Phase-contrast microscope and imaging method
A culture vessel that houses a culture liquid and a specimen is irradiated with pattern light having a pattern that is previously set for the culture vessel. Transmitted light transmitted through the culture liquid in the culture vessel because of the irradiation with the pattern light is detected. Optical characteristics of an adjusting optical system that adjusts refraction of light caused by the shape of the liquid surface of the culture liquid in the culture vessel are adjusted, on the basis of a detection signal based on the detected transmitted light. After the adjustment, the culture vessel is irradiated with illumination light for phase-contrast measurement, and the specimen irradiated with the illumination light is imaged.
1. A phase-contrast microscope, comprising:
a phase-contrast-measurement illumination-light irradiation unit that irradiates a vessel that houses a liquid and a specimen, with illumination light for phase-contrast measurement;
an imaging unit that images the specimen irradiated with the illumination light;
an adjusting optical system that has adjustable optical characteristics and adjusts refraction of the illumination light caused by a shape of a liquid surface of the liquid in the vessel in accordance with the optical characteristic;
a pattern-light irradiation unit that irradiates the liquid surface of the liquid in the vessel with pattern light having a pattern, the pattern-light irradiation unit differing from the phase-contrast-measurement illumination-light irradiation unit;
a transmitted-light detection unit that detects transmitted light transmitted through the liquid surface of the liquid in the vessel because of the irradiation with the pattern light; and
an adjusting-optical-system control unit that adjusts the optical characteristics of the adjusting optical system on the basis of a detection signal based on the transmitted light detected by the transmitted-light detection unit.
2. The phase-contrast microscope according to claim 1 , further comprising:
an image forming optical system that forms an image of the specimen caused by the irradiation with the illumination light, on the imaging unit, and is subject to automatic focus control;
a reflected-light detection unit that detects reflected light of the pattern light reflected from a bottom surface of the vessel because of the irradiation with the pattern light; and
an image-forming-optical-system control unit that executes the automatic focus control on the image forming optical system on the basis of a detection signal based on the reflected light detected by the reflected-light detection unit.
3. The phase-contrast microscope according to claim 2 , wherein the adjusting-optical-system control unit adjusts the optical characteristics of the adjusting optical system on the basis of at least one evaluation result of evenness or contrast of the detection signal based on the transmitted light.
4. The phase-contrast microscope according to claim 3 , further comprising:
a filter unit that reduces incidence on the transmitted-light detection unit of reflected light of the illumination light reflected from the liquid surface of the liquid in the vessel because of the irradiation with the illumination light.
5. The phase-contrast microscope according to claim 4 , wherein the filter unit has spectral characteristics that are changeable in accordance with a wavelength of the illumination light.
6. The phase-contrast microscope according to claim 2 , further comprising:
a filter unit that reduces incidence on the transmitted-light detection unit of reflected light of the illumination light reflected from the liquid surface of the liquid in the vessel because of the irradiation with the illumination light.
7. The phase-contrast microscope according to claim 2 , wherein the adjusting-optical-system control unit adjusts the optical characteristics of the adjusting optical system after the automatic focus control by the image-forming-optical-system control unit.
8. The phase-contrast microscope according to claim 1 , wherein the adjusting-optical-system control unit adjusts the optical characteristics of the adjusting optical system on the basis of at least one evaluation result of evenness or contrast of the detection signal based on the transmitted light.
9. The phase-contrast microscope according to claim 8 , further comprising:
a filter unit that reduces incidence on the transmitted-light detection unit of reflected light of the illumination light reflected from the liquid surface of the liquid in the vessel because of the irradiation with the illumination light.
10. The phase-contrast microscope according to claim 1 , further comprising:
a filter unit that reduces incidence on the transmitted-light detection unit of reflected light of the illumination light reflected from the liquid surface of the liquid in the vessel because of the irradiation with the illumination light.
11. The phase-contrast microscope according to claim 10 , wherein the filter unit has spectral characteristics that are changeable in accordance with a wavelength of the illumination light.
12. The phase-contrast microscope according to claim 1 , wherein the pattern light has a streak-shaped pattern.
13. The phase-contrast microscope according to claim 1 , wherein the adjusting optical system has an optical element having an adjustable focal power.
14. The phase-contrast microscope according to claim 13 , wherein the optical element has a curvature at at least one of an incidence surface or an exit surface of the illumination light.
15. The phase-contrast microscope according to claim 14 , wherein the curvature of the optical element is adjustable.
16. The phase-contrast microscope according to claim 1 , wherein the adjusting-optical-system control unit acquires an adjustment condition of the adjusting optical system, and adjusts the optical characteristics of the adjusting optical system on the basis of the adjustment condition.
17. The phase-contrast microscope according to claim 16 , wherein the adjustment condition is determined on the basis of at least one of an optical magnification of the image forming optical system that forms the image of the specimen, a type of the vessel, a type of the specimen, the number of the specimens, a type of the liquid, an amount of the liquid, an environmental temperature, an environmental humidity, an imaging position in the vessel, or a size of an imaging region in the vessel.
18. The phase-contrast microscope according to claim 1 ,
wherein the phase-contrast-measurement illumination-light irradiation unit has a light source and a slit plate provided with a slit through which light emitted from the light source passes, and
wherein the specimen is irradiated with the light passing through the slit plate, as the illumination light.
19. The phase-contrast microscope according to claim 1 ,
wherein the pattern light having a pattern is a pattern light having a streak-shaped pattern of light and shade, a pattern light having a grid pattern in which patterns of light and shade are two-dimensionally periodically arrayed, a pattern light in which patterns of light and shade are concentrically arrayed, or a pattern light in which dot patterns are two-dimensionally arrayed.
20. The phase-contrast microscope according to claim 1 , further comprising:
a reflected-light detection unit that detects reflected light of the pattern light reflected from a bottom surface of the vessel because of the irradiation with the pattern light.
21. An imaging method, comprising:
irradiating a liquid surface of a liquid in a vessel that houses the liquid and a specimen, with pattern light having a pattern;
detecting transmitted light transmitted through the liquid surface of the liquid in the vessel because of the irradiation with the pattern light;
adjusting optical characteristics of an adjusting optical system that adjusts refraction of light caused by a shape of the liquid surface of the liquid in the vessel in accordance with the optical characteristic, on the basis of a detection signal based on the detected transmitted light;
after the adjustment, irradiating the vessel with illumination light for phase-contrast measurement, the illumination light differing from the pattern light; and
imaging the specimen irradiated with the illumination light.