WO2009044902A1 - 誘電泳動装置および方法 - Google Patents
誘電泳動装置および方法 Download PDFInfo
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- WO2009044902A1 WO2009044902A1 PCT/JP2008/068114 JP2008068114W WO2009044902A1 WO 2009044902 A1 WO2009044902 A1 WO 2009044902A1 JP 2008068114 W JP2008068114 W JP 2008068114W WO 2009044902 A1 WO2009044902 A1 WO 2009044902A1
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- case
- dielectrophoresis
- small object
- electric field
- dielectric
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- 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
- B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C5/00—Separating dispersed particles from liquids by electrostatic effect
- B03C5/02—Separators
- B03C5/022—Non-uniform field separators
- B03C5/026—Non-uniform field separators using open-gradient differential dielectric separation, i.e. using electrodes of special shapes for non-uniform field creation, e.g. Fluid Integrated Circuit [FIC]
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D57/00—Separation, other than separation of solids, not fully covered by a single other group or subclass, e.g. B03C
- B01D57/02—Separation, other than separation of solids, not fully covered by a single other group or subclass, e.g. B03C by electrophoresis
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- 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
- B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C5/00—Separating dispersed particles from liquids by electrostatic effect
- B03C5/005—Dielectrophoresis, i.e. dielectric particles migrating towards the region of highest field strength
-
- 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
- B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
- B03C2201/00—Details of magnetic or electrostatic separation
- B03C2201/26—Details of magnetic or electrostatic separation for use in medical or biological applications
Definitions
- the present invention relates to a dielectrophoresis apparatus and method.
- Dielectrophoresis is a phenomenon in which a substance placed in a non-uniform electric field is driven by the interaction between the electric field and the dipole moment induced by it. Utilizing this phenomenon, research is currently being conducted in various fields, including the separation and identification of biological samples. For example, see:
- the present invention provides a dielectrophoresis apparatus capable of handling, including measurement of dielectrophoretic force, as well as moving, stopping, separating, rejecting, and classifying small target objects including a dielectric portion. aimed to.
- the present invention also provides a device suitable for the above dielectrophoresis apparatus.
- the present invention further provides a method of handling the object small object, a method of sorting, a method of measuring the electrophoretic force, and the like. These methods can be suitably carried out using the above dielectrophoresis apparatus.
- the dielectrophoresis device comprises a case for filling a small target object that is filled with a dielectric solution and includes a dielectric portion, a dielectrophoretic force acting on the small target object in the case, and a force based on gravity and buoyancy.
- An electrode device including a plurality of electrodes that form an inhomogeneous alternating electric field in the case that generates at least one stable equilibrium point with respect to the balance, and a tilt mechanism that holds and tilts the case and the electrode device. It is what it has.
- a non-uniform alternating electric field can be formed in the case.
- the dielectrophoretic force acting on a small object depends on the strength of the applied inhomogeneous alternating electric field and its gradient.
- the force based on gravity and buoyancy depends on the tilt angle of the case. This tilt angle can be obtained by the tilt mechanism.
- the dielectrophoretic force is equal to the force based on gravity and buoyancy when a small object is stationary, the dielectrophoretic force can be found by calculating the force based on gravity and buoyancy. Since the force based on gravity and buoyancy depends on the tilt angle of the case, the dielectrophoretic force can be measured by changing the tilt angle and measuring the tilt angle when a small object is stationary.
- the dielectrophoresis apparatus further includes a rotating mechanism that rotates the case and the electrode device on a plane plane tilted by the tilting mechanism.
- the dielectrophoretic force and the force based on gravity and buoyancy are not in the opposite directions, the dielectrophoretic force and the gravitational force are rotated by rotating the case in an inclined plane using the rotating mechanism. And force based on buoyancy can be balanced.
- the movement of the small object can be controlled by positively creating an imbalance by the rotation of the case and the electrode device by the rotating mechanism.
- the rotating mechanism can also be used for small object handling.
- the electrodes in the above electrode device have a plurality of (a pair of) electrodes whose sides facing each other are symmetrical with respect to the center line between the electrodes, and the electrode interval extends from one end to the other in a narrow parallel manner. Subsequently, a curve is formed so that the electrode spacing gradually increases. This pair of electrodes generates a stable equilibrium point and an unstable equilibrium point.
- the dielectrophoresis apparatus further includes an AC generator that generates an AC voltage that is applied to the plurality of electrodes and at least the frequency or voltage is variable.
- the angle of inclination by the tilt mechanism and the angle of rotation by the rotation mechanism can be measured.
- the angle may be measured visually or automatically by a sensor.
- the present invention also provides a device (apparatus) preferably used in the dielectrophoresis apparatus.
- This device includes a case for filling a small target object containing a dielectric part and filled with a dielectric solution, and the above case. k .
- a plurality of electrodes forming a non-uniform alternating electric field are provided in the four electrodes, and the sides facing each other are axisymmetric with respect to the center line between the electrodes, and the electrode spacing is narrow and parallel from one end to the other. It is then formed into a curve so that the electrode spacing gradually increases.
- the case is provided with a mouth for sucking or discharging the target small object.
- an inlet for injecting the dielectric solution into the case and an outlet for discharging the dielectric solution are opened in the case.
- Such devices can be used to separate, reject, and separate small objects.
- the case is filled with a dielectric solution and a target small object including a dielectric portion is placed, and a non-uniform alternating electric field is formed in the case.
- the dielectrophoretic force acting on the target small object in the case and the force based on gravity and buoyancy is adjusted.
- the target small object is moved in a desired direction. After moving the small object, it can be stopped at the desired position (stable equilibrium point).
- a dielectric solution is filled in a case and a plurality of different types of small objects each including a dielectric portion are placed to form a non-uniform alternating electric field in the case.
- the force based on the dielectrophoretic force acting on the small object in the case and the gravity and buoyancy This causes an imbalance between the two and the different types of target small objects to move in different directions, and stops the desired types of target small objects at the desired stable equilibrium point.
- a target small object including a dielectric part is filled in a case, a non-uniform alternating electric field is formed in the case, and a non-uniform alternating electric field is formed.
- the dielectrophoretic force acting on the target small object in the case and the force based on gravity and buoyancy are balanced.
- the dielectrophoretic force is calculated using a tilt angle in a predetermined direction of the case when the small object is stationary and the small object is stationary.
- the target small object can be applied to any dielectric including a biological cell or a small object partially including a dielectric part.
- Figure 1 shows the principle of the balance between gravitational slope component force acting on dielectric particles and dielectrophoretic force.
- Fig. 2 is a cross-sectional view showing a DEP device.
- Figure 3 is a plan view of the D E ⁇ device with the cover glass removed.
- Figure 4a shows the force acting on the particles, showing that the particles are on the center line and are balanced.
- Figure 4b is a graph showing the dielectrophoretic force profile and the gravitational slope component.
- Fig. 5a shows the force acting on the particles when the DEP device is placed upside down
- Fig. 5b shows the profile of the dielectrophoretic force and the gravitational slope component generated at this time. It is a graph.
- Figure 6 shows the force acting on the particles, and shows the state where the particles are off the center line and a resultant force is generated.
- Figure 7 shows the force acting on the particles, and the particles are off the center line. Indicates a balanced state.
- Figure 8 shows the positional relationship between the tilting plate and the rotating plate, that is, the inclination of the D D ⁇ device.
- Figure 9 shows the positional relationship between the tilting plate and the rotating plate, that is, the rotation of the device D ⁇ .
- Figure 10 shows the coordinate axes.
- Figures 11a and 11b are graphs showing the amount of particle movement over time when the angle 0 pitch is abruptly changed from 60 ° to 15 ° and from 15 ° to 60 °. is there.
- Fig. 12 is a graph showing multiple consecutive angular step responses.
- Fig. 13a shows the position of the particle in the angular step response, and
- Fig. 13b is a rough diagram showing the stable equilibrium point for each angle.
- Figure 14 is a graph showing the change in the position of a stationary particle where the gravitational slope component and the dielectrophoretic force are balanced, with the angle ⁇ pitch as the horizontal axis.
- Figures 15a and 15b are graphs showing the amount of particle movement over time when the voltage is suddenly changed from 8 Vpp to 20 Vpp and from 20 Vpp to 8 Vpp, respectively.
- Figure 16 shows a draft that shows multiple consecutive voltage step responses.
- Fig. 17a shows the position of the particles in the voltage step response, and
- Fig. 17b shows the stable equilibrium point for each voltage.
- Figure 18 shows a draft showing the change in the position of a stationary particle where the gravitational slope component and the dielectrophoretic force are balanced, with the voltage Vpp as the horizontal axis.
- Figures 19a and 19b show the amount of particle movement when the frequency f is changed rapidly from 3001: 1 ⁇ 2 to 51 ⁇ 112 and from 5 MHz to 300 KHz, respectively. It is a graph shown according to progress.
- Figure 20 is a graph showing multiple consecutive frequency step responses.
- Figure 21a shows the particle position in the frequency step response
- Figure 21b is a graph showing the stable equilibrium point for each frequency.
- Figure 22 is a graph showing the change in the position of a stationary particle where the gravitational slope component and the dielectrophoretic force are balanced, with frequency f as the horizontal axis.
- Figure 23 shows the movement of the dielectric particles (handling).
- Figure 24 shows a state in which multiple dielectric particles are separated.
- FIG. 25 is a perspective view showing the overall configuration of the dielectrophoresis apparatus.
- Fig. 26 is a perspective view showing the mounting structure of the tilting plate in the dielectrophoresis apparatus.
- Fig. 27 is a side view showing the positioning and fixing structure of the tilting plate in the dielectrophoresis apparatus.
- FIG. 28 is an enlarged sectional view taken along the line aa in FIG.
- Fig. 29 is a perspective view showing a state in which a rotating disk is provided on an XY table in the dielectrophoresis apparatus.
- FIG. 30 is a cross-sectional view showing the mounting structure of the rotating disk in the dielectrophoresis apparatus.
- Fig. 31 is a block diagram showing the electrical configuration of the dielectrophoresis apparatus.
- Figure 32 shows a modification of the DEP device, and is a plan view with a part cut away.
- Fig. 33 is a longitudinal sectional view of Fig. 32. BEST MODE FOR CARRYING OUT THE INVENTION
- R e (f CM ) R e ⁇ (-) / ( ⁇ ; + 2 ⁇ ) ⁇ Equation (2) where ⁇ ⁇ * and ⁇ m * are the complex ratio dielectric constants of the particles and the solution, respectively.
- the dielectrophoretic force differs in the direction and magnitude of the dielectrophoretic force depending on whether the dielectric constant of the particles (including the above living cells) is larger or smaller than the dielectric constant of the solution (solvent). Stop of mixed particles (including the above living cells) Handling such as separation or separation and identification of specific particles (including the above living cells) becomes possible.
- a dielectrophoresis device (apparatus) that contains particles in a solution (hereinafter referred to as a DEP device) (case).
- This DEP device fixes a frame by a spacer around a flat substrate glass (bottom plate), fills the space enclosed by the spacer with the solution, It is covered with a flat cover glass (top plate) to form a space (sealed) filled with the solution.
- An electrode that forms a non-uniform alternating electric field in the (closed) space is provided on the substrate glass, etc., details of which will be described later.
- the dielectrophoretic force F DEP is applied to the particles in the solution in an upward direction parallel to the slope, and these forces F G sin 0 pitch and F DE p If they are balanced, the particles are stationary.
- F DEP- F G Sin 9 pitch ° Equation (4)
- the state of F DEP > F G sin ⁇ piteh or F DEP ⁇ F G sin 0 piteh can be changed by changing the slope angle 0 pitch .
- the dielectric particles can be moved upward or downward along the slope, and by creating a state where Eq. (4) holds, the dielectric particles can be made stationary. (Handling of particles).
- Et al is, all parameters r representing the F G sin 0 pitch, p p , p m, g, ⁇ the pitch is assumed to be known, to the Dusko which Ri take a state to stand equation (4) is made Therefore, the dielectrophoretic force F DEP can be measured using Eq. (4) (this is called the zero method). Also, ⁇ . , ⁇ m , and E p are known, the electric field strength E can be calculated from Eqs. (1) and (2).
- This dielectrophoretic force measurement method has the following features.
- FIGS. 1-10 An example of the configuration of DEP device 10 is shown in FIGS.
- a pair of left and right electrodes (Creek-Gap Electrode) 13A and 13B forces are formed at the approximate center of the flat surface of the substrate glass 11 and are formed.
- the electrodes 13A and 13B have sides facing each other on the one side, that is, on the right side of FIG. 3 (the side that is located at the bottom when placed on the slope) (hereinafter referred to as the lower side or the terminal side).
- the length of the electrode is about 1/3 to 14
- the curves are formed so that they extend in parallel at very narrow intervals, and then increase gradually (the rate of increase increases).
- a wiring pattern extends from the lower side of the electrodes 13A and 13B, and is connected to electrode terminals 13a and 13b formed at the end of the substrate glass 11, respectively.
- a line passing through the middle of the above sides of electrodes 13A and 13B is assumed, and this is called the center line N between the electrodes.
- the electrodes 13 A and 13 B are line symmetric with respect to the center line N.
- Electrodes 13A, 13B, wiring patterns, and terminals 13a, 13b are formed by forming an aluminum thin film on the surface of the substrate glass 11 by sputtering and patterning it using photolithography technology. This comes out.
- an insulating film 14 (for example, a photosensitive polyimide with a thickness of ⁇ ⁇ or less) including the electrodes 13A and 13B and its wiring pattern is formed. Is parallel to the surface of the substrate glass 11 and is flat. The insulating film 14 is not formed on the right end portion of the substrate 11 including the terminals 13a and 13b.
- a spacer 15 with a circular hole 15A is disposed on the insulating film 14 and It is fixed. The position and size of the hole 15A are determined so that the electrodes 13A and 13B slip into the hole 15A.
- the spacer 15 can be made of, for example, silicon rubber (thickness 100 m).
- the hole 15 A of the spacer 15 is filled with a dielectric solution S (eg, distilled water) having a relative permittivity E m and has one or more dielectric particles P (eg, diameter) having a relative permittivity f p. 9.9 ⁇ m polystyrene beads).
- a dielectric solution S eg, distilled water
- P dielectric particles
- the dielectric constant of the polystyrene beads is smaller than the dielectric constant of distilled water S, so negative dielectrophoresis occurs, and the dielectrophoretic force of the polystyrene beads (particles P) toward the larger electrode spacing. If the DEP device 10 is placed horizontally, it moves in that direction (left direction in Fig. 3).
- the particle P has an upward dielectrophoretic force F DEP along the slope and a downward gravitational slope component force F c sin ⁇ pitch along the slope.
- F DEP dielectrophoretic force
- F G sin ⁇ pitch gravitational slope component force
- the intersection of these two forces F DEP and F G sin 0 pitch is two as described above. This is the point where the power of the two balances (referred to as the “equilibrium point”).
- the crank and gap electrodes There are two equilibration points for the crank and gap electrodes. This is because the dielectric swimming power F DEP has a curved profile with a peak at the center as shown in Fig. 4b. In the lower part of the electrode, the distance between the two electrodes 13A and 13B is narrow, so the electric field strength is large, but the opposing sides of the left and right electrodes 13A and 13B are parallel, so there is no electric field gradient, so dielectrophoresis There is no power. On the other hand, the distance between the electrodes is very wide above the electrodes, the electric field strength and inclination are extremely small, and the dielectrophoretic force is almost zero. As a result, the dielectrophoretic force has a peak at the center.
- the upper equilibrium point of the two equilibrium points is the stable equilibrium point. That is, the dielectrophoretic force F DEP acting on the particles below this equilibrium point is greater than the gravitational slope component Ferine piteh , and the particles rise toward the equilibrium point and above the equilibrium point. dielectric force F DEP acting on the particles in the slopes component force of gravity F c s in 0 pitch by Ri also rather small, the particles is lowered toward the equilibrium point. In any case, the particle moves toward the equilibrium point, so it is a stable equilibrium point. On the other hand, the lower equilibrium point is an unstable equilibrium point because the particles that are above or below the equilibrium point act to move the particles away from the equilibrium point.
- Fig. 5a shows the DEP device placed upside down from Fig. 4a
- Fig. 5b shows the dielectrophoretic force acting on the dielectric particles at that time. Shows the profile. In this case as well, two equilibrium points arise, one of which is the stable equilibrium point.
- a DEP device with a clique-gap electrode can secure a single stable equilibrium point for both positive and negative dielectrophoretic forces, and keep particles in a stable state. Can be stopped (stopped). The above is the case where the particle P exists on the center line N between the electrodes.
- the dielectrophoretic force F DEP can also be calculated from Eqs . (3) and (4).
- Fig. 8 To correlate with the dielectrophoresis apparatus described below, the position, angular relationship, and coordinate axes of the tilting plate 30 and the rotating table 50 that hold the DEP device 10 and rotate on the tilting surface (tilting plate 30) are shown in Fig. 8. To Figure 10 for a brief summary.
- the direction along the tilting plate 30 is the X-axis, and the direction perpendicular to the X-axis, along the surface of the tilting plate 30 is the Y-axis, and the direction perpendicular to the surface of the tilting plate 30 is This is the Z axis (the coordinate system is different from the X and Y axes of the XY table 41 described later).
- the tilt 0 pitch of the tilt plate 30 corresponds to the rotation of the Y axis.
- the rotation of the turntable 50 corresponds to the rotation 0 ya , about the Z axis. It can be understood that the above-described composite force C is canceled by rotating the turntable 50.
- the tilt angle 0 pitch of the tilting plate 30 is related to the magnitude of the dielectrophoretic force F DEP , and the rotation angle 0 ya of the turntable 50 represents the direction of the dielectrophoretic force F DEP .
- FIGS. 25 to 31 An example of a dielectrophoresis apparatus is shown in FIGS. 25 to 31.
- This dielectrophoretic device can also be used as a dielectrophoretic force measuring device, a dielectric particle handling device, and a sorting device.
- Various methods dielectrophoretic force measuring method, handling method) , Separation method can be realized.
- a tilting plate 30 is pivotally mounted on the base 20 at its lower end so as to be tiltable.
- a plurality of bearings 21 are fixed on the base 20.
- a plurality of mounting portions 32 having shaft holes are formed in the lower end portion of the tilting plate 30, and the rotary shaft 31 is inserted into and fixed to the shaft holes of these mounting portions 32.
- the rotating shaft 3 1 is rotatably received by the bearing 21.
- a rotating transducer (for example, a potentiometer) 25 for detecting the rotation angle of the rotating shaft 3 1 is attached to the base 20 (or one bearing 21).
- a fan-shaped positioning plate 22 is erected vertically on the surface of the base 20 on one side of the base 20.
- a positioning groove 23 is formed in an arc shape near the arc-shaped side edge of the positioning plate 22.
- a slide 33 in which a screw hole is formed slightly protrudes from one side of the tilting plate 30 at a position that moves along the positioning groove 23 as the tilting plate 30 tilts. Is provided.
- the fixing screw 24 with the knob 24 a enters the positioning groove 23 from the outside of the positioning plate 22 and is screwed into the screw hole of the slide 33.
- the tilting plate 30 forms the above-mentioned slope.
- 0 pi tc h is at least 0 ° to 90 °.
- the tilting plate 30 can be brought to an arbitrary angular position.
- the angle position (angle 0 pi tch ) of the tilting plate 30 can be obtained in the form of an electric signal by the output of the rotary transducer 25, and the angle scale 22A is engraved on the positioning plate 22. It is also possible to know by visual inspection.
- the angle 0 pi tch when the tilting plate 30 is in the horizontal state is 0 °.
- an XY table mechanism 40 is fixed on the tilting plate 30.
- the XY table mechanism 40 is provided with an XY table 41 at the top, and the position of the XY table 41 can be adjusted in the X and Y directions by means of knobs 42 and 43.
- the surface of the XY table 41 is parallel to the surface of the tilting plate 30.
- the X direction of the XY table mechanism 40 (with a coordinate system different from the X direction along the inclined surface shown in Fig. 1) is in the surface of the XY table 41, and the rotation axis 31 of the tilting plate 30 and The Y direction is parallel to the X direction on the surface of the XY table 41.
- a circular recess 44 is formed in the XY table 41, and a rotating disk 50 is rotatably inserted in the recess 44. That is, a rotating shaft 52 fixed to the lower center of the rotating disk 50 is rotatably received by a bearing 45 provided on the XY table 41.
- the XY table 41 is provided with a rotational transducer 46 for detecting the rotational angle position (angle ⁇ yaw ) of the rotary shaft 52 (that is, the rotary disk 50).
- the XY table 41 has a positioning groove 47 around the circular recess 44 over an angular range of 180 ° (—90 ° to + 90 °) or more.
- the positioning groove 47 is formed with a wide groove at the lower portion thereof, and a slide 55 is freely moved along the groove in the wide groove.
- a handle 53 extending radially outward is fixed to the periphery of the rotating disk 50.
- a hole is drilled in the handle 53, and a fixing screw 54 having a knob 54a passes through the hole.
- the tip of the fixing screw 54 is screwed into a screw hole ⁇ ⁇ formed in the slide 55.
- the turntable 50 can be rotated manually (within a range of at least 90 ° to 90 °) with the handle 53, and the knob 54a can be rotated at any angular position. It can be fixed to the XY table 41 by tightening the fixing screw 54.
- the angular position of the turntable 50 can be obtained from the rotary transducer 46 in the form of an electrical signal, and the scale 41A drawn (engraved) on the XY table 41 is used around the turntable 50. It can also be read visually.
- a recess 51 for accommodating the DEP device 10 is formed on the surface of the turntable 50.
- the recess 51 is sized to accommodate the DEP device 10 (the recess 51 may be larger than the DEP device 10), and the DEP device 10 contained in the recess 51 is , Does not move except for upward movement.
- the recess 51 is formed with a notch 51 A for attaching and detaching the DEP device 10 with tweezers or the like.
- a claw 58 made of a spring material is rotatably attached to the turntable 50 by pins at a plurality of locations around the recess 51. With this claw 52, the DEP device 10 housed in the recess 51 can be more firmly fixed. Furthermore, it is also possible to secure the DEP device 10 by inserting an adhesive between the bottom surface of the recess 51 and the backside of the DEP device 10.
- the surface of the turntable 50 and the bottom surface of the recess 51 are also parallel to the surface of the XY table 41 and the surface of the tilt plate 30. Therefore, the inclination angle of the DEP device 10 (substrate glass 11 and the surface of the insulating film 14) housed in the recess 51 is equal to the inclination angle of the tilting plate 30, and the tilt angle of the tilting plate 30 is measured.
- You can The center line N of the electrodes 13A and 13B of the DEP device 10 housed in the recess 51 is tilted when the angle position of the rotating disk 50 is 0 °. , It corresponds to the direction with the largest angle of the turntable 50 (Y direction of XY table 41).
- the center line The angle position of the rotating disk 50 when the direction of N coincides with the direction of the largest angle ( ⁇ ya J is assumed to be 0 °.
- a support 63 is fixed on the tilting plate 30, and the microscope 60 is supported by the support 63 in a vertical direction (a direction perpendicular to the surface of the tilting plate 30).
- the microscope 60 is arranged at a position where the particles P in the DEP device 10 set on the rotating disk 50 can be accommodated in the visual field (for example, near the rotation center of the rotating disk 50).
- the position of the field of view of the microscope 60 can be adjusted by moving the XY table 41 in the X and Y directions by the XY table mechanism 40.
- the force force of the field image of the microscope 60 can be adjusted with the knob 62.
- the microscope 60 also has a built-in CCD camera (indicated by reference numeral 61 in Fig. 31) to obtain a video signal representing the field image.
- the microscope 60 can be adjusted for magnification.
- FIG. 31 shows an outline of the electrical configuration of the dielectrophoresis apparatus.
- the processing device 70 is preferably composed of a computer system, and when measuring the electrophoretic force, etc., it accepts various inputs, which will be described later, and performs calculations according to the above-mentioned equations (1) to (4).
- the electrophoretic force F DEP and the electric field strength E are obtained and output to the output device 72 (printer, display device, etc.).
- the high-frequency generator 73 generates a high-frequency voltage applied between the electrodes 13 A and 13 B of the DEP device 10, and the frequency f and voltage V are variable.
- the generated high-frequency signal waveform can be observed with an oscilloscope 75.
- the frequency f and voltage V of the high frequency voltage are given to the processing device 70. It is also possible to visually read the frequency f and voltage V from the waveform display of the oscilloscope 75.
- the processor 70 may control the high frequency generator 73 to change the frequency f and the voltage V.
- the signals representing the tilt angle 0 pitch of the tilt plate 30 detected by the rotary transducers 25 and 45 and the angle position 0 ya of the rotary plate 50 are processed via the interface 76. _ _
- the video signal output from the CCD camera 61 of the microscope 60 mm is always displayed on the display device 74 (as a movie). If necessary, the video signal is converted to digital image data by the interface 76 and provided to the processing device 70.
- the processor 70 can be programmed to perform image processing on these digital image data and determine whether the particles P are stationary.
- Parameters necessary for the calculation of the dielectrophoretic force F DEP are input from the input device 71 to the processing device 70. If necessary, the dielectric constant ⁇ 0 of air, the relative dielectric constant im of the solution, and the relative dielectric constant £ p of the particles are also input from the input device 71.
- the processing unit 70 can control the microscope control unit 77 that adjusts the auto focus, magnification, etc. based on the input digital image data.
- Motors 78 and 79 are provided to adjust the angle ⁇ pitch of the tilting plate 30 and the angle 0 ya of the rotating plate 50 so that the particles P remain stationary under program control by the processing device 70. It can also be made. :
- the high frequency between the electrodes 13 A and 13B of the DEP device 10 Apply voltage (frequency f and voltage V are fixed values).
- the angle 0 pitch of the tilting plate 30 ie, DEP device 10.
- the direction of the dielectrophoretic force F DE p is not parallel to the center line N, but is inclined from the center line N. Since the heavy slope component force F c sin 0 pitch works in a direction parallel to the center line N, a combined force C of these forces is generated, and the particle P moves in the solution by this force C. The turntable 50 is rotated so that gravity works in the direction to cancel the resultant force C. Then, as shown in Fig. 7, the dielectrophoretic force F DEP and the gravitational slope component force F c sin0 pitch are balanced at an angular position of 0 ya , where the turntable 50 is located. At this time, Eq. (4) holds, and the dielectrophoretic force F DE p can be calculated from Eqs. (3) and (4).
- the particle P at an arbitrary position G can be moved to another desired position H and stopped at the position H in the space of the DEP device 10 mm.
- the base 20 may be tilted or rotated as described below).
- the dielectrophoretic force F DEP is a function of the dielectric constant (relative dielectric constant) and radius of the particle P, as is clear from Eqs. (1) and (2). 10), tilt angle 0 pitch , rotation angle 0 yaw) Adjust voltage, frequency, etc. (including tilting and rotation of base 20), move each particle in its own direction, and finally They can be separated as shown in Fig. 24.
- PI, P 2, and P 3 represent particles with different dielectric constants or diameters, and are positioned separately on the center line N.
- particle dielectrophoretic force F DEP measurement ranging from manual measurement to automatic control.
- the digital image data obtained from the CCD camera 61 is taken into the processing unit 70, and the movement (direction and size) of the particles P is judged in the processing unit 70 to determine the tilt of the tilting plate 30 and the rotation of the rotating plate 50. It is also possible to automatically control the particle P so that it is stationary, and the dielectrophoretic force F DEP can be calculated from the tilt angle of 0 pitch when it is stationary.
- the tilt angle may be fixed or changed at 0 pitch .
- Change the angle ⁇ ya the particle P can be moved or stopped.
- Fig. 11a, Fig. Lib, Fig. 12, Fig. 13a, Fig. 13b and Fig. 14 show the angular step response.
- the angle step response is obtained by changing the angle 0 pitch rapidly from 60 ° to 15 ° (B force, et al. A) and 15 ° to 60 ° (A to B) under the conditions of applied voltage 20Vpp and frequency 1MHz. (See Fig. 12, Fig. 13a, Fig. 13b) and obtained by measuring the amount of particle movement.
- the time constant for moving from point B to point A is 3 minutes, and the time constant from point A to point B is 1.9 minutes (see Figure 11a and Figure lib).
- Fig. 14 shows the change (movement amount) of the stationary position of particles in a state where the gravitational slope component force and the dielectrophoretic force are balanced, with the angle 0 pitch being the horizontal axis. It can be seen that the greater the angle 0, the more balanced it is at the lower position.
- the time constant for moving from point D to point C is 1 minute, and the time constant for moving from point C to point D is 5 minutes (see Figures 15a and 15b).
- the resting position moves upward as the voltage increases (see Figure 18).
- the time constant in each step response is within a few minutes, and the steady state is reached in about 15 minutes.
- the response speed can withstand practical use.
- Figures 32 and 33 show examples of DEP devices that can further expand the particle handling range.
- the DEP device 10A has a solution inlet 17A at the center of one end and a solution outlet 18A at the center of the other end. These inlet 17A and outlet 18A are connected to solution inlet pipe (tube) 17 and outlet pipe (tube) 18, respectively. These pipes 17, 18 are connected to valves (cocks or valves) 17B, 18B is provided. In addition, a particle suction port (trap port) 16A is opened in the bottom plate at the center line between the electrodes or at another appropriate position, and the suction tube (tube 16) is connected to this.
- This DEP device 10 A can be used, for example, to separate only one particle from a large number of particles (small target objects) introduced into the DEP device. That is, as described above, a specific particle is attracted by adjusting the angle 0 pilch or ⁇ ya » or the voltage or frequency (or the tilt or rotation of the base 20 if necessary). Move it to the position of the mouth 16 A and suck the particles to the suction pipe 16 (or an appropriate trough connected to it). Evacuate). In this state, the solution is injected through the injection tube 17 from the injection port 17A into the D ⁇ ⁇ device 10A, and other particles existing inside the device are discharged through the discharge port 18A and the discharge tube 18.
- Valves 17B and 18 ⁇ ⁇ are opened and closed during the injection and discharge of the solution during the above fractionation procedure.
- a valve is also provided in the suction pipe 16 as necessary.
- the particles can be moved faster by tilting the DEP device, rotating it abruptly, or changing the voltage and frequency rapidly.
- rotation about the X axis when moving if necessary rotate the base 20 itself (by rotating the X axis shown in FIG. 10 at an angle 0 pitch opposite directions on a base 20 0 r. u) It may be possible to rotate the base 20 in the 0 pitch direction.
- a rotating disk is used to place and rotate the D E P device, but it is not necessarily a disk.
- a rotating body having a shape suitable for the support may be used.
- a tilting body that is not plate-shaped can be used instead of the tilting plate of the tilting mechanism.
- a pair of electrodes 13A and 13B are used in the above embodiment, a plurality of pairs of electrodes may be arranged vertically (vertically) at intervals or horizontally. . When arranged horizontally, two adjacent electrodes (belonging to different pairs) may be integrated.
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- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Electrochemistry (AREA)
- Engineering & Computer Science (AREA)
- Environmental & Geological Engineering (AREA)
- Microelectronics & Electronic Packaging (AREA)
- General Health & Medical Sciences (AREA)
- Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
- Electrostatic Separation (AREA)
- Electrochromic Elements, Electrophoresis, Or Variable Reflection Or Absorption Elements (AREA)
Abstract
Description
Claims
Priority Applications (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020107007173A KR101162434B1 (ko) | 2007-10-05 | 2008-09-26 | 유전 영동 장치 |
| JP2009536128A JP5120968B2 (ja) | 2007-10-05 | 2008-09-26 | 誘電泳動装置および方法 |
| CN2008801103224A CN101820978B (zh) | 2007-10-05 | 2008-09-26 | 介电电泳装置及方法 |
| US12/681,616 US8864972B2 (en) | 2007-10-05 | 2008-09-26 | Dielectrophoresis apparatus and method |
| EP08835862A EP2204228A1 (en) | 2007-10-05 | 2008-09-26 | Dielectrophoresis device and method |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2007262058 | 2007-10-05 | ||
| JP2007-262058 | 2007-10-05 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2009044902A1 true WO2009044902A1 (ja) | 2009-04-09 |
Family
ID=40526317
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/JP2008/068114 Ceased WO2009044902A1 (ja) | 2007-10-05 | 2008-09-26 | 誘電泳動装置および方法 |
Country Status (6)
| Country | Link |
|---|---|
| US (1) | US8864972B2 (ja) |
| EP (1) | EP2204228A1 (ja) |
| JP (1) | JP5120968B2 (ja) |
| KR (1) | KR101162434B1 (ja) |
| CN (1) | CN101820978B (ja) |
| WO (1) | WO2009044902A1 (ja) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100116665A1 (en) * | 2008-11-13 | 2010-05-13 | University Industry Cooperation Foundation Korea Aerospace University | System and method for high throughput particle separation |
| WO2021157060A1 (ja) * | 2020-02-07 | 2021-08-12 | 日本電信電話株式会社 | 粒子配列運搬デバイスおよび粒子配列運搬方法 |
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| US20150306599A1 (en) | 2014-04-25 | 2015-10-29 | Berkeley Lights, Inc. | Providing DEP Manipulation Devices And Controllable Electrowetting Devices In The Same Microfluidic Apparatus |
| US11192107B2 (en) | 2014-04-25 | 2021-12-07 | Berkeley Lights, Inc. | DEP force control and electrowetting control in different sections of the same microfluidic apparatus |
| US20150306598A1 (en) * | 2014-04-25 | 2015-10-29 | Berkeley Lights, Inc. | DEP Force Control And Electrowetting Control In Different Sections Of The Same Microfluidic Apparatus |
| CN104165079B (zh) * | 2014-08-22 | 2017-11-21 | 阮海生 | 一种汽车尾气净化系统 |
| US9744533B2 (en) | 2014-12-10 | 2017-08-29 | Berkeley Lights, Inc. | Movement and selection of micro-objects in a microfluidic apparatus |
| AU2016250689B2 (en) | 2015-04-22 | 2021-07-08 | Berkeley Lights, Inc. | Microfluidic cell culture |
| US10799865B2 (en) | 2015-10-27 | 2020-10-13 | Berkeley Lights, Inc. | Microfluidic apparatus having an optimized electrowetting surface and related systems and methods |
| AU2017271673B2 (en) | 2016-05-26 | 2022-04-14 | Berkeley Lights, Inc. | Covalently modified surfaces, kits, and methods of preparation and use |
| DE102018105931A1 (de) * | 2018-03-14 | 2019-09-19 | Technische Universität Darmstadt | Mikrowellenporator |
| CN111973737B (zh) * | 2019-05-23 | 2022-08-26 | 比欧泰克生物技术服务(北京)有限公司 | 一种荚膜多糖疫苗蛋白核酸去除装置及去除工艺 |
| EP4640818A1 (de) * | 2024-04-24 | 2025-10-29 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Vorrichtung, system und verfahren zum charakterisieren, separieren, vermehren und/oder kryokonservieren mindestens einer biologischen zelle |
| EP4640819A1 (de) * | 2024-04-24 | 2025-10-29 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Vorrichtung, system und verfahren zum charakterisieren, separieren, vermehren und/oder kryokonservieren mindestens einer bio-logischen zelle |
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- 2008-09-26 US US12/681,616 patent/US8864972B2/en active Active
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- 2008-09-26 KR KR1020107007173A patent/KR101162434B1/ko not_active Expired - Fee Related
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Also Published As
| Publication number | Publication date |
|---|---|
| CN101820978A (zh) | 2010-09-01 |
| US20100219076A1 (en) | 2010-09-02 |
| US8864972B2 (en) | 2014-10-21 |
| JP5120968B2 (ja) | 2013-01-16 |
| CN101820978B (zh) | 2013-01-09 |
| KR20100051865A (ko) | 2010-05-18 |
| EP2204228A1 (en) | 2010-07-07 |
| JPWO2009044902A1 (ja) | 2011-02-17 |
| KR101162434B1 (ko) | 2012-07-04 |
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