# US 11,906,431 B2 · Microscope apparatus > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,906,431 B2 - Document ID: US11906431B2 - Status: Granted - Kind: B2 - Application number: 17/178,957 - Filed: 2021-02-18 - Published: 2021-06-10 - Granted: 2024-02-20 - Earliest claimed: 2015-11-27 - Assignees: Nikon Corporation - Applicants: Nikon Corporation - Inventors: Ichiro Sase; Yasutoshi Kaneko; Tatsuo Fukui - Web page: https://patentgrep.com/patent/US11906431B2 ## CPC - G01N 21/6458: {Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16)} - G01N 21/64: Fluorescence; Phosphorescence - G02B 21/0076: {arrangements using fluorescence or luminescence} - G02B 21/16: adapted for ultraviolet illumination {; Fluorescence microscopes (G02B21/0076 takes precedence)} - G02B 21/365: {Control or image processing arrangements for digital or video microscopes (G02B21/361, G02B21/362 take precedence)} - 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} - H04N 23/60: Control of cameras or camera modules - G01N 2201/06113: Coherent sources; lasers - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - G02B: OPTICAL ELEMENTS, SYSTEMS OR APPARATUS - H04N: PICTORIAL COMMUNICATION, e.g. TELEVISION ## Abstract A microscope apparatus including: an illumination optical system that radiates activation light to activate some of fluorescent materials included in a sample and excitation light to excite at least some of the activated fluorescent materials; an image forming optical system that: has an objective lens and an astigmatic optical system that generates astigmatism to at least part of fluorescence from the fluorescent materials; and forms an image of the fluorescence; an image-capturing unit that captures an image formed by the image forming optical system; a drive unit that moves an image-capturing position in the sample along an optical axis-direction of the objective lens; and a control unit, wherein the control unit causes the image-capturing unit to capture images in a plurality of numbers of frames respectively at a first image-capturing position and at a second image-capturing position different from the first image-capturing position. ## Claims ### Claim 1 A microscope apparatus comprising: an illumination optical system that radiates activation light to activate some of fluorescent materials included in a sample and excitation light to excite at least some of the activated fluorescent materials; an image forming optical system that has an objective lens and an astigmatic optical system, which generates astigmatism to at least part of fluorescence from the fluorescent materials, and that forms an image of the fluorescence; an image-capturing unit that captures an image formed by the image forming optical system; a drive unit that moves an image-capturing position in the sample along an optical axis-direction of the objective lens; and a control unit, wherein the control unit causes the image-capturing unit to capture images in a plurality of numbers of frames respectively at a first image-capturing position and at a second image-capturing position different from the first image-capturing position along the optical axis-direction, the control unit causes the image-capturing unit to capture images in a first number of frames at the first image-capturing position and capture images in a second number of frames different from the first number of frames at the second image-capturing position, and the first number of frames and the second number of frames are identified by image-capturing conditions set before the control unit causes the image-capturing unit to capture images at the first and second image-capturing positions. ### Claim 2 (depends on 1) The microscope apparatus according to claim 1, wherein the drive unit moves the image-capturing position by moving at least one of (i) a stage holding the sample and (ii) the objective lens in the optical axis-direction of the objective lens. ### Claim 3 (depends on 2) The microscope apparatus according to claim 2, wherein a distance between the second image-capturing position and a cover glass held by the stage is longer than a distance between the first image-capturing position and the cover glass, and wherein the control unit controls the image-capturing unit so that the second number of frames for the second image-capturing position is larger than the first number of frames for the first image-capturing position. ### Claim 4 (depends on 1) The microscope apparatus according to claim 1, further comprising a display apparatus, wherein the control unit: generates a three-dimensional image at the first image-capturing position based on a result of image-capturing at the first image-capturing position obtained through the image-capturing in the first number of frames, generates a three-dimensional image at the second image-capturing position based on a result of image-capturing at the second image-capturing position obtained through the image-capturing in the second number of frames, and displays, on the display apparatus, the three-dimensional image captured at the first image-capturing position and the three-dimensional image captured at the second image-capturing position in different display colors. ### Claim 5 (depends on 4) The microscope apparatus according to claim 4, wherein the control unit causes to be displayed on the display apparatus only a specified region in the optical axis-direction of the objective lens. ### Claim 6 (depends on 4) The microscope apparatus according to claim 4, wherein the control unit causes to be displayed on the display apparatus only an overlapping region. ### Claim 7 (depends on 4) The microscope apparatus according to claim 4, wherein the three-dimensional images include three-dimensional position information about the fluorescent materials. ### Claim 8 (depends on 1) The microscope apparatus according to claim 1, wherein the control unit causes the image-capturing unit to perform multiple times an image-capturing action of capturing images at the first image-capturing position and of capturing images at the second image-capturing position. ### Claim 9 (depends on 8) The microscope apparatus according to claim 8, further comprising a display apparatus, wherein the control unit: generates a three-dimensional image at the first image-capturing position based on a result of image-capturing at the first image-capturing position obtained through the image-capturing action performed multiple times, generates a three-dimensional image at the second image-capturing position based on a result of image-capturing at the second image-capturing position obtained through the image-capturing action performed multiple times, and displays, on the display apparatus, the three-dimensional image captured at the first image-capturing position and the three-dimensional image captured at the second image-capturing position in different display colors. ### Claim 10 (depends on 9) The microscope apparatus according to claim 9, wherein the control unit: generates a three-dimensional image for each image-capturing action based on results of image-capturing at the first image-capturing position and the second image-capturing position obtained through each image-capturing action; and switches the display apparatus between (i) a first display mode in which the three-dimensional image captured at the first image-capturing position and the three-dimensional image captured at the second image-capturing position are displayed on the display apparatus in different display colors and (ii) a second display mode in which the three-dimensional image for each image-capturing action is displayed on the display apparatus in a different display color from those for three-dimensional images for other image-capturing actions. ### Claim 11 (depends on 8) The microscope apparatus according to claim 8, further comprising a display apparatus, wherein a three-dimensional image for each image-capturing action is generated based on results of image-capturing at the first image-capturing position and the second image-capturing position obtained through each image-capturing action, and the three-dimensional image for each image-capturing action is displayed on the display apparatus in a different display color from those for three-dimensional images for other image-capturing actions. ### Claim 12 (depends on 1) The microscope apparatus according to claim 1, wherein the control unit calculates three-dimensional position information about the fluorescent materials based on the astigmatism. ### Claim 13 An image-capturing method performed at a microscope apparatus having: an illumination optical system that radiates activation light to activate some of fluorescent materials included in a sample and excitation light to excite at least some of the activated fluorescent materials; an image forming optical system that has an objective lens and an astigmatic optical system, which generates astigmatism to at least part of fluorescence from the fluorescent materials, and that forms an image of the fluorescence; an image-capturing unit that captures an image formed by the image forming optical system; and a drive unit that moves an image-capturing position in the sample along an optical axis-direction of the objective lens, the image-capturing method comprising: causing the image-capturing unit to capture images in a plurality of numbers of frames respectively at a first image-capturing position and at a second image-capturing position different from the first image-capturing position along the optical axis-direction such that images in a first number of frames at the first image-capturing position and images in a second number of frames different from the first number of frames at the second image-capturing position are captured, wherein the first number of frames and the second number of frames are identified by image-capturing conditions set before the image-capturing unit is caused to capture images at the first and second image-capturing positions. ### Claim 14 A non-transitory computer-readable medium storing a computer program product having computer instructions that are executed by a computer to control a microscope apparatus having: an illumination optical system that radiates activation light to activate some of fluorescent materials included in a sample and excitation light to excite at least some of the activated fluorescent materials; an image forming optical system that has an objective lens and an astigmatic optical system, which generates astigmatism to at least part of fluorescence from the fluorescent materials, and that forms an image of the fluorescence; an image-capturing unit that captures an image formed by the image forming optical system; and a drive unit that moves an image-capturing position in the sample along an optical axis-direction of the objective lens, the computer instructions, upon being executed by the computer, enabling the computer to perform operations comprising: executing a procedure of causing the image-capturing unit to capture images in a plurality of numbers of frames respectively at a first image-capturing position and at a second image-capturing position different from the first image-capturing position along the optical axis-direction such that images in a first number of frames at the first image-capturing position and images in a second number of frames different from the first number of frames at the second image-capturing position are captured, wherein the first number of frames and the second number of frames are identified by image-capturing conditions set before the image-capturing unit is caused to capture images at the first and second image-capturing positions.