Electron microscopic observation method for observing biological sample in shape as it is, and composition for evaporation suppression under vacuum, scanning electron microscope, and transmission electron microscope used in the method

Patent metadata

Field	Value
Publication number	US-9557253-B2
Application number	US-201214343537-A
Country	US
Kind code	B2
Filing date	Sep 7, 2012
Priority date	Sep 9, 2011
Publication date	Jan 31, 2017
Grant date	Jan 31, 2017

Abstract

Provided is an observation method by an electron microscope, in which a biological sample can be observed as it is alive and a situation that the biological sample is moving can be observed using an electron microscope, and a composition for evaporation suppression under vacuum, a scanning electron microscope, and a transmission electron microscope used in the method.
The sample observation method by an electron microscope according to the invention includes applying a composition for evaporation suppression containing at least one kind selected from an amphiphilic compound, oils and fats, and an ionic liquid to the surface of a sample to form a thin film, and covering the sample with the thin film, and displaying an electron microscopic image of the sample, which is covered with the thin film and accommodated in a sample chamber under vacuum, on a display device.

First claim

The invention claimed is:

1. A sample observation method by an electron microscope, the method comprising:
applying a composition for evaporation suppression including at least one kind selected from an amphiphilic compound, oils and fats, and an ionic liquid to a surface of a sample;
irradiating the sample applied with the composition for evaporation suppression with an electron beam or plasma to form a polymerized film on the surface of the sample as a thin film and covering the sample with the polymerized film; and
displaying an electron microscopic image of the sample covered with the polymerized film and accommodated in a sample chamber under vacuum on a display device.

2. The sample observation method by an electron microscope according to claim 1 , wherein the polymerized film is formed on a surface of a sample as a thin film by a polymerization reaction by irradiating the sample with an electron beam for sample observation in the sample chamber of the electron microscope.

3. The sample observation method by an electron microscope according to claim 1 , wherein the polymerized film is formed on a surface of a sample as a thin film by a polymerization reaction by irradiating the sample with an electron beam or plasma other than the electron beam for the sample observation of the electron microscope before the sample observation by the electron microscope.

4. The sample observation method by an electron microscope according to claim 1 , wherein the composition for evaporation suppression includes at least one kind selected from an amphiphilic compound, a metal compound, and a saccharide.

5. The sample observation method by an electron microscope according to claim 1 , wherein an electron microscopic image of a hydrated sample in a wet state is displayed on the displaying device without accompanying collapse of the hydrated sample.

6. The sample observation method by an electron microscope according to claim 1 , the method comprising:
applying the composition for evaporation suppression to a body surface of a living biological sample to form a thin film, and covering the sample with the thin film; and
displaying an electron microscopic image of movement of the biological sample covered with the thin film and accommodated in a sample chamber under vacuum as it is alive on a display device.

7. The sample observation method by an electron microscope according to claim 6 , wherein the biological sample is provided with motility by suppressing decrease in temperature associated with evaporation from the inside of the biological sample body, and the morphology of the biological sample is preserved as it is, by the thin film.

8. The sample observation method by an electron microscope according to claim 6 , wherein an internal body temperature of the biological sample, which enables the biological sample to be active, is maintained even under vacuum by the thin film.

9. The sample observation method by an electron microscope according to claim 1 , wherein the electron microscopic image of a sample is displayed on a display device without causing charge up of the sample using a scanning electron microscope.

10. A composition for evaporation suppression under vacuum, which is used to form a thin film on a surface of a sample including an evaporable substance under vacuum and cover the sample, and thus to impart barrier performance suppressing evaporation of the evaporable substance under vacuum to the evaporable substance, the composition comprising at least one kind selected from an amphiphilic compound, oils and fats, and an ionic liquid, wherein a polymerized film is formed on a surface of a sample as a thin film by irradiating the sample applied with the composition for evaporation suppression with an electron beam or plasma, and the sample is covered with the polymerized film.

11. The composition for evaporation suppression under vacuum according to claim 10 , the composition being used to form a thin film on a surface of a living biological sample body and cover the biological sample, and thus to impart barrier performance suppressing evaporation of an evaporable substance under vacuum to the evaporable substance in the biological sample body.

12. The composition for evaporation suppression under vacuum according to claim 10 , the composition comprising at least one kind selected from an amphiphilic compound, a metal compound, and a saccharide.

13. A scanning electron microscope, the scanning electron microscope comprising a preliminary exhaust chamber capable of being introduced with a sample to be mounted in a sample chamber in a microscope body, and an exhauster deaerating the sample chamber and the preliminary exhaust chamber,
wherein the scanning electron microscope is used in a method comprising the steps of
applying a composition for evaporation suppression including at least one kind selected from an amphiphilic compound, oils and fats, and an ionic liquid to a surface of a sample;
irradiating the sample applied with the composition for evaporation suppression with an electron beam or plasma to form a polymerized film on the surface of the sample as a thin film and covering the sample with the polymerized film; and
displaying an electron microscopic image of the sample covered with the polymerized film and accommodated in a sample chamber under vacuum on a display device.

14. The scanning electron microscope according to claim 13 , wherein the preliminary exhaust chamber is equipped with a glove box.

15. The scanning electron microscope according to claim 13 , wherein the sample chamber or the preliminary exhaust chamber is equipped with a plasma irradiation device or an electron beam irradiation device.

16. The scanning electron microscope according to claim 13 , wherein the sample chamber or the preliminary exhaust chamber is equipped with a three dimensional manipulator capable of operating on a sample.

17. The scanning electron microscope according to 13 , wherein the sample chamber is equipped with a detection position regulatory mechanism capable of three dimensionally regulating a relative position of a secondary electron detector and a sample.

18. The scanning electron microscope according to claim 13 , wherein the sample chamber is equipped with a high speed color camera capable of acquiring color information of a sample.

19. The scanning electron microscope according to claim 13 , the scanning electron microscope comprising a temperature regulator capable of regulating a temperature of a sample stub in a sample chamber.

20. The scanning electron microscope according to claim 13 , wherein the sample chamber is equipped with at least one sensor selected from an electrical sensor, a light sensor, a gas sensor, a water sensor, and a temperature sensor.

21. A transmission electron microscope, the transmission electron microscope comprising a preliminary exhaust chamber capable of being introduced with a sample to be mounted in a sample chamber in a microscope body, and an exhauster deaerating the sample chamber and the preliminary exhaust chamber,
wherein the transmission electron microscope is used in a method comprising the steps of
applying a composition for evaporation suppression including at least one kind selected from an amphiphilic compound, oils and fats, and an ionic liquid to a surface of a sample;
irradiating the sample applied with the composition for evaporation suppression with an electron beam or plasma to form a polymerized film on the surface of the sample as a thin fi

Assignees

• Hariyama Takahiko
• TAKAKU Yasuharu
• Suzuki Hiroshi
• Muranaka Yoshinori
• Ohta Isao
• Shimomura Masatsugu
• Ishii Daisuke
• Japan Science & Tech Agency

Inventors

• Hariyama Takahiko
• TAKAKU Yasuharu
• Suzuki Hiroshi
• Muranaka Yoshinori
• Ohta Isao
• Shimomura Masatsugu
• Ishii Daisuke

Classifications

• G01N2223/612

  biological material · CPC title

• H01J2237/2004

  Biological samples · CPC title

• G01N23/04 Primary

  and forming images of the material · CPC title

• G01N1/2853

  Shadowing samples · CPC title

• G01N23/225

  using electron or ion · CPC title