3D imaging microscope
View Patent ↗A laser microscope that efficiently performs 3D imaging irrespective of the shape of a specimen, includes an area segmenting section that segments an observation range of a specimen in the direction perpendicular to the optical axis of an objective lens into many areas; a surface-position storing section that stores motorized stage positions in association with the specimen surface positions, for the many areas; a surface-shape estimating section that estimates the specimen surface shape from the motorized stage positions and the specimen surface positions for the many areas; a z-scanning condition determining section that determines the specimen surface positions at desired positions of the motorized stage from the specimen surface shape; and a light detecting section that detects light from the specimen over certain ranges specified with reference to the specimen surface positions, in the optical axis direction of the objective lens.
1. A microscope comprising:
a stage on which a specimen is placeable;
a scanner that performs two-dimensional scanning of laser light on the specimen;
an objective lens that focuses the laser light used for the two-dimensional scanning performed by the scanner onto the specimen and that collects light from the specimen;
a first driving section that moves the stage relative to the objective lens in directions perpendicular to an optical axis direction of the objective lens;
a second driving section that moves the stage relative to the objective lens in the optical axis direction of the objective lens;
a control unit configured to operate as:
an area segmenting section that segments an observation range of the specimen, in the directions perpendicular to the optical axis direction of the objective lens, into a plurality of two-dimensional areas in which the scanner performs scanning;
a surface-position storing section that stores a plurality of positions of the stage in association with surface positions of the specimen for a plurality of the two-dimensional areas;
a surface-shape estimating section that estimates a surface shape of the specimen from the plurality of positions of the stage and the associated surface positions of the specimen stored in the surface-position storing section; and
an image generating section; and
a light detecting section that, for a plurality of light-collecting positions of the objective lens in the segmented plurality of two-dimensional areas, detects light from the specimen over certain ranges specified with reference to surface positions of the specimen estimated by the surface-shape estimating section in the optical axis direction of the objective lens; and
wherein the image generating section is configured to generate a 3D image of the specimen from the light from the specimen detected by the light detecting section at the plurality of light-collecting positions of the objective lens in the segmented plurality of two-dimensional areas.
2. The microscope according to claim 1 , wherein the control unit is further configured to operate as:
an imaging condition storing section that stores the positions of the stage in association with imaging conditions for the specimen in the optical axis direction of the objective lens; and
an imaging condition determining section that uses the imaging conditions stored in the imaging condition storing section to determine imaging conditions in the optical axis direction of the objective lens at the surface positions of the specimen estimated by the surface-shape estimating section.
3. The microscope according to claim 1 , wherein the surface-shape estimating section estimates the surface shape of the specimen by determining an equation of a plane passing through three points from among the positions of the stage and coordinates of the associated surface positions of the specimen for the three points.
4. The microscope according to claim 1 , wherein the control unit is further configured to operate as a surface-position detecting section that detects the associated surface positions of the specimen to be stored in the surface-position storing section.
5. The microscope according to claim 4 , wherein the surface-position detecting section detects the associated surface positions of the specimen based on luminance information about the light from the specimen, detected by the light detecting section.