Microscope apparatus
A microscope apparatus includes an illumination optical system that illuminates a sample, an observation optical system that guides light from the sample, and an intensity modulator that is provided in a pupil of the observation optical system or a position optically conjugate with the pupil and reduces light incident on the intensity modulator. The light utilization rate distribution as an intensity transmittance distribution of the intensity modulator in the pupil or in an image of the pupil monotonously increases or monotonously decreases in a first direction. The light utilization rate distribution varies on both sides of a center of an optical axis.
1. A microscope apparatus comprising:
an illumination optical system that illuminates a sample;
an observation optical system that guides light from the sample;
an intensity modulator that is provided in a pupil of the observation optical system or a position optically conjugate with the pupil and reduces light incident on the intensity modulator at an intensity transmittance corresponding to a phase gradient of the sample; and
an image acquisition section that acquires image data of the sample based on the light guided by the observation optical system from the sample, the image acquisition section including an image sensor,
wherein a light utilization rate distribution as an intensity transmittance distribution of the intensity modulator in the pupil or in an image of the pupil monotonously increases or monotonously decreases in a first direction, and varies on both sides of a center of an optical axis,
wherein the observation optical system and the intensity modulator are configured such that light beams, which constitute a light flux which is incident on the sample and is refracted by the phase gradient of the sample, pass through different positions of a pupil plane of the pupil or an optically conjugate pupil plane of the pupil where the intensity modulator is arranged, depending on the phase gradient occurring in regions through which the light beams have passed in the sample, whereby the light beams are respectively reduced at different intensity transmittances in the intensity modulator, and
wherein the observation optical system guides the light beams reduced by the intensity modulator into the image sensor to form an optical image having a light intensity corresponding to the phase gradient of the sample on a light receiving surface of the image sensor to thereby generate a phase gradient image of the sample.
2. The microscope apparatus according to claim 1 , further comprising
a contrast enhancement section that performs image processing for enhancing a contrast of the phase gradient image of the sample which is to be displayed on a display device based on the image data acquired by the image acquisition section.
3. The microscope apparatus according to claim 2 , further comprising the display device.
4. The microscope apparatus according to claim 2 , further comprising
a chroma enhancement section that performs processing for enhancing a chroma of the phase gradient image of the sample to be displayed on the display device based on the image data.
5. The microscope apparatus according to claim 1 , wherein
the observation optical system forms the optical image of the sample by the light flux to be emitted from the observation optical system and corresponding to a smaller numerical aperture than a numerical aperture on an image side of the observation optical system.
6. The microscope apparatus according to claim 5 , wherein
a numerical aperture of the illumination optical system is smaller than a numerical aperture on an object side of the observation optical system.
7. The microscope apparatus according to claim 6 , wherein
the numerical aperture of the illumination optical system is 90% or less of the numerical aperture on the object side of the observation optical system.
8. The microscope apparatus according to claim 6 , further comprising
an aperture stop arranged in an illumination optical system,
wherein
the intensity modulator is arranged on an optical path of the observation optical system, and
the aperture stop is arranged at a position which is optically conjugate with the intensity modulator along the optical path and so that a center of an opening of the aperture is positioned on an optical axis which is a center of the pupil of the observation optical system, such that an image of the center of the opening of the aperture stop is projected onto the optical axis.
9. The microscope apparatus according to claim 5 , further comprising an aperture stop.
10. The microscope apparatus according to claim 9 , wherein
the aperture stop has a structure in which an opening position is nonconcentric with respect to an optical axis.
11. The microscope apparatus according to claim 1 , wherein
a second-order differential value of the light utilization rate distribution in the pupil or in the image of the pupil with respect to a position in the first direction is positive.
12. The microscope apparatus according to claim 11 , wherein
the light utilization rate distribution in the pupil or in the image of the pupil is an exponential function of the position in the first direction.
13. The microscope apparatus according to claim 1 ,
wherein the light utilization rate distribution in the pupil or in the image of the pupil has an increase rate or a decrease rate that differs in a positive orientation of the first direction depending on a wavelength.
14. The microscope apparatus according to claim 13 , wherein
the intensity modulator includes a plurality of intensity modulation elements respectively having different spectral light utilization rate distributions.
15. The microscope apparatus according to claim 1 , wherein
a first light utilization rate distribution corresponding to a first wavelength of the intensity modulator in the pupil or in the image of the pupil monotonously increases in a positive orientation of the first direction,
a second light utilization rate distribution corresponding to a second wavelength of the intensity modulator in the pupil or in the image of the pupil monotonously increases in a negative orientation of the first direction, and
the first wavelength and the second wavelength differ from each other.
16. The microscope apparatus according to claim 1 , wherein
a first light utilization rate distribution corresponding to a first wavelength of the intensity modulator in the pupil or in the image of the pupil monotonously increases or monotonously decreases in the first direction,
a second light utilization rate distribution corresponding to a second wavelength of the intensity modulator in the pupil or in the image of the pupil monotonously increases or monotonously decreases in a direction different from the first direction, and
the first wavelength and the second wavelength differ from each other.
17. The microscope apparatus according to claim 1 , further comprising
a change device for changing the light utilization rate distribution in the pupil or in the image of the pupil.
18. The microscope apparatus according to claim 17 , wherein
the change device rotates the intensity modulator.
19. The microscope apparatus according to claim 17 , wherein
the intensity modulator includes a plurality of intensity modulation elements,
the plurality of intensity modulation elements respectively have light utilization rate distributions that differ from one another in orientation that monotonically increases or monotonically decreases, and
the change device moves the plurality of intensity modulation elements in a direction intersecting an optical axis of the observation optical system or an optical axis of the illumination optical system.
20. The microscope apparatus according to claim 17 , wherein
the change device changes an angle of the intensity modulator with respect to an optical axis of the observation optical system or an optical axis of the illumination optical system.
21. The microscope apparatus according to claim 17 , wherein
the intensity modulator is arranged on a detection optical path, and
the change device is a variable focus optical system included in the observation optical system and arranged on an optical path between the intensity modulator and the sample.
22. The microscope apparatus according to claim 1 , further comprising
a movement device for moving the intensity modulator in a direction along an optical axis of the observation optical system or a direction along an optical axis of the illumination optical system.
23. The microscope apparatus according to claim 1 , wherein
the intensity modulator includes a spatial light modulator in which a plurality of pixels are arranged in a lattice shape.
24. The microscope apparatus according to claim 1 , wherein
the intensity modulator includes a gradation filter having the intensity transmittance distribution.
25. A microscope apparatus comprising:
an illumination optical system that illuminates a sample;
an observation optical system that guides light from the sample; and
an intensity modulator that is provided in a pupil of the observation optical system or a position optically conjugate with the pupil and reduces light incident on the intensity modulator, at an intensity reflectance corresponding to a phase gradient of the sample; and
an image acquisition section that acquires image data of the sample based on the light guided by the observation optical system from the sample, the image acquisition section including an image sensor,
wherein a light utilization rate distribution as an intensity reflectance distribution of the intensity modulator in the pupil or in an image of the pupil monotonously increases or monotonously decreases in a first direction, and varies on both sides of a center of an optical axis,
wherein the observation optical system and the intensity modulator are configured such that light beams, which constitute a light flux which is incident on the sample and is refracted by the phase gradient of the sample, pass through different positions of a pupil plane of the pupil or an optically conjugate pupil plane of the pupil where the intensity modulator is arranged, depending on the phase gradient occurring in regions through which the light beams have passed in the sample, whereby the light beams are respectively reduced at different intensity reflectances in the intensity modulator, and
wherein the observation optical system guides the light beams reduced by the intensity modulator into the image sensor to form an optical image having a light intensity corresponding to the phase gradient of the sample on a light receiving surface of the image sensor to thereby generate a phase gradient image of the sample.
26. The microscope apparatus according to claim 25 , further comprising
a contrast enhancement section that performs image processing for enhancing a contrast of the phase gradient image of the sample which is to be displayed on a display device based on the image data acquired by the image acquisition section.
27. The microscope apparatus according to claim 26 , further comprising
the display device.
28. The microscope apparatus according to claim 26 , further comprising
a chroma enhancement section that performs processing for enhancing a chroma of the phase gradient image of the sample to be displayed on the display device based on the image data.
29. The microscope apparatus according to claim 25 , wherein
the observation optical system forms the optical image of the sample by the light flux to be emitted from the observation optical system and corresponding to a smaller numerical aperture than a numerical aperture on an image side of the observation optical system.
30. The microscope apparatus according to claim 29 , wherein
a numerical aperture of the illumination optical system is smaller than a numerical aperture on an object side of the observation optical system.
31. The microscope apparatus according to claim 30 , wherein
the numerical aperture of the illumination optical system is 90% or less of the numerical aperture on the object side of the observation optical system.
32. The microscope apparatus according to claim 30 , further comprising
an aperture stop arranged in an illumination optical system,
wherein
the intensity modulator is arranged on an optical path of the observation optical system, and
the aperture stop is arranged at a position which is optically conjugate with the intensity modulator along the optical path and so that a center of an opening of the aperture is positioned on an optical axis which is a center of the pupil of the observation optical system, such that an image of the center of the opening of the aperture stop is projected onto the optical axis.
33. The microscope apparatus according to claim 29 , further comprising
an aperture stop.
34. The microscope apparatus according to claim 33 , wherein
the aperture stop has a structure in which an opening position is nonconcentric with respect to an optical axis.
35. The microscope apparatus according to claim 25 , wherein
a second-order differential value of the light utilization rate distribution in the pupil or in the image of the pupil with respect to a position in the first direction is positive.
36. The microscope apparatus according to claim 35 , wherein
the light utilization rate distribution in the pupil or in the image of the pupil is an exponential function of the position in the first direction.
37. The microscope apparatus according to claim 25 , wherein the light utilization rate distribution in the pupil or in the image of the pupil has an increase rate or a decrease rate that differs in a positive orientation of the first direction depending on a wavelength.
38. The microscope apparatus according to claim 37 , wherein
the intensity modulator includes a plurality of intensity modulation elements respectively having different spectral light utilization rate distributions.
39. The microscope apparatus according to claim 25 , wherein
a first light utilization rate distribution corresponding to a first wavelength of the intensity modulator in the pupil or in the image of the pupil monotonously increases in a positive orientation of the first direction,
a second light utilization rate distribution corresponding to a second wavelength of the intensity modulator in the pupil or in the image of the pupil monotonously increases in a negative orientation of the first direction, and
the first wavelength and the second wavelength differ from each other.
40. The microscope apparatus according to claim 25 , wherein
a first light utilization rate distribution corresponding to a first wavelength of the intensity modulator in the pupil or in the image of the pupil monotonously increases or monotonously decreases in the first direction,
a second light utilization rate distribution corresponding to a second wavelength of the intensity modulator in the pupil or in the image of the pupil monotonously increases or monotonously decreases in a direction different from the first direction, and
the first wavelength and the second wavelength differ from each other.
41. The microscope apparatus according to claim 25 , further comprising
a change device for changing the light utilization rate distribution in the pupil or in the image of the pupil.
42. The microscope apparatus according to claim 41 , wherein
the change device rotates the intensity modulator.
43. The microscope apparatus according to claim 41 , wherein
the intensity modulator includes a plurality of intensity modulation elements, the plurality of intensity modulation elements respectively have light utilization rate distributions that differ from one another in orientation that monotonically increases or monotonically decreases, and
the change device moves the plurality of intensity modulation elements in a direction intersecting an optical axis of the observation optical system or an optical axis of the illumination optical system.
44. The microscope apparatus according to claim 41 , wherein
the change device changes an angle of the intensity modulator with respect to an optical axis of the observation optical system or an optical axis of the illumination optical system.
45. The microscope apparatus according to claim 41 , wherein
the intensity modulator is arranged on a detection optical path, and
the change device is a variable focus optical system included in the observation optical system and arranged on an optical path between the intensity modulator and the sample.
46. The microscope apparatus according to claim 25 , further comprising
a movement device for moving the intensity modulator in a direction along an optical axis of the observation optical system or a direction along an optical axis of the illumination optical system.
47. The microscope apparatus according to claim 25 , wherein
the intensity modulator includes a spatial light modulator in which a plurality of pixels are arranged in a lattice shape.
48. The microscope apparatus according to claim 25 , wherein
the intensity modulator includes a gradation mirror having the intensity reflectance distribution.