26a and 26b, and the floating cells continuously flow into these branch flow channels.
  • the nucleus-free red blood cells 27 continuously flow into the branch channel 26a.
  • the non-nucleated red blood cells 27 in the mixed liquid MX are hydraulically classified in the flow channel part 25b. Classification is continuously performed on the side of the flow of the mixed liquid MX that is not in contact with the flow of the clarified liquid CL.
  • the branch channel 26a functions as a channel for removing nucleated red blood cells 27.
  • the inscribed diameter of the narrow channel of the branch channel 26a is 12 to 19 ⁇ m.
  • the inscribed diameter may be any of 13, 14, 15, 16, 17, and 18 ⁇ m.
  • nucleated cells 29a-c continuously flow into the branch channel 26b.
  • the nucleated cells 29a-c in the mixed liquid MX are hydrodynamically classified. Classification is continuously performed on the side of the flow of the mixed liquid MX that is not in contact with the flow of the clarified liquid CL.
  • the cell suspension CS is continuously obtained from the branch channel 26b.
  • the branch channel 26b functions as a recovery channel for nucleated cells 29a-c.
  • the inscribed diameter of the narrow channel of the branch channel 26b is 20 to 30 ⁇ m.
  • the inscribed diameter may be any of 21, 22, 23, 24, 25, 26, 27, 28 and 29 ⁇ m.
  • the size of nucleated cells including nucleated red blood cells is considered to be 11 to 13 ⁇ m.
  • the inscribed diameter of each of the narrow flow paths of the branch flow paths 26a and 26b shown in FIG. 4 is the diameter of an inscribed circle in a cross section orthogonal to the fine flow paths.
  • the cross section of each narrow channel is square.
  • the cross section of the narrow channel may be another polygon or a circle. The same applies to other branch channels.
  • FIG. 5 shows the details of the fine circuit while focusing on the flow path part 25c.
  • the value of the inscribed diameter of the narrow channel is not always equal to the maximum value of the diameter of the floating cells to be classified. Therefore, the hydraulic classification according to the present embodiment is different from simple filtration. The hydraulic classification according to the present embodiment will be described below.
  • FIG. 5 shows the flow path part 25c. Further, a branch channel 26a is shown. In order to simplify the explanation, only one narrow channel constituting the branch channel 26a is provided. In the description of FIG. 5, the narrow channel forming the branch channel 26a is simply referred to as a branch channel 26a.
  • the liquid flow LF is continuously introduced into the flow path part 25c.
  • the liquid stream LF includes the above-described mixed liquid MX and the clarified liquid CL. These liquids are partially mixed in the liquid stream LF.
  • nucleated cells 29a as large cells and nucleated red blood cells 27 as small cells are suspended.
  • the nucleated cell 29a is drawn as a representative of other nucleated cells.
  • a part of the liquid flow LF that is introduced into the branch flow path 26a is referred to as a liquid flow LE.
  • a portion of the liquid flow LF that is not introduced into the branch flow path 26a and flows downstream is referred to as a liquid flow LG.
  • the flow rate of the liquid flow LE is smaller than a certain value.
  • the flow rate is proportional to the cross section of the liquid flow LE.
  • the liquid flow LE is located on the inner wall of the flow path part 25c on the side of the branch flow path 26a.
  • the flow rate of the liquid flow LE is also proportional to the flow velocity of the liquid flow LE in the branch flow path 26a.
  • the nucleus-free red blood cells 27 flowing in the liquid flow LE are introduced into the branch channel 26a.
  • more than half of the volume of the nucleated cell 29a belongs to the liquid flow LG side.
  • the nucleated cells 29a only partially contact the liquid flow LE. Therefore, the nucleated cells 29a are not introduced into the branch channel 26a.
  • the diameter of the nucleated cell 29a may be smaller than the inscribed diameter of the branch channel 26a. If the flow rate of the liquid flow LE increases, the cross section of the liquid flow LE increases. In this case, it is conceivable that the nucleated cell 29a is swallowed by the liquid flow LE and is guided to the branch channel 26a.
  • the nucleated cells 29a are carried further downstream on the liquid flow LG. As described above, a fluid that does not contain floating cells of a certain size or more can be collected from the branch channel 26a. Nucleated cells 29a and other nucleated cells undergo classification downstream.
  • FIG. 6 shows the fluid device 10 in a stereoscopic view.
  • the fluid device 10 includes a case 55.
  • the case 55 fixes the position and orientation of each component of the fluid device 10. The operation of the case will be described later.
  • floating cells in the mixed solution MX can fall toward the microcircuit 20 by gravity over the entire path 13. It is preferable that the inclination with respect to the horizontal direction is 80 degrees or more and 90 degrees or less at any point in the route.
  • the slope may be any of 81, 82, 83, 84, 85, 86, 87, 88 and 89 degrees. It is preferable to target 90 degrees.
  • the fine circuit 20 is installed so that each flow path located inside the fine circuit 20 is horizontal.
  • the cell suspension CS when the cell suspension CS is sent from the microcircuit 20 to the outlet 40, the cell suspension CS passes through the fifth conduit 16.
  • the nucleated cells in the cell suspension CS can fall toward the outlet 40 by gravity throughout the fifth conduit 16.
  • the inclination with respect to the horizontal direction is preferably 80 degrees or more and 90 degrees or less.
  • the slope may be any of 81, 82, 83, 84, 85, 86, 87, 88 and 89 degrees. It is preferable to target 90 degrees.
  • the drainage reservoir 46 is sealed.
  • the upper part of the drainage container 47a and the upper part of the drainage container 47b communicate with each other.
  • the drainage reservoir 46 may be composed of a single container instead of having the drainage container 47a and the drainage container 47b.
  • the drainage container 47a and the drainage container 47b may be box-shaped or bag-shaped. Further, the outlet 45a and the outlet 45b may be integrated into one outlet by joining the sixth conduit 17a and the seventh conduit 17b.
  • the drain reservoir 46 need not be sealed.
  • drainage absorbents 48a and 48b are arranged in the drainage reservoir 46. These drainage absorbents are enclosed in a drainage reservoir 46.
  • the drainage absorbent 48a is disposed in the drainage container 47a.
  • the drainage absorbent 48b is disposed in the drainage container 47b.
  • the fluid device 10 includes a ventilation tube 43.
  • the ventilation pipe 43 is used as an air vent hole.
  • One end of the ventilation pipe 43 is connected to the upper part of the drainage reservoir 46.
  • the other end of the ventilation pipe 43 is open to the outside of the drainage reservoir 46. As the drainage in the drainage reservoir 46 increases, the air in the drainage reservoir 46 is pushed out from the ventilation pipe 43.
  • the fluid device 10 does not need to include the ventilation tube 43.
  • the ventilation pipe 43 includes a filter 44 as shown in FIG.
  • the presence of the filter 44 prevents the drainage in the drainage reservoir 46 from leaking outside the drainage reservoir 46. With such a configuration, the sealed state of the drainage in the drainage reservoir 46 is maintained.
  • the filter 44 is installed on the opposite side of the drainage reservoir 46 with the vent pipe 43 interposed therebetween.
  • the ventilation pipe 43 does not have to include the filter 44.
  • FIG. 7 shows the case 55 in a stereoscopic view.
  • the case 55 includes a front member 56a and a back member 56b.
  • the front member 56a and the back member 56b may be formed by vacuum foam.
  • the case 55 may be formed as a blister pack.
  • a fluid circuit including the syringe 30, the microcircuit 20, and other members is sandwiched between the front member 56a and the back member 56b.
  • Such a configuration is one mode of the case 55, and the case 55 can have various parts configurations.
  • a blood collection tube holder 60 is attached to the blood collection tube 51. Whole blood collected in advance is stored in the blood collection tube 51. The whole blood in the blood collection tube 51 is guided to the inlet through the blood collection tube holder 60.
  • the blood collection tube holder 60 has a flange 63 at the upper end of the cylinder. The flange 63 serves to fix the blood collection tube 51 and the blood collection tube holder 60 to the case 55.
  • the syringe 30 includes a piston 32.
  • the syringe 30 has a flange 33.
  • the flange 33 serves to fix the syringe 30 to the case 55.
  • the syringe 30 further includes a cylinder 34.
  • a discharge port 31 is provided at a lower end of the cylinder 34.
  • the cylinder 34 has a flange 33 at its upper end. Piston 32 is inserted into cylinder 34. A plunger is attached to the piston 32.
  • the syringe 35 has a piston 37.
  • the syringe 35 has a flange 38.
  • the flange 38 serves to fix the syringe 35 to the case 55.
  • the syringe 35 further includes a cylinder 39.
  • a discharge port 36 is provided at the lower end of the cylinder 39.
  • the cylinder 39 has a flange 38 at its upper end.
  • the piston 37 is inserted into the cylinder 39.
  • a plunger is attached to the piston 37.
  • the case 55 includes a retainer 58a.
  • the retainer 58a restrains the blood collection tube holder 60 so that the blood collection tube holder 60 cannot move in the horizontal direction. Further, the contact between the flange 63 and the upper side of the retainer 58a fixes the blood collection tube holder 60 in the vertical direction. Specifically, the blood collection tube holder 60 is prevented from lowering below a predetermined height. By fixing the blood collection tube holder 60, the blood collection tube 51 attached thereto is also fixed.
  • the case 55 includes a retainer 58b.
  • the retainer 58b restrains the syringe 30 from moving in the horizontal direction.
  • the case 55 includes a retainer 58c.
  • the retainer 58c restrains the syringe 35 from moving in the horizontal direction.
  • the blood collection tube holder 60, the syringe 30, and the syringe 35 be arranged in a plane by the retainers 58a to 58c.
  • a blood collection tube holder 60, a syringe 30, and a syringe 35 are arranged in order from the left as viewed from the left. These orders can be interchanged. Since the blood collection tube holder 60 and the syringe 30 are adjacent to each other, intersection and contact between the branch 15 and the surrounding conduit and the fourth conduit 12 can be avoided.
  • the case 55 includes a retainer 58d.
  • the retainer 58d restrains the collection container 41 from moving in the horizontal direction. It is preferable that the retainer 58d is further restrained so that the collection container 41 cannot move in the vertical direction.
  • the retainer 58d may hold the collection container 41.
  • the sampling container 41 may be placed with a seating surface provided on the retainer 58d.
  • a flange may be provided on the collection container 41, and the retainer 58d may support the flange.
  • the fine circuit 20, the discharge port 45a, the discharge port 45b, and the drain reservoir 46 are embedded in a case 55.
  • the case 55 has a compartment 57a.
  • the microcircuit 20 is located in the compartment 57a.
  • the case 55 has a compartment 57b.
  • the drain reservoir 46 is located in the compartment 57b. In other embodiments, the compartment 57b itself may function as the drainage reservoir 46.
  • the entire sixth conduit 17a is also embedded in the case 55. It is preferable that the entire seventh conduit 17b is also embedded in the case 55. Further, a filter 44 may be provided at an end of the case. Further, the entire ventilation pipe 43 may be embedded in the case 55.
  • the front member 56a and the back member 56b may be bonded or thermally welded to embed the above members in the respective compartments.
  • the joining of the front member 56a and the back member 56b may be a method of fitting one to the other.
  • FIG. 8 shows the blood collection tube holder 60 as viewed from the front. However, only the cross section of the case 55 is illustrated.
  • the blood collection tube holder 60 is fixed to the retainer 58a.
  • the first conduit 11a and the blood collection tube 51 are connected via a blood collection tube holder 60.
  • the blood collection tube holder 60 has a discharge port 65.
  • the intake port 50 of the first conduit 11a is attached to the discharge port 65.
  • the blood collection tube 51 has a cap 52.
  • the cap 52 is a rubber stopper.
  • the blood collection tube 51 is pushed into the blood collection tube holder 60 with the cap 52 of the blood collection tube 51 facing downward.
  • the blood collection tube holder 60 has a needle-like plug 66.
  • the plug 66 pierces the cap 52 and is pushed into the blood collection tube 51.
  • the plug 66 communicates with the discharge port 65.
  • the blood collection tube holder 60 can rotate around the vertical direction in the retainer 58a.
  • the discharge port 54 and the intake port 50 may be threaded.
  • the first conduit 11a and the blood collection tube holder 60 may be connected by fitting these screws.
  • the blood collection tube holder 60 and the inlet 50 may be formed integrally with each other.
  • the plug 66 plays the role of the inlet 50.
  • the blood collection tube holder 60 and the first conduit 11a may be integrated. That is, the intake port 50 and the discharge port 65 may be connected to each other. Further, the blood collection tube holder 60 and the ventilation needle 68 may be integrated.
  • an access window 59a is provided around the inlet 50.
  • the access port 50 and one end of the blood collection tube holder 60 can be accessed from outside through the access window 59a. For this reason, the fitting of the screws is facilitated by the access window 59a.
  • the access window 59a may form an access window connected to at least one of the access window 59b and the access window 59c.
  • a ventilation needle 68 may be further attached to the blood collection tube 51 as shown in FIG.
  • the ventilation needle 68 passes through the cap 52.
  • the air outside the blood collection tube 51 is led into the lumen of the blood collection tube 51.
  • the ventilation needle 68 may further penetrate the blood collection tube holder 60.
  • the second conduit 11b is connected to the discharge port 31 of the syringe 30 corresponding to the syringe A. As shown in FIG. 7, the second conduit 11b may be attached to the syringe 30 after fixing the syringe 30 to the retainer 58b. In the retainer 58b, the syringe 30 can rotate around the vertical direction as an axis. The discharge port 31 and one end of the second conduit 11b may be threaded. The second conduit 11b and the syringe 30 may be connected by fitting these screws.
  • an access window 59b is provided around the discharge port 31.
  • the discharge port 31 and one end of the second conduit 11b can be accessed from outside through the access window 59b. For this reason, the fitting of the screws is facilitated by the access window 59b.
  • the access window 59b may form an access window connected to at least one of the access window 59a and the access window 59c.
  • the fourth conduit 12 is connected to a discharge port 36 of a syringe 35 corresponding to the syringe B. As shown in FIG. 7, the fourth conduit 12 may be attached to the syringe 35 after fixing the syringe 35 to the retainer 58c. The syringe 35 can rotate around the vertical direction in the retainer 58c. The outlet 36 and one end of the fourth conduit 12 may be threaded. The fourth conduit 12 and the syringe 35 may be connected by fitting these screws.
  • an access window 59c is provided around the discharge port 36.
  • the discharge port 31 and one end of the fourth conduit 12 can be accessed from outside through the access window 59c. For this reason, the fitting of the screws is facilitated by the access window 59c.
  • the access window 59c may form an access window connected to at least one of the access window 59a and the access window 59b.
  • FIG. 9 schematically shows the driving device 70.
  • the driving device 70 is a device for driving the fluid device 10.
  • the drive 70 comprises an intake valve 71 and a start valve 76.
  • the driving device 70 includes a reciprocating mechanism 72a and a pressing mechanism 72b. These members are drawn separately, but may be connected to a control device (not shown) by wire or wirelessly.
  • the reciprocating mechanism 72a shown in FIG. 9 reciprocates vertically.
  • the reciprocating mechanism 72a holds the plunger of the piston 32.
  • the reciprocating mechanism 72a can pull the piston 32 and push it further.
  • the flange 33 may be pressed from above so that the body of the syringe 30 does not lift upward.
  • the pressing mechanism 72b moves at least downward.
  • the pressing mechanism 72b presses the plunger of the piston 37 in accordance with the movement of the reciprocating mechanism 72a pressing the piston 32.
  • the pressing mechanism 72b may be a reciprocating mechanism equivalent to the reciprocating mechanism 72a.
  • the first conduit 11a can be closed and opened by an external force.
  • the center of the first conduit 11a has rubber properties.
  • the second conduit 11a may be a silicone tube.
  • the intake valve 71 is a pinch valve.
  • the first conduit 11a can be freely closed and opened by the combination of the pinch valve and the rubber conduit.
  • the central part of the first conduit 11a is exposed in the access window 59a.
  • the intake valve 71 accesses the first conduit 11a through the access window 59a.
  • the third conduit 11c can be closed and opened by an external force.
  • the center of the third conduit 11c has rubber properties.
  • the start valve 76 is a pinch valve.
  • the third conduit 11c can be freely closed and opened by a combination of the pinch valve and the rubber conduit.
  • the central portion of the third conduit 11c is exposed in the access window 59b.
  • the start valve 76 accesses the third conduit 11c through the access window 59b.
  • the driving device 70 may further include a fastener 74 as shown in FIG.
  • the fastener 74 detachably supports the fluid device 10.
  • the fastener 74 supports the case 55.
  • the fastener 74 holds the fluid device 10 so as to hold it.
  • the shape, position, size, and type of the fastener 75 can be appropriately changed.
  • the fastener 74 may not be provided.
  • a stirrer 75 may be arranged in the syringe 30 in advance. By moving the stirrer 75, the mixed liquid MX taken into the syringe 30 can be stirred.
  • the stirrer 75 has a ferromagnetic material.
  • the driving device 70 moves the stirrer 75 by magnetic force.
  • the driving device 70 may reciprocate the stirrer 75 up and down by a permanent magnet or an electromagnet.
  • the stirrer 75 has a size that does not fit into the discharge port 31.
  • each component of the driving device 70 is separated from the fluid device 10.
  • the fluid circuit required for classification is mounted on the fluid device 10.
  • the power required for classification is provided by the drive unit 70.
  • the fluidic device 10 can be attached to the driving device 70 as a kind of cartridge. Further, the fluid device 10 after classification can be removed from the driving device and discarded. By disposing the fluid device 10 for each classifying operation, contamination between different blood samples can be prevented.
  • the stirrer 75 may be disposable together with the fluid device 10.
  • FIG. 10 shows a flowchart of the classification operation.
  • a predetermined preparation operation is performed on the fluid device.
  • the inside of the fine circuit and the peripheral conduit is immersed in the diluent DL or the clarifying liquid CL.
  • the mixed liquid MX is taken into the syringe A of the fluid device.
  • FIG. 11 shows a state where the inside of the fluid device 10 is immersed in the diluent DL and the clarifying liquid CL.
  • the mixed liquid MX is collected from the human or animal body into the blood collection tube 51 as a preparatory operation.
  • the mixed liquid MX is whole blood.
  • the blood collection tube 51 storing the mixed liquid MX is attached to the inlet 50.
  • the ventilation needle 68 is attached to the blood collection tube 51 as shown in FIG.
  • the diluent DL is injected into the syringe 30 in advance.
  • the syringe 30 may be a pre-filled syringe in which a diluent is filled in the syringe in advance.
  • the clarified liquid CL is injected into the syringe 35 in advance.
  • the syringe 35 may be a prefilled syringe in which a clarified liquid CL is previously filled in a syringe.
  • the collection container 41 is arranged so that the collection container 41 can receive the cell suspension from the outlet 40.
  • the collection container 41 is preferably an unused product. As soon as the preparation operation is completed, the classification operation is started by flowing each liquid into the fluid device 10 in a predetermined order.
  • Step S81 in FIG. 11 is executed, and the insides of the second conduit 11b, the third conduit 11c, and the fine circuit 20 are immersed in the diluent DL.
  • the diluent DL is guided to the second conduit 11b, the branch 15, the third conduit 11c, and the fine circuit 20.
  • the intake valve 71 is closed and the start valve 76 is opened. Thereby, the air in the second conduit 11b, the branch 15, the third conduit 11c, and the microcircuit 20 is evacuated.
  • the inside of the fourth conduit 12 and the fine circuit 20 is further immersed in the clarifying liquid CL.
  • the clarified liquid CL is guided to the fourth conduit 12 and the fine circuit 20.
  • the air in the fourth conduit 12 and the fine circuit 20 is evacuated.
  • the diluent DL and the clarified liquid CL may pass through the microcircuit 20 and reach the fifth conduit 16, the sixth conduit 17a, and the seventh conduit 17b, and may reach further ends thereof. ing.
  • step S82 the mixed liquid MX is taken into the syringe A of the fluid device.
  • FIG. 12 shows the fluid device 10 during the intake of the mixed liquid MX.
  • the fluid device 10 takes in the mixed liquid MX from the blood collection tube 51.
  • the intake valve 71 is opened and the start valve 76 is closed.
  • the mixed liquid MX is taken into the syringe 30 along this route by pulling the piston of the syringe 30. That is, a predetermined amount of the mixed liquid MX is taken into the syringe 30 from the intake port 50 via the branch 15 with the expansion of the volume of the syringe 30.
  • the mixed liquid MX when the mixed liquid MX is taken into the syringe 30, the mixed liquid MX is diluted in the syringe 30 by the diluent DL.
  • the stirrer 75 stirs the diluted mixture MX so that the floating cells do not settle in the syringe 30. Further, by the stirring by the stirrer 75, the mixed liquid MX and the diluted liquid DL before the dilution may be sufficiently mixed.
  • step S83 a pressure is applied to the mixed liquid MX already diluted in the syringe A (hereinafter, simply referred to as mixed liquid MX). Further, a pressure is applied to the clarified liquid CL in the syringe B.
  • FIG. 13 shows the fluid device 10 applying pressure to the mixed liquid MX.
  • the piston of the syringe 30 is pressed in step S83. Thereby, pressure is applied not only to the syringe 30 but also to the mixed liquid MX in the second conduit 11b.
  • the piston of the syringe 30 may be pushed while the start valve 76 is closed and the intake valve 71 is open. Thereby, the mixed liquid MX is extruded at least to the branch 15. In the figure, the mixed liquid MX is pushed back to the vicinity of the intake valve 71. As a result, the undiluted mixed liquid MX remaining between the branch 15 and the syringe 30, in particular, remaining in the second conduit 11 b can be pushed back toward the inlet 50.
  • step S84 the start valve is opened to pump the mixed liquid MX toward the fine circuit.
  • FIG. 14 shows a fluid device during classification of floating cells in the mixed solution MX.
  • step S84 the start valve 76 is released.
  • the pressure accumulated in the mixed liquid MX is transmitted to the fine circuit 20 via the third conduit 11c.
  • the intake valve 71 is closed to direct all pressure to the microcircuit 20.
  • a route from the syringe 30 to the microcircuit 20 via the branch 15 is formed.
  • the mixed liquid MX is sent from the syringe 30 to the fine circuit 20 via the branch 15 as the volume of the syringe 30 is reduced.
  • the mixed liquid MX is continuously poured into the fine circuit 20 by the pressure generated by the syringe 30.
  • the piston of the syringe 35 is kept pressed to keep sending the clarified liquid CL to the fine circuit 20.
  • the fine circuit 20 continuously classifies the floating cells in the mixed solution MX.
  • the cell suspension CS is continuously obtained from the outlet 40.
  • the nucleated cells in the cell suspension CS are collected in the collection container 41.
  • a suspension containing nucleated red blood cells, the remainder of the mixed solution MX, and a used clarified solution CL are continuously stored.
  • step S85 the application of the pressure in the syringe A and the syringe B is stopped. Further, in step S86, the cell suspension CS is taken out.
  • Step S85 is executed by stopping pushing the pistons of the syringe 30 and the syringe 35 in FIG. This stops applying pressure to the mixed liquid MX and the clarified liquid CL. In other words, the pressure returns to atmospheric pressure.
  • the discharge of the cell suspension CS from the outlet 40 stops.
  • the collection container 41 is removed from the fluid device 10.
  • the nucleated cells collected in the cell suspension CS are obtained.
  • the nucleated cells contained in the cell suspension CS are more concentrated than whole blood. In other words, the removal of the nucleated red blood cells increases the proportion of nucleated cells in all the floating cells in the cell suspension CS.
  • the mixed liquid MX collected in the blood collection tube may be pregnant woman's blood.
  • fetal nucleated erythrocytes which are a kind of nucleated cells, may be collected in the cell suspension CS. Collected fetal nucleated red blood cells can be used for prenatal diagnosis of fetuses and pregnant women.
  • the mixed liquid MX collected in the blood collection tube may be blood of a cancer patient or a potential cancer patient.
  • circulating tumor cells which are a kind of nucleated cells, may be collected in the cell suspension CS.
  • the collected circulating tumor cells (CTC) can be used for diagnosis and treatment of cancer.
  • the present invention is not limited to the above embodiments and examples, and can be appropriately modified without departing from the gist.
  • whole blood was used as the mixture MX.
  • the mixture MX may be a suspension of other floating cells. Examples of such suspensions are lymph and bone marrow fluid.
  • the mixture MX may be other body fluids collected from humans or other animals.
  • the mixture MX need not be diluted in the syringe A regardless of the type of cells contained in the mixture MX.
  • the mixed liquid MX may be diluted before being taken in from the inlet 50.
  • FIG. 15 is a front view of the fluid device 80 and the driving device 70.
  • the fluid device has the same configuration as the fluid device 10 except as described below.
  • the fluid device 80 includes a case 85 instead of the case 55 shown in FIGS.
  • the case 85 has a series of access windows 89.
  • the access window 89 has all the functions of the access windows 59a to 59c shown in FIGS.
  • the access window 89 may be applied to the case 55 described above.
  • the case 85 includes retainers 58b and 58c.
  • the retainer 58b restrains the lower part of the cylinder 34.
  • the retainer 58c restrains the lower part of the cylinder 39.
  • the upper portions of these cylinders are restrained by the drive device 70. The mode in which the upper part of the cylinder is restrained by the driving device 70 may be applied to the case 55 described above.
  • the piston 32 included in the fluid device 80 includes a piston head 82a, a piston rod 82b, and a flange 82c.
  • the piston head 82a and the piston rod 82b can be freely inserted into and removed from the cylinder 34.
  • the piston 37 includes a piston head 87a, a piston rod 87b, and a flange 87c.
  • the piston head 87a and the piston rod 87b can be freely inserted into and removed from the cylinder 39.
  • the piston 32 and the piston 37 may be applied to the fluid device 10 described above.
  • the driving device 70 includes a support 78a.
  • the support 78a supports the lower surface of the flange 33 of the cylinder 34 so as to prevent the cylinder 34 from falling.
  • the support 78a preferably supports the lower surface of the flange 33 on both left and right sides of the cylinder 34. Further, the cylinder 34 is restrained from moving in the left-right direction in the figure. The support 78a restrains the left-right movement of the cylinder 34 near the flange 33 in the cylinder 34.
  • the driving device 70 includes a holding member 77a.
  • the presser 77a presses the upper surface of the flange 33 of the cylinder 34 to restrain the cylinder 34 from jumping out of the support 78b.
  • the presser 77a presses the cylinder 34.
  • the flange 33 is sandwiched between the lower surface of the holding member 77a and the upper surface of the support 78a. It is preferable that the presser 77a presses the upper surface of the flange 33 on both left and right sides of the piston 32 inserted into the cylinder 34. It is preferable that the lower surface of the presser 77a and the upper surface of the support 78a sandwich the flange 33 on both left and right sides of the cylinder 34.
  • the driving device 70 includes a support 78b.
  • the support 78b supports the lower surface of the flange 38 of the cylinder 39 so that the cylinder 39 does not fall.
  • the support 78b preferably supports the lower surface of the flange 38 on both left and right sides of the cylinder 39. Further, the cylinder 39 is restrained from moving in the left-right direction in the figure.
  • the support 78b restricts the left and right movement of the cylinder 39 near the flange 38 in the cylinder 39.
  • the driving device 70 includes a holding member 77b.
  • the presser 77b presses the upper surface of the flange 38 provided in the cylinder 39 to restrain the cylinder 39 from jumping out of the support 78b.
  • the presser 77b presses the cylinder 39.
  • the flange 38 is sandwiched between the lower surface of the holding member 77b and the upper surface of the support 78b. It is preferable that the presser 77b presses the upper surface of the flange 38 on both left and right sides of the piston 37 inserted into the cylinder 39. It is preferable that the lower surface of the presser 77b and the upper surface of the support 78b sandwich the flange 38 on both left and right sides of the cylinder 39.
  • the reciprocating mechanism 72a vertically sandwiches the flange 82c of the piston 32.
  • the reciprocating mechanism 72a preferably sandwiches the flange 82c on both sides of the piston rod 82b.
  • the reciprocating mechanism 72a moves the piston 32 up and down by applying a force to the flange 82c.
  • the reciprocating mechanism 72a may move the piston 32 up and down by gripping the flange 82c.
  • the pressing mechanism 72b may vertically sandwich the flange 87c of the piston 37. It is preferable that the pressing mechanism 72b sandwich the flange 87c on both sides of the piston rod 87b.
  • the pressing mechanism 72b moves the piston 37 downward by applying force to at least the upper surface of the flange 87c.
  • the pressing mechanism 72b may be a reciprocating mechanism that moves the piston 37 up and down by gripping the flange 87c.
  • the reciprocating mechanism 72a and the pressing mechanism 72b shown in FIG. 15 may be applied to the fluid device 10 described above.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Microbiology (AREA)
  • Sustainable Development (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

La présente invention concerne un dispositif fluidique (10) muni : d'un orifice d'entrée (50) à travers lequel un mélange liquide (MX) contenant des cellules flottantes ayant une distribution de tailles de particules doit être aspiré ; une seringue (30) qui correspond à une seringue A ; un fin passage (20) pour le tamisage de manière hydrodynamique des cellules flottantes ; un orifice d'évacuation (40) à travers lequel une suspension de cellules liquide (CS) contenant les cellules nucléées tamisées à travers le fin passage (20) doit être évacuée ; et une ramification (15) à travers laquelle trois constituants, c'est-à-dire, l'orifice d'entrée (50), la seringue (30) et le fin passage (20), peuvent être mis en communication l'un avec l'autre. Le mélange liquide (MX) est prélevé dans la seringue (30) depuis l'orifice d'entrée (50) par l'intermédiaire de la ramification (15). Le mélange liquide (MX) est alimenté au fin passage (20) depuis la seringue (30) par l'intermédiaire de la ramification (15). Les cellules flottantes dans le mélange liquide (MX) peuvent être tamisées de manière continue par écoulement continuellement du mélange liquide (MX) vers l'intérieur du fin passage (20) à l'aide d'une pression générée à l'aide de la seringue (30). Simultanément, la suspension de cellules liquide (CS) est séparée par lavage de manière continue à travers l'orifice d'évacuation (40).
PCT/JP2019/031309 2018-08-14 2019-08-08 Dispositif fluidique WO2020036115A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2018-152646 2018-08-14
JP2018152646 2018-08-14

Publications (1)

Publication Number Publication Date
WO2020036115A1 true WO2020036115A1 (fr) 2020-02-20

Family

ID=69525481

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2019/031309 WO2020036115A1 (fr) 2018-08-14 2019-08-08 Dispositif fluidique

Country Status (1)

Country Link
WO (1) WO2020036115A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020203510A1 (fr) * 2019-03-29 2020-10-08 株式会社 ＴＬ Ｇｅｎｏｍｉｃｓ Appareil pour commander une combinaison d'un microcircuit et d'une seringue
WO2024024967A1 (fr) * 2022-07-29 2024-02-01 日機装株式会社 Système de culture

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004503745A (ja) * 2000-07-07 2004-02-05 コールター　インターナショナル　コーポレイション 生物試料の調製と分析のための装置及び方法
JP2007175684A (ja) * 2005-12-26 2007-07-12 Minoru Seki 微粒子の濃縮・分級のための流路構造および方法
JP2009213460A (ja) * 2008-03-12 2009-09-24 Tokyo Women's Medical College 粒子処理用マイクロ流路システムおよび粒子処理方法
JP2012076016A (ja) * 2010-10-01 2012-04-19 Chiba Univ 連続的２次元粒子分離装置および粒子分離方法
JP2013017429A (ja) * 2011-07-12 2013-01-31 Konica Minolta Holdings Inc 血球溶解後のサイズ分離により血液から希少な目的細胞を回収する方法
WO2019078277A1 (fr) * 2017-10-19 2019-04-25 株式会社 ＴＬ Ｇｅｎｏｍｉｃｓ Puce pour classification de cellules

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2004503745A (ja) * 2000-07-07 2004-02-05 コールター　インターナショナル　コーポレイション 生物試料の調製と分析のための装置及び方法
JP2007175684A (ja) * 2005-12-26 2007-07-12 Minoru Seki 微粒子の濃縮・分級のための流路構造および方法
JP2009213460A (ja) * 2008-03-12 2009-09-24 Tokyo Women's Medical College 粒子処理用マイクロ流路システムおよび粒子処理方法
JP2012076016A (ja) * 2010-10-01 2012-04-19 Chiba Univ 連続的２次元粒子分離装置および粒子分離方法
JP2013017429A (ja) * 2011-07-12 2013-01-31 Konica Minolta Holdings Inc 血球溶解後のサイズ分離により血液から希少な目的細胞を回収する方法
WO2019078277A1 (fr) * 2017-10-19 2019-04-25 株式会社 ＴＬ Ｇｅｎｏｍｉｃｓ Puce pour classification de cellules

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
MATSUDA, MIYUKI, YAMADA, MASUMI, SEKI, MINORU: "Blood Cell Classification Utilizing Hydrodynamic Filtration", THE TRANSACTIONS OF THE INSTITUTE OF ELECTRICAL ENGINEERS OF JAPAN, vol. 128, no. 10, 2008, pages 396 - 401, XP055687339 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020203510A1 (fr) * 2019-03-29 2020-10-08 株式会社 ＴＬ Ｇｅｎｏｍｉｃｓ Appareil pour commander une combinaison d'un microcircuit et d'une seringue
WO2024024967A1 (fr) * 2022-07-29 2024-02-01 日機装株式会社 Système de culture

Similar Documents

Publication Publication Date Title
RU2539989C2 (ru) Система и способ фильтрации частиц
Dykes et al. Efficient removal of platelets from peripheral blood progenitor cell products using a novel micro-chip based acoustophoretic platform
Nivedita et al. A high throughput microfluidic platform for size-selective enrichment of cell populations in tissue and blood samples
EP0693182B1 (fr) Procede de separation de particules a partir d'un filtre
JP2004503745A (ja) 生物試料の調製と分析のための装置及び方法
KR101855490B1 (ko) 점탄성유체와 점성유체의 병행층류를 이용한 미소 입자 분리 및 세정 방법
WO2020036115A1 (fr) Dispositif fluidique
WO2010140706A1 (fr) Systèmes d'opérations biologiques et industrielles
JP6311609B2 (ja) 希少細胞の回収方法および検出方法
CA2787135A1 (fr) Separation magnetique de cellules rares
JP6308525B2 (ja) 微粒子分離用チップ、該微粒子分離用チップを用いた微粒子分離用システム及び微粒子分離方法
JP6573175B2 (ja) 分離装置、流体デバイス、分離方法及び混合方法
AU2013204820B2 (en) A System and Method for Particle Filtration
JPWO2014061631A1 (ja) 微粒子分離用マイクロ流路チップ、該チップを用いた微粒子分離用システム及び微粒子分離方法
JP4509607B2 (ja) 細胞分析装置および方法
US20220128549A1 (en) Method for Magnetically Labeling Particles and Labeling Apparatus
JP6737185B2 (ja) 相互作用する分子を有する血中細胞の同時検出方法
JP5825460B1 (ja) 細胞展開用デバイスの前処理方法、細胞展開用デバイス、および細胞展開用デバイスの前処理システム
JP6582486B2 (ja) 血液中の稀少細胞検出方法
CN115698254A (zh) 靶标粒子的分离方法及系统
JP6244589B2 (ja) 微粒子分離用マイクロ流路チップ、移流集積ユニット、微粒子分離用システム及び微粒子分離方法
JP2017181096A (ja) 希少細胞を捕捉する方法
JPWO2015093116A1 (ja) 細胞の蛍光免疫染色方法ならびにそのためのシステムおよびキット
US20180348098A1 (en) Cell-spreading method and cell-spreading kit for observing rare cells
WO2020203510A1 (fr) Appareil pour commander une combinaison d'un microcircuit et d'une seringue

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19849986

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19849986

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: JP