Microscope and microscope image acquisition method

Patent metadata

Field	Value
Publication number	US-2016124203-A1
Application number	US-201514922757-A
Country	US
Kind code	A1
Filing date	Oct 26, 2015
Priority date	Nov 4, 2014
Publication date	May 5, 2016
Grant date	—

Abstract

To prevent unwanted exposure of a specimen with excitation light despite magnification switching, a microscope of the present invention includes: a detection optical system that detects fluorescence emitted from the specimen; a light-sheet illumination optical system, serving as an excitation light source, that causes planar excitation light to be incident along a direction intersecting an optical axis of the detection optical system; and a control section that, when the magnification of the detection optical system is raised, controls the light-sheet illumination optical system so as to reduce a radiation width of the excitation light made incident on the specimen by the light-sheet illumination optical system.

First claim

1 . A microscope comprising:
a detection optical system that detects fluorescence emitted from a specimen; a light-sheet illumination optical system that causes planar excitation light to be incident along a direction intersecting an optical axis of the detection optical system; and a control section that, when a magnification of the detection optical system is raised, controls the light-sheet illumination optical system so as to reduce a radiation width of the excitation light that is incident on the specimen from the light-sheet illumination optical system.

2 . The microscope according to claim 1 , wherein the detection optical system includes an imaging device that acquires fluorescence, and
the control section calculates the radiation width of the excitation light that is incident on the specimen from the light-sheet illumination optical system on the basis of the magnification of the detection optical system and a size of the imaging device.

3 . The microscope according to claim 1 , wherein the light-sheet illumination optical system includes:
an excitation-light-forming optical system that forms planar excitation light having a predetermined width; and a scanning section that scans the planar excitation light formed by the excitation-light-forming optical system in a width direction of the planar excitation light.

4 . The microscope according to claim 3 , wherein the scanning section includes:
a galvanometer mirror that swivels in the width direction of the excitation light, and the control section controls a swivel angle of the galvanometer mirror.

5 . A microscope comprising:
a detection optical system that detects fluorescence emitted from a specimen to acquire a fluorescence image; a light-sheet illumination optical system that causes planar excitation light to be incident along a direction intersecting an optical axis of the detection optical system; and an image processing section that processes the image acquired by the detection optical system, wherein the light-sheet illumination optical system moves a focal position of the excitation light along an incidence direction thereof, and the image processing section combines a plurality of the fluorescence images that are acquired by the detection optical system each time the excitation light is made incident on a different focal position by the light-sheet illumination optical system, to generate a composite image.

6 . The microscope according to claim 5 , wherein the light-sheet illumination optical system can set a movement start position, a number of movement steps, and a displacement of each movement step for the focal position of the excitation light.

7 . The microscope according to claim 6 , wherein the light-sheet illumination optical system can switch a wavelength of the excitation light and can set the movement start position and the displacement for each wavelength of the excitation light.

8 . The microscope according to claim 5 , wherein the light-sheet illumination optical system can cause the excitation light to be incident alternately from two or more different directions from outside the specimen.

9 . The microscope according to claim 8 , wherein the light-sheet illumination optical system can cause the excitation light to be incident from both sides with respect to the optical axis of the detection optical system.

10 . A microscope image acquisition method comprising:
a radiation step of causing planar excitation light extending along one direction to be incident on a specimen; a movement step of moving a focal position of the excitation light along an incidence direction thereof; a detection step of detecting fluorescence that is emitted from the specimen each time the focal position of the excitation light is moved in the movement step to acquire a fluorescence image; and a combination step of combining a plurality of the fluorescence images acquired in the detection step to generate a composite image.

11 . The microscope according to claim 5 , further comprising a control section that, when a magnification of the detection optical system is raised, controls the light-sheet illumination optical system so as to reduce a radiation width of the excitation light to be made incident on the specimen by the light-sheet illumination optical system.

12 . The microscope according to claim 11 , wherein the light-sheet illumination optical system includes:
an excitation-light-forming optical system that forms planar excitation light having a predetermined width; and a scanning section that scans the planar excitation light formed by the excitation-light-forming optical system in a width direction of the planar excitation light, and the control section controls a swivel angle of the scanning section.

13 . The microscope according to claim 8 , wherein the light-sheet illumination optical system includes:
an illumination-direction switching section that alternately switches the incidence direction between two or more different directions from outside the specimen to sequentially cause the excitation light to be incident along a plane intersecting the optical axis of the detection optical system; and an image processing section that combines a plurality of the fluorescence images acquired by the detection optical system when the excitation light is incident along a different incidence direction, and the light-sheet illumination optical system switches the incidence direction of the excitation light in such an order that the excitation light is made incident starting from a direction equal to an identical incidence direction between before and after combining processing by the image processing section.

14 . The microscope according to claim 8 , further comprising:
an illumination-direction switching section that alternately switches the incidence direction between two or more different directions from outside the specimen to sequentially cause the planar excitation light to be incident along a plane intersecting the optical axis of the detection optical system; an image processing section that combines a plurality of the fluorescence images acquired by the detection optical system when the excitation light is incident along a different incidence direction; a movement mechanism that moves the specimen in an optical-axis direction of the detection optical system; and a control section that controls the illumination-direction switching section and the movement mechanism, wherein when the fluorescence image based on the planar excitation light that is incident along each of the different directions is acquired, the control section controls the movement mechanism so as to move the specimen to a different position in the optical-axis direction and controls the illumination-direction switching section so as to switch the incidence direction of the excitation light in such an order that the excitation light is made incident starting from a direction equal to an identical incidence direction between before and after movement of the specimen by the movement mechanism.

15 . A microscope comprising:
a detection optical system that detects fluorescence emitted from a specimen to acquire a fluorescence image; a light-sheet illumination optical system that alternately switches an incidence direction between two or more different directions from outside the specimen to sequentially cause planar excitation light to be incident along a plane intersecting an optical axis of the detection optical system; and an image processing section that combines a plurality of the fluorescence images acquired by the detection optical syst

Assignees

• Olympus Corp

Inventors

• Ryu Go

Classifications

• G02B26/105

  with one or more pivoting mirrors or galvano-mirrors (G02B26/101 takes precedence) · CPC title

• G02B21/16

  adapted for ultraviolet illumination {; Fluorescence microscopes (G02B21/0076 takes precedence)} · CPC title

• G02B21/06

  Means for illuminating specimens · CPC title

• G02B21/367 Primary

  providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison · CPC title

• G02B21/0048

  scanning mirrors, e.g. rotating or galvanomirrors, MEMS mirrors · CPC title