JP2011000079A - Method for operating particle and micro fluid device - Google Patents

Abstract

A method for reliably manipulating particles under mild conditions is provided.
In the method according to the present invention, particles to be operated are captured in a plurality of microchambers each having at least one of an electrolysis anode and cathode (S1), and the plurality of micros By selectively applying a voltage to the electrodes of some of the microchambers in the chamber, water is electrolyzed to generate bubbles (S2), and the actuators that operate as the bubbles are generated are used to generate the bubbles. In this method, the particles are selectively extruded from the microchamber (S3).
[Selection] Figure 1

Description

  The present invention relates to a method for manipulating particles and a microfluidic device, and more particularly to a method and device for manipulating particles using the generation of bubbles by electrolysis.

  Microfluidic systems that analyze a large number of cells at high speed and analyze them with high sensitivity have played an important role as analytical tools in the field of basic science in addition to industrial applications such as medical diagnosis and new drug development.

  As a further developed microfluidic system, for example, after analyzing a large number of arrayed cells, in order to analyze in detail the genes and proteins of the cells that showed a specific reaction, Development of a system having a function of selectively extracting specific cells from the cell is desired.

  Therefore, conventionally, for example, in Non-Patent Document 1, one microbead is captured in each of a plurality of microchambers in which aluminum patches are arranged, and the aluminum patch in a specific microchamber is irradiated with a laser to locally distribute the solution. A technique has been proposed in which bubbles are generated by boiling and the microbeads are selectively released from the specific microchamber.

  For example, Non-Patent Document 2 proposes a technique for taking out microbeads and cells from a microwell by electrophoretic force.

Wei-Heong Tan and Shoji Takeuchi (2007) Proc Natl Acad Sci USA, 104,1146-1151. Ching-Yu Chang et al. (2009) Lab Chip, 9, 1185-1192.

  However, in the technique described in Non-Patent Document 1, for example, (1) there is a possibility of damaging cells because the solution is partially boiled, (2) an apparatus for generating a laser is expensive, (3) There is a problem that high technical skill is required to accurately irradiate the target microchamber with the laser.

  Further, the technique described in Non-Patent Document 2 has a problem in that relatively large cells such as a fertilized egg cannot be reliably manipulated because the electrophoretic force is small.

  The present invention has been made in view of the above problems, and an object thereof is to provide a method and a microfluidic device for reliably manipulating particles under mild conditions.

  A method according to an embodiment of the present invention for solving the above problem is to capture particles to be manipulated in a plurality of microchambers each having at least one of an electrode for electrolysis and a cathode, and By applying a voltage to the electrodes of some of the plurality of microchambers, water is electrolyzed to generate bubbles, and an actuator that operates along with the generation of the bubbles causes the some microchambers And selectively extruding the particles. According to the present invention, it is possible to provide a method for reliably manipulating particles under mild conditions.

  Moreover, the said actuator is good also as being the liquid in the said micro chamber which flows with the generation | occurrence | production of the said bubble which floats, or the said bubble. The actuator may be a stretchable film that is provided between the electrode and the particle in the micro chamber and deforms so as to push out the particle as the bubble is generated.

  The particles are cells, and the cells should be captured and cultured in each of the plurality of microchambers, and a part of the plurality of cells cultured in the plurality of microchambers should be collected. A cell is determined, and by applying a voltage to the electrodes of a part of the plurality of microchambers in which the cells to be collected are captured, water is electrolyzed to generate bubbles, and the bubbles are generated. The cells may be selectively pushed out from the partial microchambers by the actuator that operates in accordance with the occurrence of In this case, the cell is a fertilized egg, and the fertilized egg selectively pushed out from the partial microchamber may be recovered in a live state.

  A microfluidic device according to an embodiment of the present invention for solving the above-described problem includes a microfluidic device including a plurality of microchambers for capturing particles and a common channel in which the plurality of microchambers are open. The apparatus is characterized in that at least one of an anode for electrolysis and a cathode is disposed in each of the plurality of micro chambers. According to the present invention, it is possible to provide a microfluidic device that can reliably manipulate particles under mild conditions.

  A first member provided with the plurality of micro-chambers; and a second member laminated on the first member. Each of the plurality of micro-chambers includes one of the anode and the cathode. An electrode may be disposed, and the other electrode of the anode and the cathode may be disposed on the second member. Further, both of the anode and the cathode may be disposed in each of the plurality of micro chambers.

  Each of the plurality of microchambers includes a storage chamber that opens to the common flow path and stores the particles, and an electrolysis chamber that is provided in communication with the storage chamber and in which the electrode is disposed. It is good as well. Each of the plurality of microchambers includes a storage chamber that opens in the common flow path and stores the particles, an electrolysis chamber in which the electrodes are disposed, and a space between the storage chamber and the electrolysis chamber. And a stretchable film that is deformed so as to extrude the particles from the storage chamber as bubbles are generated by electrolysis of water in the electrolysis chamber.

  According to the present invention, it is possible to provide a method and a microfluidic device for reliably manipulating particles under mild conditions.

It is explanatory drawing which shows the main processes included in the method which concerns on one Embodiment of this invention. It is a top view about an example of the microfluidic device concerning one embodiment of the present invention. It is sectional drawing of the microfluidic device cut | disconnected by the III-III line | wire shown in FIG. It is explanatory drawing which shows an example of the process of forming the microchamber which concerns on one Embodiment of this invention. It is explanatory drawing which shows each process in the method using the microfluidic device shown in FIG.2 and FIG.3. It is a partial top view of the other example of the microfluidic device concerning one embodiment of the present invention. It is sectional drawing of the microfluidic device cut | disconnected by the VII-VII line shown in FIG. It is a partial top view of the other example of the microfluidic device concerning one embodiment of the present invention. It is sectional drawing of the microfluidic device cut | disconnected by the IX-IX line shown in FIG. It is a partial sectional view of other examples of the microfluidic device concerning one embodiment of the present invention. It is explanatory drawing which shows each process in the method using the microfluidic device shown in FIG. It is a partial sectional view of other examples of the microfluidic device concerning one embodiment of the present invention. It is explanatory drawing which shows each process in the method using the microfluidic device shown in FIG. It is a partial sectional view of other examples of the microfluidic device concerning one embodiment of the present invention. It is explanatory drawing which shows each process in the method using the microfluidic device shown in FIG. It is a partial top view of the other example of the microfluidic device concerning one embodiment of the present invention. It is sectional drawing of the microfluidic device cut | disconnected by the XVII-XVII line | wire shown in FIG. It is a partial sectional view of other examples of the microfluidic device concerning one embodiment of the present invention. It is explanatory drawing which shows each process in the method using the microfluidic device shown in FIG. It is a partial top view of the other example of the microfluidic device concerning one embodiment of the present invention. It is explanatory drawing which shows each process in the method using the microfluidic device shown in FIG. It is explanatory drawing which shows an example of the microscope picture of the microfluidic device which concerns on one Embodiment of this invention. It is explanatory drawing which shows the other example of the microscope picture of the microfluidic device which concerns on one Embodiment of this invention. It is explanatory drawing which shows an example of the time lapse video image which image | photographed selective release | release of the microbead in the microfluidic device which concerns on one Embodiment of this invention. It is explanatory drawing which shows an example of the result of having evaluated the influence which generation | occurrence | production of the bubble by electrolysis has on the survival rate of a cell in the microfluidic device which concerns on one Embodiment of this invention.

  Hereinafter, an embodiment of the present invention will be described. Note that the present invention is not limited to this embodiment.

  First, a method according to the present embodiment (hereinafter referred to as “the present method”) will be described. FIG. 1 is an explanatory view showing main steps included in the present method.

  As shown in FIG. 1, in this method, particles to be manipulated are captured in a plurality of microchambers each having at least one of an anode for electrolysis and a cathode (step S1). By applying a voltage to the electrodes of some of the microchambers, water is electrolyzed to generate bubbles (step S2), and the actuators that operate as the bubbles are generated are used to generate the bubbles. The particles are selectively extruded from the microchamber (step S3).

  That is, in the first step S1, first, a plurality of microchambers having electrodes for electrolysis are prepared. The microchamber will be described in detail later, but each of the plurality of microchambers has an electrode for electrolysis capable of applying a voltage independently of each other.

  Next, particles to be operated (hereinafter referred to as “target particles”) are placed in each microchamber. Here, the target particle is not particularly limited as long as it is a particle that can be carried on a liquid flow, and for example, a cell or a microbead can be used.

  The kind of cell is not particularly limited, and for example, a human or non-human animal cell can be preferably used. More specifically, for example, various differentiated cells and undifferentiated cells (stem cells, ES cells, iPS cells, etc.) derived from humans or non-human animals, and fertilized eggs can also be used. As the cell, a single cell can be used, and a cell tissue formed by assembling a plurality of cells can also be used.

  As the microbeads, for example, resin beads or gel beads can be used. In addition, microcapsules formed by coating cells or cell tissues with a gel can also be used. In addition, although spherical bodies, such as these cells and a microbead, can be used preferably as object particle | grains, the shape is not restricted to spherical shape.

  The size of the target particle is not particularly limited as long as it is in a range that can be suspended in a liquid and extruded from the microchamber, but when the target particle is a spherical body, its diameter is, for example, 1 μm or more. And preferably in the range of several tens to several hundreds of μm.

  In the subsequent second step S2, a voltage is selectively applied to the electrodes of some of the microchambers in which the target particles are captured, and water is electrolyzed in the some of the microchambers. To generate bubbles.

  In other words, bubbles are generated from the electrodes in the partial microchambers by connecting only the electrodes of the selected partial microchambers to the power source and energizing them. Note that hydrogen gas is generated at the cathode and oxygen gas is generated at the anode by electrolysis of water.

  The magnitude of the voltage to be applied is not particularly limited as long as water can be electrolyzed to generate bubbles, but when cells are used as target particles, the survival of the cells can be appropriately maintained. A relatively small range is preferable.

  Specifically, for example, it can be in the range of 3 to 10 V, and is preferably in the range of 4 to 6 V. However, the magnitude of the voltage to be applied is not limited to this, and can be appropriately set according to conditions such as the structure of the microfluidic device used in the present method and the composition of the aqueous solution to be electrolyzed.

  In the third step S3, the target particles are selectively released from some of the microchambers by an actuator that operates in association with the generation of bubbles. The actuator will be described in detail later. For example, the actuator can be a floating bubble generated by electrolysis or a liquid in a microchamber that flows along with the generation of the bubble.

  That is, in this method, for example, by using bubbles generated and floating in the micro chamber as an actuator, the target particles can be pushed out of the micro chamber by the bubbles. In the present method, for example, by using a liquid that flows as bubbles are generated in the microchamber as an actuator, the target particles can be pushed out of the microchamber by being put on the flow of the liquid.

  In addition, the actuator in the present method may be, for example, a stretchable film that is provided between the electrode and the target particle in the microchamber and deforms so as to push out the target particle as bubbles are generated. Although this membrane will be described in detail later, in this method, a membrane that is deformed so as to push out the target particles from the microchamber by the pressure accompanying the generation of bubbles can also be used as an actuator.

  According to such a method, selective release of target particles from a specific microchamber among a plurality of microchambers can be reliably realized under mild conditions.

  That is, since this method uses the generation of bubbles by electrolysis, for example, (1) the generated heat is very small and the damage to the target particles (particularly cells) is small, and (2) a special apparatus such as a laser irradiation apparatus Highly versatile because it is not necessary, (3) The advantage that bubbles can be generated easily and reliably only in the targeted micro-chamber by arranging and wiring electrodes using general-purpose electronics technology Have

  Moreover, since this method uses the pressure accompanying generation | occurrence | production of a bubble, even if it is a target particle which cannot be manipulated with an electrophoretic force, it also has the advantage that it can be manipulated reliably.

  Therefore, in this method, for example, a relatively large cell that is sensitive to environmental conditions and susceptible to damage, such as a fertilized egg of a mammal, can be efficiently and reliably operated intact.

  Specifically, for example, in the field of livestock, the development of livestock techniques is being promoted in which fertilized eggs of Japanese beef are placed in Holstein womb and the Holsteins are born. In this case, the fertilized egg is cultured in vitro until a blastocyst is formed, and then the fertilized egg that has become a blastocyst is transplanted into the womb of the mother's womb.

  In general, a plurality of fertilized eggs are prepared and cultured, and among the plurality of fertilized eggs that have formed blastocysts, those in good condition suitable for transplantation are selectively collected and used for actual transplantation. Whether or not a fertilized egg is in a good state suitable for transplantation can be determined visually by its size and shape in observation under a microscope. On the other hand, fertilized eggs are sensitive to environmental conditions and susceptible to damage, and therefore selective recovery must be performed under mild conditions.

  Therefore, according to the present method having the advantages as described above, a fertilized egg is cultured in each of a plurality of microchambers until becoming a blastocyst, and then transplanted among a plurality of fertilized eggs that have formed a blastocyst. A specific fertilized egg determined to be suitable can be selectively recovered in a good state.

  That is, in this case, in this method, one fertilized egg is captured and cultured in each of the plurality of microchambers (step S1), and one of the plurality of fertilized eggs cultured in the plurality of microchambers. The portion is determined as a fertilized egg to be recovered, and water is electrolyzed by applying voltage to the electrodes of some of the microchambers in which the fertilized eggs to be recovered are captured. (Step S2), the actuator that operates in association with the generation of the bubble selectively pushes out the fertilized egg from the partial microchamber, and collects the extruded fertilized egg in a live state (step) S3).

  Specifically, in the first step S1, a plurality of fertilized eggs and a plurality of microchambers are prepared, and the fertilized eggs are put into each of the plurality of microchambers and cultured. By this culture, blastocysts are formed from each fertilized egg.

  In the second step S2, first, the form of a fertilized egg in which a blastocyst is formed in each microchamber is observed, and a fertilized egg to be recovered is determined from a plurality of fertilized eggs based on the result of the observation. .

  Next, by selectively applying a voltage to an electrode of a specific microchamber in which the fertilized egg to be collected is cultured, electrolysis is performed in the specific microchamber to generate bubbles.

  Then, in the third step S3, the target fertilized egg is selectively selected from the specific microchamber by the flow of bubbles generated by electrolysis or the culture solution in the specific microchamber accompanying the generation of the bubbles. To release. Thereafter, the released fertilized eggs are collected and used for transplantation.

  As described above, according to the present method, after fertilized eggs of a large amount of Japanese beef are arrayed and cultured using a microchamber, only highly fertilized eggs of the large amount of fertilized eggs are selectively recovered intact. be able to.

  That is, it is possible to realize a microfluidic system that selectively collects only preferred fertilized eggs from a large amount of fertilized eggs at high speed. In such a microfluidic system, it is also possible to automate the culture and selective recovery of fertilized eggs. In this method, any one kind or two or more kinds of cells as described above can be used instead of a fertilized egg. In other words, a single cell can be used as the cell, and a cell tissue (cell mass) formed by aggregating a plurality of cells can also be used. In addition, a microcapsule that encloses a single cell or cell tissue can also be used. In addition, when cells are used as target particles including fertilized eggs, cells pushed out from the microchamber can be collected, but the cells do not necessarily need to be collected in a living state. Even if it is not recovered in a living state, it is possible to analyze characteristics such as expression of cellular genes and proteins.

  Next, the microfluidic device (hereinafter referred to as “the present device”) according to the present embodiment will be described. In addition, although this method mentioned above can be implemented preferably by using this apparatus demonstrated below, it is not restricted to this, It can implement also using another apparatus.

  The apparatus includes a plurality of microchambers for capturing target particles, and a common flow path in which the plurality of microchambers are opened. Each of the plurality of microchambers includes an anode and a cathode for electrolysis. At least one of the electrodes is disposed. That is, in this apparatus, each of the plurality of micro chambers has an electrode for electrolysis that can apply a voltage independently of each other.

  FIG. 2 is a plan view of an example of the apparatus 1. FIG. 3 is a cross-sectional view of the device 1 taken along line III-III shown in FIG.

  As shown in FIGS. 2 and 3, the apparatus 1 according to this example is laminated on the first member 20 provided with a plurality of (9 in FIG. 2) microchambers 10 and the first member 20. Each of the plurality of microchambers 10 is provided with one electrode 40 of an anode and a cathode for electrolysis, and the second member 30 includes the anode and the cathode. Of these, the other electrode 41 is arranged.

  In addition, in FIG. 2, the top view of this apparatus 1 of the state which removed the 2nd member 30 is shown for convenience of explanation. Further, in the following description, when the apparatus 1 has a plurality of similar portions, the symbols of the plurality of portions may be distinguished by attaching a lower case alphabet (for example, micro chambers 10a, 10b, If 10c) does not need to distinguish between them, the lower case alphabet is omitted.

  As shown in FIG. 3, the first member 20 has a common flow path between the first substrate 21, the coating layer 22 laminated on the first substrate 21, and the first member 20 and the second member 30. And a spacer 23 for forming 24.

  All of the first substrate 21, the coating layer 22, and the spacer 23 are formed of a non-conductive material. The non-conductive material is not particularly limited, and for example, glass or non-conductive resin can be preferably used. That is, for example, the first substrate 21 can be a glass substrate, the covering layer 22 can be a photoresist layer made of a non-conductive resin, and the spacer 23 can be a silicone rubber molded body.

  The microchamber 10 is formed as a hole with a bottom opening on the surface of the coating layer 22. FIG. 3 shows an example in which the first member 20 is a laminated body having the first substrate 21 and the covering layer 22. For example, the first member 20 includes a single non-conductive substrate. In this case, the microchamber 10 is formed to open on the surface of the non-conductive substrate.

  That is, the microchamber 10 is not limited to the example shown in FIG. 3 and other drawings as long as it opens on the surface of the non-conductive first member 20. When cells are used as the target particles, the inner surface of the microchamber 10 is preferably a so-called non-cell-adhesive surface to which the cells do not adhere.

  2 and 3 show an example in which the bottom and the opening of the microchamber 10 are circular, but the shape of the bottom and the opening is not limited to this, for example, an oval or polygonal shape. You can also

  The size of the microchamber 10 can be arbitrarily set according to the size of the target particles to be captured. For example, the microchamber 10 can have an appropriate size for capturing the target particles one by one.

Specifically, the bottom area and the opening area of the micro-chamber 10, for example, can range from 25～90000Myuemu ^2, preferably in the range of 100-400 ^2. That is, when the microchamber 10 is a circular bottomed hole, the diameters of the bottom and the opening can be, for example, in the range of 5 to 300 μm, and preferably in the range of 80 to 250 μm. it can. Moreover, the depth of the micro chamber 10 can be made into the range of 5-300 micrometers, for example, Preferably it can be set as the range of 80-250 micrometers.

  When the size of the microchamber 10 is too large compared to the size of the target particle, two or more target particles enter one microchamber 10. In this case, the bubbles generated in the microchamber 10 easily pass through the inner surface of the microchamber 10 and the captured target particles and leak out of the microchamber 10. It may not be used effectively as an actuator. Therefore, for example, when using a fertilized egg of a human or a non-human mammal as the target particle, in order to capture the fertilized egg one by one in each microchamber 10, the diameter of the microchamber 10 is, for example, 80 It is preferable to set it as the range of -250 micrometers, and it is preferable that the depth shall be the range of 80-250 micrometers, for example.

  One electrode 40 for electrolysis is arranged on the bottom of the microchamber 10 one by one. In the example shown in FIGS. 2 and 3, the entire bottom portion of the microchamber 10 is the electrode 40.

  The plurality of electrodes 40 formed in the apparatus 1 are formed so that voltages for electrolysis can be applied independently of each other. That is, the plurality of electrodes 40 can be connected to a power source independently of each other. Specifically, in the example shown in FIG. 2, the three electrodes 40a, 40b, and 40c are disposed in the three micro chambers 10a, 10b, and 10c, respectively, and are electrically connected to each other. 42a, 42b and 42c are connected to each other.

  Therefore, for example, by connecting only one wiring 42a of the three wirings 42a, 42b, and 42c to the power source, a voltage can be selectively applied only to one electrode 40a connected to the wiring 42a. . In this case, water can be electrolyzed only in one microchamber 10a having an electrode 40a to which a voltage is applied to generate bubbles.

  The electrode 40 and the wiring 42 are formed from a conductive material. The conductive material is not particularly limited, and for example, ITO (Indium Oxide Tin), gold, platinum, and aluminum can be used. In particular, ITO that can easily and reliably form the transparent electrode 40 and the wiring 42 is preferably used. Can do.

  The microchamber 10 having such an electrode 40 can be formed using, for example, a known semiconductor manufacturing technique. FIG. 4 is an explanatory diagram showing an example of a process for forming the microchamber 10.

  First, as shown in FIG. 4A, a conductive film 27 made of ITO is patterned on a first substrate 21 made of a glass substrate, and electrodes 40a, 40b, 40c (see FIG. 4B) of the conductive film 27 are further patterned. The positive photoresists 28a, 28b, and 28c are patterned in regions corresponding to each of the above.

  Next, as shown in FIG. 4B, the conductive film 27 is etched using a predetermined etchant to form electrodes 40a, 40b, and 40c. Further, as shown in FIG. 4C, a non-conductive layer 29 made of a negative photoresist is coated on the electrodes 40a, 40b, and 40c.

  Then, as shown in FIG. 4D, microchambers 10a, 10b, and 10c are pattern-formed at positions corresponding to the electrodes 40a, 40b, and 40c in the non-conductive layer 29. In this way, the microchambers 10a, 10b, and 10c having openings on the surface of the non-conductive covering layer 22 laminated on the first substrate 21 and having the electrodes 40a, 40b, and 40c at the bottom can be formed.

  On the other hand, the second member 30 includes a second substrate 31 and an electrode 41 for electrolysis formed on the surface of the second substrate 31. The second substrate 31 is made of a nonconductive material. The non-conductive material is not particularly limited, and for example, glass or non-conductive resin can be preferably used. That is, for example, the second substrate 31 can be a glass substrate.

  In the example shown in FIG. 3, the electrode 41 is formed at a position facing the electrodes 40 of the plurality of microchambers 10 via the common flow path 24. That is, the apparatus 1 includes a plurality of electrodes 40 disposed on the bottom surfaces of the plurality of microchambers 10 and a common electrode 41 disposed to face the plurality of electrodes 40.

  The electrode 41 is formed from a conductive material. The conductive material is not particularly limited, and for example, ITO, gold, platinum, and aluminum can be used. In particular, ITO that can easily and reliably form the transparent electrode 41 can be preferably used.

  And this apparatus 1 is comprised by bonding these 1st member 20 and the 2nd member 30 together. The common flow path 24 is formed as a gap between the plurality of micro chambers 10 and the second member 30.

  In the example shown in FIG. 2, the apparatus 1 includes an inlet 25 for allowing liquid to flow into the interior and an outlet 26 for allowing liquid to flow out from the interior. That is, in the present apparatus 1, the liquid can be caused to flow into the common channel 24 and the microchamber 10 from the inlet 25, and the liquid in the common channel 24 and the microchamber 10 is allowed to flow out from the outlet 26. be able to. FIG. 2 shows an example in which the inflow port 25 and the outflow port 26 open at the longitudinal ends of the apparatus 1. However, the present invention is not limited to this example. For example, the inflow port 25 and the outflow port 26 At least one of them can be provided so as to open on the outer surface of the second member 30.

  FIG. 5 is an explanatory diagram showing each step in the case of carrying out the method using the apparatus 1 shown in FIGS. 2 and 3. In this case, in the first step S1 of the present method, the target particles 50 are captured in each of the plurality of microchambers 10, as shown in FIG. 5A.

  That is, for example, the aqueous solution in which the target particles 50 are dispersed is injected into the common channel 24 from the inlet 25 (see FIG. 2) of the apparatus 1. As a result, in each microchamber 10, the target particle 50 is placed on the electrode 40 which is the bottom.

  Each microchamber 10 and common channel 24 are filled with an aqueous solution. 5 shows an example in which one target particle 50 is captured in one microchamber 10. However, the present invention is not limited to this, and a plurality of target particles 50 can also be captured in one microchamber 10. In this case, the microchamber 10 is formed in a size that can accommodate a plurality of target particles 50.

  Next, in the second step S2, as shown in FIG. 5B, water is discharged by selectively applying a voltage to the electrode 40a of one microchamber 10a among the plurality of microchambers 10a, 10b, 10c. Decomposes to generate bubbles 60.

  That is, the target electrode 40a of one microchamber 10a and the electrode 41 of the second member 30 facing the electrode 40a are connected to a power source to serve as an anode and a cathode, and the aqueous solution in the microchamber 10a is electrically A voltage of a magnitude that can be decomposed to generate bubbles 60 is applied.

  As a result, it is possible to generate bubbles 60 by electrolysis of the aqueous solution only in one target micro chamber 10a among the plurality of micro chambers 10a, 10b, and 10c.

  In the third step S3, as shown in FIG. 5B, at least one of the bubbles 60 generated by electrolysis and the aqueous solution in the microchamber 10a that flows along with the generation of the bubbles 60 is used as an actuator. The target particle 50a is extruded from the microchamber 10a.

  That is, the target particle 50a captured in one target microchamber 10a is an aqueous solution that leaks out of the microchamber 10a due to the bubble 60 generated on the electrode 40a and floating. Is pushed out to the common flow path 24 by the flow of. Normally, both the bubble 60 and the aqueous solution flow act on the target particle 50a.

  Thereafter, by passing a new aqueous solution through the common channel 24, one target particle 50a released from one target micro chamber 10a can be selectively recovered. On the other hand, as shown in FIG. 5C, in the other microchambers 10b and 10c where the electrolysis was not performed, the target particles 50b and 50c can be maintained in a captured state.

  Thus, in this apparatus 1, since the electrode 40 is disposed behind the position in the microchamber 10 where the target particle 50 is captured and held, by the generation of bubbles on the electrode 40, The target particles 50 can be reliably pushed out from the microchamber 10 into the common flow path 24.

  FIG. 6 is a plan view of one peripheral portion of a plurality of microchambers 10 in another example of the apparatus 1. FIG. 7 is a cross-sectional view of the device 1 taken along the line VII-VII shown in FIG.

  As shown in FIGS. 6 and 7, the device 1 according to this example includes a first member 20 provided with a plurality of microchambers 10 and a second member 30 stacked on the first member 20. Each of the plurality of micro chambers 10 is provided with one electrode 40 of an anode and a cathode for electrolysis, and the second member 30 is provided with the other electrode 41 of the anode and the cathode. Has been. That is, the electrode 40 is disposed at the bottom of the microchamber 10.

  On the other hand, the electrode 41 of the second member 30 is not disposed at a position facing the electrode 40 of the microchamber 10 but is disposed at a position shifted from the facing position. That is, in this apparatus 1 according to this example, the electrode 41 is disposed at a position shifted from the position facing the electrodes 40 disposed in the plurality of microchambers 10 in the second member 30. In this case, the degree of freedom of wiring in the device 1 is relatively high.

  FIG. 8 is a plan view of one peripheral portion of a plurality of microchambers 10 in another example of the apparatus 1. FIG. 9 is a cross-sectional view of the device 1 taken along the line IX-IX shown in FIG.

  In this apparatus 1 according to this example, as shown in FIGS. 8 and 9, both the anode and cathode electrodes 40 and 41 for electrolysis are arranged in each of the plurality of microchambers 10. That is, one electrode 40 of the anode and the cathode is disposed on a part of the bottom of the microchamber 10, and the other electrode 41 of the anode and the cathode is disposed on the other part of the bottom. . Therefore, as shown in FIG. 9, the second member 30 is not provided with electrodes.

  In this apparatus 1, the microchamber is applied by selectively connecting one electrode 40 and the other electrode 41 arranged in one target microchamber 10 to a power source and applying a voltage. In 10, water can be electrolyzed to generate bubbles.

  In this case, since the electrodes 40 and 41 for performing electrolysis are all disposed in one microchamber 10, electrolysis can be performed only in the microchamber 10. In particular, as shown in FIG. 9, since both electrodes 40 and 41 are disposed at the bottom of the microchamber 10, an electrolysis reaction can be performed only in the vicinity of the bottom. Therefore, it is easy to control the generated bubbles.

  In addition, since the pair of electrodes 40 and 41 necessary for electrolysis can be disposed close to each other, the applied voltage can be effectively reduced. Further, the structure of the second member 30 can be simplified, and the assembly of the apparatus 1 is facilitated.

  In the example shown in FIGS. 8 and 9, one electrode 40 (for example, the anode) is formed so as to cover the central portion of the bottom of the microchamber 10. That is, the area of one electrode 40 is enlarged and disposed in the central portion of the microchamber 10. Therefore, it is possible to reliably push out the target particles using the generation of bubbles in one electrode 40.

  Further, the other electrode 41 (for example, a cathode) is formed so as to surround a portion of the one electrode 40 that covers the central portion of the bottom of the microchamber 10. Therefore, it is possible to reliably push out the target particles while reducing the magnitude of the voltage applied to the pair of electrodes 40 and 41. The electrode 40 can be a cathode and the electrode 41 can be an anode.

  FIG. 10 is a cross-sectional view of one peripheral portion of a plurality of microchambers 10 in another example of the apparatus 1. FIG. 11 is an explanatory diagram showing each step when the present method 1 is performed using the apparatus 1 shown in FIG.

  In this apparatus 1 according to this example, as shown in FIGS. 10 and 11, each of the plurality of microchambers 10 includes a storage chamber 11 that opens to a common flow path 24 and stores target particles 50, and the storage An electrolysis chamber 12 provided in communication with the chamber 11 and in which electrodes 40 and 41 for electrolysis are arranged.

  That is, in the example shown in FIGS. 10 and 11, the storage chamber 11 and the electrolysis chamber 12 are connected via the communication path 13. More specifically, the electrolysis chamber 12 is provided directly below the storage chamber 11. In the electrolysis chamber 12, both one electrode 40 and the other electrode 41 of the anode and cathode for electrolysis are arranged.

  The communication path 13 is formed in a size that prevents the target particles 50 from passing therethrough. That is, the opening 15 of the communication path 13 in the bottom 14 (see FIG. 10) of the storage chamber 11 is formed in a size smaller than the target particle 50 (see FIG. 11). That is, the opening 15 of the communication path 13 is formed in a part of the bottom 14 of the storage chamber 11 with a size smaller than the bottom 14.

  For this reason, as shown in FIG. 11A, the target particles 50 are held in the storage chamber 11 without falling into the electrolysis chamber 12. At this time, the target particles 50 can also be held at the bottom 14 of the storage chamber 11 so as to close the opening 15 of the communication passage 13.

  10 and 11 show an example in which the electrodes 40 and 41 are arranged at the bottom of the electrolysis chamber 12, but the present invention is not limited to this. For example, at least one of the electrodes 40 and 41 is connected to the electric chamber. It can also be arranged in other parts of the decomposition chamber 12.

  In the first step S1 of this method using the present apparatus 1, first, the target particles 50 are captured in the storage chambers 11 of the plurality of microchambers 10, as shown in FIG. 11A. Next, in the second step S2, as shown in FIGS. 11B and 11C, by selectively applying a voltage to the electrodes 40 and 41 of the target microchamber 10 (see FIG. 5B), the electrical Water is electrolyzed in the decomposition chamber 12 to generate bubbles 60.

  In the third step S3, as shown in FIG. 11B and FIG. 11C, at least one of the bubbles 60 generated by electrolysis and the aqueous solution in the microchamber 10 that flows along with the generation of the bubbles 60 is used as an actuator. The target particles 50 are pushed out from the microchamber 10 by using the same.

  That is, for example, as shown in FIG. 11B, due to the generation of bubbles 60 in the electrolysis chamber 12, the aqueous solution filled in the electrolysis chamber 12 is pushed out to the storage chamber 11 through the communication path 13, and further the storage chamber 11 is pushed out to the common channel 24. The target particles 50 can be pushed out of the microchamber 10 by the flow of the aqueous solution from the electrolysis chamber 12 toward the common flow path 24.

  Further, for example, as shown in FIG. 11C, when the generated bubble 60 passes through the communication path 13 and rises to the storage chamber 11, the target particle 50 is pushed out to the common channel 24 by the rising bubble 60. You can also

  FIG. 12 is a cross-sectional view of one peripheral portion of a plurality of microchambers 10 in another example of the apparatus 1. FIG. 13 is an explanatory diagram showing each step in the case of carrying out the method using the apparatus 1 shown in FIG.

  In this apparatus 1 according to this example, as shown in FIGS. 12 and 13, each of the plurality of microchambers 10 includes a storage chamber 11 that opens into a common flow path 24 and stores target particles 50, and the storage And an electrolysis chamber 12 provided in communication with the chamber 11 and in which the electrodes 40 and 41 are arranged. The electrolysis chamber 12 is provided at a position shifted from directly below the storage chamber 11, and the electrolysis chamber 12 and the storage chamber 11 are connected by a bent communication path 13.

  In the first step S1 of this method using the present apparatus 1, first, the target particles 50 are captured in the storage chambers 11 of the plurality of microchambers 10, as shown in FIG. 13A. Next, in the second step S2, as shown in FIG. 13B, by selectively applying a voltage to the electrodes 40 and 41 of the target microchamber 10 (see FIG. 5B), the electrolysis chamber 12 thereof. The water is electrolyzed to generate bubbles 60.

  And in 3rd process S3, as shown to FIG. 13B, the target particle 50 is extruded from the said micro chamber 10 with the aqueous solution in the micro chamber 10 which flows with generation | occurrence | production of the bubble 60. FIG.

  That is, in the present apparatus 1 according to this example, the bubbles generated in the electrolysis chamber 12 are held in the electrolysis chamber 12, so that the bubbles mainly flow out from the microchamber 10 to the common flow path 24 as the bubbles are generated. The flow of aqueous solution is used as an actuator.

  Of course, similarly to the example shown in FIG. 11C described above, when the bubble 60 passes through the communication path 13 and rises to the storage chamber 11, the bubble 60 may be used as an actuator to push out the target particle 50. it can.

  FIG. 14 is a cross-sectional view of one peripheral portion of a plurality of microchambers 10 in another example of the apparatus 1. FIG. 15 is an explanatory diagram showing each step when the present method 1 is carried out using the present apparatus 1 shown in FIG.

  In this apparatus 1 according to this example, as shown in FIGS. 14 and 15, each of the plurality of microchambers 10 includes an accommodation chamber 11 that opens into a common flow path 24 and accommodates target particles 50, and electrolysis. With the generation of bubbles 60 due to the electrolysis of water in the electrolysis chamber 12 provided between the electrolysis chamber 12 in which the electrodes 40 and 41 are disposed, the storage chamber 11 and the electrolysis chamber 12. And a stretchable film (hereinafter referred to as “extruded film 70”) that is deformed so as to extrude the target particles 50 from the 11 storage chamber.

  That is, in the example shown in FIGS. 14 and 15, the extruded film 70 is provided so as to partition the storage chamber 11 and the electrolysis chamber 12 and constitutes the bottom of the storage chamber 11. The extruded film 70 has elasticity that can be flexibly deformed by the generation of bubbles 60 in the electrolysis chamber 12. The extruded film 70 can also be formed so as to seal the electrolysis chamber 12.

  The material constituting the extruded film 70 is not particularly limited as long as it can realize the above-described stretchability. For example, a polymer such as polydimethylsiloxane (PDMS), polystyrene, polycarbonate, thick photoresist can be used. Moreover, the thickness of the extrusion film | membrane 70 can be made into the range of 10-200 micrometers, for example.

  The microchamber 10 having such an extruded film 70 can be produced, for example, by spin-coating PDMS on a mold on a silicon wafer. In this case, the thickness of the extruded film 70 can be arbitrarily controlled by the number of rotations during spin coating.

  In the first step S1 of the present method using the present apparatus 1, first, the target particles 50 are captured in the storage chambers 11 of the plurality of microchambers 10, as shown in FIG. 15A. That is, the target particles 50 are allowed to settle on the extruded film 70 in the storage chamber 11.

  Next, in the second step S2, as shown in FIG. 15B, by selectively applying a voltage to the pair of electrodes 40, 41 of the target microchamber 10, water is discharged in the electrolysis chamber 12. Decomposes to generate bubbles 60.

  And in 3rd process S3, as shown to FIG. 15B, the target particle 50 is extruded from the micro chamber 10 with the extrusion film | membrane 70 deform | transformed so that it may protrude upwards with generation | occurrence | production of the bubble 60. FIG.

  That is, since the extruded film 70 covers the upper part of the electrolysis chamber 12, the bubbles 60 generated in the electrolysis chamber 12 rise and push up the extruded film 70 from below. Since the extruded film 70 has elasticity, a part thereof is deformed so as to be displaced upward.

  As a result, the target particles 50 are pushed upward by the extruded film 70 and pushed out to the common flow path 24. Thus, the target particle 50 can be reliably extruded from the microchamber 10 by using the extruded film 70 as an actuator.

  FIG. 16 is a plan view of one peripheral portion of a plurality of microchambers 10 in another example of the apparatus 1. FIG. 17 is a cross-sectional view of the present apparatus 1 cut along the line XVII-XVII shown in FIG.

  This apparatus 1 according to this example has an extruded film 70 as in the examples shown in FIGS. 14 and 15 described above. However, as shown in FIG. 16, the extruded film 70 according to this example connects a central portion 71 constituting the central portion, a part of the central portion 71, and the inner wall (coating layer 22) of the microchamber 10. And a plurality of (four in FIG. 16) connection portions 72 that partially support the central portion 71. That is, in this example, the storage chamber 11 and the electrolysis chamber 12 are separated by the extruded film 70 while being in communication.

  In the example shown in FIG. 17, the central portion 71 of the extruded film 70 is formed so that its lower surface 73 has a concave shape that is recessed toward the common flow path 24. For this reason, the foam 60 generated in the electrolysis chamber 12 can be efficiently collected in the depression of the central portion 71 of the extruded film 70, and the extruded film 70 can be efficiently deformed. The extruded film 70 shown in FIG. 16 can also be formed in a flat shape as shown in FIG. 14 without having such a depression.

  FIG. 18 shows a cross-sectional view of one peripheral portion of a plurality of microchambers 10 in another example of the apparatus 1. FIG. 19 is an explanatory diagram showing each step when the present method 1 is carried out using the apparatus 1 shown in FIG.

  As shown in FIGS. 18 and 19, the extruded film 70 provided in the apparatus 1 according to this example is formed in a hollow bellows shape and can be expanded and contracted toward the common flow path 24, and the expansion and contraction portion 74. And a base portion 75 that connects the inner wall (the coating layer 22) of the microchamber 10 and supports the stretchable portion 74.

  The extruded film 70 can be manufactured using the same material as the extruded film 70 according to the example shown in FIGS. The stretchable part 74 and the base part 75 can be formed using a mold having a shape corresponding to the stretchable part 74 and the base part 75. That is, for example, when using a polymer such as PDMS as a material, an outer frame having a bellows-like portion corresponding to the stretchable portion 74 and an inner frame having the same shape slightly smaller than the outer frame are overlapped, The extruded film 70 having the bellows-like stretchable part 74 can be produced by pouring and curing the polymer between the outer frame and the inner frame.

  In the first step S1 of this method using the present apparatus 1, first, the target particles 50 are captured in the storage chambers 11 of the plurality of microchambers 10, as shown in FIG. 19A. That is, the target particles 50 are allowed to settle on the stretchable part 74 of the extruded film 70 in the storage chamber 11.

  Next, in the second step S2, as shown in FIG. 19B, by selectively applying a voltage to the pair of electrodes 40, 41 of the target microchamber 10, water is discharged in the electrolysis chamber 12. Decomposes to generate bubbles 60.

  And in 3rd process S3, as shown to FIG. 19B, the target particle 50 is extruded from the micro chamber 10 by the extrusion film | membrane 70 to which the expansion-contraction part 74 displaces upwards with generation | occurrence | production of the bubble 60. FIG.

  That is, since the extruded film 70 covers the upper part of the electrolysis chamber 12, the foam 60 generated in the electrolysis chamber 12 rises and pushes the stretchable portion 74 of the extruded film 70 from below. At this time, the stretchable part 74 of the extruded film 70 efficiently extends upward utilizing the bellows shape. As a result, the target particle 50 is pushed upward by the expansion / contraction part 74 of the extruded film 70 and pushed out to the common flow path 24.

  FIG. 20 is a plan view of a peripheral portion of three microchambers 10a, 10b, and 10c among a plurality of microchambers 10 in another example of the apparatus 1. FIG. 21 is an explanatory diagram showing each step when the present method 1 is performed using the apparatus 1 shown in FIG.

  As shown in FIGS. 20 and 21, the device 1 according to this example includes a common flow path 24 that meanders in a zigzag manner. In addition, a plurality of microchambers 10 are formed so as to cross from a part of the common channel 24 arranged in parallel to each other to another part.

  That is, as shown in FIG. 20, each microchamber 10 is provided so as to open in the middle portion of the common flow path 24, and an electrode 40 for electrolysis is disposed in the back thereof, and further in the back of the electrode 40. Is formed with a narrowed passage 16 having a reduced cross-sectional area, and the narrowed passage 16 communicates with the other middle portion of the common flow path 24.

  In the first step S1 of this method using this apparatus 1, first, as shown in FIG. 21A, target particles 50a, 50b, and 50c are captured in each of a plurality of microchambers 10a, 10b, and 10c.

  For example, as described in Non-Patent Document 1, the trapping of the target particles 50 is a meandering flow along the common flow path 24 in a state where the target particles 50 are not trapped (solid arrows in FIG. 20). The flow length and cross-sectional area of the flow path are such that the pressure loss of the flow indicated by (2) is larger than the pressure loss of the bypass flow (flow indicated by the broken arrow in FIG. 20) via the microchamber 10 and the narrow path 16. It can be realized by adjusting.

  Next, in the second step S2, as shown in FIG. 21B, by selectively applying a voltage to the electrode 40a of the target microchamber 10a among the plurality of microchambers 10a, 10b, 10c, Water is electrolyzed in the microchamber 10a to generate bubbles 60.

  Then, in the third step S3, as shown in FIG. 21B, at least one of the bubbles 60 generated by electrolysis and the aqueous solution in the microchamber 10a that flows along with the generation of the bubbles 60 is used as an actuator. The target particles 50a are selectively extruded from the microchamber 10a. Thereafter, by flowing a new aqueous solution through the common channel 24, the target particles 50a pushed into the common channel 24 can be selectively recovered.

  Moreover, in this apparatus 1 which concerns on this example, the object particle | grains 50a released from some micro chambers 10a among several micro chambers 10a, 10b, 10c can also be capture | acquired by the other micro chamber 10b.

  That is, for example, after a voltage is applied to the electrode 40b of one microchamber 10b among the plurality of microchambers 10a, 10b, and 10c to release the target particle 50b from the microchamber 10b, another microchamber further upstream. By applying a voltage to the electrode 40a of 10a to release the target particle 50a from the microchamber 10a and causing the common flow path 24 to flow downstream, the target particle 50a is made to flow based on the above-described pressure loss balance. It can be captured in the microchamber 10b on the downstream side that has been previously evacuated.

  In this way, by selectively capturing and releasing the plurality of target particles 50, the plurality of target particles 50 can be rearranged or replaced arbitrarily. 20 and FIG. 21 show an example in which one electrode 40 of the anode and cathode for electrolysis is arranged in each microchamber 10. However, the present invention is not limited to this, and the anode is placed in each microchamber 10. Both cathode and cathode electrodes can also be provided.

  Next, specific examples according to the present embodiment will be described.

[Production of Microfluidic Device] The device 1 as shown in FIG. 3 was manufactured in the same process as the example shown in FIG. That is, first, an ITO layer was formed on a glass substrate. Next, a resist layer made of a positive photoresist S1818 was patterned by standard lithography at a position corresponding to the electrode arranged at the bottom of the microchamber (see FIG. 4A). Then, the ITO layer was etched using an etchant (HCl: H ₂ O: HNO ₃ = 1: 1: 0.16) (3.20 minutes at 50 ° C.) to form an electrode on the glass substrate (FIG. 4B).

  Thereafter, this electrode was coated with a negative-type thick film photoresist SU8 to form a resist layer made of SU8 (see FIG. 4C). Then, this resist layer was patterned to form 48 (6 × 8) microchambers each having an electrode at the bottom (see FIG. 4D).

  The diameter of the microchamber was 150 μm and the depth was 150 μm. The size of the microchamber was adopted as an appropriate size for capturing one fertilized egg of a mammal.

  Furthermore, a silicone rubber spacer was placed around the coating layer (SU8 sheet) on which the microchamber was formed. And this apparatus 1 was produced by covering this 1st member with the 2nd member by which the electrode which consists of an ITO layer is formed on the glass substrate.

  [Microscope Observation] The produced apparatus 1 was observed with a bright field microscope. Since the ITO constituting the electrode provided in the apparatus 1 was transparent, it could be observed with an inverted microscope (Olympus IX71). The electrode of the microchamber was connected to the plus of the DC power supply (Agilent E3641A) via a conductive line tape, and the electrode of the second member was connected to the minus of the DC power supply. That is, the electrode of the microchamber was used as the anode, and the electrode of the second member facing was used as the cathode.

  FIG. 22 shows an enlarged part of a micrograph of the apparatus 1. As shown in FIG. 22A, it was confirmed that a plurality of microchambers were regularly arranged in the coating layer made of SU8. In addition, an electrode formed by an ITO film pattern was disposed at the bottom of each microchamber. In addition, since ITO which comprises an electrode and wiring was transparent and it was difficult to visually recognize under a microscope, the pattern (ITO) of the said electrode and wiring was shown in FIG. 22B.

  [Selective extrusion of microbeads] Polystyrene microbeads having a diameter of 100 μm were used as target particles. Further, a phosphate buffered solution (PBS) was used as the electrolyte aqueous solution.

  First, the inside of the apparatus 1 was filled with PBS. Next, microbeads dispersed in PBS were injected into the microchamber using a micropipette. As a result, one microbead could be put in each of 44 microchambers out of 48 microchambers. Microbeads that did not enter the microchamber were washed away with PBS. FIG. 23 shows an example of a micrograph in which the device 1 captures one microbead captured in each microchamber.

  The method was then performed to selectively release microbeads from one of the plurality of microchambers. FIG. 24 shows an image of a part of the device 1 in the course of carrying out the method taken with time lapse video. 24A shows an image immediately before applying a voltage (0 second), FIG. 24B shows an image 1.1 seconds after applying the voltage, and FIG. 24C shows 4.3 seconds after applying the voltage. The image of is shown.

  Here, the microbead in the central microchamber indicated by the arrow shown in FIG. 24A was selected as a target. The electrode arranged at the bottom of the selected microchamber was selectively connected to a DC power supply.

  A voltage of 5.2 V is applied between the electrode of the selected microchamber and the electrode of the second member facing the electrode, and a current is applied between the bottom ITO electrode and the upper ITO electrode. And electrolysis of PBS was performed. As a result, as shown in FIG. 24B, immediately after the voltage application, a bubble was generated from the electrode disposed at the bottom only in the target microchamber.

  Thereafter, as shown in FIG. 24C, the microbeads captured in the target microchamber were pushed out of the microchamber by bubbles and completely released. On the other hand, as shown in FIG. 24C, the microbeads captured in other microchambers to which no voltage was applied were not released and remained held.

  In this way, the target microbead could be selectively released from the microchamber. Even when polystyrene microbeads having a diameter of 10 μm or 50 μm were used, selective release could be realized in the same manner.

  [Evaluation of the Effect of Electrolysis on the Survival Rate of Mammalian Cells] As the device 1, the same microfluidic device as that prepared in Example 1 was used. Bovine arterial endothelial cells were used as the cells.

  First, the culture medium in which the cells were dispersed was injected into the apparatus 1 to capture the cells in the microchamber and cultured for a predetermined time. Next, a voltage of 0 to 8 V was applied to the microchamber for 10 seconds. Immediately after applying the voltage, a commercially available reagent (Live / Dead Assay (registered trademark) Viability / Cytotoxicity reagent) for evaluating cell viability was injected into the microchamber.

  Thereafter, the treated cells were observed under a fluorescence microscope. With the above reagent, dead cells were stained red and live cells were stained green. The number of dead cells was counted under a microscope. Then, the ratio (%) of the number of dead cells to the total number of cells (live cell number + dead cell number) was calculated.

  FIG. 25 shows the evaluation result. In FIG. 25, the horizontal axis indicates the voltage (V) applied to the microchamber, and the vertical axis indicates the ratio (%) of the number of dead cells. In addition, when a voltage (0V, 2V) on the left side of the broken line shown in FIG. 25 is applied, no bubbles are generated (no bubbles), and when a voltage (4V, 6V, 8V) on the right side of the broken line is applied. Bubbles formed.

  As shown in FIG. 25, it was found that a voltage higher than 6V causes stronger damage to the cells. On the other hand, most cells survived at voltages lower than 6V (2V, 4V). Therefore, the electrolysis condition of applying a voltage less than 6V was confirmed to be mild enough for selective release of cells. Even when 8 V was applied, the proportion of dead cells was less than 10%, and many cells remained viable.

  [Measurement of temperature before and after electrolysis] The temperature of the apparatus 1 while applying a voltage of 5.2 V between the ITO electrodes was measured using a thermograph device (Neo Thermo TVS-600, NEC Avio Infrared Technology Co., Ltd.). It measured using.

  As a result, there was no change in the temperature in the apparatus 1 before and after the voltage was applied. Further, even after the voltage was applied, the temperature of the device 1 did not increase. Therefore, it was confirmed that this method can be carried out without damaging the cells due to heat generation.

  DESCRIPTION OF SYMBOLS 1 Microfluidic device, 10 Micro chamber, 11 Accommodating chamber, 12 Electrolysis chamber, 13 Communication path, 14 Bottom part, 15 Opening part, 16 Constriction path, 20 1st member, 21 1st board | substrate, 22 Coating layer, 23 Spacer, 24 common flow path, 25 inlet, 26 outlet, 27 ITO film, 28 positive photoresist layer, 29 negative photoresist layer, 30 second member, 31 second substrate, 40, 41 electrode, 42 wiring, 50 Target particles, 60 foam, 70 extruded film, 71 center part, 72 connection part, 73 lower surface, 74 stretchable part, 75 base part.

Claims (10)

Each of the particles to be manipulated is captured in a plurality of microchambers each having at least one electrode of an anode and a cathode for electrolysis,
Water is electrolyzed by applying a voltage to the electrodes of some of the plurality of microchambers to generate bubbles,
A method of selectively extruding the particles from the partial microchamber by an actuator that operates as the bubbles are generated. The method according to claim 1, wherein the actuator is the bubble that floats or the liquid in the microchamber that flows with the generation of the bubble. 2. The actuator according to claim 1, wherein the actuator is a stretchable film that is provided between the electrode and the particle in the microchamber and is deformed so as to push out the particle as the bubble is generated. Method. The particles are cells;
Capturing and culturing the cells one by one in each of the plurality of microchambers;
A part of the plurality of cells cultured in the plurality of microchambers is determined as a cell to be collected;
By applying a voltage to the electrodes of some of the microchambers in which the cells to be collected are captured among the plurality of microchambers to generate bubbles by electrolyzing water,
The method according to any one of claims 1 to 3, wherein the cells are selectively pushed out from the partial microchamber by the actuator that operates in association with the generation of the bubbles. The cell is a fertilized egg;
The method according to claim 4, wherein the fertilized eggs selectively extruded from the partial microchamber are collected in a live state. A plurality of microchambers for capturing particles;
A common flow path in which the plurality of microchambers are opened;
A microfluidic device comprising:
Each of the plurality of micro chambers is provided with at least one of an anode for electrolysis and a cathode. A first member provided with the plurality of microchambers;
A second member laminated to the first member;
With
In each of the plurality of microchambers, one of the anode and the cathode is disposed,
The microfluidic device according to claim 6, wherein the second member is provided with the other electrode of the anode and the cathode. The microfluidic device according to claim 6, wherein both the anode and the cathode are arranged in each of the plurality of microchambers. Each of the plurality of microchambers includes a storage chamber that opens to the common channel and stores the particles, and an electrolysis chamber that is provided in communication with the storage chamber and in which the electrode is disposed. The microfluidic device according to any one of claims 6 to 8, characterized in that Each of the plurality of microchambers is provided between a storage chamber that opens to the common channel and stores the particles, an electrolysis chamber in which the electrodes are disposed, and the storage chamber and the electrolysis chamber. And a stretchable film that is deformed so as to push out the particles from the storage chamber as bubbles are generated by electrolysis of water in the electrolysis chamber. The microfluidic device described in 1.