WO2016047829A1

WO2016047829A1 - Electron microscope holder for sample observation in gas or liquid phase

Info

Publication number: WO2016047829A1
Application number: PCT/KR2014/009012
Authority: WO
Inventors: 정종만; 천성식; 김윤중; 김진규
Original assignee: 한국기초과학지원연구원
Priority date: 2014-09-26
Filing date: 2014-09-26
Publication date: 2016-03-31

Abstract

The present invention relates to an electron microscope holder equipped with a micro-device (a wet cell), for allowing an electron microscope, requiring high vacuum, to observe, in real time, a reaction occurring under a complex dynamic environment, by injecting a gas or liquid into a sample. According to the present invention, a sample containing moisture, such as an organism or mineral, can be simply and economically observed in the natural state as it is, in which an air atmosphere or water atmosphere is maintained, through the electron microscope.

Description

Electron microscope holder for sample observation in gas phase or liquid phase

The present invention relates to an electron microscope holder equipped with a wet-cell for injecting gas or liquid into a sample so that a reaction occurring under a complex dynamic environment can be observed in real time in an electron microscope requiring high vacuum. .

Observation of the sample by electron microscope is performed under high vacuum. Therefore, because the sample mounted inside the electron microscope is exposed to high vacuum atmosphere, the electron microscope sample holder equipped with a special device called a wet-cell is used to observe the moisture in the sample or inject gas or liquid into the sample. Should be used.

Japanese Patent JP 2781320 is a sample holder having a sample stand, in which a sample is embedded between a pair of sheet meshes having a thin film sealing material, and a sample paper placed on the sample stand and a mesh paper for pressing the sample mesh from above . A sample holder comprising a sealing member for sealing between a strip and a sheet mesh periphery-sample stage, a sheet mesh-mesh support, and a sample stand-mesh support is disclosed.

According to such a prior art, it is possible to maintain a liquid state or a gaseous state, so that samples of living organisms, minerals, and the like in a state containing water can be observed with an electron microscope. However, according to this method, it is difficult to manufacture sheet mesh (Prior art document 0009; attaching a programmable computer grid having a hole diameter of several μm to a metallic sheet mesh having a hole size of about 100 μm, and placing a polyvinyl thin film device thereon. The process of mounting the sample sealed to the holder is complicated (see Fig. 1). ③ Above all, it is difficult to use a special dedicated holder (see Fig. 2).

U.S. Patent No. 7807979 is a sample kit in which a pair of substrates having a thin portion of an electron beam passing through (observation window) are disposed to face each other, and a sample is embedded therein by interposing a gap material and a sealing material in the periphery therebetween. (cell) is disclosed (FIGS. 3 and 4).

This makes it possible to maintain a liquid or gaseous state more conveniently and simply than the above-described Japanese patent. However, there is a disadvantage in that it is not versatile because a special structure substrate having an observation window is essential.

In the international application invented by the inventors of the present invention (PCT / KR2013 / 10219, a micro device mounted on the TEM holder to observe a sample containing water in the TEM), a predetermined number of through holes are formed in the center of (A). Lower grid; (B) a window film seated to cover all the through holes of the lower grid, the sample film is loaded on the upper surface; (C) a closed disk seated on the outer periphery of the lower disc; (D) an upper grid is formed in the through hole corresponding to the through hole of the lower disc and seated on the closed disk; And (E) a window film attached to a lower surface of the lower grid so as to cover all of the through holes, and a micro device is mounted on the TEM holder to observe a water-containing sample in the TEM. (FIG. 5).

According to this, a sample set can be produced quickly and easily by using a simple material for observing the TEM, and a sample containing moisture can be observed in the TEM, but a change in the sample due to the injection of gas or liquid cannot be observed. .

US Patent Publication US 2013/0264476 proposes an observation cell in which a gas or a liquid can flow while having a cell structure as shown in US Pat. No. 7,077,979 (Fig. 6).

Such a device allows electron microscope to observe the change of a sample while supplying gas or liquid, but it is not universal because it requires a substrate having a special structure having a viewing window and a holder having a corresponding structure. Although not shown, considerable skill is required to set up the cell and mount it in the holder because it must allow gas or liquid to flow while maintaining the space between the substrates. In addition, a silicon O-ring is used for sealing, and grooves are formed in the corresponding portions, and in case of repeated use, fine silicon particles that are dropped may affect the sample, and fine impurities may be caught in the O-ring grooves. There is a lot of room for care.

The present invention uses a material required for normal electron microscope observation and has a very simple structure, so that a sample set-up and maintenance is easy to configure a wet-cell to maintain a liquid state or a gaseous state or a gas or a liquid state. To provide an electron microscope holder to observe the sample by flowing into the cell.

The present invention for achieving the above object is (A) a cylindrical accommodating space in which the step is formed so that the following cell is inserted into the tip portion in one direction; Electron microscope holder tip including an inlet and outlet formed toward the receiving space: (B) ① the diameter corresponding to the receiving space is seated on the stepped, a predetermined number of beam through holes formed in the center Lower grid ; A lower window film seated on the upper surface of the lower grid to cover all of the through holes; ③ The outer diameter corresponds to the receiving space, the hollow portion is formed so that the through hole of the lower grid is exposed, the through passage corresponding to the inflow passage and the outlet passage is formed on the side, it is seated on the outer periphery of the lower grid Spacer ; ④ an upper window film seated on the lower surface of the upper grid to cover all of the through holes; ⑤ and a diameter corresponding to the receiving space, the upper grid, which is formed through-holes corresponding to the through hole of the lower grid, at the center; A: wet-cell containing; fixing cap for fixing the pressure on the upper grid ⑥ a diameter corresponding to the receiving space and the and the upper grid portion a hollow so that the through-hole is exposed is formed of, wherein the lower grid rests - - spacer The branch relates to a wet-cell holder for electron microscopy.

As described above, according to the present invention, it is possible to observe samples such as biological matters and minerals containing water in an electron microscope in a simple and economical manner in a natural state in which an air atmosphere or a water atmosphere is maintained.

In addition, according to the present invention, it is possible to observe not only the state containing the gas or liquid but also the change state of the sample due to the change of the input of the gas or the liquid with an electron microscope, so that the bio, chemical, medicine, In the field of materials, research and development can be made from a new perspective.

1 to 6 is a view showing the structure of a wet-cell holder according to the prior art.

7 is a conceptual perspective view of the end portion of a wet-cell holder for an electron microscope according to the present invention.

8 is a conceptual partial cross-sectional perspective view showing a receiving space area in the wet-cell holder for electron microscope according to the present invention.

9 is a conceptual partial cross-sectional exploded perspective view showing a receiving space area in the wet-cell holder for electron microscope according to the present invention.

10 is a conceptual vertical cross-sectional view of a receiving space region in a wet-cell holder for an electron microscope according to the present invention.

Description of the sign

10. Holder tip

11. receiving space 12. step 12a. 2nd step

20. wet-cell

21. Lower grid 21a. Through hole

22. Lower Window Film 23. Spacer

23a. Throughway 24. Upper Window Film

25. Upper grid 25a. Through hole

26. Fixed cap

Hereinafter, the present invention will be described in more detail with reference to the accompanying drawings. However, the accompanying drawings are only examples for easily describing the content and scope of the technical idea of the present invention, and thus the technical scope of the present invention is not limited or changed. It will be apparent to those skilled in the art that various modifications and variations are possible within the scope of the present invention based on these examples.

As described above, the present invention relates to an electron microscope holder for including a holder tip 10 and a wet-cell 20 to observe a sample in a gas phase or a liquid phase. 7 is a conceptual perspective view of an electron microscope holder end portion according to the present invention. A conceptual partial cross-sectional perspective view showing an area of the accommodation space 11 in FIG. 8, a conceptual partial cross-sectional exploded perspective view showing an area of the accommodation space 11 in FIG. 9, and a conceptual vertical cross-sectional view of an area of the accommodation space 11 in FIG. 10, respectively. Shown.

In the present invention, the holder tip 10 is a part for introducing gas or liquid into the wet-cell 20 while the wet-cell 20 is hermetically sealed, and the tip of the wet-cell 20 at the tip portion thereof. And a cylindrical accommodating space 11 having a stepped portion 12 formed therein so that the component part is inserted and mounted in one direction, and an inflow path and an outflow path for inflow and outflow of gas or liquid formed toward the accommodating space 11. . The inflow path and the outflow path are preferably formed horizontally in the center of the holder tip 10 when viewed in a vertical section, and are in communication with a predetermined connection pipe, a pump and a storage tank. In the drawings, the inlet and outlet, the connection pipe, the pump and the storage tank are not shown.

In the present invention, the wet-cell 20 is in the order to be accommodated in the receiving space 11, the lower grid 21, the lower window film 22, the spacer 23, the upper window film 24, the upper grid 25 and a fixed cap 26 are included.

The lower grid 21 has a diameter corresponding to the accommodation space 11 and is seated on the step 12, and a predetermined number of beam passing through holes 21a are formed in the center thereof. The material of the lower grid 21 may be a conductive and nonmagnetic material such as copper, gold, nickel, aluminum, molybdenum, titanium, berliumlium, carbon, or a mixture or alloy thereof. It is preferable that it is a copper material normally used by an electron microscope. Such a material may be used as a material of other components except for the lower window film 22 and the upper window film 24. It is preferable that the diameter of the lower grid 21 is generally about 3.0 mm and the thickness is about 50 µm (see http://www.tedpella.com/grids_html/Pelco-TEM-Grids.htm ). The number of the through holes 21a of the lower grid 21 is not limited, but is formed in the center, which is an area through which the electron beam can pass. The diameter of the through hole 21a should be small enough that the lower window film 22 does not suck out by the vacuum pressure, and should be large enough not to affect the straightness of the electron beam, but preferably around 100 μm.

While the lower window film 22 is in contact with the lower grid 21 and passes through the electron beam and prevents leakage of air or moisture in the wet-cell 20, the lower window film 22 is in contact with the lower grid 21. It serves to settle (attach) the sample on the opposite surface (ie, the upper surface). As the lower window film 22, various kinds of synthetic resin films that do not affect the straightness of the electron beam may be applied. However, in order to observe the TEM, the lower window film 22 is preferably a carbon film. There is no big restriction | limiting in the thickness of a film, and about 5-10 micrometers is enough. The size of the lower window film 22 is larger than the size enough to cover all of the through holes 21a formed in the lower grid 21. In the drawing showing the electron microscope holder according to the present invention, although the lower window film 22 covers all of the through holes 21a but is not sized in contact with the spacer 23, the bottom window film 22 is larger than the bottom of the spacer 23. Even if it comes in contact with, it is obvious.

When the wet-cell 20 according to the present invention is mounted and observed on the holder tip 10, a strong adhesion force naturally occurs on the lower grid 21 and the lower window film 22 because the outside (electron microscope barrel) is a vacuum. do. Therefore, in the present invention, the lower grid 21 and the lower window film 22 need not be adhered to each other by an adhesive or the like, but only enough to be in close contact with each other so as not to be separated during the assembly process.

In the present invention, the spacer 23 is interposed on the outer periphery between the lower grid 21 and the upper grid 25 to be mounted oppositely to maintain a gap between the lower grid 21 and the upper grid 25. While serving to seal the inside and at the same time serves to communicate the inlet and outlet of the holder tip 10 and the inside of the wet-cell (20). That is, the spacer 23 has an outer diameter corresponding to the accommodating space 11, and a hollow part is formed to expose the through hole 21a of the lower grid 21. Through passages (23a) is formed, it is seated on the outer periphery of the lower grid (21). The through passage may be formed in a size corresponding to a position corresponding to the inflow passage and the outflow passage. In this case, the accommodation space 11 and the spacer 23 may be easily matched with the inflow passage, the outflow passage and the through passage. It is preferable to form predetermined positioning means (not shown) in between. Alternatively, the through passage may be formed on the side of the spacer 23 so that the through passage and the inflow passage and the outflow passage may be in contact with each other, or three or more passage passages may be provided so that any passage may communicate with the inflow passage and the outflow passage. It is also good. In FIG. 10, six through paths are illustrated. The thickness of the spacer 23 eventually determines the distance between the lower grid 21 and the upper grid 25 (the height of the space in which the sample is seated). The thickness of the spacer 23 is about 50 to 500 μm. It is good to be. The material of the spacer 23 may be a metal, such as the material of the lower grid 21, or may be a synthetic resin having appropriate strength.

The upper window film 24 is mounted on the lower surface of the upper grid 25 so as to cover all the through holes 25a, and the upper grid 25 has a diameter corresponding to the receiving space 11, and the center portion of the

upper window

25 is The through hole 25a corresponding to the through hole 21a of the lower grid 21 is formed. The size, material, and characteristics thereof are the same as those of the lower window film 22 and the lower grid 21 described above. May be the same.

In the micro apparatus according to the present invention, the lower grid 21 and the upper grid 25 have the lower window film 22 and the upper window film 24 attached to the inner surface thereof, respectively, and the through

holes

21a and 25a are vertically attached. In agreement with each other, the electron beams are opposed to each other so as to pass through

holes

21a and 25a formed in the lower grid 21 and the upper grid 25. Since the components according to the present invention have a very fine size (the lower grid 21 and the upper grid 25 are about 3 mm), the through holes 21a, when the lower grid 21 and the upper grid 25 are opposed to each other, are formed. It may not be easy to vertically match 25a). In order to solve this inconvenience, it is also possible to form a predetermined position (direction) determining means (not shown) in contact with the lower grid 21, the spacer 23, the upper grid 25 and the receiving space (11). good. For example, a portion of the outer circumference of the lower grid 21, the spacer 23, and the upper grid 25 may be a straight line, or a groove is formed on the outer circumference, and a portion of the accommodating space 11 corresponding thereto is a straight line. Or it may be easy to determine the position (direction) of the lower grid 21, the spacer 23, the upper grid 25 when mounting the wet-cell (20) assembly.

In the present invention, the fixing cap 26 serves to press and fix the lower grid 21, the spacer 23, and the upper grid 25 seated in order, and the diameter of the accommodation space ( 11) and has a hollow portion in which a hollow part is formed to expose the through hole 25a of the upper grid 25. For safe fixing and sealing, the outer periphery of the fixing cap 26 and the receiving space 11 corresponding thereto have a screw structure (not shown) corresponding to each other, so that the screw can be fixed in a screwing manner.

On the other hand, when the sealing force is reduced at the vertical contact surface between the receiving space 11 and each of the components of the wet-cell 20, gas or liquid inside the wet-cell 20 may leak to the outside (electron microscope tube). There is concern. In order to prevent the possibility of such a leak in the vertical contact surface, as shown in Figs. 7 to 10, the second step 12a is formed in the portion corresponding to the upper grid 25 of the accommodation space 11, Side of the upper grid 25 may be a structure corresponding to the second step (12a). As a result, the upper grid 25 is pressed down by the fixing cap 26 and is strongly adhered to the receiving space 11 at the second step 12a so that there is no fear of leakage of gas or liquid therein.

Electron microscope wet-cell holder 20 according to the present invention made of the above components, for example, can be assembled in a state where the sample is seated through the following process.

(A) the lower window film 22 or the upper window film 24 is floated on a liquid such as distilled water, and (B) the lower grid 21 or the upper grid 25 is held with a forceps or the like. The lower window film 22 or the upper window film 24 is lifted up from the bottom of the lower window 21 or the upper window 25 on the lower window film 22 or the upper window film 24, through-holes It is settled and dried so that all (21a, 25a) may be covered. Subsequently, (C) the lower window film 22 having the lower window film 22 seated on the upper surface is inserted into the receiving space 11 of the holder tip 10 and seated on the step 12. 23 is inserted into the receiving space 11 to be seated in the lower grid 21 while positioning the through passage (23a) to correspond to the inlet and outlet of the holder tip (10). Subsequently, (E) the upper grid 25 is inserted into the receiving space 11 so that the upper window film 24 faces downward, and penetrates the lower grid 21 and the upper grid 25 on the spacer 23. The

seats

21a and 25a are seated vertically, and then (F) the fixing cap 26 is inserted into the receiving space 11 so that the lower grid 21-spacer 23-upper grid 25 ) Press to fix it. A partial cross-sectional perspective view of the assembled state is illustrated in FIGS. 8 and 9 (sample not shown).

On the other hand, loading the sample on the surface of the lower window film 22 before or after the step (C); may be further added.

The holder for the electron microscope wet-cell 20 according to the present invention thus assembled is used to observe the change over time of the sample while being inserted into the barrel of the electron microscope and applying a predetermined gas or liquid.

Claims

(A) a cylindrical accommodating space in which a step is formed so that the following cell is inserted and mounted in one direction at a tip portion; Electron microscope holder tip including; inlet and outlet formed toward the receiving space:

(B) a lower grid having a diameter corresponding to the receiving space and seated on the stepped portion, and having a predetermined number of through holes for beam passing therethrough;

A lower window film seated on the upper surface of the lower grid to cover all of the through holes;

③ The outer diameter corresponds to the receiving space, the hollow portion is formed so that the through hole of the lower grid is exposed, the through passage corresponding to the inflow passage and the outlet passage is formed on the side, it is seated on the outer periphery of the lower grid Spacer ;

④ an upper window film seated on the lower surface of the upper grid to cover all of the through holes;

⑤ and a diameter corresponding to the receiving space, the upper grid, which is formed through-holes corresponding to the through hole of the lower grid, at the center;

⑥ a diameter corresponding to the receiving space, the hollow portion is formed so that the through hole of the upper grid is exposed, a fixed cap for pressing and fixing the seated lower grid-spacer-upper grid; wet-cell comprising:

Wet-cell holder for electron microscopes.
The method of claim 1,

The second stepped portion is formed in a portion corresponding to the upper grid of the receiving space, the side of the upper grid wet-cell holder for the electron microscope, characterized in that the structure corresponding to the second step.
The method according to claim 1 or 2,

The lower window film 22 is a wet-cell holder for an electron microscope, characterized in that the carbon film.
(A) floating the lower window film or the upper window film in distilled water;

(B) mounting and drying the lower window film and the upper window film on the lower grid or the upper grid;

(C) inserting the lower grid seated on the upper surface of the lower window film in the receiving space of the holder tip and seated on the step;

(D) inserting the spacer into the receiving space to be fitted to the lower grid to correspond to the inlet and outlet of the holder tip;

(E) inserting the upper grid into the receiving space so that the upper window film faces downward;

(F) pressing and fixing the lower grid-spacer-upper grid with the fixing cap;

Method for assembling the micro device according to any one of claims 1 to 3, characterized in that it comprises a.
The method of claim 4, wherein

The method of assembling the micro device according to any one of claims 1 to 3, further comprising the step of loading a sample on the surface of the lower window film 22 before or after the step (C). .