JP2019072698A - Lipid double membrane forming device, and lipid double membrane forming method using the same - Google Patents

Abstract

To provide a lipid double membrane forming device and a forming method of lipid double membrane using the same, capable of forming a lipid double membrane with a simple operation at a service space.SOLUTION: A lipid double membrane forming device comprises: a substrate; a first well having a flat opening; a first lipid single membrane formable droplet filled in the first well; a moving membrane relatively movable to the substrate; a second well having the flat opening; a second lipid single membrane formable droplet filled in the second well; and one or two seal member coating the first droplet and the second droplet at same time or individually. Before use of the device, the first well and the second well at positions apart from each other, during the use of the device, the moving member is moved relatively to the substrate, the opening of the first well and the opening of the second well are overlapped, the first droplet and the second droplet are in contact, and a lipid double membrane is formed at the contact part.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a lipid bilayer membrane forming apparatus and a lipid bilayer membrane forming method using the same.

  The cells constituting the organism, various organelles such as mitochondria, Golgi body, endoplasmic reticulum, etc. existing in the cells, cell nuclei, etc. are covered with a living membrane on the outside, and this living membrane is basically a lipid bilayer membrane. It consists of Various proteins having physiological activity, ie, receptors, enzymes, etc., are retained on the lipid bilayer membrane in such a manner as to penetrate the lipid bilayer membrane. These transmembrane proteins play an important role in vivo. In particular, various receptors present on cell membranes are known to trigger various physiological responses by binding to ligands present in vivo. Therefore, various ligands that enhance the function of the receptor, inhibitors that inhibit the function of the receptor, and the like are used as pharmaceuticals, and natural or artificial ligands and inhibitors that can be used as new pharmaceuticals are studied. ing.

  Moreover, it is also known to hold | maintain a protein on a lipid bilayer membrane, and to utilize as a sensor. For example, a lipid bilayer membrane holds an olfactory receptor protein to form an odor sensor, or a specific binding property to specifically bind to a test substance in a droplet when the lipid bilayer membrane is formed by a droplet contact method A sensor that contains a substance, while retaining an ion channel protein in a lipid bilayer membrane, and in the presence of a test substance, causes the test substance to bind to a specific binding substance and occlude the ion channel. It is also known (patent document 1). Patent Document 1 also describes that one droplet to be contacted in the droplet contact method is a hydrogel in order to facilitate detection of a test substance present in air.

  On the other hand, Patent Document 2 mechanically separates two droplets brought into contact by a droplet contact method, and then moves one droplet to recontact with different droplets to form a lipid bilayer membrane. A lipid bilayer membrane forming device capable of exchanging droplets to be brought into contact has been described.

JP 2017-83210 A JP, 2014-100672, A

  The sensor having a lipid bilayer membrane described in Patent Document 1 or the like is also used for detection of odorous substances contained in the atmosphere. If a droplet is prepared in the laboratory (or in a manufacturing plant when producing a lipid bilayer membrane forming instrument commercially) to form a lipid bilayer membrane, an instrument comprising a lipid bilayer membrane is used. It needs to be transported to the place of use, and there is a risk that the lipid bilayer membrane may be broken during transportation or storage. In order to solve this problem, it is conceivable to form a lipid bilayer membrane at the place of use, but using an experimental instrument at a place of use such as outdoors, drop droplets into the wells to form a lipid bilayer membrane Is unrealistic.

  Therefore, an object of the present invention is to provide a lipid bilayer membrane-forming apparatus and a method of forming a lipid bilayer membrane using the same, which can form a lipid bilayer membrane by a simple operation at the place of use. .

  As a result of intensive studies, the inventors of the present invention transport the solution to the use place without contact between the two droplets to be subjected to the droplet contact method, and contact the two droplets by simple operation at the use location to perform lipid doublets. It was conceived that forming the membrane could achieve the above-mentioned purpose, and a lipid bilayer membrane-forming device was devised which makes this possible.

That is, the present invention provides the following.
1. A lipid bilayer membrane forming device,
A substrate,
A first well provided in the substrate and having a flat opening;
A first lipid monolayer forming droplet filled in the first well;
A movable member movable relative to the substrate;
A second well provided in the moving member and having a flat opening;
A second lipid monolayer forming droplet filled in the second well;
And one or two seal members that simultaneously or separately cover the first lipid single-film-forming droplet and the second lipid single-film-forming droplet,
Before use of the device, the first well and the second well are spaced apart from each other, and in use, the moving member is moved relative to the substrate to The opening of the well and the opening of the second well overlap so that the first lipid single film forming droplet and the second lipid single film forming droplet are in contact with each other. A lipid bilayer is formed at the contact site,
Lipid Bilayer Forming Device.

2. The device according to 1, wherein the moving member slides and moves in the substrate.
3. The device according to 2, wherein the moving member is slidably inserted in a groove provided in the substrate.
4. 3. The device according to 3, wherein the moving member comprises a protrusion that protrudes out of the substrate.
5. The sealing member is a single sealing member that simultaneously covers the first lipid single-film-forming droplet and the second lipid single-film-forming droplet, according to any one of 1 to 4, Fixtures.
6. The seal member according to any one of 1 to 5, wherein the seal member is disposed at a position not in contact with each of the first lipid monolayer-forming droplet and the second lipid monolayer-forming droplet. Fixtures.
7. The device according to any one of 1 to 6, wherein the sealing member is an adhesive tape.
8. In the opening of the first well or the opening of the second well, a partition having a through hole is disposed, and the first lipid single film-forming droplet and the second lipid single film are provided. The device according to any one of 1 to 7, wherein the forming droplets are in contact via the through holes.
9. The device according to any of 1 to 8, wherein the first lipid monolayer forming droplet and the second lipid monolayer forming droplet are in the form of a hydrogel.
10. The device according to any one of 1 to 9, wherein the first lipid monolayer forming droplet and the second lipid monolayer forming droplet are in a frozen state.
11. 10. The device according to 10, wherein the organic solvent contained in the first lipid monolayer forming droplet and the second lipid monolayer forming droplet comprises n-hexadecane.
12. The device according to 11, wherein the organic solvent contained in the first lipid monolayer-forming droplet and the second lipid monolayer-forming droplet is a mixed solvent of n-hexadecane and n-decane.
13. The device according to any one of the preceding claims, wherein an electrode is disposed at the bottom of each of the first and second wells.

14. Providing the device according to any one of 1 to 13;
Transporting the device to its place of use;
At the place of use, when the first lipid monolayer-forming droplet and the second lipid monolayer-forming droplet are in a frozen state, after melting these droplets, the first well The moving member is moved relative to the substrate to a position where the opening and the opening of the second well overlap, so that the first lipid single-film-forming droplet and the second lipid are moved. Contacting a single membrane-forming droplet and forming a lipid bilayer membrane at the contact site.

Further, the present invention provides a device of the present invention as described above,
Transporting the device to its place of use, and where the first lipid single membrane forming droplet and the second lipid single membrane forming droplet are in a frozen state at the location of use After melting the droplet, the moving member is moved relative to the substrate to a position where the opening of the first well and the opening of the second well overlap, to thereby form the first lipid Contacting a single membrane-forming droplet with the second lipid single-film-forming droplet, and forming a lipid bilayer membrane at the contact site, provides a method for forming a lipid bilayer membrane.

  The present invention provides a lipid bilayer membrane forming device and a method of forming a lipid bilayer membrane using the same, which can form a lipid bilayer membrane by a simple operation at the place of use. By using the device of the present invention, a lipid bilayer membrane can be formed in situ by a simple operation without using any experimental instrument at the place of detection of a test substance such as outdoors. There is no risk of being destroyed during transportation or storage.

It is a typical disassembled perspective view of one embodiment of a lipid bilayer membrane formation instrument of the present invention. FIG. 5 is a schematic exploded perspective view of another embodiment of the lipid bilayer membrane formation device of the present invention, showing the state during storage and transportation of the device. It is a figure which shows the state at the time of use of the lipid bilayer membrane formation instrument shown in FIG. It is a figure which shows the result of the current measurement performed in the following reference example and the Example. It is a figure which shows the result of the electric current measurement in reciprocation test performed in the following Example.

  The lipid bilayer membrane formation device of the present invention is to form a lipid bilayer membrane by a lipid bilayer membrane formation method by a droplet contact method. Droplet contact methods themselves are well known in the art and are also described in the above-mentioned U.S. Pat. That is, this is a method of forming a lipid bilayer membrane on the contact surface by bringing droplets enclosed in a lipid single membrane into contact with each other. A lipid single membrane-enclosed droplet can be easily formed according to known methods. For example, a lipid bilayer membrane can be easily formed by adding water or an aqueous solution (such as a buffer solution) to a lipid bilayer-forming lipid solution. Here, the lipid bilayer membrane-forming lipid may be a known phospholipid used in the preparation of a liposome, and in order to simulate a reaction in a biological membrane, one that is the same as or similar to a biological membrane is preferable, Phospholipids conventionally widely used in this field, such as diphytanoyl phosphatidyl choline (DPhPC), dipalmitoyl phosphatidyl choline, palmitoyl oleoyl phosphatidyl choline (1- Palmitoyl 2-Oleoyl phosphatidylcholine (POPC), dioleoylphosphatidylcholine (DOPC), etc. can be mentioned as a preferred example. Since many of these are commercially available, commercially available products can be preferably used. The concentration of the phospholipid in the solution used to form the lipid bilayer membrane is not particularly limited as long as it is a concentration at which the lipid bilayer membrane can be formed, but generally 5 g / L to 30 g / L or so, preferably 10 g / L. It is about L to 20 g / L. Also, the solvent for the phospholipid solution is not particularly limited, but an organic solvent is preferable, and an aliphatic hydrocarbon solvent such as n-decane is preferable. When a lipid bilayer membrane retains an ion channel protein such as α-hemolysin or a functional protein such as an olfactory receptor, as is well known, the protein is dissolved in at least one of the droplets. Keep it. Although the final concentration of the protein at this time is not particularly limited, it is usually about 0.1 nM to about 10 nM, preferably about 0.5 nM to about 2 nM.

  The droplets may be liquid or may be in the form of a hydrogel, and in the present invention, the term "droplet" is used in the sense to include also those in the form of hydrogel. Even when the droplet is in the form of a hydrogel, a lipid bilayer membrane can be formed, and a lipid bilayer membrane can be formed by contacting with another droplet on which the lipid monolayer membrane is formed. Patent Document 1 describes the above. In the present invention, the droplet is in the form of a hydrogel from the viewpoint of preventing the droplet from coming into contact with the seal member (described later) by enhancing the stability during transportation or storage and by vibration etc. during transportation. Is preferred. The hydrogel can be easily formed simply by including agarose, agar, gelatin or the like (hereinafter, "hydrogelating agent") in the droplet. Although the final concentration of the hydrogel forming agent in the droplets is not particularly limited as long as it can form a hydrogel, it is usually about 0.05% by mass to about 20% by mass, preferably about 0.1% by mass to about 15% by mass, more preferably And about 0.3% by mass to 3.0% by mass.

  The droplets may be in a frozen state to enhance the stability of the droplets during storage and transport. Freezing can be carried out by holding the device in a conventional freezer (usually -20 ° C). Droplets in the form of hydrogels can also be frozen and stored and transported. In this case, the frozen droplet is thawed immediately before use to form a lipid bilayer membrane. When the droplets are in the form of a hydrogel, “melting” does not have to dissolve the hydrogel itself, as long as the water retained in the hydrogel is melted. This is true if the device is left at room temperature. When the droplets are stored in a frozen state and transported, the organic solvent contained in the droplets preferably contains n-hexadecane, and in particular, a mixture of n-hexadecane and widely used n-decane It is preferably a solvent. The organic solvent containing n-hexadecane, in particular a mixed solvent of n-hexadecane and n-decane, facilitates formation of a lipid bilayer membrane rapidly and reliably after melting. When a mixed solvent of n-hexadecane and n-decane is used, the ratio (n-hexadecane: n-decane) is preferably about 4: 6 to 8: 2 on a volume basis.

  In the lipid bilayer film formation tool of the present invention, the first droplet is filled in the first well formed in the substrate, and the first well is provided in the movable member movable relative to the substrate. Fill the second well into the second well. The first well and the second well each have a flat opening. During storage and transportation, the first well and the second well are separated from each other. In use, the moving member is moved relative to the substrate, and the opening of the first well and the opening of the second well overlap to form the first single lipid membrane. The liquid droplet and the second lipid single film-forming droplet come into contact, and a lipid bilayer membrane is formed at the contact site.

  Hereinafter, preferred embodiments of the present invention will be described with reference to the drawings.

  FIG. 1 is a schematic exploded perspective view of a preferred embodiment of the lipid bilayer membrane formation device of the present invention. The lipid bilayer film forming apparatus comprises a substrate 10 in which a first well 12 is provided. The first well 12 has a flat opening 12a. The first well 12 shown in FIG. 1 is in a C shape because it has a cylindrical shape with a flat opening 12a, but the wells (including the second well described later) are not necessarily C. The shape does not have to be a letter, and may be a rectangular parallelepiped or a cube as shown in Patent Document 2. At the bottom of the first well 12, a through hole 12b for connecting an electrode is formed. The first well 12 is filled with a first lipid monolayer-forming droplet (not shown) (hereinafter, “first droplet”).

  The lipid bilayer membrane forming apparatus comprises a moving member 14 movable relative to the substrate 10. In the moving member 14, a second well 16 is provided. Similar to the first well 12, the second well 16 has a flat opening and is C-shaped. A partition 18 having a through hole 18 a is disposed at the opening of the second well 16, and the second well 16 is opened only by the through hole 18 a. Although the partition 18 having such a through hole 18a is not necessarily required, the lipid bilayer formed rapidly by forming the lipid bilayer by reducing the area of the formed lipid bilayer. Help to be held stable. The diameter of the through hole is not particularly limited, but is usually about 0.1 mm to 1 mm. As a material which constitutes this partition, as it is indicated also in patent documents 1, paraxylene system polymer (brand name parylene) can be used preferably. The moving member 14 is slidably inserted in a groove 20 provided in the substrate 10. The bottom surface of the groove 20 constitutes the bottom surface of the second well 16 and a through hole for connecting an electrode to a portion constituting the bottom surface of the second well 16 when using the lipid bilayer membrane formation device 20a is provided. The second well 16 is filled with a second lipid single-film-forming droplet (not shown) (hereinafter, “second droplet”).

  At the time of storage and transportation of the device, the first well 12 and the second well 16 are respectively covered by seal members (not shown). The sealing member may be provided separately for covering the first well 12 and for covering the second well 16, but may cover both wells with a single sealing member. The latter is preferable because it is simple and the structure is simple. Covering both droplets with the seal member prevents drying of the droplets and contamination of the droplets. As shown in FIG. 1, a step 22 is provided between the top surface of the substrate 10 and the top of the first well 12, while the height of the second well 16 is also usually the first well. Since it is the same as the height of 12, by covering the upper surface of the substrate 10 with a single seal member, the seal member is disposed at a position where the first droplet and the second droplet do not contact the seal member. be able to. The seal member is preferably made of an adhesive tape. It is convenient and preferable to form a seal member with an adhesive tape and to stick one adhesive tape on the upper surface of the substrate 10. When using the device, the sealing member can be removed simply by peeling off a piece of adhesive tape.

  When using the device, remove the seal member (if the seal member is an adhesive tape, peel off the adhesive tape), and the opening of the first well 12 and the opening of the second well 16 overlap The moving member 14 is moved by a finger or the like (sliding in the groove 20). Thereby, the first droplet and the second droplet come in contact with each other through the through hole 18a, and a lipid bilayer membrane is formed in the through hole 18a. When the droplets are in a frozen state, the droplets are melted before contacting the two droplets. Melting of the droplets can be performed simply by leaving the device at room temperature. For example, it can be carried out by leaving at room temperature for about 30 minutes to 6 hours. Or you may heat at 50 degreeC for about 10 to 30 minutes. Further, even when the device is stored under refrigeration (for example, 4 ° C.), it is preferable to form a lipid bilayer membrane after leaving the liquid droplets at room temperature for about 5 to 30 minutes at room temperature. As was confirmed experimentally in the examples described later, by leaving the first droplet and the second droplet in contact with each other through the through hole 18a, the lipid in the through hole 18a can be reduced. A heavy membrane is formed, and the functional protein (in the example, α-hemolysin, which is an ion channel protein) contained in the droplet is functionally retained by the lipid bilayer membrane.

  FIGS. 2 and 3 show an embodiment of a preferred lipid bilayer membrane forming device of the present invention, wherein the transfer member has a finger pinch for movement. In FIGS. 2 and 3, the same parts as in the embodiment shown in FIG. FIG. 2 shows the state of storage and transportation of the device, and FIG. 3 shows the state of use of the device.

  In the embodiment shown in FIGS. 2 and 3, the through hole 26 is provided on the side surface of the substrate 10, while the movable member 14 has a projection 14 a that penetrates the through hole 26 and protrudes to the outside of the substrate 10. Is provided. Further, a step 24 is provided between the upper surface of the moving member 14 and the upper portion of the second well 16. In this case, the first well 12 and the second well 16 are covered at the same time as covering the first well 12 and the second well 16 by using one adhesive tape as a seal member and sticking it on the upper surface of the substrate 10 and the upper surface of the movable member 14. The position with respect to the substrate 10 can be fixed, and the moving member 14 can be prevented from moving unintentionally during storage and transportation.

  During storage and transportation of the device, as shown in FIG. 2, the first well 12 and the second well 16 are at mutually separated positions. When using the device, as shown in FIG. 3, the finger 14 pinches and moves the protrusion 14 a so that the openings of the first well 12 and the second well 16 are in the overlapping position. Then, a lipid bilayer membrane is formed in the through hole 18a, as described in the embodiment shown in FIG.

  In the embodiment shown in FIGS. 2 and 3, the protrusion 14 a is a protrusion that protrudes in the horizontal direction, but may be a protrusion that protrudes in the vertical direction. Alternatively, a recess may be provided on the upper surface of the moving member 14 and the moving member 14 may be moved by hooking the recess.

  In each of the embodiments shown in FIGS. 1 to 3, the movable member 14 slides in the horizontal direction in the groove 20 provided in the substrate 10. However, the substrate 10 has a guide extending in the vertical direction. It is also possible to slide the moving member 14 vertically in this guide. In this case, the moving member 14 is disposed above the substrate 10 so that the first well 12 and the second well 16 do not contact each other when storing and transporting the tool, and in use, the moving member 14 Can be pushed downward with a finger to overlap the openings of the first well 12 and the second well 16.

  Hereinafter, the present invention will be specifically described based on examples. However, the present invention is not limited to the following examples.

Reference Example 1, Example 1
1. Lipid Bilayer Forming Device A lipid bilayer device shown in FIG. 1 was produced. The thickness of the substrate 10 is 4 mm, the thickness of the moving member 14 is 3 mm, the diameter of each of the first well 12 and the second well 16 is 4 mm, and each height is 3 mm (thus, the step 22 is 1 mm). The diameter of the through hole 18a was 0.6 mm. In addition, silver-silver chloride electrodes were disposed in the through holes 12 b and the through holes 20 a.

2. Preparation of lipid, gel and aqueous solution
(1) A lipid dispersed decane solution was prepared. Lipids (DPhPC, Avanti Polar Lipids) were dispersed in n-decane: hexadecane (Sigma) at a weight ratio of 1: 1 to make 20 mg / mL (lipid solution).
(2) 45 mg of agarose gel (Agarose Type IX-A, Ultra-low Gelling Temperature, Sigma-Aldrich) was weighed and placed in a centrifuge tube.
(3) 1 M KCl (10 mM phosphate buffer, pH 7.4) separately prepared to have an agarose concentration of 1.5% was added.
(4) While paying attention to bumping, it was boiled in a microwave oven (500 W) to melt the agarose. In order to make the gel uniform, it was sufficiently dissolved while stirring with an appropriate vortex mixer.
(5) After dissolution, aliquots were placed in an Eppendorf tube and held on a heat block (40 ° C.) (agarose gel solution).
(6) A 100 nM α-hemolysin (Sigma-Aldrich) aqueous solution was separately prepared.

3. Drop and seal lipid and gel solution to wells
(1) 100 nM α-Hemolysin (1 μL) was added to a gel solution (99 μL) warmed by heat block (40 ° C.) (final concentration 1 nM, hemolysin gel solution). Hold on heat block until dripping into wells.
(2) The moving member was moved so that the wells were separated from each other and were not in contact with each other.
(3) 3 μL of lipid solution was added to two wells. Oil spread on the bottom of the well.
(4) Agarose gel solution (27 μL) was added to the first well.
(5) Subsequently, hemolysin gel solution (27 μL) was added to the second well.
(6) As a sealing member, an adhesive tape (polyolefin micro sealing tape, 3 M) was attached to the upper surface of the substrate so that air did not enter (Example 1). As described above, since there is a step 22 of 1 mm, the gel solution (upper well) and the upper surface of the moving member 14 do not touch the adhesive tape. In Reference Example 1, the seal member was not attached.

4. Storage The prepared devices were stored refrigerated (4 ° C.) overnight (15 hours) (Example 1). In addition, in the reference example 1, the following measurement was performed immediately after preparation of the droplet without storing.

5. Lipid Bilayer Formation and Measurement In Example 1, the device after refrigerated storage was allowed to stand at room temperature for 5 to 30 minutes to return to room temperature, and the adhesive tape was peeled off. In both reference example 1 and example 1, the electrode disposed in the through hole 12b is grounded, and the electrode disposed in the through hole 20a is connected to a known patch amplifier circuit (patent document 1) to measure the current flowing between both wells. did. The applied voltage was 50 to 100 mV, and the ion current was observed at an amplification rate of 1 mV / pA or 10 mV / pA and a sampling frequency of 5 kHz. The obtained signal was subjected to a Bessel low pass filter 1 kHz. The moving member 14 is moved with a finger so that the first droplet and the second droplet are in contact with each other through the through hole 18a. The current measurement was continued after this.

6. Measurement Results The measurement results of the current are shown in FIG. 4 (a) (Reference Example 1) and FIG. 4 (b) (Example 1). Stepped currents were measured as shown in (a) and (b) of FIG. As a result, lipid bilayers were formed, and α-hemolysin was functionally retained in the lipid bilayer, and migration of ions occurred through nanopores (ion channels) possessed by α-hemolysin. It is shown that. As the number of molecules of α-hemolysin retained on the lipid bilayer increases, the measured current increases, resulting in a step-like current. As shown in (a) of FIG. 4, in Reference Example 1, the time until the first step current is measured (ie, the time until the lipid bilayer membrane retaining α-hemolysin is formed) is It was 24 ± 28 seconds, and a lipid bilayer membrane was rapidly formed. Moreover, also in Example 1, the time until the first step current is measured is approximately 10 seconds to 5 minutes, and a lipid bilayer membrane is similarly formed rapidly.

  Furthermore, as in Patent Document 2, a 50-mer single-stranded polythymidylate (ssDNA) is contained in the second droplet, and the same operation as above is performed, and the result of measuring the current is shown in FIG. Shown in. When ssDNA passes through the nanopore of α-hemolysin, a drop in current occurs because the ion channel is blocked and ions do not flow. As shown in (c) of FIG. 4, such a current drop was observed, and it was confirmed that the device of this example can detect ssDNA.

Example 2
The device prepared in Example 1 is stored frozen (-20 ° C.) for various times (about 15 hours, about 40 hours, about 90 hours, 7 days or 1 month) or frozen frozen stool (-10 ° C.) After carrying out the transportation test (total 2 days), the current measurement was performed in the same manner as in Example 1.

  As a result, after moving the transfer member 14, a lipid bilayer membrane was rapidly formed, and a signal indicating nanopore formation derived from α-hemolysin was observed. The time until the lipid bilayer membrane was formed was approximately 10 seconds to 5 minutes. The lipid bilayer formation success rate within 2 minutes for the delivery test was 5/6. The current measurement result at that time is shown in FIG. It was shown that a lipid bilayer membrane can be formed by the lipid bilayer membrane forming device of the present invention under any of the above storage conditions and transport conditions.

Reference Signs List 10 substrate 12 first well 12 a opening 12 b through hole 14 moving member 14 a protrusion 16 second well 18 partition 18 a through hole 20 groove 20 a through hole 22 step 24 step 24 step 26 through hole

Claims (14)

A lipid bilayer membrane forming device,
A substrate,
A first well provided in the substrate and having a flat opening;
A first lipid monolayer forming droplet filled in the first well;
A movable member movable relative to the substrate;
A second well provided in the moving member and having a flat opening;
A second lipid monolayer forming droplet filled in the second well;
And one or two seal members that simultaneously or separately cover the first lipid single-film-forming droplet and the second lipid single-film-forming droplet,
Before use of the device, the first well and the second well are spaced apart from each other, and in use, the moving member is moved relative to the substrate to The opening of the well and the opening of the second well overlap so that the first lipid single film forming droplet and the second lipid single film forming droplet are in contact with each other. A lipid bilayer is formed at the contact site,
Lipid Bilayer Forming Device.   The apparatus according to claim 1, wherein the moving member slides and moves in the substrate.   The apparatus according to claim 2, wherein the moving member is slidably inserted in a groove provided in the substrate.   The apparatus according to claim 3, wherein the moving member comprises a protrusion protruding out of the substrate.   The seal member is a single seal member that simultaneously covers the first lipid single-film-forming droplet and the second lipid single-film-forming droplet. The equipment described in.   The seal member is disposed at a position not in contact with each of the first lipid single-film-forming droplet and the second lipid single-film-forming droplet. The equipment described in.   The device according to any one of claims 1 to 6, wherein the sealing member is an adhesive tape.   In the opening of the first well or the opening of the second well, a partition having a through hole is disposed, and the first lipid single film-forming droplet and the second lipid single film are provided. The device according to any one of claims 1 to 7, wherein the forming droplet is in contact via the through hole.   The device according to any one of the preceding claims, wherein the first lipid monolayer forming droplet and the second lipid monolayer forming droplet are in the form of a hydrogel.   10. The device of any of claims 1-9, wherein the first lipid monolayer forming droplet and the second lipid monolayer forming droplet are in a frozen state.   The device according to claim 10, wherein the organic solvent contained in the first lipid monolayer-forming droplet and the second lipid monolayer-forming droplet comprises n-hexadecane.   The device according to claim 11, wherein the organic solvent contained in the first lipid monolayer-forming droplet and the second lipid monolayer-forming droplet is a mixed solvent of n-hexadecane and n-decane.   13. The device according to any of the preceding claims, wherein electrodes are arranged at the bottom of each of the first and second wells. Providing the device according to any one of claims 1 to 13.
Transporting the device to its place of use;
At the place of use, when the first lipid monolayer-forming droplet and the second lipid monolayer-forming droplet are in a frozen state, after melting these droplets, the first well The moving member is moved relative to the substrate to a position where the opening and the opening of the second well overlap, so that the first lipid single-film-forming droplet and the second lipid are moved. Contacting a single membrane-forming droplet and forming a lipid bilayer membrane at the contact site.