MICROSCOPE DEVICE, VIEWING METHOD, AND CONTROL PROGRAM

• US 20180017774A1
• Filed: 09/26/2017
• Published: 01/18/2018
• Est. Priority Date: 03/27/2015
• Status: Active Grant

• Current Assignee: Nikon Corporation
• Original Assignee: Nikon Corporation
• Inventors: TOMOSUGI, Wataru, TAKATSUKA, Kenji
• Granted Patent: US 11,086,115 B2
• Time in Patent Office: Days
• CPC Class Codes: G01N 2021/178 Methods for obtaining spatial resolution of the property being measured
• CPC Class Codes: G01N 2021/1791 stroboscopic; pulse gated; time range gated
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G02B 21/0076 arrangements using fluorescence or luminescence
• CPC Class Codes: G02B 21/06 Means for illuminating specimens

Abstract

A microscope apparatus includes an illumination optical system that emits activating light and exciting light, an image capturing unit that captures an image formed by an imaging optical system, an image processing unit that carries out image processing using an image capturing result from the image capturing unit, and a controller . The controller provides a plurality of image generation periods and interrupt periods; causes, in the image generation periods, the activated fluorescent material to be irradiated with the exciting light and causes the image capturing unit to capture an image of the fluorescent light from the activated fluorescent material in a plurality of frame periods; and sets an intensity of the exciting light in the interrupt periods to be lower than an intensity in the image generation periods or causes the emission of the exciting light to stop in the interrupt periods. The image processing unit generates one picture image from at least some of image capturing results obtained in the plurality of frame periods.

Claims

1. A microscope apparatus comprising:

an illumination optical system configured to emit activating light that activates a fluorescent material contained in a sample and exciting light that excites the activated fluorescent material;

an imaging optical system configured to form an image of fluorescent light from the fluorescent material;

an image capturing unit configured to capture the image formed by the imaging optical system;

an image processing unit configured to carry out image processing using an image capturing result from the image capturing unit; and

a controller,

the controller being configured to;

provide a plurality of image generation periods and interrupt periods;

cause, in the image generation periods, the activated fluorescent material to be irradiated with the exciting light and cause the image capturing unit to capture the image of the fluorescent light from the activated fluorescent material in a plurality of frame periods; and

set an intensity of the exciting light in the interrupt periods to be lower than an intensity in the image generation periods or causing the emission of the exciting light to stop in the interrupt periods, and

the image processing unit being configured to generate one picture image from at least some of image capturing results obtained in the plurality of frame periods.

2. The apparatus according to claim 1, wherein the controller is configured to alternately provide the image generation periods and the interrupt periods.

3. The apparatus according to claim 1, wherein the interrupt periods are longer than any of the plurality of frame periods.

4. The apparatus according to claim 1, wherein the interrupt periods are longer than the image generation periods.

5. The apparatus according to claim 1, wherein the controller is configured to:

provide a first period and a second period in at least one of the plurality of image generation periods;

cause, in the first period, the exciting light of a first wavelength to be emitted and cause the image capturing unit to capture the image of the fluorescent light from the activated fluorescent material in a plurality of frame periods; and

cause, in the second period, the exciting light of a second wavelength to be emitted and causes the image capturing unit to capture the image of the fluorescent light from the activated fluorescent material in a plurality of frame periods, and

the image processing unit is configured to form at least one picture image from at least some of image capturing results from the image capturing unit in the first period and at least some of image capturing results from the image capturing unit in the second period.

6. The apparatus according to claim 5,

wherein the image processing unit is configured to generate one picture image from at least some of the image capturing results from the image capturing unit in the first period and at least some of the image capturing results from the image capturing unit in the second period.

7. The apparatus according to claim 5,

wherein the image processing unit is configured to form a first picture image from at least some of the image capturing results from the image capturing unit in the first period and form a second picture image from at least some of the image capturing results from the image capturing unit in the second period.

8. The apparatus according to claim 7,

wherein the image processing unit is configured to generate one picture image from the first picture image and the second picture image.

9. The apparatus according to claim 5,

wherein the controller is configured to execute, a plurality of times, a process in which the illumination optical system emits auxiliary light for detecting a position of a fiducial marker and the image capturing unit captures an image of light from the fiducial marker.

10. The apparatus according to claim 9,

wherein an optical intensity of the auxiliary light is lower than an optical intensity of the exciting light.

11. The apparatus according to claim 9,

wherein the image processing unit is configured to correct the image capturing results from the image capturing unit in the first period and correct the image capturing results from the image capturing unit in the second period using at least some of image capturing results based on the light from the fiducial marker.

12. The apparatus according to claim 9,

wherein the image processing unit corrects a positional relationship between a picture image obtained from the image capturing results from the image capturing unit in the first period and a picture image obtained from the image capturing results from the image capturing unit in the second period.

13. The apparatus according to claim 1,

wherein the controller is configured to:

provide a plurality of first periods and second periods in at least one of the plurality of image generation periods;

cause, in the first periods, the exciting light of a first wavelength to be emitted and cause the image capturing unit to capture an image of light from the sample in one frame period; and

cause, in the second period, the exciting light of a second wavelength to be emitted and cause the image capturing unit to capture an image of light from the sample in one frame period, and

the image processing unit is configured to form at least one picture image from at least some of image capturing results from the image capturing unit in the plurality of first periods and at least some of image capturing results from the image capturing unit in the plurality of second periods.

14. The apparatus according to claim 13,

wherein the controller is configured to alternately provide the first periods and the second periods in the image generation period.

15. The apparatus according to claim 13,

wherein the image processing unit is configured to generate one picture image from at least some of image capturing results from the image capturing unit in the plurality of first periods and at least some of image capturing results from the image capturing unit in the plurality of second periods.

16. The apparatus according to claim 13,

wherein the image processing unit is configured to form a first picture image from the image capturing results from the image capturing unit in the plurality of first periods and form a second picture image from the image capturing results from the image capturing unit in the plurality of second periods.

17. The apparatus according to claim 16,

wherein the image processing unit is configured to generate one picture image from the first picture image and the second picture image.

18. An observation method comprising:

emitting activating light that activates a fluorescent material contained in a sample and exciting light that excites the activated fluorescent material;

forming an image of fluorescent light from the fluorescent material;

capturing the image formed by an imaging optical system;

carrying out image processing using a result of the capturing;

providing a plurality of image generation periods and interrupt periods;

causing, in the image generation periods, the activated fluorescent material to be irradiated with the exciting light and causing the image of the fluorescent light from the activated fluorescent material to be captured in a plurality of frame periods; and

setting an intensity of the exciting light in the interrupt periods to be lower than an intensity in the image generation periods or causing the emission of the exciting light to stop in the interrupt periods, and

the image processing comprising generating one picture image from at least some of image capturing results obtained in the plurality of frame periods.

19. A storage medium storing therein a control program causing a computer to execute control of a microscope apparatus configured to emit activating light that activates a fluorescent material contained in a sample and exciting light that excites the activated fluorescent material, capture an image of fluorescent light from the fluorescent material, and carry out image processing using a result of the capturing,

the control comprising:

providing a plurality of image generation periods and interrupt periods;

causing, in the image generation periods, the activated fluorescent material to be irradiated with the exciting light and causing the image of the fluorescent light from the activated fluorescent material to be captured in a plurality of frame periods; and

setting an intensity of the exciting light in the interrupt periods to be lower than an intensity in the image generation periods or causing the emission of the exciting light to stop in the interrupt periods, and

the image processing comprising generating one picture image from at least some of image capturing results obtained in the plurality of frame periods.