JP6877028B2 - Lipid bilayer film forming apparatus, lipid bilayer film forming method, and evaluation system - Google Patents

Description

The present invention relates to a lipid bilayer membrane forming apparatus, a lipid bilayer membrane forming method, and an evaluation system.

A cell is generally covered with a cell membrane formed of a lipid bilayer membrane, which forms a boundary between the cell and the outside world.

The cell membrane not only has a function of preventing the substance from freely diffusing from the outside world into the cell, but also has a function of selectively taking in the substance from the outside world into the cell. Such functions of the cell membrane are expressed by lipids forming the cell membrane and various membrane proteins embedded in the cell membrane.

In recent years, attention has been focused on various functions possessed by cell membranes, and attempts have been made to analyze the functions.

For example, since the cell membrane has a function of preventing the free diffusion of substances from the outside world into the cell, it can affect the diffusion of the drug in the body. Therefore, in the development of drugs, attempts have been made to analyze the permeability of the cell membrane of the drug.

For example, although the cell membrane has the function of preventing the free diffusion of substances from the outside into the cell, some toxic compounds can diffuse through the cell membrane. Therefore, in evaluating the toxicity of a toxic compound, an attempt has been made to analyze the permeability of the cell membrane of the compound.

For example, membrane proteins may denature when separated from the cell membrane. Therefore, in order to analyze the function of membrane proteins, attempts have been made to artificially implant the membrane proteins in the cell membrane.

In order to analyze various functions of cell membranes, a technique for artificially producing a lipid bilayer membrane is required, and the development of such a technique has been promoted.

As a technique for artificially producing a lipid bilayer membrane, for example, a method in which a droplet covered with a lipid single membrane and another droplet covered with the lipid single membrane are brought into contact with each other by a mechanical external force is used. (Patent Document 1).

In addition, as a technique for artificially producing a lipid bilayer membrane, a lipid bilayer membrane is formed by forming droplets covered with a lipid bilayer membrane in advance in oil and manipulating the droplets with a manipulator or the like. (Non-Patent Document 1).

Also,

Japanese Unexamined Patent Publication No. 2014-100672 (published on June 5, 2014)

Masayuki Iwamoto & Shigetoshi Oiki, Sci. Rep. 5, 9110, 2015

However, the prior art as described above requires many problems, such as requiring a large-scale device for producing a lipid bilayer membrane and a skilled technique for producing a lipid bilayer membrane. have.

The present invention has been made in view of the above problems, and an object of the present invention is a lipid bilayer membrane forming apparatus and a lipid bilayer membrane forming method capable of easily forming a lipid bilayer membrane, and the lipid. The purpose is to realize an evaluation system including a bilayer film forming apparatus.

One embodiment of the present invention has the following configuration.

[1] A first accommodating portion for accommodating oil and a plurality of second accommodating portions provided at the bottom of the first accommodating portion and for accommodating droplets covered with a lipid bilayer are provided. The inner surface of the first accommodating portion has a second accommodating portion so that when an aqueous solution is added to the oil contained in the first accommodating portion, the droplets formed from the aqueous solution are guided to the second accommodating portion. The shape converges toward the accommodating portion, and the plurality of second accommodating portions are arranged so that the droplets accommodating in the second accommodating portion come into contact with each other. Lipid bilayer film forming apparatus.

[2] When the shape of the droplet is _{a sphere having a diameter of r 3} and the diameter of the maximum inscribed circle of the space in the second accommodating portion is r ₂ , r ₃ ≤ r ₂ is satisfied. , [1]. The lipid bilayer film forming apparatus.

[3] The lipid bilayer film forming apparatus according to [1] or [2], wherein _{the depth H 2} of the second accommodating portion satisfies 0 <H ₂ <(r _{3) / 2.} ..

[4] The depth H ₂ of the second accommodating portion satisfies (r ₃ ) / 2 ≦ H _2, and a hole penetrating the wall surface is formed on the wall surface separating the second accommodating portions. The lipid bilayer film forming apparatus according to [1] or [2].

[5] A method for forming a lipid bilayer film using the lipid bilayer film forming apparatus according to any one of [1] to [4], wherein the oil contained in the first storage portion is used. A method for forming a lipid bilayer membrane, which comprises a step of adding an aqueous solution.

[6] An evaluation system comprising the lipid bilayer film forming apparatus according to any one of [1] to [4].

[7] The evaluation system according to [6], wherein the evaluation system further includes electrodes.

According to the lipid bilayer film forming apparatus according to the embodiment of the present invention, there is an effect that the lipid bilayer film can be easily formed.

(A) It is a perspective view of the lipid bilayer film forming apparatus which concerns on one Embodiment of this invention. (B) is a plan view of the lipid bilayer film forming apparatus of (a) seen from above vertically. (C) It is sectional drawing in AA of the double film forming apparatus of (b). (D) It is sectional drawing in BB of the double film forming apparatus of (b). (E) It is a top view from the vertical view of the lipid bilayer film forming apparatus which concerns on one Embodiment of this invention. It is sectional drawing in the longitudinal direction of the 2nd accommodating part of the lipid bilayer membrane apparatus which concerns on one Embodiment of this invention before and after accommodating a droplet. (A) It is a schematic diagram of the 2nd accommodating part of the lipid bilayer film forming apparatus which concerns on one Embodiment of this invention. (B) It is sectional drawing in the longitudinal direction of the 2nd accommodating part of the lipid bilayer film forming apparatus which concerns on one Embodiment of this invention. (C) It is sectional drawing in the short side of the 2nd accommodating part of the lipid bilayer film forming apparatus which concerns on one Embodiment of this invention. It is the schematic of the 2nd accommodating part of the lipid bilayer film forming apparatus which concerns on one Embodiment of this invention (d), (e) and (f). (G) It is sectional drawing in the longitudinal direction of the 2nd accommodating part of the lipid bilayer film forming apparatus which concerns on one Embodiment of this invention. (H) It is sectional drawing in the short side of the 2nd accommodating part of the lipid bilayer film forming apparatus which concerns on one Embodiment of this invention. (I) and (j) are schematic views of the 2nd accommodating part of the lipid bilayer film forming apparatus which concerns on one Embodiment of this invention. It is a perspective view of the lipid bilayer film forming apparatus which concerns on one Embodiment of this invention. It is sectional drawing in the longitudinal direction of the 2nd accommodating part of the lipid bilayer film forming apparatus included in the evaluation system which concerns on one Embodiment of this invention.

An embodiment of the present invention will be described below, but the present invention is not limited thereto. The present invention is not limited to the configurations described below, and various modifications can be made within the scope of the claims. The present invention also includes embodiments and examples obtained by appropriately combining the technical means disclosed in different embodiments and examples within the technical scope of the present invention. All academic documents, non-patent documents and patent documents described in the present specification are incorporated as references in the present specification. Further, unless otherwise specified in the present specification, "A to B" representing a numerical range is intended to be "A or more (including A and larger than A) and B or less (including B and smaller than B)".

[Lipid bilayer membrane forming device]
The lipid bilayer film forming apparatus according to the embodiment of the present invention is provided at the first accommodating portion in which oil is accommodating and the bottom of the first accommodating portion, and accommodates droplets covered with a lipid monolayer. A plurality of second accommodating portions for the purpose, and the inner surface of the first accommodating portion provides the droplets formed from the aqueous solution when the aqueous solution is added to the oil contained in the first accommodating portion. The shape converges toward the second accommodating portion so as to lead to the second accommodating portion, and the plurality of second accommodating portions are in contact with each other of the droplets contained in the second accommodating portion. It is characterized by being arranged so as to do so.

As described above, the technique described in Patent Document 1 required a large-scale device for producing a lipid bilayer membrane. In addition, the technique described in Non-Patent Document 1 requires skillful techniques, such as the ability to produce a lipid bilayer membrane only after advanced training.

On the other hand, the lipid bilayer film forming apparatus according to the embodiment of the present invention can automatically prepare a lipid bilayer film only by adding an aqueous solution to oil, and therefore, it is a person who is new to the technical field. Even if there is, it has an advantage that it is possible to easily prepare a lipid bilayer membrane.

Further, the lipid bilayer film forming apparatus according to the embodiment of the present invention does not require a special apparatus such as a manipulator, and therefore has an advantage that it can be compact and lightweight.

Further, the lipid bilayer membrane forming apparatus according to the embodiment of the present invention has an advantage that it can be carried and the lipid bilayer membrane can be produced at an arbitrary place (for example, outdoors).

Further, the lipid bilayer film forming apparatus according to the embodiment of the present invention has an advantage that it can be made into a disposable apparatus because it has a simple structure and can be manufactured at low cost.

Further, the lipid bilayer film forming apparatus according to the embodiment of the present invention can easily adjust the amount of the aqueous solution to be added to the oil contained in the first accommodating portion, and can easily adjust the amount of the aqueous solution formed from the aqueous solution. The size can be easily adjusted. If the size of the droplets can be adjusted, the size of the area where the droplets are in contact with each other can be easily adjusted. If the area where the droplets are in contact with each other is too large, noise may be mixed in the data when acquiring the data for measuring the function of the lipid bilayer membrane. Therefore, the lipid bilayer film forming apparatus according to the embodiment of the present invention has an advantage that when data for measuring the function of the lipid bilayer membrane is acquired, noise mixed in the data can be reduced. Have.

The lipid bilayer film forming apparatus according to the embodiment of the present invention will be described with reference to FIGS. 1 to 5.

FIG. 1A is a perspective view of a lipid bilayer membrane forming apparatus according to an embodiment of the present invention. FIG. 1B is a plan view of the lipid bilayer film forming apparatus of FIG. 1A as viewed vertically from above. FIG. 1 (c) is a cross-sectional view taken along the line AA of the double film forming apparatus of FIG. 1 (b). FIG. 1D is a cross-sectional view taken along the line BB of the double film forming apparatus of FIG. 1B. FIG. 1 (e) is a plan view of the lipid bilayer film forming apparatus according to the embodiment of the present invention as viewed from above vertically.

As shown in FIG. 1, the lipid bilayer film forming apparatus according to the embodiment of the present invention includes a substrate 1, a first accommodating portion 2 provided on the substrate 1, and a plurality of second accommodating portions 3. And. The first accommodating portion 2 is formed on one of the surfaces of the substrate 1, and the plurality of second accommodating portions 3 are formed on the surface of the first accommodating portion 2 (more specifically, the bottom portion of the first accommodating portion 2). ) Is formed. The first accommodating portion 2 may be provided on a plurality of surfaces of the substrate 1. In this case, if the configurations of the plurality of second accommodating portions 3 generated in each of the first accommodating portions 2 are different, it is possible to provide a plurality of types of lipid bilayer film forming apparatus by one substrate. it can.

FIG. 2 shows one of the present inventions before and after the oil 4 is contained in the first accommodating portion 2 and the droplet 5 is accommodated in the second accommodating portion 3 formed at the bottom of the first accommodating portion 2. It is sectional drawing in the longitudinal direction of the lipid bilayer film forming apparatus which concerns on embodiment.

As shown in FIG. 2, in the lipid bilayer film forming apparatus according to the embodiment of the present invention, a first accommodating portion 2 for accommodating the oil 4 and a lipid monolayer on the bottom of the first accommodating portion 2. A plurality of second accommodating portions 3 for accommodating the droplet 5 covered by the above are provided. Since the second accommodating portion 3 is provided at the bottom of the second accommodating portion 2, when the oil 4 is accommodating in the first accommodating portion 2, until the droplet 5 is accommodating in the second accommodating portion 3. During that time, the second accommodating portion 3 is filled with oil 4. Further, even after the droplet 5 is accommodated in the second accommodating portion 3, the space not occupied by the droplet 5 in the second accommodating portion 3 is filled with the oil 4. Further, in FIG. 2, the first accommodating portion 2 is filled with oil, but the first accommodating portion 2 does not need to be filled with oil. The amount of oil contained in the first accommodating portion 2 is not particularly limited as long as the amount of the droplet 5 contained in the second accommodating portion 3 can be immersed.

The shape of the substrate 1 is not particularly limited to a quadrangular prism (for example, a rectangular parallelepiped and a cube) as shown in FIG. 1, as long as the first accommodating portion 2 can be formed, and is, for example, a cylinder, a polygonal prism, or the like. You may.

FIG. 1 will be described below with reference to the configuration of the substrate 1 by taking as an example a case where the shape of the substrate 1 is a quadrangular prism and the substrate 1 includes one first accommodating portion 2. Is not particularly limited.

Let X be the length in the longitudinal direction of the substrate 1, Y be the length in the lateral direction, and Z be the height. The length of the substrate 1 in the longitudinal direction X is preferably 5 mm to 100 mm, more preferably 8 mm to 75 mm, further preferably 12 mm to 50 mm, and particularly preferably 20 to 30 mm. The length in the lateral direction Y is preferably 3 mm to 50 mm, more preferably 4 mm to 35 mm, further preferably 6 mm to 25 mm, and particularly preferably 10 mm to 15 mm. The length of the height Z is preferably 0.5 mm to 8 mm, more preferably 0.6 mm to 6 mm, further preferably 1 mm to 4 mm, and 1.5 mm to 2.5 mm. Is particularly preferred. With this configuration, it is possible to realize a lipid bilayer film forming apparatus that is easy to carry.

The material of the substrate 1 is not particularly limited, but a material that is not deformed or corroded by the oil 4 is preferable. For example, glass, plastic, acrylic, Teflon (registered trademark) and the like can be mentioned. Among these, glass is preferable because it has excellent chemical resistance and optical observation is performed.

The inner surface of the first accommodating portion 2 has a shape capable of accommodating (1) oil 4, and (2) when an aqueous solution is added to the oil 4 contained in the first accommodating portion 2, from the above aqueous solution. The second accommodating portion 3 so as to guide the formed droplet 5 covered with the lipid monolayer to the second accommodating portion 3 (more specifically, automatically to the second accommodating portion 3). It has a shape that converges toward. With the above configuration, the droplet 5 can be automatically guided to the second accommodating portion 3, and the droplet 5 can be automatically accommodated in the second accommodating portion 3.

The shape of the inner surface of the first accommodating portion 2 is not particularly limited as long as it satisfies the above (1) and (2). The shape of the inner surface of the first accommodating portion 2 may be, for example, a shape corresponding to a part of the surface of the sphere (for example, a hemisphere), a conical shape, a polygonal thrust shape, or a combination thereof. In this case, if the second accommodating portion 3 is provided, for example, at or near the apex of the first accommodating portion 2, the droplet 5 can be automatically guided to the second accommodating portion 3.

An example of the shape of the inner surface of the first accommodating portion 2 is a groove, which is a shape recessed from the surrounding surface. When a groove is provided on the surface of the inner surface of the first accommodating portion 2, the droplet 5 can be guided to the second accommodating portion 3 along the groove and is accommodated in the second accommodating portion 3. It is possible to prevent the droplets 5 from coming into contact with each other before. In FIG. 1 (e), the surface of the inner surface of the first accommodating portion 2 has the same width as the diameter of the second accommodating portion 3 from the outer circumference of the first accommodating portion 2 toward the second accommodating portion 3. A linear groove is provided, but the shape of the groove is not particularly limited. The shape of the groove may be, for example, a shape that spreads in a fan shape from the second accommodating portion 3 toward the outer periphery of the first accommodating portion 2. When a groove having a fan shape as described above is provided on the surface of the inner surface of the first accommodating portion 2, the droplets 5 formed in various places are placed along the groove in the second. It can be stably guided to the accommodating portion 3. The height of the groove is not particularly limited, may be greater than the diameter r ₃ of the droplets 5 which will be described later, it may be small. For example, the height of the groove may be greater than 1/1000 of a diameter _{r 3} of the droplet 5, may be higher than 1/100, may be higher than 1/10. The higher the height of the groove, the more stably the droplet 5 can be guided to the second accommodating portion 3 along the groove.

The shape of the surface of the inner surface of the first accommodating portion 2 is not particularly limited, and may be a flat surface without unevenness, or may be provided with unevenness. The unevenness is intended to be the roughness of the surface of the inner surface of the first accommodating portion 2, and is distinguished from the above-mentioned groove.

When the unevenness is provided, it may be provided on the entire surface of the inner surface of the first accommodating portion 2, or may be provided on a part of the surface of the inner surface of the first accommodating portion 2. More specifically, when the above-mentioned groove is provided on the inner surface of the first accommodating portion 2, the surface of the groove may be provided with irregularities.

If the surface of the inner surface of the first accommodating portion 2 is flat, the droplet 5 can be quickly accommodated in the second accommodating portion 3, while the surface of the inner surface of the first accommodating portion 2 is uneven. For example, the time required for accommodating the droplet 5 in the second accommodating portion 3 can be lengthened. That is, by providing the unevenness, a lipid single film is formed over a long period of time, or by providing a difference in the shape of the unevenness so that the formation time of the lipid single film is different, a plurality of droplets are formed. The formation order can be specified. The shape of the unevenness is not particularly limited, and the depth of the concave portion or the height of the convex portion is not particularly limited. For example, the height of the depth and the convex portion of the concave portion, the diameter _{r 3} of the droplet 5, may be 1/10 or less, may be 1/100 or less, in 1 / 1,000 or less There may be. The larger the depth of the concave portion and the height of the convex portion, the longer the time required for accommodating the droplet 5 in the second accommodating portion 3 can be increased.

The shape of the inner surface of the first accommodating portion 2 is also more preferably the shape of the surface of a sphere having a radius of curvature. The radius of curvature is not particularly limited, but is preferably 3 mm to 50 mm, more preferably 4 mm to 35 mm, further preferably 6 mm to 25 mm, or particularly preferably 8 mm to 16 mm. ..

When the first accommodating portion 2 is viewed from above vertically, the shape of the outer circumference of the first accommodating portion 2 is not particularly limited to a perfect circle as shown in FIG. 1, for example, an ellipse, a quadrangle, and many. It may be a quadrangle or the like.

As shown in FIG. 1, when viewed first housing part 2 from the vertically upward, when the shape of the outer circumference of the first housing section 2 is true circle, defining the diameter of the perfect circle as r ₁ and, the shape of the outer periphery of the first housing section 2, oval, square, and in the case of polygonal defines the diameter of those of the inscribed circle as r _1. As shown in FIG. 1, when the first accommodating portion 2 is provided with two second accommodating portions 3, the diameter r ₁ of the first accommodating portion 2 is not particularly limited, but is 2.5 mm or more. It is preferably 40 mm, more preferably 3 mm to 30 mm, further preferably 5 mm to 20 mm, and particularly preferably 7.5 mm to 15 mm.

_{The depth H 1} of the first accommodating portion 2 will be described. _{The depth H 1} of the first accommodating portion 2 can be set based on the relationship with the _{depth H 2} of the second accommodating portion 3 described later. For example, the sum of the depth of the first housing part 2 H ₁ and the depth H ₂ of the second housing part 3 _(H 1 ₊ H _2), as not to exceed the height Z of the substrate 1 can be set.

When the diameter of the droplets 5 and _{r 3,} the sum of the depth of the first housing part 2 _{H 1} and the depth _{H 2} of the second housing part 3 _(H 1 + H ₂₎ is larger than the diameter _{r 3} Can be set as such. As used herein, the term "droplet 5 diameter r ₃ " is intended to mean that _{the diameter of the sphere is r 3} , assuming that the droplet 5 is a perfect sphere. The diameter r ₃ can be calculated from the volume of the aqueous solution added to the oil 4. The droplet 5 contained in the second accommodating portion 3 cannot have a perfect sphere shape due to gravity or a force received from the oil 4 or the like around the droplet 5, and has a substantially sphere shape. Become.

_{The depth H 1} of the first accommodating portion 2 is preferably 0.3 mm to 6 mm, more preferably 0.5 mm to 4.5 mm, further preferably 0.75 mm to 3 mm, and 1 mm. It is particularly preferably about 2 mm.

The shape of the inner surface of the second accommodating portion 3 is not particularly limited as long as it can accommodate the droplet 5 covered with the lipid single film. The shape of the inner surface of the second accommodating portion 3 may be, for example, a shape that converges vertically downward. The shape of the inner surface of the second accommodating portion 3 may be, for example, a shape corresponding to a part of the surface of the sphere (for example, a hemisphere), a cylindrical shape, a polygonal prism shape, or a combination thereof.

The shape of the inner surface of the second accommodating portion 3 is preferably the shape of the surface of a sphere having a radius of curvature. The radius of curvature is not particularly limited, but is preferably 0.3 mm to 5 mm, more preferably 0.4 mm to 3.5 mm, further preferably 0.6 mm to 2.5 mm, and 0. It is particularly preferably 0.8 mm to 1.6 mm.

The radius of curvature of the diameter r ₃ of the droplet 5, more preferably preferably less large and 4 times higher than 0.5 times or less larger and 3 times than 0.5 times, 0. It is more preferably 55 times or more and 2 times or less, and particularly preferably 0.6 times or more and 1 time or less. Within the above range relationship between the diameter r ₃ of the curvature radius of the droplet 5, it is possible to perform the clearance process easily. The radius of curvature is preferably set so that the volume of the sphere having the radius of curvature is larger than 1 times and 35 times or less of the volume of the droplet 5, more preferably 1.2 times or more. It is set to be 25 times or less, more preferably 1.4 times or more and 15 times or less, and particularly preferably 1.7 times or more and 8 times or less.

When the second accommodating portion 3 is viewed from above vertically, the shape of the outer circumference of the second accommodating portion 3 is not particularly limited to a perfect circle as shown in FIG. 1, for example, an ellipse, a quadrangle, and many. It may be a quadrangle or the like.

The lipid bilayer membrane forming apparatus according to an embodiment of the present invention, the shape of the droplet 5 is a sphere of diameter r _3, if the diameter of the largest inscribed circle of the space in the second container 3 was r ₂ It is preferable that r ₃ ≤ r _{2 is satisfied.} With the above configuration, it is possible to keep the contact surface between the droplets 5 small. Specifically, even when the droplet 5 is deformed by gravity or the like, the contact surface between the droplets 5 can be kept small. Here, in the present specification, the space in the second accommodating portion 3 means a space vertically above the inner side surface of the second accommodating portion 3. _{Therefore, the diameter r 2} of the maximum inscribed circle of the space in the second accommodating portion 3 is the diameter of the second accommodating portion 3 when the second accommodating portion 3 is viewed from vertically above, as shown in FIG. It may be equal to r _2. Here, the diameter r _{2 of the second accommodating portion 3 defines the diameter of the perfect circle as r 2} when the outer peripheral shape of the second accommodating portion 3 is a perfect circle, and the diameter r 2 of the second accommodating portion 3 the shape of outer circumference, an oval, a square, and in the case of polygonal defines the diameter of those of the inscribed circle and r _2. More specifically, in the lipid bilayer film forming apparatus according to the embodiment of the present invention, when the shape of the droplet 5 is _{a sphere having a diameter of r 3} , the r ₃ and the second accommodating portion 3 are vertically above. It is preferable that _{the diameter r 2} of the second accommodating portion 3 when viewed from the above satisfies r ₃ ≦ r _2.

_{The diameter r 2} of the maximum inscribed circle of the space in the second accommodating portion 3 is not particularly limited, but is preferably 0.3 mm to 5 mm, more preferably 0.4 mm to 3.5 mm, and 0. It is more preferably 0.6 mm to 2.5 mm, and particularly preferably 0.8 mm to 1.6 mm.

The lowermost portion of the second accommodating portion 3 does not have to be arranged at the center of the second accommodating portion 3 when the second accommodating portion 3 is viewed from vertically above. Here, the lowermost portion of the second accommodating portion 3 is the most vertically lower portion of the second accommodating portion 3. Further, the center of the second accommodating portion 3 is intended to be the center of the outer circumference when the second accommodating portion 3 is viewed from vertically above, and when the outer peripheral shape is a perfect circle, it is the center of the perfect circle. , When the outer peripheral shape is elliptical, polygonal, etc., the center of their inscribed circle is intended. When the lowermost part of the second accommodating portion 3 is not arranged in the center of the second accommodating portion 3 when the second accommodating portion 3 is viewed from vertically above, the lowermost portion of the second accommodating portion 3 is When the second accommodating portion 3 is viewed from vertically above, from the center of the second accommodating portion 3 to the side of the adjacent second accommodating portion 3 (more specifically, the lowermost part of the adjacent second accommodating portion 3). It is preferable that they are arranged so as to be offset from each other. With the above configuration, even when the amount of the aqueous solution to be added is small, the droplets 5 can be brought into contact with each other to form a lipid bilayer film.

The distance between the second housing portion 3 adjacent is not particularly limited, preferably greater than the diameter r ₃ of the droplets 5. The "adjacent second accommodating portion 3" is a second accommodating portion 3 in which two droplets 5 accommodating one by one in each second accommodating portion 3 can come into contact with each other to form a lipid bilayer membrane. Two are intended. Further, the distance between the adjacent second accommodating portions 3 is intended to be the distance between the lowermost portions of the adjacent second accommodating portions 3. The droplet 5 contained in the second accommodating portion 3 is such that the center of gravity of the droplet 5 and the lowermost portion of the second accommodating portion 3 substantially coincide with each other in the second accommodating portion 3 when viewed from the vertical direction. In addition, it can be stably fastened. The distance between the adjacent second accommodating portions 3 is more preferably 1.05 times or more, more preferably 1.1 times or more, and particularly preferably 1.2 times or more of _{r 3.} preferable. With the above configuration, it is possible to keep the contact surface between the droplets 5 small even when the droplets 5 are deformed by their own weight.

_{The depth H 2} of the second accommodating portion 3 will be described with reference to FIG. (A), (d), (e), (f), (i) and (j) of FIG. 3 are schematic views of a second accommodating portion of the lipid bilayer membrane forming apparatus according to the embodiment of the present invention. Is. (B) and (g) of FIG. 3 are sectional views in the longitudinal direction of the second accommodating portion of the lipid bilayer film forming apparatus according to the embodiment of the present invention. (C) and (h) of FIG. 3 are cross-sectional views in the lateral direction of the second accommodating portion of the lipid bilayer film forming apparatus according to the embodiment of the present invention.

More specifically, (a) to (e) of FIG. 3 is an embodiment of the present invention in the case where the _{depth H 2} of the second accommodating portion 3 satisfies 0 <H ₂ <(r _{3) / 2.} The second accommodating portion of the lipid bilayer film forming apparatus is shown, and (f) to (j) of FIG. 3 show the case where _{the depth H 2} of the second accommodating portion 3 satisfies (r ₃ ) / 2 ≦ H _2. The second accommodating portion of the lipid bilayer membrane forming apparatus according to the embodiment of the present invention is shown.

_{The depth H 2} of the second accommodating portion 3 may satisfy 0 <H ₂ <(r ₃ ) / 2. In the above configuration, since the height of the inner surface (in other words, the wall) of the second accommodating portion 3 that separates the second accommodating portions 3 is less than the radius of the droplet 5, the height of the droplet 5 is shown in FIG. _{As shown, even when a wall having the same height as the depth H 2} of the second accommodating portion 3 is installed between the adjacent second accommodating portions 3, the droplets 5 are said to be the same. It is possible to easily make contact above the wall.

Further, when the depth H ₂ of the second accommodating portion 3 satisfies 0 <H ₂ <(r ₃ ) / 2, as shown in FIG. 3A, the adjacent second accommodating portion 3 A hole (specifically, a U-shaped notch) may be provided so as to penetrate the wall separating the two. According to the above configuration, the droplets 5 contained in the second accommodating portion can come into contact with each other through the holes, so that the contact area between the droplets 5 is adjusted (for example, increased). It becomes possible.

Further, when the depth H ₂ of the second accommodating portion 3 satisfies 0 <H ₂ <(r ₃ ) / 2, as shown in FIG. 3 (e), the adjacent second accommodating portion 3 Holes (specifically, openings) that penetrate the walls that separate them from each other may be installed. According to the above configuration, the droplets 5 contained in the second accommodating portion can come into contact with each other through the holes, so that the contact area between the droplets 5 is adjusted (for example, increased). It becomes possible.

_{The depth H 2} of the second accommodating portion 3 may satisfy (r ₃ ) / 2 ≦ H _2. In the above configuration, since the height of the inner surface (in other words, the wall) of the second accommodating portion 3 that separates the second accommodating portions 3 is equal to or larger than the radius of the droplet 5, the height of the droplet 5 is shown in FIG. _{As shown, when a wall having the same height as the depth H 2} of the second accommodating portion 3 is installed between the adjacent second accommodating portions 3, the wall causes the droplets 5 to each other. It is possible to prevent easy contact. In this case, as the shape of the droplet 5 changes (for example, with the passage of time, the lipid capable of forming a lipid double film from the oil 4 moves to the interface of the droplet 5, and as a result, the droplet 5 The droplets 5 can come into contact with each other (for example, when the area of the lipid double film increases with time due to a decrease in interfacial tension).

Further, when the depth H ₂ of the second accommodating portion 3 satisfies (r ₃ ) / 2 ≦ H ₂ , as shown in FIG. 3 (f), the adjacent second accommodating portions 3 are connected to each other. A hole (specifically, a U-shaped notch) penetrating the separating wall may be provided. According to the above configuration, the droplets 5 contained in the second accommodating portion can come into contact with each other through the holes, so that the contact area between the droplets 5 is adjusted (for example, increased). It becomes possible.

Further, when the depth H ₂ of the second accommodating portion 3 satisfies (r ₃ ) / 2 ≦ H ₂ , as shown in FIG. 3 (j), the adjacent second accommodating portions 3 are connected to each other. A hole (specifically, an opening) that penetrates the separating wall may be formed. According to the above configuration, the droplets 5 contained in the second accommodating portion can come into contact with each other through the holes, so that the contact area between the droplets 5 is adjusted (for example, increased). It becomes possible.

The size and number of the above-mentioned holes are not particularly limited, and can be appropriately set to a desired size and a desired number. For example, when three or more second accommodating portions 3 are arranged in series, one hole is formed in each of the two second accommodating portions 3 arranged at the ends, and the two second accommodating portions 3 are formed. It is also possible to form a plurality of (for example, two) holes in the second accommodating portion 3 arranged between the two. Of course, the present invention is not limited to this configuration.

4 (a) to 4 (e) are perspective views of a lipid bilayer film forming apparatus according to an embodiment of the present invention.

As shown in FIG. 4 (e), the lipid bilayer film forming apparatus according to the embodiment of the present invention may have two or more first accommodating portions 2 per substrate 1. In this case, each first accommodating portion 2 has two or more second accommodating portions 3. In FIG. 4 (e), one substrate 1 includes six first accommodating portions 2, but the present invention is not limited to this, and one substrate 1 has six or more first accommodating portions 2. It may have 12 or more, 24 or more, 48 or more, or 96 or more. With the above configuration, it is possible to form a plurality of lipid bilayer membranes at the same time by one lipid bilayer membrane forming device. Therefore, when the evaluation system according to the embodiment of the present invention described later includes the lipid bilayer film forming apparatus having the above-described configuration, it is possible to evaluate a plurality of lipid bilayer membranes at the same time. ..

The lipid bilayer film forming apparatus according to the embodiment of the present invention also has three or more second accommodating portions per one first accommodating portion 2, as shown in FIGS. 4 (a) to 4 (d). May have 3. When the lipid bilayer membrane forming apparatus has three second accommodating portions 3 per one first accommodating portion 2 ((b) in FIG. 4), two (for example, two types) lipid bilayers are simultaneously formed. It becomes possible to form a film. Therefore, when the evaluation system described later is provided with the lipid bilayer film forming apparatus, it is possible to evaluate two lipid bilayer membranes at the same time. For example, if the three droplets contained in the three second containments 3 are configured to have the constituent compositions of lumen, cytoplasm, and blood, respectively, in order, the epithelium. It is possible to reproduce the tissue.

Further, when the lipid bilayer film forming apparatus has four second accommodating portions 3 per one first accommodating portion 2 ((c) in FIG. 4), four (for example, four types) lipids are simultaneously formed. It is possible to form a bilayer film. Therefore, when the evaluation system described later is provided with the lipid bilayer film forming apparatus, it is possible to evaluate four lipid bilayer membranes at the same time. For example, the four droplets contained in the four second containments 3, in a clockwise direction, are the components of endolymph, stria vascularis, perilymph, and hair cell, respectively. It is possible to reproduce the organ if it is configured to have.

The number of the second accommodating portions 3 included in one first accommodating portion 2 is not particularly limited as long as it is two or more, and may be three, four or more, and nine or more. There may be.

[Method for forming lipid bilayer membrane]
The method for forming a lipid bilayer membrane according to an embodiment of the present invention is a lipid bilayer using the lipid bilayer membrane forming apparatus according to the embodiment of the present invention described in the above [lipid bilayer membrane forming apparatus]. It is a film forming method, and is characterized by having a step of adding an aqueous solution to the oil stored in the first storage portion.

In the lipid bilayer forming method according to the embodiment of the present invention, there is no upper limit to the number of lipid bilayers formed as long as one or more lipid bilayers are formed. That is, as described above, when the lipid bilayer membrane forming apparatus according to the embodiment of the present invention has a plurality of first accommodating portions and / or, three or more second accommodating portions per one first accommodating portion. When having a portion, two or more lipid bilayer membranes can be formed.

In the above-mentioned "step of adding an aqueous solution to the oil contained in the first containing portion", the method of adding the aqueous solution to the oil finally forms droplets from the aqueous solution in the oil contained in the first containing portion. The result is not particularly limited. The above method may be, for example, a method of dropping an aqueous solution into oil using a micropipette or a syringe (hereinafter, also referred to as a dropping method), or the tip of the micropipette or syringe is inserted into the oil. A method of injecting an aqueous solution into oil (hereinafter, also referred to as an injection method) may be used.

The aqueous solution dropped onto the oil by the dropping method enters the oil and forms droplets. Depending on the injection method, the aqueous solution injected into the oil forms droplets in the oil. More specifically, in the aqueous solution placed in the oil by a method such as a dropping method or an injection method, the desired lipid contained in the oil or the aqueous solution is then moved to the interface of the aqueous solution, and the lipid causes the desired lipid. A lipid monolayer is formed. As a result, droplets in which the lipid single film is arranged on the surface of the aqueous solution are formed. The droplet covered with the lipid bilayer is then contained in the second storage section 3, where it comes into contact with other droplets covered with the lipid bilayer and automatically interacts with the lipid bilayer. Form a lipid bilayer membrane.

The time for forming the lipid monolayer on the surface of the droplet (in other words, the time for forming the lipid monolayer) is the concentration of lipids that can form the lipid monolayer, which is contained in the oil or the aqueous solution in advance, and the lipid. It depends on the type of oil and / or other components of the lipid that can form a monolayer. In the second accommodating portion, a lipid single film needs to be formed on the surface of these droplets before the droplets come into contact with other droplets. Therefore, by appropriately adjusting the concentration of the lipid capable of forming the lipid monolayer, the type of the oil of the lipid capable of forming the lipid monolayer, and / or other components contained in the oil or the aqueous solution in advance. , The lipid monolayer formation time may be adjusted.

In the lipid bilayer forming method according to the embodiment of the present invention, the position of the oil to which the aqueous solution is added can be arbitrarily determined in order to change the lipid bilayer forming time. Specifically, the distance from the second accommodating portion (specifically, the distance from the second accommodating portion when the lipid bilayer film forming apparatus is viewed from above vertically) is changed as the position of the oil to which the aqueous solution is added. Thereby, it is possible to change the lipid monolayer formation time. More specifically, when the aqueous solution is added to the oil at a position far from the second accommodating portion, the lipid is single as compared with the case where the aqueous solution is added to the oil at a position close to the second accommodating portion. The membrane layer formation time can be lengthened.

The oil used in the lipid bilayer forming method according to the embodiment of the present invention is not particularly limited, and examples thereof include phospholipids, hexadecanes, decanes, squalene, silicon oils, and mineral oils. Among these, hexadecane is preferable because it does not penetrate into the formed lipid monolayer and the formed lipid monolayer has a membrane structure close to that of a biological membrane and has low volatility.

The oil may contain lipids capable of forming a lipid monolayer. As another configuration, the aqueous solution may contain a lipid capable of forming a lipid monolayer. Alternatively, both the oil and the aqueous solution may contain lipids capable of forming a lipid monolayer. When the oil contains lipids capable of forming a lipid monolayer, the droplets can be covered with the lipid monolayer even if the aqueous solution does not contain lipids capable of forming a lipid monolayer. Become.

The lipid capable of forming the lipid single membrane is not particularly limited as long as it can form a lipid bilayer membrane, and examples thereof include phospholipids and cholesterol. Examples of the phospholipid include, but are not limited to, phosphatidylcholine, difitanoylphosphatidylcholine, dipalmitoylphosphatidylcholine, palmitoyloleoil phosphatidylcholine, dioleoil phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, sphingomyelin, and phosphatidylglycerol. ..

When the oil contains a lipid capable of forming a lipid monolayer, the concentration of the lipid is not particularly limited, but is preferably 10 mg / ml to 200 mg / ml, and more preferably 15 mg / ml to 150 mg / ml. , 25 mg / ml to 100 mg / ml, more preferably 40 mg / ml to 60 mg / ml. With the above configuration, even when the aqueous solution does not contain a lipid capable of forming a lipid monolayer, a short-time (for example, less than 10 seconds) lipid monolayer formation time can be realized.

The aqueous solution used in the lipid bilayer forming method according to the embodiment of the present invention is not particularly limited as long as it is a solution containing water as a main component, and the aqueous solution has various components (for example, salts, bases, sugars, lipids). , And proteins, etc.) may be included. The composition of the aqueous solution can be appropriately set, and may be a composition that imitates a specific environmental component in a living body such as a specific cytosol component, a blood component, or a lymph fluid component, for example.

The salt contained in the aqueous solution is not particularly limited, but is appropriately selected from salts contained in the living body, for example, metal salts such as sodium salt, potassium salt, magnesium salt, calcium salt, and phosphate, and amino acids. Examples include salt.

The base contained in the aqueous solution is not particularly limited, and examples thereof include guanine, adenine, thymine, and cytosine.

The lipid contained in the aqueous solution is not particularly limited, and examples thereof include cholesterol, glycolipid, phospholipid, and complex lipid. Some of these lipid components can move to the interface between the droplet and the oil in the formed droplets and become a component of the lipid monolayer. Such lipids that can be components of the lipid monolayer include cholesterol, glycolipids, phospholipids, and complex lipids. That is, in one embodiment of the present invention, the lipid monolayer covering the droplets may contain lipids such as cholesterol, glycolipids, phospholipids, and complex lipids.

The aqueous solution may contain lipids capable of forming a lipid monolayer. When the aqueous solution contains lipids capable of forming a lipid monolayer, the droplets can be covered with the lipid monolayer even if the oil does not contain lipids capable of forming a lipid monolayer. Become. Examples of the lipid capable of forming the lipid single membrane contained in the aqueous solution include the same substances as the lipid capable of forming the lipid single membrane contained in the oil, as described above.

When the aqueous solution contains a lipid capable of forming a lipid monolayer, the concentration of the lipid is not particularly limited, but is preferably 10 mg / ml to 200 mg / ml, and more preferably 15 mg / ml to 150 mg / ml. , 25 mg / ml to 100 mg / ml, more preferably 40 mg / ml to 60 mg / ml. With the above configuration, even when the aqueous solution does not contain a lipid capable of forming a lipid monolayer, a short-time (for example, less than 10 seconds) lipid monolayer formation time can be realized.

The protein contained in the aqueous solution is not particularly limited, and examples thereof include an enzyme protein, a transport protein, and a membrane protein. These proteins are embedded within the lipid bilayer membrane, which allows the reproduction of cell membranes.

Membrane proteins include, but are not limited to, various ion channels, transmembrane adhesion proteins, transporters, transmembrane transport proteins, and the like. Further, the protein may be a protein whose function is unknown, a protein whose function is unknown even though it is a membrane protein, or a protein whose function is unknown.

The method for forming a lipid bilayer according to an embodiment of the present invention also forms a lipid bilayer, and at the same time or after the lipid bilayer is formed, for example, free diffusion and / or a micropipette, etc. It is possible to embed a membrane protein in a lipid bilayer using.

In the method for forming a lipid bilayer membrane according to an embodiment of the present invention, it is also possible to add an arbitrary component into the droplet forming the lipid bilayer membrane after the lipid bilayer membrane is formed. Alternatively, it is also possible to recover any component from the droplet. In this case, for example, an arbitrary component may be injected into the droplet or an arbitrary component in the droplet may be sucked by using a micropipette or the like.

[Evaluation system]
The evaluation system according to the embodiment of the present invention is characterized by including the lipid bilayer film forming apparatus according to the embodiment of the present invention described in the above [lipid bilayer film forming apparatus].

The evaluation system according to the embodiment of the present invention preferably further includes electrodes (in other words, sensors capable of measuring various changes). It is more preferable that the electrode is arranged in the second accommodating portion. With the above configuration, the movement of substances between the droplets via the lipid bilayer formed between the plurality of droplets 5 or the function of the membrane protein contained in the lipid bilayer can be electrically controlled. It becomes possible to analyze. For example, it is possible to electrically analyze the inflow and outflow of various ions between droplets via various ion channels contained in the lipid bilayer membrane.

The method of arranging the electrode in the second accommodating portion is not particularly limited, and the electrode may be inserted into the second accommodating portion from the periphery of the lipid bilayer film forming apparatus by a manipulator, or may be inserted into the second accommodating portion in advance. It may be arranged. When the electrode is inserted into the second housing by a manipulator, the electrode is movable, so that the electrode can be inserted into the second housing at any position and time. On the other hand, when the electrodes are arranged in advance in the second accommodating portion, (1) the electrodes can be arranged in the droplet 5 accommodated in the second accommodating portion without the need for a manipulator or the like. And (2) it is possible to carry the lipid bilayer film forming apparatus together with the electrode.

The arrangement form of the electrodes is not particularly limited, but it is preferable that the electrodes are arranged at the bottom of the second accommodating portion of the lipid bilayer film forming apparatus.

FIG. 5 is a cross-sectional view in the longitudinal direction of the second accommodating portion of the lipid bilayer film forming apparatus included in the evaluation system according to the embodiment of the present invention.

In FIG. 5, in the lipid bilayer film forming apparatus included in the evaluation system according to the embodiment of the present invention, the electrode 6 is arranged at the bottom of the second accommodating portion 3 provided in the first accommodating portion 2 of the substrate 1. ing.

According to the above configuration, after the droplet 5 is accommodated in the second accommodating portion 3, the electrode 6 penetrates the single film formed on the surface of the droplet 5 by the weight of the droplet 5 and is inside the droplet 5. Can be placed in.

The configuration of the electrode 6 is not particularly limited, and a commercially available electrode can be used as appropriate.

The electrodes 6 may be arranged in each of the second accommodating portions 3, or may be arranged in two or more second accommodating portions 3. For example, when three or more second accommodating portions 3 are arranged in series, the electrodes 6 can be arranged at least in the two second accommodating portions 3 arranged at both ends. According to this configuration, the movement of substances through a plurality of lipid bilayer membranes can be electrically analyzed.

The power supply or measuring device connected to the electrode 6 may be included in the evaluation system. Alternatively, a power source or measuring device not included in the evaluation system may be used.

The electrode 6 and the power supply or the measuring device may be connected at all times, or may be connected at any time and time. If the electrode 6 and the power supply or measuring device are connected at any time and time, the electrode 6 can have removable contacts with the power supply or measuring device. With the above configuration, a plurality of lipid bilayer film forming devices having electrodes 6 can be connected to a power source or a measuring device in any order, time and time.

The evaluation system according to the embodiment of the present invention is not particularly limited as long as it includes the lipid bilayer film forming apparatus according to the embodiment of the present invention, but specifically, (1) an electrical change measuring apparatus, ( 2) Radioisotope evaluator, (3) Smell sensor device, and (4) DNA sequencer (eg Byley H., Sequencing single molecules of DNA, Curr. Opin. Chem. Biol., 10 (6), 628 -See 637, 2006).

In the case of an electrical change measuring device including a lipid bilayer film forming device according to an embodiment of the present invention, the device is separated by a lipid bilayer film by the above electrode (in other words, a sensor capable of measuring various changes). Various changes between the aqueous solutions (eg, changes in the amount of ions and / or the charge of the ions) may be measured.

In the case of the radioisotope evaluation device including the lipid bilayer film forming device according to the embodiment of the present invention, the device is separated by the lipid bilayer film by the above electrode (in other words, a sensor capable of measuring various changes). Various changes between aqueous solutions (eg, changes in the amount of radioisotopes) may be measured.

In the case of the odor sensor device including the lipid bilayer film forming device according to the embodiment of the present invention, the aqueous solutions separated by the lipid bilayer film by the above electrode (in other words, a sensor capable of measuring various changes). Various changes (eg, changes in the amount of odorous substances) may be measured.

In the case of a DNA sequencer including a lipid bilayer forming apparatus according to an embodiment of the present invention, various types of aqueous solutions separated by the lipid bilayer by the above electrodes (in other words, sensors capable of measuring various changes). Changes (eg, changes in ion amount and / or ion charge) may be measured.

One embodiment of the present invention can also be configured as follows.

[1] A first accommodating portion for accommodating oil and a plurality of second accommodating portions provided at the bottom of the first accommodating portion and for accommodating droplets covered with a lipid bilayer are provided. The inner surface of the first accommodating portion has a second accommodating portion so that when an aqueous solution is added to the oil contained in the first accommodating portion, the droplets formed from the aqueous solution are guided to the second accommodating portion. The shape converges toward the accommodating portion, and the plurality of second accommodating portions are arranged so that the droplets accommodating in the second accommodating portion come into contact with each other. Lipid bilayer film forming apparatus.

According to the above configuration, the aqueous solution added to the oil forms droplets covered with a single lipid membrane. In the droplet, the hydrophobic region of the lipid is arranged so as to face the oil, and the hydrophilic region of the lipid is arranged so as to face the aqueous solution. The droplet is accommodated in the second accommodating portion after moving toward the second accommodating portion along the inner surface of the first accommodating portion. The droplets housed in the plurality of second accommodating portions come into contact with each other, and at this time, the lipid single membranes arranged outside the droplets also come into contact with each other. As described above, in the lipid monolayer, the hydrophobic region of the lipid is arranged so as to face the oil, and the hydrophilic region of the lipid is arranged so as to face the aqueous solution, so that the lipid monolayers come into contact with each other. In this region, the hydrophobic regions of the lipid are arranged so as to face each other, and the hydrophilic regions of the lipid are arranged so as to face the aqueous solution. As a result, a lipid bilayer membrane similar to the structure of the cell membrane is formed.

[2] When the shape of the droplet is _{a sphere having a diameter of r 3} and the diameter of the maximum inscribed circle of the space in the second accommodating portion is r ₂ , r ₃ ≤ r ₂ is satisfied. , [1]. The lipid bilayer film forming apparatus.

The droplet contained in the second accommodating portion can change from a spherical shape to a flat shape due to gravity. With the above configuration, a flat-shaped droplet can be accommodated in the second accommodating portion.

[3] The lipid bilayer film forming apparatus according to [1] or [2], wherein _{the depth H 2} of the second accommodating portion satisfies 0 <H ₂ <(r _{3) / 2.} ..

According to the above configuration, the wide area on the upper side of the droplet contained in the second accommodating portion is exposed on the second accommodating portion, and the exposed droplet portions can easily come into contact with each other.

[4] The depth H ₂ of the second accommodating portion satisfies (r ₃ ) / 2 ≦ H _2, and a hole penetrating the wall surface is formed on the wall surface separating the second accommodating portions. The lipid bilayer film forming apparatus according to [1] or [2].

According to the above configuration, (i) the narrow area above the droplets contained in the second containment is exposed above the second containment, or (ii) is contained in the second containment. Even if the upper region of the droplets is not exposed above the second accommodating portion, the droplets can easily come into contact with each other through the holes.

[5] A method for forming a lipid bilayer film using the lipid bilayer film forming apparatus according to any one of [1] to [4], wherein the oil contained in the first storage portion is used. A method for forming a lipid bilayer membrane, which comprises a step of adding an aqueous solution.

According to the above configuration, the aqueous solution added to the oil forms droplets covered with a single lipid membrane. In the droplet, the hydrophobic region of the lipid is arranged so as to face the oil, and the hydrophilic region of the lipid is arranged so as to face the aqueous solution. The droplet is accommodated in the second accommodating portion after moving toward the second accommodating portion along the inner surface of the first accommodating portion. The droplets housed in the plurality of second accommodating portions come into contact with each other, and at this time, the lipid single membranes arranged outside the droplets also come into contact with each other. As described above, in the lipid monolayer, the hydrophobic region of the lipid is arranged so as to face the oil, and the hydrophilic region of the lipid is arranged so as to face the aqueous solution, so that the lipid monolayers come into contact with each other. In this region, the hydrophobic regions of the lipid are arranged so as to face each other, and the hydrophilic regions of the lipid are arranged so as to face the aqueous solution. As a result, a lipid bilayer membrane similar to the structure of the cell membrane is formed.

[6] An evaluation system comprising the lipid bilayer film forming apparatus according to any one of [1] to [4].

According to the above configuration, since the lipid bilayer membrane can be easily formed, the function of the lipid bilayer membrane itself and the function of the membrane protein embedded in the lipid bilayer membrane are related to the lipid bilayer membrane. Various functions can be easily evaluated (eg, using a convenient device and / or in the field rather than in the laboratory).

[7] The evaluation system according to [6], wherein the evaluation system further includes electrodes.

With the above configuration, it is possible to arrange the electrodes inside each of the droplets contained in the second accommodating portion. Therefore, by measuring the voltage and / or the voltage between the electrodes, the movement of substances (for example, ions, drugs, toxic substances, etc.) through the lipid bilayer formed between the plurality of droplets can be evaluated. Can be done.

The lipid bilayer film forming apparatus according to one embodiment of the present invention includes (1) an electrical change measuring apparatus, (2) a radioisotope evaluation apparatus, (3) an odor sensor apparatus, and (4) a DNA sequencer. It can be used.

1 Substrate 2 1st accommodating part 3 2nd accommodating part 4 Oil 5 Droplet 6 Electrode r ₁ Diameter of 1st accommodating part r ₂ Diameter of maximum inscribed circle in space of 2nd accommodating part r ₃ Diameter of droplet H ₁ Depth of 1st containment H ₂ Depth of 2nd containment

Claims (7)

The first storage part where oil is stored and
A plurality of second accommodating portions provided at the bottom of the first accommodating portion and for accommodating droplets covered with a single lipid membrane are provided.
The inner surface of the first accommodating portion is such that when an aqueous solution is added to the oil contained in the first accommodating portion, the droplets formed from the aqueous solution are guided to the second accommodating portion. It has a shape that converges toward the containment part,
The shape of the inner surface of the first accommodating portion is a shape corresponding to a part of the surface of the sphere, a conical shape, a polygonal pyramid shape, or a combination thereof.
A lipid bilayer film forming apparatus, wherein the plurality of second accommodating portions are arranged so that the droplets accommodating in the second accommodating portion come into contact with each other. The claim is characterized in that r ₃ ≤ r ₂ is satisfied when the shape of the droplet is _{a sphere having a diameter of r 3} and the diameter of the maximum inscribed circle of the space in the second accommodating portion is r _2. The lipid bilayer film forming apparatus according to 1. The lipid bilayer film forming apparatus according to claim 1 or 2, wherein _{the depth H 2} of the second accommodating portion satisfies 0 <H ₂ <(r _{3) / 2. The depth H 2} of the second accommodating portion satisfies (r ₃ ) / 2 ≦ H _2.
The lipid bilayer film forming apparatus according to claim 1 or 2, wherein a hole penetrating the wall surface is formed on a wall surface separating the second accommodating portions. A method for forming a lipid bilayer membrane using the lipid bilayer membrane forming apparatus according to any one of claims 1 to 4.
Have a step of adding an aqueous solution to the oil contained in the first housing portion,
A method for forming a lipid bilayer membrane, wherein at least one selected from the group consisting of the above oil and the above aqueous solution contains a lipid capable of forming a lipid single membrane . An evaluation system comprising the lipid bilayer film forming apparatus according to any one of claims 1 to 4. The evaluation system according to claim 6, wherein the evaluation system further includes electrodes.

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