Laser scanning microscope, and laser scanning microscope control method

• US 10,642,012 B2
• Filed: 05/30/2017
• Issued: 05/05/2020
• Est. Priority Date: 06/06/2016
• Status: Active Grant

• Current Assignee: Evident Corporation
• Original Assignee: Olympus Corporation
• Inventors: Tamano, Shingo, Hosono, Kazuhiko
• Primary Examiner(s): King, George G
• Application Number:
  US15/608,401
• Publication Number:
  US 20170351074A1
• Time in Patent Office: 1,071 Days
• Field of Search: 359380, 359381, 359388
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 2201/06 Illumination; Optics
• CPC Class Codes: G01N 2201/06113 Coherent sources; lasers
• CPC Class Codes: G02B 19/0085 for use with both a detector and a source , e.g. in a transceiver, G02B19/009, G02B19/0095 take precedence
• CPC Class Codes: G02B 21/0032 Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers G02B21/0036 - G02B21/008; means for illumination of specimens in general G02B21/06
• CPC Class Codes: G02B 21/0048 scanning mirrors, e.g. rotating or galvanomirrors, MEMS mirrors
• CPC Class Codes: G02B 21/006 focusing arrangements; selection of the plane to be imaged
• CPC Class Codes: G02B 21/0064 multi-spectral or wavelength-selective arrangements, e.g. wavelength fan-out, chromatic profiling G02B21/0076 takes precedence
• 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 21/0088 Inverse microscopes
• CPC Class Codes: G02B 21/086 for transillumination only
• CPC Class Codes: G02B 21/16 adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence
• 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 5/02 Diffusing elements; Afocal elements

Abstract

A laser scanning microscope includes: an objective that irradiates a specimen with a laser beam; a detection lens that condenses the laser beam that passes through the specimen, the detection lens being arranged so as to face the objective; an optical element that is removably arranged between an image plane on which the detection lens forms an image of the specimen and a first surface that is a lens surface closest to the specimen of the detection lens, the optical element converting the laser beam made incident on the optical element into diffused light or deflecting a portion of the laser beam made incident on the optical element; and a photodetector that detects detection light emitted from the optical element arranged between the image plane and the first surface to the image plane.

Claims

1. A laser scanning microscope comprising:

a laser light source that emits a laser beam;

an objective that irradiates a specimen with the laser beam;

a detection lens that condenses the laser beam that passes through the specimen, the detection lens being arranged so as to face the objective with the specimen interposed between the detection lens and the objective;

a light diffusion element that is removably arranged between an image plane on which the detection lens forms an image of the specimen and a first surface that is a lens surface of the detection lens closest to the specimen, the light diffusion element converting the laser beam made incident on the light diffusion element into diffused light;

a photodetector that detects detection light emitted from the detection lens to the image plane; and

a controller that controls at least one of (i) an amplification factor of the photodetector so that the amplification factor of the photodetector increases when the light diffusion element is inserted into an optical path and decreases when the light diffusion element is removed from the optical path, and (ii) an output of the laser light source so that the output of the laser light source increases when the light diffusion element is inserted into the optical path and decreases when the light diffusion element is removed from the optical path,

wherein the objective is one of a plurality of objectives that are switchable, and

wherein the controller controls insertion of the light diffusion element into the optical path when the objective is switched to an objective having a magnification lower than 10 times, and controls removal of the light diffusion element when the objective is switched to an objective having a magnification equal to or higher than 10 times.

2. The laser scanning microscope according to claim 1, further comprising:

a light shielding member that is arranged between the detection lens and the photodetector, the light shielding member including an aperture, and partially shielding an incident light flux.

3. The laser scanning microscope according to claim 1, wherein the controller generates a new image of the specimen from a plurality of scanning images including a first scanning image of the specimen that is generated according to the output from the photodetector in a state in which a focal plane of the detection lens is located closer to the detection lens than a specimen surface is and a second scanning image of the specimen that is generated according to the output from the photodetector in a state in which the focal plane of the detection lens is located closer to the objective than the specimen surface is.

4. The laser scanning microscope according to claim 1, further comprising:

a scanner that scans the specimen with the laser beam.

5. The laser scanning microscope according to claim 1, further comprising:

a relay optical system that is arranged between the light diffusion element arranged between the image plane and the first surface, and the photodetector.

6. A control method of a laser scanning microscope having a plurality of objectives that are switchable, the method comprising:

controlling insertion and removal of a light diffusion element between an image plane on which a detection lens forms an image of a specimen irradiated with a laser beam and a first surface that is a lens surface of the detection lens closest to the specimen so that the light diffusion element is (i) inserted into an optical path when switching to an objective having a magnification lower than 10 times, and (ii) removed from the optical path when switching to an objective having a magnification equal to or higher than 10 times, the light diffusion element converting the laser beam into diffused light, and the detection lens being arranged so as to face the objective with the specimen interposed between the detection lens and the objective; and

controlling at least one of (i) an amplification factor of a photodetector so that the amplification factor of the photodetector increases when the light diffusion element is inserted into the optical path and decreases when the light diffusion element is removed from the optical path, and (ii) an output of a laser light source so that the output of the laser light source increases when the light diffusion element is inserted into the optical path and decreases when the light diffusion element is removed from the optical path, the photodetector detecting detection light emitted from the detection lens to the image plane, and the laser light source emitting the laser beam.