A microscope is provided that enables a phase object or surface pits and projections to be observed at a relatively low image-formation magnification of 4 or lower over a wide viewing range yet in a relatively narrow spatial frequency distribution range. The microscope comprises a light source, an i
A microscope is provided that enables a phase object or surface pits and projections to be observed at a relatively low image-formation magnification of 4 or lower over a wide viewing range yet in a relatively narrow spatial frequency distribution range. The microscope comprises a light source, an illumination optical system, a partial aperture located at the pupil position of the illumination optical system, an image-formation optical system, and an eyepiece optical system or an image pickup optical system, wherein the diameter of the image of a partial aperture at the pupil position of the image-formation optical system is set smaller than the pupil diameter of the image-formation optical system, and at the pupil position of the image-formation optical system there is located an element for introducing in the pupil position of the image-formation optical system a wavefront varying in size with the pupil diameter.
대표청구항▼
What we claim is: 1. A method for implementing microscopy, comprising: providing a partial aperture at a pupil position of an illumination optical system in a microscope; providing an absorption film at a pupil position of an objective included in an image-formation optical system: illuminating an
What we claim is: 1. A method for implementing microscopy, comprising: providing a partial aperture at a pupil position of an illumination optical system in a microscope; providing an absorption film at a pupil position of an objective included in an image-formation optical system: illuminating an object under observation by passing light through said partial aperture; and introducing a substantially spherical wavefront at a pupil position of the image-formation optical system or at near a position conjugate thereto; wherein said object under observation is visualized on the basis of said introduced substantially spherical wavefront; and wherein the wavefront to be introduced is based on a displacement of a focal position of said objective. 2. The method for implementing microscopy according to claim 1, characterized in that said image-formation optical system has an image-formation magnification of 4 or lower. 3. The method for implementing microscopy according to claim 1, characterized in that said partial aperture comprises a circular aperture or a zonal aperture, with satisfaction of condition (10): D0/10≦D1 ≦D0/3 (10) where D1 is an aperture width of an image at the pupil position of said image-formation optical system, and D0 is a diameter of the pupil of said image-formation optical system. 4. The method for implementing microscopy according to claim 1, characterized in that the absorption film conforms in shape to the image of said partial aperture or a substantial point light source at the pupil position of said image-formation optical system and is slightly larger than said shape is located at the pupil of said image-formation optical system or at or near a position conjugate thereto, with satisfaction of condition (11) Δ≦D0/10 (11) where Δ is a margin width on an inner or outer peripheral side of said absorption film with respect to the image of said partial aperture or said substantial point light source at the pupil position of said image-formation optical system, and D0 is a diameter of the pupil of said image-formation optical system. 5. The method for implementing microscopy according to claim 1, characterized in that while the wavefront introduced in the pupil position of said image-formation optical system is substantially symmetrically changed, two images are taken of the same object under observation, and a subtract operation is applied between the two images to form a subtraction image. 6. The method for implementing microscopy according to claim 1, characterized in that the object under observation comprises cells, and further characterized in that while the wavefront introduced in the pupil position of said image-formation optical system is substantially symmetrically changed, two images with inverted contrasts are taken of the same object under observation, a subtract operation is applied between the two images to form an image with the subtract operation applied thereto, and at least one of the number of cells, the area taken up by cells, the abundance distribution of cells and the abundance density of cells in a viewing range is measured from the obtained image with the subtract operation applied thereto. 7. The method for implementing microscopy according to claim 6, characterized in that a portion having an intensity value of zero or nearly zero is extracted out of an intensity distribution of the image subjected to the subtract operation to take said portion as being a cell-free portion and other portion as being a cell-containing portion, thereby making a separation between the portions containing cells and no cells. 8. The method for implementing microscopy according to claim 6, characterized in that a portion having an intensity value of zero or nearly zero is extracted out of an intensity distribution of the image subjected to the subtract operation to assign an intensity value to one value and assign the other portion to another one intensity value for binarization. 9. The method for implementing microscopy according to claim 1, characterized in that the object under observation comprises cells having a thickness, and further characterized in that while the wavefront introduced in the pupil position of said image-formation optical system is substantially symmetrically changed, two images with inverted contrasts are taken of the same object under observation, and a subtract operation and an add operation are applied between the two images to form two images wherein the image subjected to the subtract operation is divided by the image subjected to the add operation to measure information about the thickness of cells in a viewing range. 10. A microscope comprising a light source, an illumination optical system for guiding light from the light source to an object under observation, an image-formation optical system for forming an image-formation plane of an image of the object under observation illuminated by light passing through a partial aperture disposed substantially at a pupil position of the illumination optical system, an eyepiece optical system or an image pickup optical system for viewing the image formed on the image-formation plane, and a wavefront introduction means for introducing a substantially spherical wavefront at a pupil position of the image-formation optical system or at or near a position conjugate thereto, characterized in that: a diameter of said partial aperture at a pupil position of said image-formation optical system is set smaller than a pupil diameter of said image-formation optical system; and said wavefront introduction means comprises means for displacement and adjustment of the focus position of an objective of said image-formation optical system in an optical axis direction. 11. The microscope according to claim 10, characterized in that said image-formation optical system has an image-formation magnification of 4 or lower. 12. The microscope according to claim 10, characterized in that said partial aperture comprises a circular aperture or a zonal aperture, with satisfaction of condition (10): D0/10≦D0/3 (10) where D1 is an aperture width of an image at the pupil position of said image-formation optical system, and D0 is a diameter of the pupil of said image-formation optical system. 13. The microscope according to claim 12, characterized in that an absorption film that conforms in shape to the image of said partial aperture or a substantial point light source at the pupil position of said image-formation optical system and is slightly larger than said shape is located at the pupil of said image-formation optical system or at or near a position conjugate thereto, with satisfaction of condition (11) Δ≦D0/10 (11) where Δ is a margin width on an inner or outer peripheral side of said absorption film with respect to the image of said partial aperture or said substantial point light source at the pupil position of said image-formation optical system, and D0 is the diameter of the pupil of said image-formation optical system. 14. The microscope according to claim 10, characterized in that the object under observation comprises cells, and further characterized by further comprising a processing unit for capturing two images comprising contrast images having a phase difference of opposite sign by means of said microscope and applying a subtract operation to the captured two images to form an image subjected to the subtract operation, so that at least one of the number of cells, the area taken up by cells, the abundance distribution of cells and the abundance density of cells in a viewing range is measured from the obtained image with the subtract operation applied thereto. 15. The microscope according to claim 14, characterized by further comprising a separation unit for extracting a portion having an intensity value of zero or nearly zero out of an intensity distribution of the image subjected to the subtract operation to take said portion as being a cell-free portion and other portion as being a cell-containing portion, thereby making a separation between the portions containing cells and no cells. 16. The microscope according to claim 14, characterized by further comprising a binarization unit for extracting a portion having an intensity of zero or nearly zero out of an intensity distribution of the image subjected to the subtract operation to assign an intensity value to one value and assign the other portion to another one intensity value for binarization. 17. The microscope according to claim 10, characterized in that the object under observation comprises cells having a thickness, and further characterized by further comprising an operation unit for capturing two images comprising contrast images having a phase difference of opposite sign and applying a subtract operation and an add operation to the captured two images, and a thickness measurement unit for dividing the image subjected to the subtract operation by the image subjected to the add operation to measure information about the thickness of cells in a viewing range. 18. A method for implementing microscopy, comprising: introducing a substantially spherical wavefront in a pupil plane of a microscope image-formation optical system, so that an object under observation is visualized on the basis of said introduced substantially spherical wavefront; wherein the substantially spherical wavefront is introduced by a displacement of a focal position of an objective. 19. The method for implementing microscopy according to claim 18, wherein while the wavefront introduced in the pupil plane of said image-formation optical system is substantially symmetrically changed with respect to the pupil plane, two images are taken of the same object under observation, and a subtraction operation is applied between the two images to form a subtraction image. 20. A microscope comprising a light source, an illumination optical system for guiding light from the light source to an object under observation, an image-formation optical system for forming an image-formation plane of an image of the object under observation illuminated by said illumination optical system, an eyepiece optical system or an image pickup optical system for viewing the image formed on the image-formation plane, and a substantially spherical wavefront introduction means for introducing a substantially spherical wavefront in a pupil plane of the image-formation optical system by a displacement of a focal position of an objective.
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