SPECIMEN OBSERVATION APPARATUS

• US 20150316757A1
• Filed: 04/23/2015
• Published: 11/05/2015
• Est. Priority Date: 04/30/2014
• Status: Active Grant

• Current Assignee: Evident Corporation
• Original Assignee: Olympus Corporation
• Inventors: KITAGAWA, Hisao
• Granted Patent: US 10,642,013 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G02B 21/0044 moving apertures, e.g. Nipkow disks, rotating lens arrays
• CPC Class Codes: G02B 21/0048 scanning mirrors, e.g. rotating or galvanomirrors, MEMS mirrors
• CPC Class Codes: G02B 21/0076 arrangements using fluorescence or luminescence
• CPC Class Codes: G02B 21/008 Details of detection or image processing, including general computer control
• CPC Class Codes: G02B 21/361 Optical details, e.g. image relay to the camera or image sensor G02B21/364 takes precedence; illumination details G02B21/06 and subgroups
• CPC Class Codes: G02B 21/367 providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison
• CPC Class Codes: G02B 27/58 Optics for apodization or superresolution; Optical synthetic aperture systems

Abstract

A superior superresolution image can be generated. Provided is a specimen observation apparatus including an objective lens that radiates excitation laser light emitted from a light source onto a specimen; and a main controller that obtains a plurality of sets of image data of the same region on the specimen by repeatedly detecting fluorescence from the same region on the specimen irradiated with the excitation laser light by the objective lens, and that emphasizes high-frequency components in an addition image data obtained by adding the obtained plurality of sets of image data of the same region on the specimen.

Claims

1. A specimen observation apparatus comprising:

an objective lens that radiates laser light emitted from a light source onto a specimen;

an image-data obtaining portion that detects return light from the specimen irradiated with the laser light by the objective lens and that obtains image data;

a control portion that repeatedly detects the return light from the same region on the specimen with the image-data obtaining portion and that obtains a plurality of sets of the image data for the same region;

an image-data processing portion that adds the plurality of sets of the image data for the same region, obtained by the image-data obtaining portion; and

an image computational processing portion that emphasizes high-frequency components in the addition image data obtained by the image-data processing portion.

2. A specimen observation apparatus according to claim 1, further comprising:

a confocal aperture portion that has an opening at a position that is optically conjugate with a focal position of the objective lens; and

a confocal lens that focuses return light from the specimen irradiated with the laser light by the objective lens and that projects a spot of the return light at the opening position of the confocal aperture portion,

wherein the opening of the confocal aperture portion has a diameter smaller than the diameter of the spot of return light projected by the confocal lens.

3. A specimen observation apparatus according to claim 1, further comprising:

a scanning portion that scans the laser light, irradiated onto the specimen by the objective lens, on the specimen in a main scanning direction and a sub-scanning direction that intersects therewith,

wherein the control portion repeats an operation in which, after the laser light is repeatedly scanned on the same scanning line in the main scanning direction by the scanning portion for a prescribed number of times, the position thereof in the sub-scanning direction is changed so as to move to the next scanning line, and a plurality of sets of image data for the same scanning line are obtained by the image-data obtaining portion.

4. A specimen observation apparatus according to claim 1, further comprising:

the light source that emits line-shaped laser light,

wherein the image-data obtaining portion includes an image-acquisition device formed in the shape of a line, which acquires an image of the return light that returns from the specimen irradiated with the line-shaped laser light.

5. A specimen observation apparatus according to claim 1, further comprising:

a mask that has openings in the shape of slits or pinholes arranged periodically and that restricts the laser light incident on the objective lens; and

a driving portion that rotates the mask at high speed about a center axis thereof,

wherein the image-data obtaining portion includes a two-dimensional image-acquisition device that acquires an image of the return light returning from the specimen.

6. A specimen observation apparatus comprising:

a scanning mirror that scans laser light emitted from a light source on a specimen;

an objective lens that radiates the laser light scanned by the scanning mirror onto the specimen;

a light detector that detects return light from the specimen irradiated with the laser light and that outputs a signal according to a light level thereof; and

a controller that generates image data of the specimen by using the signal output from the light detector,

wherein the controller performs processing for generating a plurality of sets of the image data for the same position by controlling the scanning mirror so as to repeatedly scan the laser light a plurality of times at the same position on the specimen, processing for adding the generated plurality of sets of image data for the same position, and processing for emphasizing high-frequency components in an addition image data obtained by the adding.

7. A specimen observation apparatus according to claim 6, wherein the scanning mirror includes at least one galvanometer mirror that deflects the laser light in a main scanning direction and a sub-scanning direction, which intersect each other,

wherein the controller controls the scanning mirror so as to repeatedly perform an operation in which, after the laser light is repeatedly scanned on the same scanning line in the main scanning direction a prescribed number of times, the position thereof in the sub-scanning direction is changed so as to move to the next scanning line.

8. A specimen observation apparatus comprising:

an objective lens that radiates excitation light emitted from a light source onto a specimen;

a mask that is disposed on the light source side of the objective lens and that has openings in the shape of slits or pinholes which are periodically arranged;

a motor that rotates the mask about a prescribed rotation axis;

a two-dimensional image-acquisition device that acquires an image of return light from the specimen irradiated with the excitation light via the rotating mask; and

a controller that performs processing for repeatedly obtaining and adding the images of the same region on the specimen, obtained by the two-dimensional image-acquisition device, and processing for emphasizing high-frequency components in addition image data obtained by the adding.