Specimen support device for transmission electron microscope, specimen support device for scanning electron microscope or electron beam microanalyzer, microscope device, and method for imaging liquid specimen

Abstract

To provide a specimen support device for a transmission electron microscope that can confine a liquid specimen in a liquid pocket highly reproducibly and reliably.SOLUTION: A specimen support device for a transmission electron microscope is configured to include: (a) a transmission electron microscope grid 10 having an observation area; (b) a support film 12 attached to the transmission electron microscope grid 10; (c) a hydrophilic bottom film 14 attached to a top of the support film 12; (d) a hydrophobic top film 16 attached to the hydrophilic bottom film 14 so as to include and cover the observation area; and (e) a liquid specimen that is encapsulated in a region formed between the hydrophilic bottom film 14 and the hydrophobic top film 16. Therein the hydrophilic bottom film 14 is graphene oxide, and the hydrophobic top film 16 is graphene.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to a sample support device for a transmission electron microscope, a sample support device for a scanning electron microscope or an electron probe microanalyzer, a microscope device, and a method for imaging a liquid sample. [Background technology]

[0002] Introducing a sample sealed in a liquid cell into the vacuum of an electron microscope makes it possible to observe a liquid sample or a sample present in a liquid. This liquid cell must have a diaphragm that is strong enough to withstand the pressure difference between the vacuum outside the cell and the liquid inside the cell, yet thin enough to allow electron beams to pass through. A commonly used liquid cell is one in which a liquid sample is sandwiched between two silicon chips with silicon nitride membrane windows formed in the diaphragm using semiconductor microfabrication technology (see, for example, Patent Document 1).

[0003] There is a type of ultimate liquid cell that uses graphene (see, for example, Patent Documents 2 and 3, and Non-Patent Document 1). This uses a liquid pocket that is accidentally formed by sandwiching a liquid between two sheets of graphene. Graphene hardly scatters electrons, so there is almost no degradation in the resolution of the electron microscope; its high electrical conductivity reduces sample damage; it is flexible and has high mechanical strength; and it is chemically stable, making it an excellent material for liquid cells in electron microscopes. Well-type liquid cells can create chambers with controlled liquid volumes, allowing for the highly reproducible definition of environmental conditions for the behavior of samples in liquid. They also enable the observation of soft materials, whose shape is easily changed when sandwiched between two thin films (see, for example, Non-Patent Document 1). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Special Publication No. 2010-527123 (WO2008 / 141147) [Patent Document 2] Chinese Patent No. 106769287 [Patent Document 3] Special Publication No. 2023-507397 (WO2021 / 123458) [Non-patent literature]

[0005] [Non-Patent Document 1] Jungjae Park et al.; Graphene Liquid Cell Electron Microscopy: Progress, Applications, and Perspectives, ACS Nano(2021) 15, 288-308. Summary of the Invention [Problem to be solved by the invention]

[0006] However, in liquid cells in which a liquid sample is sandwiched between two silicon chips with silicon nitride film windows, the silicon nitride film is approximately 30 to 50 nm thick, so the contrast of the liquid sample (or the sample within the liquid) is insufficient, and the resolution of the electron microscope is inferior to that of observations under normal vacuum. Furthermore, graphene-based liquid cells have the problem of low reproducibility in fabrication, since graphene is hydrophobic and it is not easy to accidentally trap a liquid pocket containing a sample. On the other hand, graphene oxide, which is made by treating the graphene surface to give it oxygen functional groups, exhibits hydrophilicity but has the drawback of poor electrical conductivity, and has not been used in liquid cells for electron microscopes.

[0007] When graphene is used in a well-type liquid cell, there is a problem in that the narrow well repels water due to the hydrophobicity of graphene, making it difficult for water to enter the well.

[0008] The present invention has been made to solve the above-mentioned problems of the prior art, and aims to provide a sample support device for a transmission electron microscope, a sample support device for a scanning electron microscope or an electron beam microanalyzer, a microscope device, and a method for imaging a liquid sample, which are capable of confining a liquid sample in a liquid pocket with high reproducibility and reliability. [Means for solving the problem]

[0009] [1] The sample support device for a transmission electron microscope of the present invention is, for example, as shown in FIG. (a) a transmission electron microscope grid 10 having an observation area; (b) a support film 12 attached to a transmission electron microscope grid 10; (c) a hydrophilic bottom membrane 14 attached to the top of the support membrane 12; (d) a hydrophobic top membrane 16 attached to cover the observation area of ​​the hydrophilic bottom membrane 14; (e) A liquid sample 17 is encapsulated in an area formed between a hydrophilic bottom film 14 and a hydrophobic top film 16, the hydrophilic bottom film 14 being graphene oxide and the hydrophobic top film 16 being graphene.

[0010] [2] In the sample support device for a transmission electron microscope [1] of the present invention, the graphene oxide is preferably modified with an epoxy group or a hydroxyl group. [3] In the sample support device for a transmission electron microscope [1] of the present invention, the thickness of the graphene oxide preferably corresponds to a thickness of 1 to 10 atomic layers, and may also be a thickness of 1 to 100 atomic layers. [4] In the sample support device for a transmission electron microscope [1] of the present invention, the thickness of the graphene preferably corresponds to a thickness of 1 to 10 atomic layers, and may also be a thickness of 1 to 100 atomic layers. [5] In the sample support device for a transmission electron microscope [1] to [4] of the present invention, it is preferable to further include (f) a well-type liquid pocket 20 that includes the observation area, is attached to the area formed between the hydrophilic bottom membrane 14 and the hydrophobic top membrane 16, and has a well-shaped liquid pocket 20 whose periphery is covered with the hydrophilic bottom membrane. When a micro-sized well is introduced into the gap between the hydrophilic bottom membrane 14 and the hydrophobic top membrane 16, water is retained around the well-shaped liquid pocket, making it possible to obtain a liquid pocket of any thickness more efficiently. [6] The microscope apparatus of the present invention may include the sample support device according to [1] mounted in a manner that allows microscopic imaging of a sample on the sample support device.

[0011] [7] The method for imaging a liquid sample of the present invention is a method for imaging a liquid sample using a transmission electron microscope device, in which a hydrophilic bottom film 14 is attached to a support film 12 attached to a transmission electron microscope grid 10 having an observation area, a liquid sample 17 is dropped onto the observation area on the hydrophilic bottom film 14, a hydrophobic top film 16 is attached to the hydrophilic bottom film 14 and the dropped liquid sample 17, and the liquid sample 17 is imaged using the transmission electron microscope. [8] The method for imaging a liquid sample of the present invention is a method for imaging a liquid sample using a transmission electron microscope device, in which a hydrophilic bottom membrane 14 is attached to a support membrane 12 attached to a transmission electron microscope grid 10 having an observation area, a well-type liquid pocket 20 whose well periphery is covered with the hydrophilic bottom membrane 14 is placed on the hydrophilic bottom membrane 14 in an area including the observation area, a liquid sample 17 is dropped into the well-type liquid pocket, a hydrophobic top membrane 16 is attached to the well-type liquid pocket 20 and the dropped liquid sample 17, and the liquid sample 17 is imaged using the transmission electron microscope.

[0012] [9] The sample support device for a scanning electron microscope or an electron beam microanalyzer of the present invention is, for example, as shown in FIG. (a) A sample stage 30; (b) a support film 32 attached to a sample stage 30; (c) a hydrophilic bottom membrane 34 attached to the top of the support membrane 32; (d) a hydrophobic top membrane 36 attached to cover the observation area of ​​the hydrophilic bottom membrane 34; (e) A liquid sample is encapsulated in an observation area formed by a hydrophilic bottom film 34 and a hydrophobic top film 36, where the hydrophilic bottom film 34 is made of graphene oxide and the hydrophobic top film 36 is made of graphene.

[0013]

[10] In the sample support device [9] for a scanning electron microscope or an electron probe microanalyzer of the present invention, the graphene oxide is preferably modified with an epoxy group or a hydroxyl group.

[11] In the sample support device [9] for a scanning electron microscope or an electron beam microanalyzer of the present invention, the thickness of the graphene oxide preferably corresponds to a thickness of 1 to 10 atomic layers, and may also correspond to a thickness of 1 to 100 atomic layers.

[12] In the sample support device for a scanning electron microscope or an electron probe microanalyzer [9] of the present invention, the thickness of the graphene preferably corresponds to a thickness of 1 to 10 atomic layers, and may also correspond to a thickness of 1 to 100 atomic layers.

[13] In the sample support device [9] to

[12] of the present invention for a scanning electron microscope or an electron beam microanalyzer, it is preferable to further include (f) a well-shaped liquid pocket attached to the region formed between the hydrophilic bottom membrane 34 and the hydrophobic top membrane 36, the periphery of which is covered with the hydrophilic bottom membrane. When a micro-sized well is introduced between the hydrophilic bottom membrane 34 and the hydrophobic top membrane 36, water is retained around the well-shaped liquid pocket, making it possible to obtain a liquid pocket of any thickness more efficiently.

[14] The microscope apparatus of the present invention may include a sample support device according to [9] mounted in a manner that allows microscopic imaging of a sample on the sample support device.

[0014]

[15] The method for imaging a liquid sample of the present invention is a method for imaging a liquid sample using a scanning electron microscope or an electron beam microanalyzer, in which a hydrophilic bottom film 34 is attached to a support film 32 attached to a sample stage 30, a liquid sample is dropped onto the hydrophilic bottom film 34, a hydrophobic top film 36 is attached to the hydrophilic bottom film 34 and the dropped liquid sample, and the liquid sample is imaged using a scanning electron microscope or an electron beam microanalyzer.

[16] The method for imaging a liquid sample of the present invention is a method for imaging a liquid sample using a scanning electron microscope or an electron beam microanalyzer, in which a hydrophilic bottom membrane 34 is attached to a support membrane 32 attached to a sample stage 30, a well-shaped liquid pocket whose well periphery is covered with the hydrophilic bottom membrane 34 is placed on the hydrophilic bottom membrane 34, a liquid sample is dropped into the well-shaped liquid pocket, a hydrophobic top membrane 36 is attached to the well-shaped liquid pocket and the dropped liquid sample, and the liquid sample is imaged using a scanning electron microscope or an electron beam microanalyzer.

[0015] The sample support device for a transmission electron microscope of the present invention uses a combination of graphene and graphene oxide to efficiently and reproducibly fabricate thin-film liquid cells. Applying graphene oxide to foreign matter or wells allows for the creation of targeted liquid pockets due to its hydrophilicity. By using graphene as the other membrane, the excellent electrical conductivity of graphene can be maintained, minimizing sample damage. According to the sample support device for a transmission electron microscope of the present invention, the combination of graphene and graphene oxide makes it easy to observe liquid samples (or samples submerged in liquid) under an electron microscope, thereby providing insight into chemical reactions and structural changes at the nanoscale and atomic scale of the liquid sample (or sample submerged in liquid). The well-type liquid cell of the present invention uses graphene oxide, allowing water to be contained even in small wells. Simply put, a circular or rectangular doughnut-shaped thin film is placed on graphene oxide to form a well, and a liquid sample is dropped onto the well, allowing the liquid sample to fill the well. By attaching graphene on top of the well, it is possible to fabricate a well-type liquid cell with good reproducibility. [Brief explanation of the drawings]

[0016] [Figure 1] FIG. 1 is a cross-sectional view of a liquid cell (liquid pocket) having graphene oxide and graphene according to an embodiment of the present invention, for use in a transmission electron microscope. [Figure 2] Annular dark-field scanning transmission electron microscope (ADF-STEM) images of liquid cells made from graphene oxide and graphene are shown. [Figure 3] Annular dark-field scanning transmission electron microscope (ADF-STEM) image of a liquid cell of water containing gold colloids is shown. [Figure 4] 1 shows an annular dark-field scanning transmission electron microscope (ADF-STEM) image at high magnification of gold colloid in water. [Figure 5] Shown is an annular dark-field scanning transmission electron microscope (ADF-STEM) image of a calcium carbonate particle inclusion and the surrounding liquid pocket. [Figure 6] FIG. 10 is a cross-sectional view of the configuration of a liquid cell (liquid pocket) having graphene oxide and graphene according to a second embodiment of the present invention, for use in a scanning electron microscope. [Figure 7] FIG. 10 is a cross-sectional view of a well-type liquid cell (liquid pocket) having graphene oxide and graphene according to a third embodiment of the present invention, for use in a transmission electron microscope. [Figure 8] FIG. 10 is a cross-sectional view of a well-type liquid cell (liquid pocket) having graphene oxide and graphene according to a fourth embodiment of the present invention, for use in a scanning electron microscope. DETAILED DESCRIPTION OF THE INVENTION

[0017] The present invention will be described below with reference to the drawings. The present invention relates to a sample support for a transmission electron microscope, and to a device for confining a liquid sample to be observed. The device generally includes a membrane region and a frame region. The device is placed in the sample tip region of a sample holder.

[0018] The terms used in this specification will be defined below. "Specimen" means an object that is at least partially electron transparent, such as a nanoparticle, catalyst, thin section, etc., that is examined in an electron microscope. "Liquid cell" means a system (eg, one device, two devices, or a system of three or more devices) for containing and controlling the environment around a sample. "Sample holder" means a precision-machined piece of equipment used to hold and secure one or more devices arranged individually, as a group, or as a liquid cell, and to provide an interface between the devices and / or liquid cells and the outside world.

[0019] A "window device" refers to a device used to create a physical, electron-transparent barrier at one boundary between the liquid cell and the vacuum environment of an electron microscope. For example, a TEM grid is used to place a sample for observation using a transmission electron microscope. A TEM grid is a metal or other plate with a diameter of 3 mm and a thickness of 20 to 50 μm on which a sample for observation using a transmission electron microscope is placed. Grids come in a variety of shapes, including square, round, and slit, and the appropriate shape is selected depending on the purpose of observation. Grid materials include copper, molybdenum, gold, and titanium. For elemental analysis, grids made of materials that do not contain the target element are used. Among these, lattice-shaped grids are commonly used. For microparticles, a support film is attached to the surface of the grid, and the sample is placed on top of it for observation.

[0020] "Frame" refers to a rigid area around the device used to provide mechanical support for the entire device structure, such as a silicon frame. In the case of a TEM grid, this refers to the frame structure of the grid. In the case of a sample stage, this refers to the outer frame structure of the sample stage. A sample stage 30 is used for scanning electron microscopes. The sample stage is a metal or carbon base used to stably hold a sample. Since it is difficult to attach relatively small or powder-like samples directly to a sample holder, the sample is first fixed to the sample stage and then attached to the sample holder. The sample holder is a component that mechanically holds the sample stage and attaches it to the sample stage. The sample stage is a unit that provides stable support for the sample and allows smooth horizontal and vertical movement, rotation, tilt, and other sample movements. Since it only has a sample support mechanism and movement function, the entire sample stage is usually replaced when applying external force or temperature changes to the sample.

[0021] The membrane region is the portion of the liquid cell structure that is not supported by the frame. The membrane region can consist of one or more thin films, including other deposited films such as graphene. The membrane region is less than about 1 micron thick, but need not be of uniform thickness. The membrane region can serve to create a physical barrier between the environment at the sample and the environment within the electron microscope, or as a support for placing the sample, or both. One or more membrane regions can be placed in the device, generally separated by thicker demarcating regions between them, where the demarcating regions are above, below, and / or to the sides of the membrane region. The membrane region can be continuous or have perforations of any shape or size.

[0022] The membrane regions of each device are produced from membrane materials that are less than about 1 micron thick, are robust, and can be insulating or conductive, and can be constructed using a variety of membrane fabrication techniques in combination with other deposition and float-down techniques. Generally, however, the membrane material is affixed to a frame material and has a tensile stress profile to keep the subsequently formed membrane region pulled tight across the frame. One embodiment of the membrane region provides a surface upon which a sample can be placed. Membrane materials that can be used to create the membrane region include graphene and graphene oxide. The membrane regions may or may not include additional elements integrated into or disposed directly on their top or bottom surfaces.

[0023] FIG. 1 is a cross-sectional view of the structure of a liquid cell (liquid pocket) having graphene oxide and graphene according to one embodiment of the present invention, and shows a structure for a transmission electron microscope. A support film 12 on a TEM grid 10 is prepared with graphene oxide 14 attached to it, a liquid sample 17 (or a liquid-submerged sample) is dropped onto it, and graphene 16 is attached on top of it to obtain a liquid pocket 18. Carbon materials with a graphene skeleton, such as graphene and graphene oxide, are sp 2 Although there are no particular limitations on the material as long as it has carbon (C) bonded by bonds and the carbon is arranged in a plane, it is preferably a material having carbon bonded to oxygen (O). More preferably, it is graphene oxide, in which oxygen is bonded to the carbon of graphene. Generally, graphene is a 2 A graphene refers to a sheet consisting of a single layer in which carbon atoms bonded together are arranged in a plane, and a stack of many graphene sheets is called graphite, but the carbon material having a graphene skeleton in the present invention and the graphene oxide in the present invention include not only sheets consisting of a single layer but also those having a structure in which several to about 100 layers are stacked. The same applies to graphene crystals described below. Graphene oxide having such a stacked structure can be obtained, for example, by treating graphite with a known oxidizing agent. For example, a graphene oxide composite composition can be produced by the method disclosed in Japanese Patent No. 6499450. The carbon material having a graphene skeleton may further have functional groups such as a carboxyl group, a hydroxyl group, or a sulfur-containing group.

[0024] Figure 2 shows an annular dark-field scanning transmission electron microscope (ADF-STEM) image of the graphene oxide and graphene-based liquid cell. Holes 12a are regularly arranged in the carbon support film 12, and films of graphene oxide 14 and graphene 16 are bonded to the holey support film 12. The bright spots are liquid pockets 18 (the liquid is water). The outer diameter of the liquid pockets 18 is, for example, 0.2 to 0.3 μm. The carbon support film 12 is not present in the holes 12a; the liquid pockets 18 are supported only by the graphene oxide 14 and graphene 16. The holes 12a are elliptical, with a major axis of approximately 3 μm and a minor axis of approximately 2.5 μm. A unit shape of the holey support film 12 is a hexagon with six holes 12a at its vertices and one at its center. The side of this hexagon is, for example, approximately 10 μm, but this is not limited to this.

[0025] Figure 3 shows an annular dark-field scanning transmission electron microscope (ADF-STEM) image of a liquid cell of water containing gold colloids. Gold colloids 22 are clearly seen to be concentrated at the edges of the liquid pockets 18 and in the surrounding wrinkles 21 (containing water).

[0026] Figure 4 is a high-magnification image of the gold colloid in the wrinkled area of ​​Figure 3, enlarged approximately 20 times compared to Figure 3. It shows that the atomic structure and single atoms, each about 0.2 nm in size, have been captured.

[0027] FIG. 5 shows calcium carbonate as a foreign body, with a liquid pocket 18 formed around it. The tail 24 of the liquid pocket 18 is attached to the right side of the calcium carbonate. The calcium carbonate as the foreign body 23 has an outer diameter of 20 to 40 nm. The outer diameter of the liquid pocket 18 is about 0.2 μm. The extension of the tail 24 is about 0.2 μm, which is almost the same as the outer diameter of the liquid pocket 18. When the foreign body 23 is present in this way, the solution pocket 18 adheres to the periphery of the foreign body 23.

[0028] FIG. 6 is a cross-sectional view of the configuration of a liquid cell (liquid pocket) having graphene oxide and graphene according to a second embodiment of the present invention, and shows a configuration for a scanning electron microscope. A support film 32 on a sample stage 30 is prepared with graphene oxide 34 attached thereon, a liquid sample 37 (or a liquid-submerged sample) is dropped onto it, and graphene 36 is attached on top of it to obtain a liquid pocket 38. This liquid pocket 38 is of a type called a veil-type liquid pocket 39 (veil-type graphene liquid cell (GLC)) defined in Non-Patent Document 1.

[0029] FIG. 7 is a cross-sectional view of the configuration of a well-type liquid cell (liquid pocket) having graphene oxide and graphene according to a third embodiment of the present invention, and shows a transmission electron microscope. A support film 12 on a TEM grid 10 is prepared with graphene oxide 14 attached thereon, and a well-type liquid pocket 20 is placed on top of the support film 12. The well-type liquid pocket 20 is defined in Non-Patent Document 1 (well-type graphene liquid cell (GLC)). A liquid sample 17 (or a liquid-containing sample) is dropped into a recess provided in a well-type liquid pocket 20, and graphene 16 is attached to the top surface of the recess of the well-type liquid pocket 20 containing the liquid sample 17, thereby obtaining a liquid pocket 18.

[0030] FIG. 8 is a cross-sectional view of the structure of a well-type liquid cell (liquid pocket) having graphene oxide and graphene according to a fourth embodiment of the present invention, and shows a scanning electron microscope. A support film 32 on a sample stage 30 is prepared with graphene oxide 34 attached thereon, and a well-shaped liquid pocket 40 is placed on top of it. A liquid sample 37 (or a liquid-containing sample) is dropped into a recess provided in a well-type liquid pocket 40, and graphene 36 is attached to the top surface of the recess of the well-type liquid pocket 40 containing the liquid sample 37, thereby obtaining a liquid pocket 38.

[0031] Although the above embodiment of the present invention shows a sample support device for a scanning electron microscope, this sample support device can also be applied to surface spectroscopic analysis devices such as an electron probe microanalyzer (EPMA). An electron probe microanalyzer (EPMA) is capable of observing the structure and morphology of a surface and performing local trace element analysis. By using detectors such as a wavelength dispersive X-ray spectrometer (WDS) or a soft X-ray spectrometer (SXES), more detailed measurement results can be obtained than with the energy dispersive detector (EDS) used in ordinary scanning electron microscopes (SEM). In addition, scanning electron microscopes use a variety of detectors, including secondary electron detectors, backscattered electron detectors, direct exposure detectors, phosphor (scintillator) detectors, EDS (energy dispersive X-ray spectroscopy) detectors, and wavelength-dispersive X-ray spectroscopy (WDS) detectors. [Industrial Applicability]

[0032] The sample support device for a transmission electron microscope of the present invention can confine a liquid sample within a liquid pocket with high reproducibility and reliability, making it possible to create a chamber with a controlled liquid volume, a characteristic of a well-type liquid cell, and also has the characteristic of being able to define environmental conditions for the behavior of a sample in liquid with high reproducibility, making it suitable for observing soft materials whose shape easily changes when sandwiched between two thin films. Like the sample support device for a transmission electron microscope of the present invention, the sample support device for a scanning electron microscope or electron beam microanalyzer of the present invention is also suitable for observing soft materials whose shape is easily changed when sandwiched between two thin films. [Explanation of symbols]

[0033] 10 Transmission Electron Microscope Grids 12 Support membrane 12a hole 14 Hydrophilic bottom membrane (graphene oxide) 16 Hydrophobic top film (graphene) 17 Liquid samples 18 Liquid pocket (liquid cell part) 20 well-type graphene liquid cell (GLC) 30 Sample stage 32 Support membrane 34 Hydrophilic bottom membrane 36 Hydrophobic upper membrane 37 Liquid samples 38 Liquid pocket (liquid cell part) 39 Veil-type graphene liquid cell (GLC) 40 well-type graphene liquid cell (GLC)

Claims

1. (a) a transmission electron microscope grid having an observation area; (b) a support film attached to the transmission electron microscope grid; (c) a hydrophilic bottom membrane attached to the top of the support membrane; (d) a hydrophobic top membrane that is attached to the hydrophilic bottom membrane so as to include and cover the observation area of ​​the hydrophilic bottom membrane; (e) a liquid sample is encapsulated in an area formed between the hydrophilic bottom membrane and the hydrophobic top membrane; the hydrophilic bottom membrane is graphene oxide; the hydrophobic top film is graphene; A specimen support device for a transmission electron microscope.

2. The graphene oxide is modified with an epoxy group or a hydroxyl group.

2. A sample support device for a transmission electron microscope according to claim 1.

3. The thickness of the graphene oxide corresponds to the thickness of 1 to 10 atomic layers.

2. A sample support device for a transmission electron microscope according to claim 1.

4. The thickness of the graphene corresponds to the thickness of 1 to 10 atomic layers.

2. A sample support device for a transmission electron microscope according to claim 1.

5. and (f) a well-shaped liquid pocket including the observation area, attached to the area formed between the hydrophilic bottom membrane and the hydrophobic upper membrane, the periphery of which is covered with the hydrophilic bottom membrane.

5. A sample support device for a transmission electron microscope according to claim 1, comprising:

6. 10. A microscope apparatus including the sample support of claim 1 mounted in a manner that allows for microscopic imaging of a sample on the sample support.

7. 1. A method for imaging a liquid sample using a transmission electron microscope apparatus, comprising: A hydrophilic bottom membrane is attached onto a support membrane that is attached to a transmission electron microscope grid having an observation area; Dropping a liquid sample onto the observation area on the hydrophilic bottom membrane; applying a hydrophobic top membrane to the hydrophilic bottom membrane and the dropped liquid sample; imaging the liquid sample with a transmission electron microscope; A method for imaging liquid samples.

8. 1. A method for imaging a liquid sample using a transmission electron microscope apparatus, comprising: A hydrophilic bottom membrane is attached onto a support membrane that is attached to a transmission electron microscope grid having an observation area; a well-shaped liquid pocket, the periphery of which is covered with a hydrophilic bottom membrane, is disposed on the hydrophilic bottom membrane in a region including the observation region; A liquid sample is dropped into the well-shaped liquid pocket, a hydrophobic upper membrane is attached to the well-shaped liquid pocket and the dropped liquid sample; imaging the liquid sample with a transmission electron microscope; A method for imaging liquid samples.

9. (a) a sample stage; (b) a support film attached to the sample stage; (c) a hydrophilic bottom membrane attached to the top of the support membrane; (d) a hydrophobic top membrane attached to the bottom membrane so as to include and cover the observation area of ​​the bottom membrane; (e) a liquid sample is encapsulated in an area formed between the hydrophilic bottom membrane and the hydrophobic top membrane; the hydrophilic bottom membrane is graphene oxide; the hydrophobic top film is graphene; A sample support device for a scanning electron microscope or electron beam microanalyzer.

10. The graphene oxide is modified with an epoxy group or a hydroxyl group.

10. The sample support device for a scanning electron microscope or an electron beam microanalyzer according to claim 9.

11. The thickness of the graphene oxide corresponds to the thickness of 1 to 10 atomic layers.

10. The sample support device for a scanning electron microscope or an electron beam microanalyzer according to claim 9.

12. The thickness of the graphene corresponds to the thickness of 1 to 10 atomic layers.

10. The sample support device for a scanning electron microscope or an electron beam microanalyzer according to claim 9.

13. Furthermore, (f) a well-shaped liquid pocket attached to the region formed between the hydrophilic bottom membrane and the hydrophobic upper membrane, the well periphery being covered with the hydrophilic bottom membrane; 13. The sample support device for a scanning electron microscope or an electron beam microanalyzer according to claim 9, comprising:

14. 10. A microscope apparatus including a sample support device according to claim 9 mounted in a manner that allows for microscopic imaging of a sample on the sample support device.

15. A method for imaging a liquid sample using a scanning electron microscope or an electron beam microanalyzer, comprising: A hydrophilic bottom membrane is attached onto the support membrane attached to the sample stage; Dropping a liquid sample onto the hydrophilic bottom membrane; applying a hydrophobic top membrane to the hydrophilic bottom membrane and the dropped liquid sample; imaging the liquid sample with a scanning electron microscope or electron microanalyzer; A method for imaging liquid samples.

16. A method for imaging a liquid sample using a scanning electron microscope or an electron beam microanalyzer, comprising: A hydrophilic bottom membrane is attached onto the support membrane attached to the sample stage; A well-shaped liquid pocket having a periphery covered with a hydrophilic bottom membrane is placed on the hydrophilic bottom membrane; A liquid sample is dropped into the well-shaped liquid pocket, a hydrophobic upper membrane is attached to the well-shaped liquid pocket and the dropped liquid sample; imaging the liquid sample with a scanning electron microscope or electron microanalyzer; A method for imaging liquid samples.

Images