WO2016132563A1 - Dispositif d'observation d'échantillon et procédé d'observation d'échantillon - Google Patents

Dispositif d'observation d'échantillon et procédé d'observation d'échantillon Download PDF

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Publication number
WO2016132563A1
WO2016132563A1 PCT/JP2015/055493 JP2015055493W WO2016132563A1 WO 2016132563 A1 WO2016132563 A1 WO 2016132563A1 JP 2015055493 W JP2015055493 W JP 2015055493W WO 2016132563 A1 WO2016132563 A1 WO 2016132563A1
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WIPO (PCT)
Prior art keywords
objective lens
pupil
light
image
transmission part
Prior art date
Application number
PCT/JP2015/055493
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English (en)
Japanese (ja)
Inventor
鈴木　良政
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オリンパス株式会社
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Publication date
Application filed by オリンパス株式会社 filed Critical オリンパス株式会社
Priority to PCT/JP2015/055493 priority Critical patent/WO2016132563A1/fr
Publication of WO2016132563A1 publication Critical patent/WO2016132563A1/fr

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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens

Definitions

  • ml is the magnification of the projection lens
  • d1 is the outer diameter of the ring-shaped opening
  • d2 is the inner diameter of the ring-shaped opening
  • D is the size (diameter) of the pupil of the objective lens.
  • an image of a ring-shaped opening is formed at the pupil position of the objective lens.
  • mld1 1.05D
  • a part of the image of the ring-shaped opening is located outside the pupil of the objective lens.
  • the image of the ring-shaped opening represents the light that has passed through the sample.
  • the fact that part of the image of the ring-shaped opening is located outside the pupil of the objective lens indicates that part of the light that has passed through the sample does not pass through the objective lens. Therefore, in the apparatus of cited document 1, the sample image becomes dark.
  • R 1 ′ is the maximum length of the length from the optical axis of the illumination optical system to the outermost light in the illumination optical system
  • R ob is the radius of the pupil of the objective lens
  • ⁇ ′ is a value obtained by dividing the focal length of the objective lens by the focal length of the illumination optical system, It is.
  • FIG. 1 is a diagram illustrating a configuration of a sample observation apparatus according to an embodiment.
  • 2A and 2B are diagrams showing the configuration of the opening member, in which FIG. 2A is a view showing the opening member made of an opaque member, and FIG. 2B is a view showing the opening member made of a transparent member.
  • FIGS. 3A and 3B are diagrams showing the relationship between the pupil of the objective lens and the image of the aperture member when no specimen is present, in which FIG. 3A shows the state of light refraction at the specimen position, and FIG. 3B shows the objective lens. It is a figure which shows the relationship between this pupil and the image of an aperture member.
  • the specimen observation apparatus and specimen observation method of the embodiment will be described.
  • the specimen observation apparatus and specimen observation method of each of the following embodiments are used in a bright field observation state.
  • a fluorescence mirror unit including an excitation filter, a dichroic mirror, and an absorption filter is not used.
  • the optical system is arranged so as to include the optical axis, the transmission part is located outside the outer edge of the first light quantity restriction part, the second light quantity restriction part is located outside the transmission part, and the objective lens
  • An image of the inner edge of the transmissive part is formed inside the outer edge of the pupil of the lens, and an image of the outer edge of the transmissive part is formed at a position that coincides with the outer edge of the pupil of the objective lens or inside the outer edge of the pupil of the objective lens.
  • the opening member 5 has a first light quantity limiting unit and a second light quantity limiting unit.
  • Examples of the light quantity limiting unit include a light shielding unit and a light reduction unit.
  • description will be made using the light shielding portion.
  • the light shielding part 5a1 and the light shielding part 5′a1 correspond to the first light quantity limiting part
  • the light shielding part 5a2 and the light shielding part 5′a2 correspond to the second light quantity limiting part.
  • the opening member 5 includes a light shielding part 5a1, a light shielding part 5a2, and a transmission part 5b.
  • the light shielding portions 5a1 and 5a2 are made of an opaque member, for example, a metal plate.
  • the transmission part 5b is a space (hole) formed in the metal plate.
  • three connection parts 5a3 are formed between the light shielding part 5a1 and the light shielding part 5a2. Therefore, the transmission part 5b is divided into three.
  • Each shape of the transmission part 5b is substantially fan-shaped (discrete ring zone shape).
  • the number of the connection parts 5a3 is not limited to three.
  • the light shielding portion 5a1 is disposed so as to include the optical axis of the illumination optical system.
  • a circle (circumference) indicated by reference numeral 9 is the outer edge of the objective lens pupil, and the inside of the circle (circumference) is the pupil of the objective lens.
  • the shape of the image 20 of the transmission part is a ring
  • the shape of the image 21 of the light shielding part is a circle
  • the shape of the image 22 of the light shielding part is a ring
  • the pupil 9 of the objective lens is a circle.
  • the image 20 of the transmission part, the image 21 of the light shielding part, the image 22 of the light shielding part, and the pupil 9 of the objective lens are concentric.
  • the image 20a of the outer edge of the transmission part may be formed so as to coincide with the outer edge of the pupil 9 of the objective lens.
  • the sample exists, as shown in FIG. 4A, the light traveling direction differs between the light incident on the holding member 6 and the light emitted from the sample.
  • the image of the aperture member formed at the pupil position of the objective lens is as shown in FIG. 4B, the circle (circumference) indicated by reference numeral 9 is the outer edge of the objective lens pupil, and the inside of the circle (circumference) is the objective lens pupil.
  • FIG. 4B the circle (circumference) indicated by reference numeral 9 is the outer edge of the objective lens pupil
  • the inside of the circle (circumference) is the objective lens pupil.
  • the shape of the image 20 of the transmission part is a ring
  • the shape of the image 21 of the light shielding part is a circle
  • the shape of the image 22 of the light shielding part is a ring
  • the pupil 9 of the objective lens is a circle.
  • the image 20 of the transmission part, the image 21 of the light shielding part, the image 22 of the light shielding part, and the pupil 9 of the objective lens are not concentric.
  • the center of the image 20 of the transmission part, the center of the image 21 of the light shielding part, and the center of the image 22 of the light shielding part do not coincide with the center of the pupil 9 of the objective lens.
  • the center of the image 20 of the transmission part, the center of the image 21 of the light shielding part, and the center of the image 22 of the light shielding part are shifted leftward in the drawing with respect to the center of the pupil 9 of the objective lens.
  • the image 20a of the outer edge of the transmission part is located inside the outer edge of the pupil 9 of the objective lens. This is because the inclination of the sample surface is small.
  • the image 21 of the light shielding part is located inside the outer edge of the pupil 9 of the objective lens. For this reason, even when a sample is present, if the inclination of the surface of the sample is small, the brightness of the sample image is the same as when the sample is not present.
  • FIG. 5B is a diagram showing a deviation of the image of the aperture member with respect to the pupil of the objective lens. In FIG. 5B, the center of the image of the aperture member is deviated from the center of the pupil of the objective lens.
  • FIG. 7 is a graph showing the relationship between the amount of deviation of the image of the aperture member with respect to the pupil of the objective lens and the amount of light beam passing through the pupil of the objective lens. 1 ⁇ ⁇ ) / R ob It is a graph when the value of is changed.
  • R 1 ′ Is the maximum length of the length from the optical axis of the illumination optical system to the outermost light in the illumination optical system
  • R ob Is the pupil radius of the objective lens
  • ⁇ ′ is a value obtained by dividing the focal length of the objective lens by the focal length of the illumination optical system, It is.
  • the sample observation apparatus of the present embodiment does not require the use of a modulator unlike the modulation contrast method. Therefore, it is not necessary to adjust the position of the opening member with respect to the modulator. As a result, the position adjustment of the opening member is simplified.
  • the objective lens can be a bright field observation objective lens. Therefore, various observation methods (for example, observation methods such as bright field observation, fluorescence observation, and polarization observation) can be easily performed with the same objective lens.
  • the area of the image of the first light quantity limiting unit at the pupil position of the objective lens is 20% or more of the area of the pupil of the objective lens.
  • the size of the loss area does not depend on the size of the area of the entire image of the transmission part. Therefore, when the amount of inclination of the surface of the specimen is the same, the ratio of the loss area to the area of the image of the transmissive portion decreases as the area of the image of the transmissive portion increases. That is, as the area of the image of the transmission part is larger, the contrast in the sample image is lowered.
  • the area of the image of the first light quantity limiting part is smaller than 20% of the area of the pupil of the objective lens, the area of the image of the transmission part becomes too large. Therefore, the contrast in the sample image is lowered.
  • R 0 Is the minimum length from the optical axis of the illumination optical system to the inner edge of the transmission part
  • R 1 is the maximum length of the length from the optical axis of the illumination optical system to the outer edge of the transmission part
  • R ob Is the pupil radius of the objective lens
  • ⁇ is the value obtained by dividing the focal length of the objective lens by the focal length of the condenser lens. It is. If the upper limit value of conditional expression (2) is exceeded, the area of the image of the transmission part becomes too small. Therefore, the brightness of the sample image becomes dark.
  • FIG. 9 is a graph showing the relationship between the amount of deviation of the image of the aperture member with respect to the pupil of the objective lens and the amount of light beam passing through the pupil of the objective lens. 0 ⁇ ⁇ ) / R ob It is a graph when the value of is changed.
  • conditional expression (2) 0.7 ⁇ R ob ⁇ (R 0 ⁇ ⁇ ) ⁇ 0.98 ⁇ (R 1 ⁇ ⁇ ) (2 ') It is preferable to satisfy the following conditional expression (2 ′′) instead of conditional expression (2). 0.7 ⁇ R ob ⁇ (R 0 ⁇ ⁇ ) ⁇ 0.97 ⁇ (R 1 ⁇ ⁇ ) (2 ”) Further, the lower limit value of conditional expression (2) may be 0.9. Further, in the sample observation device of the present embodiment, it is preferable that the entire inner side of the outer edge of the pupil of the objective lens is a transparent region.
  • an image of the transmission part of the aperture member is formed at the pupil position of the objective lens, and the change in the shape of the specimen is detected as a change in light and darkness using the movement of the image of the transmission part. ing.
  • the change in the movement of the image of the transmission part with respect to the change in the shape of the sample has a correct relationship.
  • a modulator is disposed at the pupil position. For this reason, when such an objective lens is used in the sample observation apparatus of the present embodiment, a part of the image of the transmission part is affected by the modulator.
  • the phase difference objective lens used for phase difference observation a phase plate is arranged at the pupil position. Therefore, there is a possibility that the change in the movement of the image of the transmission part with respect to the change in the shape of the specimen does not have a correct relationship. For this reason, in the objective lens used in the sample observation apparatus of the present embodiment, it is preferable that the entire inner side of the outer edge of the pupil of the objective lens is a transparent region. As such an objective lens, for example, there is an objective lens for bright field observation.
  • the light shielding unit 31 has a region where the transmittance changes in a step shape in the circumferential direction and a region where the transmittance is constant. In the region where the transmittance changes in a step-like manner, a region where the transmittance is 100% and a region where the transmittance is 0% are alternately formed along the circumference around the center of the transmitting portion 31.
  • the shape of the light shielding part 32 is a ring.
  • the shape of the transmission part 33 is such that the inside of the circle is cut off by the light shielding part 31. Therefore, at the boundary portion with the light shielding portion 31, similarly to the light shielding portion 31, the transmittance changes in a step shape in the circumferential direction.
  • the transmission part 33 has a region where the transmittance changes in a step shape and a region where the transmittance is constant.
  • the region where the transmittance is constant is formed outside the region where the transmittance changes stepwise, and the transmittance is 100%.
  • the boundary between the region where the transmittance is 100% and the region where the transmittance is 0% is rectangular, but it may be sawtooth or sinusoidal. Further, the transmittance of each region is not limited to the above value.
  • the uneven portion 34 of the region where the transmittance changes in a step shape corresponds to the inner edge of the transmission portion 33.
  • the intersection between the line passing through the center of the opening member 30 and perpendicular to the recess 35 and the recess 35 is the closest position to the optical axis of the illumination optical system. Therefore, R 0 Is the length from the optical axis of the illumination optical system to this intersection.
  • the circumference 36 corresponds to the outer edge of the transmission part 33. At the circumference 36, the distance from each point on the circumference to the optical axis of the illumination optical system is the same. Therefore, R 1 Is a length from the optical axis of the illumination optical system to the circumference 36.
  • the light shielding part 32 is formed so as to include the outer edge of the pupil image 9 'of the objective lens.
  • the transmission part 33 is located inside the outer edge of the pupil image 9 ′ of the objective lens. At this time, the transmission part 33 is formed so as to satisfy at least the conditional expression (1). Therefore, even if the specimen is colorless and transparent, a shadow image with high contrast can be obtained. Further, the direction of occurrence of the shadow is not limited. Further, no flare occurs even when the surface of the specimen is flat. Furthermore, in the opening member 30, the transmittance at the transmission part varies depending on the location. Therefore, according to the observation apparatus of the present embodiment using such an opening member, it is possible to detect a minute shape change (change in inclination) in the specimen as a change in brightness. As shown in FIG.
  • the opening member 40 of Modification 2 includes a light shielding part 41, a light shielding part 42, and a transmission part 43.
  • the light shielding parts 41 and 42 and the transmission part 43 are made of a transparent member, for example, a glass plate.
  • the light shielding portions 41 and 42 are formed, for example, by applying a light shielding paint on a glass plate.
  • the shape of the light shielding part 41 is an equilateral triangle, and the shape of the light shielding part 42 is a ring.
  • the shape of the transmissive part 43 is such that the inside of the circle is cut off by the light shielding part 41.
  • the shape of the light shielding part 41 may be a polygon such as a quadrangle or a pentagon.
  • R 1 Is a length from the optical axis of the illumination optical system to the circumference 45.
  • the light shielding portion 42 is formed so as to include the outer edge of the pupil image 9 'of the objective lens. Therefore, the transmission part 43 is located inside the outer edge of the pupil image 9 ′ of the objective lens. At this time, the transmission part 43 is formed so as to satisfy at least the conditional expression (1). Therefore, even if the specimen is colorless and transparent, a shadow image with high contrast can be obtained. Moreover, when the inclination of the surface of the specimen is small, the generation direction of the shadow is not limited. Further, no flare occurs even when the surface of the specimen is flat. As shown in FIG.
  • R 0 Is the length from the optical axis of the illumination optical system to the circumference 54.
  • R 1 Is a length from the optical axis of the illumination optical system to the circumference 55.
  • the light shielding part 52 is formed so that the outer edge of the pupil image 9 ′ of the objective lens coincides with the inner edge of the light shielding part 52. Therefore, the transmission part 53 is located inside the outer edge of the pupil image 9 ′ of the objective lens. At this time, the transmission part 53 is formed so as to satisfy at least the conditional expression (1).
  • the light shielding portions 61 and 62 are formed, for example, by applying a light shielding paint on a glass plate.
  • the shape of the light shielding part 61 is an ellipse
  • the shape of the light shielding part 62 is a ring.
  • the shape of the transmission part 63 is such that the inside of the circle is cut off by the light shielding part 61 (ellipse).
  • the elliptical outer peripheral portion 64 corresponds to the inner edge of the transmission portion 63.
  • the intersection between the short axis of the ellipse and the outer peripheral portion 64 is the closest position to the optical axis of the illumination optical system. Therefore, R 0 Is the length from the optical axis of the illumination optical system to this intersection.
  • FIG. 11 shows an opening member of Modification 5.
  • 11A and 11B are diagrams showing an opening member according to the modified example 5, where FIG. 11A is an overall view, FIG. 11B is an enlarged view of a transmission portion, and a diagram when the diameters of the minute openings are the same; ) Is an enlarged view of the transmission part, and is a view when the diameter of the minute opening is changed.
  • the opening member 70 includes a light shielding part 71, a light shielding part 72, and a transmission part 73.
  • the light shielding part 71 and the light shielding part 72 are opaque members, for example, metal plates.
  • the transmission part 73 is formed between the light shielding part 71 and the light shielding part 72.
  • the transmission part 73 includes a plurality of minute openings 74a and 74b.
  • the minute openings 74a and 74b are voids (holes) formed in the metal plate.
  • the minute openings 74a and 74b can be formed by irradiating a metal plate with a laser.
  • the diameters of the minute openings 74a and 74b are the same. In this way, the transmission part 73 can be easily formed. Further, by changing the diameter and density of the minute openings 74a and 74b, it is possible to obtain the transmission part 73 having various transmittances.
  • FIG. 11 (b) the transmission part 73 includes a plurality of minute openings 74a and 74b.
  • the minute openings 74a and 74b are voids (holes) formed in the metal plate.
  • the minute openings 74a and 74b can be formed by i
  • the opening member 80 includes a light shielding part 81, a light shielding part 82, and a transmission part 83.
  • the light shielding parts 81 and 82 and the transmission part 83 are made of a transparent member, for example, a glass plate.
  • the light shielding portions 81 and 82 are formed, for example, by applying a light shielding paint on a glass plate.
  • the transmittance of the light shielding part 81 and the transmittance of the light shielding part 82 are both 0%.
  • the transmittance changes continuously. Therefore, for example, a reflection film (absorption film) whose transmittance continuously changes is formed in the transmission portion 83.
  • the opening member shown in FIG. 13 (a) is the same as the opening member shown in FIG. 2 (b).
  • the opening member 5 ' includes a light shielding part 5'a1, a light shielding part 5'a2, and a transmission part 5'b.
  • the transmittance of the light shielding portion 5'a1 and the transmittance of the light shielding portion 5'a2 are both 0%.
  • the transmittance of the transmission part 5'b is the position P. in From position P out Up to 100%.
  • the distance from the center of the opening member to the position where the transmittance is 50% is R 0 become.
  • the position where the transmittance is 50% is the position P in And position P out It is in the middle.
  • the position P from the center of the opening member out Distance to R 1 become.
  • the transmittance of the light shielding portion is 0% at any position, and the transmittance of the transmissive portion is any. The position is also 100%. Therefore, in these opening members, the boundary between the light shielding part and the transmission part is clear.
  • the transmittance of the light shielding portion is 0% at any position.
  • the transmittance of the transmissive part is in the range of 50% to 100% depending on the position, but the minimum transmittance of the transmissive part is 50%.
  • the boundary between the light shielding part and the transmission part is clear even in the opening member of the modification 6.
  • the boundary between the two regions becomes clear.
  • the boundary between the two regions becomes ambiguous. For example, even if the transmittance of the light shielding portion is 0%, the boundary between the light shielding portion and the transmission portion becomes ambiguous when the transmittance of the transmission portion is 1% or when the transmittance is continuously increased from 1%. .
  • the light-shielding part is a light-reducing part.
  • a region satisfying the following conditional expression (4) is a transmission part.
  • the first light quantity limiting unit corresponds to the light reducing unit 95 whose transmittance continuously changes
  • the light blocking unit 96 corresponds to the second light quantity limiting unit.
  • the region between the light reducing unit 95 and the light shielding unit 96 corresponds to the transmission unit.
  • the opening member 94 has a light reducing part 95, a light shielding part 96, and a transmission part 97.
  • the light reduction unit 95, the light shielding unit 96, and the transmission unit 97 are made of a transparent member, for example, a glass plate.
  • the light shielding part 96 is formed, for example, by applying a light shielding paint on a glass plate. In the light reduction part 95 and the transmission part 97, the transmittance changes continuously.
  • FIG. 15 is a diagram illustrating an aperture member according to Modification 8 and is a diagram illustrating an aperture member having a characteristic of shielding light in a predetermined wavelength range. As shown in FIG.
  • the observation with the shadow image and the observation with the bright field image can be performed by changing the wavelength of the illumination light.
  • the light source 1 and the lenses 2 and 3 can be omitted in FIG.
  • FIG. 16 shows an opening member of Modification 9.
  • FIG. 16 is a view showing the opening member of the modification 9, and shows a case where a plurality of light sources are arranged in the transmission part.
  • the opening member 110 includes a light shielding part 111, a light shielding part 112, and a transmission part 113.
  • this circumference 114 is used as the inner edge of the transmission part 113
  • the circumference of a circle circumscribing the outer periphery of each LED 113 a may be used as the inner edge of the transmission part 113.
  • the circumference connecting the centers of the respective LEDs 113 a may be the inner edge of the transmission portion 113.
  • the circumference 115 is the circumference of a circle circumscribing the outer periphery of each LED 113b.
  • this circumference 115 is used as the outer edge of the transmission part 113
  • the circumference of the circle inscribed in the outer peripheral part of each LED 113b may be used as the outer edge of the transmission part 113.
  • a shaded sample image can be obtained for a colorless and transparent sample.
  • a shadow image is formed by the annular illumination light.
  • the shaded image is not only shaded but also added with colors and shades according to staining.
  • a shadow image is not formed, but a sample image (hereinafter referred to as a “light / dark image”) having a color and lightness corresponding to the staining is formed.
  • the sample image is an overlap of the shadow image and the gray image.
  • axial light beam a light beam that exits from the center of the sample and reaches the center of the sample image
  • off-axis light beam a light beam that exits from the center of the sample and reaches the center of the sample image
  • axial light beam a light beam that exits from the center of the sample and reaches the center of the sample image
  • off-axis light beam a light beam that exits from the center of the sample and reaches the center of the sample image
  • off-axis light beam a difference in the size of the light beam reaching the light beam.
  • the light source is a monochromatic light source or the illumination optical system has a wavelength selection unit. Since the wavelength range of illumination light can be narrowed, the occurrence of chromatic aberration of the pupil can be suppressed. Therefore, the contrast of the sample image can be improved. Moreover, in the sample observation device of the present embodiment, it is preferable that the shape of the transmission part can be changed. In this way, it is possible to select an opening member having the optimal shape of the transmission part according to the specimen. In a specimen having a small slope on the surface of the specimen, for example, a Ptk2 cell, an aperture member in which the entire transmission part is brought close to the outer edge of the pupil of the objective lens is used as the aperture member.
  • FIG. 17 is a diagram showing a configuration of another specimen observation apparatus of the present embodiment.
  • the specimen observation apparatus 300 includes an opening member 5 and an opening member 301.
  • the opening member 5 and the opening member 301 are held by the moving mechanism 302.
  • an image of the inner edge of the transmission part of the first aperture member is formed inside the outer edge of the pupil of the second objective lens, and the position coincides with the outer edge of the pupil of the second objective lens, or the objective. It is preferable that an image of the outer edge of the transmission part of the second opening member be formed inside the outer edge of the pupil of the lens.
  • the second opening member may have a transmission part at the center.
  • the second aperture member has a transmission portion having a diameter smaller than the diameter of the transmission portion of the first aperture member. The structure provided may be sufficient.
  • the specimen observation aperture member including the objective lens in the present embodiment further includes a third aperture member, and the second aperture member has a transmission portion for phase difference observation, and the third aperture.
  • the member may have a transmission part for bright field observation.
  • the objective lens can be irradiated with illumination light according to various observation methods.
  • the opening member may include a transmission region variable portion that makes the transmission region of the transmission portion variable.
  • the transmissive region variable unit may be constituted by a liquid crystal shutter, for example.
  • an opening member 303 is provided at the position of the pupil 9 of the objective lens.
  • the position of the pupil 9 of the objective lens and the position of the opening member 303 are drawn apart for ease of viewing.
  • the wavelength selection element 304 may be detachable between the light path of the illumination optical system, for example, between the light source 1 and the lens 2.
  • the light source 1 is white light
  • light having a wide wavelength range is emitted from the light source 1. Therefore, by inserting the wavelength selection element 304 into the optical path, light having a narrower wavelength range than white light can be extracted as illumination light.
  • the light source 1 may be a monochromatic light source 1 '.
  • the sample observation device 300 may include an image sensor 305 and an image processing device 306.
  • the image sensor 305 is, for example, a CCD or a CMOS.
  • the image sensor 305 is disposed at the image position 11.
  • the sample image captured by the image sensor 305 is sent to the image processing device 306.
  • the image processing apparatus 306 can perform processing such as contrast enhancement, noise removal, and color conversion.
  • the shape of the opening member is preferably an axisymmetric shape. By doing so, the generation direction of the shadow is not limited. Axisymmetric shapes include, for example, circles and polygons.
  • a light ray that passes through the innermost side of the transmission part (hereinafter referred to as “light ray L in ”) Reaches the inside of the objective lens pupil.
  • the aperture member deviates from the pupil position of the condenser lens
  • the light beam L in Is directed from the inside to the outside of the pupil of the objective lens. That is, the image of the transmission part is shifted from the pupil of the objective lens.
  • changing the objective lens changes the observation range.
  • the observation range changes, the light beam L in The angle with respect to the optical axis also changes. When this angle changes, the ray L that reaches the pupil of the objective lens in The position of changes.
  • FIG. 18 is a diagram illustrating a configuration of a sample observation device including an image processing device.
  • the specimen observation device 400 includes a main body 410, an illumination optical system 420, an observation optical system 430, an imaging device 440, and an image processing device 450.
  • the main body 410 includes a light source 411, a stage 412, and a revolver 413.
  • the illumination optical system 420 includes various optical filters 421, a field stop 422, a mirror 423, a lens 424, an aperture member 425, and a condenser lens 426.
  • the observation optical system 430 includes an objective lens 431, an imaging lens 433, and an eyepiece lens 434.
  • an objective lens pupil 432 is located in the vicinity of the objective lens 431.
  • a light source 411 is connected to the main body 410. The illumination light emitted from the light source 411 enters the illumination optical system 420 and reaches the condenser lens 426.
  • the aperture member 425 is disposed at the pupil position of the condenser lens 426.
  • the opening member 5 shown in FIG. 2A is used as the opening member 425.
  • a stage 412 is disposed above the condenser lens 426.
  • a specimen 460 is placed on the stage 412. Further, a revolver 413 is positioned above the stage 412, and the objective lens 431 is held on the revolver 413.
  • the illumination light emitted from the condenser lens 426 is applied to the sample 460.
  • Light from the sample 460 enters the objective lens 431.
  • the pupil 432 of the objective lens and the aperture member 425 have a conjugate relationship. Therefore, an image of the aperture member 425 is formed at the position of the pupil 432 of the objective lens.
  • an image of the inner edge of the transmission part of the aperture member 425 is formed inside the outer edge of the pupil of the objective lens 431, or the position that matches the outer edge of the pupil of the objective lens 431, or the objective lens
  • An image of the outer edge of the transmission part of the opening member 425 is formed inside the outer edge of the pupil 431.
  • the amount of imaging light emitted from the objective lens 431 changes according to the change in shape (change in inclination) in the sample 460.
  • a change in the shape of the sample can be detected as a change in brightness.
  • a shaded sample image can be obtained even if the sample is colorless and transparent.
  • the imaging light emitted from the objective lens 431 is condensed by the imaging lens 433, and an image of the sample 460 is formed at the condensing position.
  • a prism is disposed after the imaging lens 433. A part of the imaging light is reflected by the prism toward the eyepiece 434 side. As a result, an optical image 435 of the specimen is formed in the vicinity of the eyepiece lens 434.
  • the imaging device 440 includes an imaging element 441.
  • the imaging lens 433 forms an optical image of the sample 460 on the image sensor 441, and thereby the optical image of the sample 460 is captured. In this way, a sample image with a shadow is obtained.
  • an optical system may be disposed between the imaging lens 433 and the imaging element 441. In this case, an optical image of the specimen 460 is formed on the image sensor 441 by the imaging lens 433 and this optical system.
  • a camera controller 442 and a video board 443 are connected to the imaging device 440.
  • the camera controller 442 and the video board 443 are both connected to the image processing device 450. Imaging control is performed by the camera controller 442.
  • the camera controller 442 is controlled by the image processing apparatus 450.
  • the camera controller 442 may be controlled by another device such as a computer.
  • the image signal output from the imaging device 440 is input to the image processing device 450 via the video board 443.
  • various electrical processes are performed.
  • the processing result is displayed on the display device 451. Since the specimen observation device includes an image processing device, various image processing can be performed. Next, an example of image processing will be described.
  • FIG. 19 is an electronic image of a specimen obtained by the specimen observation apparatus of the present embodiment, and is an image after image processing.

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  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

L'invention concerne un dispositif d'observation d'échantillon qui comprend un système optique d'éclairage et un système optique d'observation, le système optique d'éclairage comprenant une source de lumière 1, une lentille de condenseur 4, et un élément d'ouverture 5, et le système optique d'observation comprenant un objectif 8 et une lentille d'imagerie 10. L'élément d'ouverture 5 comprend une première partie de restriction de quantité de lumière 5a1, une seconde partie de restriction de quantité de lumière 5a2, et une partie de transmission 5b. L'élément d'ouverture 5 est agencé de telle sorte que la première partie de restriction de quantité de lumière 5a1 comprend l'axe optique du système optique d'éclairage. La partie de transmission 5b est située à l'extérieur de la périphérie externe de la première partie de restriction de quantité de lumière 5a1. La seconde partie de restriction de quantité de lumière 5a2 est située à l'extérieur de la partie de transmission 5b. Une image 20b sur la périphérie interne de la partie de transmission est formée à l'intérieur de la périphérie externe de la pupille 9 de l'objectif. Une image 20a sur la périphérie externe de la partie de transmission est formée à une position correspondant à la périphérie externe de la pupille 9 de l'objectif ou à l'intérieur de la périphérie externe de la pupille 9 de l'objectif. Le dispositif d'observation d'échantillon satisfait l'expression conditionnelle (1) suivante: 0.9 ≤ (R1 ×β) / Rob ≤ 1 (1)
PCT/JP2015/055493 2015-02-19 2015-02-19 Dispositif d'observation d'échantillon et procédé d'observation d'échantillon WO2016132563A1 (fr)

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JPH09218357A (ja) * 1996-02-09 1997-08-19 Olympus Optical Co Ltd 偏斜・暗視野検鏡装置
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JP2019144294A (ja) * 2018-02-16 2019-08-29 オリンパス株式会社 画像処理装置、顕微鏡システム、画像処理方法および画像処理プログラム
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