WO2016052743A1 - 光軸方向走査型顕微鏡装置 - Google Patents
光軸方向走査型顕微鏡装置 Download PDFInfo
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- WO2016052743A1 WO2016052743A1 PCT/JP2015/078100 JP2015078100W WO2016052743A1 WO 2016052743 A1 WO2016052743 A1 WO 2016052743A1 JP 2015078100 W JP2015078100 W JP 2015078100W WO 2016052743 A1 WO2016052743 A1 WO 2016052743A1
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- phase modulation
- optical axis
- modulation element
- axis direction
- light
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0052—Optical details of the image generation
- G02B21/0072—Optical details of the image generation details concerning resolution or correction, including general design of CSOM objectives
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0032—Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0036—Scanning details, e.g. scanning stages
- G02B21/0044—Scanning details, e.g. scanning stages moving apertures, e.g. Nipkow disks, rotating lens arrays
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0052—Optical details of the image generation
- G02B21/006—Optical details of the image generation focusing arrangements; selection of the plane to be imaged
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
- G02B27/0025—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00 for optical correction, e.g. distorsion, aberration
- G02B27/0068—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00 for optical correction, e.g. distorsion, aberration having means for controlling the degree of correction, e.g. using phase modulators, movable elements
Definitions
- the present invention relates to an optical axis direction scanning type microscope apparatus that optically scans in the optical axis direction, for example.
- Patent Document 1 and Patent Document 2 Conventionally, there is known a method of moving a focal point position of an object in a direction along the optical axis (Z-axis direction) by adjusting an optical path length at an intermediate image position (for example, Patent Document 1 and Patent Document 2). reference.).
- Patent Document 1 and Patent Document 2 since the plane mirror is disposed on the intermediate image plane, scratches and foreign matter on the surface of the plane mirror overlap the acquired final image and the illumination light projected on the object. There is an inconvenience. Further, since the method of Patent Document 2 is an optical system in which an enlarged intermediate image is interposed between the optical path length adjusting means and the object, the optical magnification is such that the vertical magnification is equal to the square of the horizontal magnification. According to the basic principle, the enlarged intermediate image greatly moves in the direction of the optical axis even if the focal point is slightly moved in the direction along the optical axis.
- the present invention has been made in view of the above-described circumstances, and prevents the intermediate image from being damaged by the optical element even if the intermediate image is formed at a position coinciding with the optical element. It is an object of the present invention to provide an optical axis direction scanning microscope apparatus that can acquire a clear and clear final image.
- One aspect of the present invention includes a plurality of imaging lenses that form a final image and at least one intermediate image, and are disposed closer to the object side than any of the intermediate images formed by the imaging lens.
- a first phase modulation element that imparts spatial disturbance to the wavefront of the light, and at least one intermediate image interposed between the first phase modulation element and the first phase modulation element
- An imaging optical system including a second phase modulation element that cancels the spatial disturbance applied to the wavefront of the light from the object, and the wavefront from the object passes through the imaging optical system.
- An optical axis direction scanning microscope apparatus comprising a scanning system for scanning an image to be imaged in the optical axis direction.
- clear image is a state in which no spatial disturbance is applied to the wavefront of light emitted from an object, or in a state where the applied disturbance is canceled and eliminated.
- a “blurred image” is an image generated through an imaging lens in a state where spatial disturbance is added to the wavefront of light emitted from an object. It means that the surface of the optical element arranged in the vicinity of the image, a scratch, a foreign object, a defect, or the like existing on the inside has a characteristic that is not substantially formed as a final image.
- the “blurred image” (or “blurred image”) formed in this way is different from simply an out-of-focus image, and is supposed to be originally imaged (ie, the spatial disturbance of the wavefront).
- the image contrast does not have a clear peak over a wide range in the direction of the optical axis, including the image at the position where the image is formed when no is applied.
- the spatial frequency band of the “unclear image” is always narrower than the spatial frequency band of the “clear image”.
- the “clear image” and the “unclear image” (or “blurred image”) in this specification are based on the above concept, and the movement of the intermediate image in the Z-axis direction is defined as the present invention. It means to move in the state of a blurred intermediate image.
- the Z-axis scanning is not limited to the movement of light in the Z-axis direction, and may be accompanied by light movement on XY as will be described later.
- the Z-axis direction means a direction along the optical axis.
- the light incident from the object side of the imaging lens is focused by the imaging lens to form a final image.
- a spatial disturbance is imparted to the wavefront of the light, and the formed intermediate image is blurred.
- the light that forms the intermediate image passes through the second phase modulation element, thereby canceling the spatial disturbance of the wavefront imparted by the first phase modulation element.
- the final image formed after the second phase modulation element becomes clear.
- the first phase modulation element and the second phase modulation element may be disposed at an optically conjugate position.
- the first phase modulation element and the second phase modulation element may be disposed in the vicinity of the pupil position of the imaging lens.
- the first phase modulation element and the second phase modulation element can be reduced in size by being arranged in the vicinity of the pupil position where the luminous flux does not vary.
- the imaging position of the final image can be easily changed in the optical axis direction by changing the optical path length between the two imaging lenses by the operation of the optical path length varying means.
- the optical path length varying means is arranged perpendicular to the optical axis and reflects the light that folds back the light forming the intermediate image, and the actuator that moves the flat mirror in the optical axis direction;
- a beam splitter that branches light reflected by the plane mirror in two directions may be provided.
- the light from the object side collected by the imaging lens on the object side is reflected by the plane mirror and folded, and then branched by the beam splitter and incident on the imaging lens on the image side.
- the actuator to move the plane mirror in the optical axis direction, the optical path length between the two imaging lenses can be easily changed, and the imaging position of the final image can be easily changed in the optical axis direction. Can be changed.
- variable space that changes the final image position in the optical axis direction by changing the spatial phase modulation applied to the wavefront of the light in the vicinity of the pupil position of any one of the imaging lenses.
- a phase modulation element may be provided. By doing so, spatial phase modulation that changes the final image position in the optical axis direction by the variable spatial phase modulation element can be applied to the wavefront of the light. By adjusting the phase modulation to be applied, the imaging position of the final image can be easily changed in the optical axis direction.
- At least one function of the first phase modulation element or the second phase modulation element may be performed by the variable spatial phase modulation element.
- the spatial phase modulation that changes the final image position in the optical axis direction and the phase modulation that blurs the intermediate image or the blur of the intermediate image are canceled by the variable spatial phase modulation element. Both phase modulation can be handled. Thereby, a simple imaging optical system can be configured with fewer components.
- the first phase modulation element and the second phase modulation element may impart phase modulation that changes in a one-dimensional direction orthogonal to the optical axis to the wavefront of light.
- phase modulation that changes in a one-dimensional direction orthogonal to the optical axis to the wavefront of the light by the first phase modulation element.
- an optical element is arranged at the intermediate image position and there are scratches, foreign matter, defects, etc. on the surface or inside of the optical element, the scratches, foreign matter, defects, etc. of these optical elements overlap the intermediate image.
- a phase modulation that cancels the phase modulation changed in the one-dimensional direction is applied to the wavefront of the light by the second phase modulation element, and a clear final image that is not blurred can be formed.
- the first phase modulation element and the second phase modulation element may impart phase modulation that changes in a two-dimensional direction orthogonal to the optical axis to the wavefront of the light beam.
- phase modulation that changes in a two-dimensional direction orthogonal to the optical axis to the wavefront of the light by the first phase modulation element.
- a phase modulation that cancels the phase modulation changed in the two-dimensional direction is applied to the wavefront of the light by the second phase modulation element, so that a clearer final image can be formed.
- first phase modulation element and the second phase modulation element may be transmission elements that give phase modulation to the wavefront when transmitting light.
- first phase modulation element and the second phase modulation element may be reflective elements that give phase modulation to a wavefront when light is reflected.
- the first phase modulation element and the second phase modulation element may have complementary shapes.
- the first phase modulation element that imparts to the wavefront spatial disturbance that blurs the intermediate image, and the second that applies phase modulation that cancels the spatial disturbance applied to the wavefront can be configured easily.
- the first phase modulation element and the second phase modulation element may impart phase modulation to the wavefront by a refractive index distribution of a transparent material.
- the above aspect may include a light source that is disposed on the object side of the imaging optical system and that generates illumination light incident on the imaging optical system.
- the illumination light emitted from the light source arranged on the object side is incident on the imaging optical system, so that the illumination object arranged on the final image side can be irradiated with the illumination light.
- the intermediate image formed by the imaging optical system is blurred by the first phase modulation element, some optical element is disposed at the intermediate image position, and the surface or the inside of the optical element is scratched. Even if foreign matter or defect exists, it is possible to prevent the occurrence of inconvenience that scratches, foreign matter or defect of these optical elements overlap with the intermediate image and are finally formed as a part of the final image. .
- the photodetector which is arrange
- the image forming optical system detects a clear final image formed by preventing an image such as a scratch, a foreign object, or a defect from overlapping the intermediate image on the surface or inside of the optical element. Can be detected.
- the photodetector may be an image sensor that is disposed at a final image position of the imaging optical system and captures the final image.
- the light source is disposed on the object side of the imaging optical system and generates illumination light to be incident on the imaging optical system; and the observation target is disposed on the final image side of the imaging optical system. You may provide the photodetector which detects the light emitted from the thing.
- the light from the light source is collected by the imaging optical system and applied to the observation object, and the light generated in the observation object is detected by the photodetector arranged on the final image side.
- a clear final image formed by preventing an image such as a scratch, a foreign object or a defect from overlapping the intermediate image on the surface or inside of the intermediate optical element can be detected by the photodetector.
- a Nipkow disc type confocal optical system disposed between the light source and the photodetector and the imaging optical system may be provided. In this way, a clear image of the observation object can be acquired at high speed by causing the observation object to scan with multiple spot lights.
- the light source may be a laser light source
- the photodetector may include a confocal pinhole and a photoelectric conversion element.
- the photodetector which detects the light emitted from the observation target object illuminated by the said light source may be provided, and a pulse laser light source may be sufficient as the said light source.
- a pulse laser light source may be sufficient as the said light source.
- the first phase modulation element and the second phase modulation element may be a combination of cylindrical lenses disposed at optically non-conjugated positions. That is, by arranging a cylindrical lens having an appropriate power at an appropriate location, even if the first phase modulation element and the second phase modulation element are optically non-conjugated, the first phase modulation element causes It is possible to form an image without causing astigmatism by canceling the disturbance of the wave front of the light by the second phase modulation element. As a result, even in an optical system in which the first phase modulation element and the second phase modulation element cannot be optically conjugate with each other due to spatial restrictions, the intermediate image is blurred. To prevent the occurrence of inconvenience that scratches, foreign matters or defects existing on the surface or inside of the optical element disposed at the intermediate image position overlap with the intermediate image and are finally formed as a part of the final image. Can do.
- At least one of the first phase modulation element and the second phase modulation element may be disposed in the vicinity of the pupil position of the imaging lens.
- optical path length varying means capable of changing the optical path length between the two imaging lenses arranged at positions sandwiching any one of the intermediate images.
- the optical path length varying means is a plane mirror that is arranged orthogonal to the optical axis and reflects the light that forms the intermediate image so as to be folded back, the actuator that moves the plane mirror in the optical axis direction, and the plane mirror
- the beam splitter which branches the light reflected by 2 in two directions may be provided.
- variable spatial phase modulation that changes the final image position in the optical axis direction by changing the spatial phase modulation applied to the wavefront of light near the pupil position of any of the imaging lenses
- An element may be provided.
- at least one function of the first phase modulation element or the second phase modulation element may be carried by the variable spatial phase modulation element.
- first phase modulation element and the second phase modulation element may be transmission elements that impart phase modulation to the wavefront when transmitting light.
- first phase modulation element and the second phase modulation element may be reflective elements that give phase modulation to the wavefront when light is reflected.
- first phase modulation element and the second phase modulation element may have complementary shapes.
- first phase modulation element and the second phase modulation element may impart phase modulation to the wavefront by a refractive index distribution of a transparent material.
- the above aspect may further include a light source for generating illumination light that is disposed on the object side of the imaging optical system and is incident on the imaging optical system.
- the optical system may further include a photodetector that is disposed on the final image side of the imaging optical system and detects light emitted from the observation object.
- the photodetector may be an image sensor that is disposed at a final image position of the imaging optical system and captures the final image.
- the light source is disposed on the object side of the imaging optical system and generates illumination light to be incident on the imaging optical system, and is disposed on the final image side of the imaging optical system. You may further provide the photodetector which detects the emitted light.
- a Nipkow disc type confocal optical system disposed between the light source and the photodetector and the imaging optical system may be provided.
- the light source may be a laser light source
- the photodetector may include a confocal pinhole and a photoelectric conversion element.
- the photodetector which detects the light emitted from the observation target object illuminated by the said light source may be provided, and the said light source may be a pulse laser light source.
- an optical scanner is provided, and the optical scanner is disposed at an optically conjugate position with respect to the first phase modulation element, the second phase modulation element, and the pupil of the imaging lens. It is good to be.
- the present invention even if the intermediate image is formed at a position that coincides with the optical element, it is possible to prevent a scratch, a foreign object, a defect, or the like of the optical element from overlapping the intermediate image and obtain a clear final image. There is an effect that can be done.
- the intermediate image is moved by focusing or the like in a magnifying optical system such as a microscope, even if the intermediate image moved in the Z-axis direction overlaps with the lens located before and after the intermediate image,
- the present invention solves a problem that could not be solved for many years in the optical axis direction scanning type microscope apparatus because no noise image is generated in which scratches on the surface, foreign matter or defects in the lens are reflected in the final image. There is a special effect that can be done.
- FIG. 6 is an enlarged view showing a position from a pupil position on the object side to a wavefront recovery element.
- FIG. 6 is an enlarged view showing a position from a pupil position on the object side to a wavefront recovery element.
- FIG. 6 is an enlarged view showing a position from a pupil position on the object side to a wavefront recovery element.
- FIG. 6 is an enlarged view showing a position from a pupil position on the object side to a wavefront recovery element.
- FIG. 6 is an enlarged view showing a position from a pupil position on the object side to a wavefront recovery element.
- FIG. 6 is an enlarged view showing a position from a pupil position on the object side to a wavefront recovery element.
- FIG. 6 is an enlarged view showing a position from a pupil position on the object side to a wavefront recovery element.
- FIG. 6 is an enlarged view showing a position from a pupil position on the object side to a wavefront recovery element.
- FIG. 6 is an
- FIG. 31A shows an example of the lens arrangement
- FIG. 40 is a transverse cross-sectional view showing a passage position of a light beam by a scanning operation in the wavefront recovery element of FIG. 39.
- FIG. 40 is a transverse cross-sectional view showing a light beam passage position by a scanning operation at a pupil position of the objective lens of FIG. 39; It is an enlarged schematic diagram which shows a part of illuminating device which concerns on one Example of this invention.
- the imaging optical system 1 used in the microscope apparatus (optical axis direction scanning microscope apparatus) of the present invention will be described below with reference to the drawings.
- the imaging optical system 1 includes a pair of imaging lenses 2 and 3 arranged at intervals, and an intermediate between these imaging lenses 2 and 3. a field lens 4 arranged on the imaging plane, the pupil position PP O vicinity disposed wavefront confusion element (first phase modulation element) 5 of the imaging lens 2 on the object O side, the imaging of the image I side lens 3 of the pupil position PP I vicinity disposed wavefront recovery device and a (second phase modulation element) 6.
- reference numeral 7 denotes an aperture stop.
- the wavefront confusion element 5 imparts disturbance to the wavefront when transmitting the light emitted from the object O and collected by the imaging lens 2 on the object O side. By imparting disturbance to the wavefront by the wavefront confusion element 5, the intermediate image formed on the field lens 4 is blurred.
- the wavefront recovery element 6 imparts phase modulation to the light so as to cancel the disturbance of the wavefront imparted by the wavefront confusion element 5 when transmitting the light collected by the field lens 4. .
- the wavefront recovery element 6 has a phase characteristic opposite to that of the wavefront confusion element 5, and forms a clear final image I by canceling the disturbance of the wavefront.
- the imaging optical system 1 has a telecentric arrangement with respect to the object O side and the image I side. Further, the wavefront confusion element 5 is arranged spaced a distance a F from the field lens 4 on the object O side, the wavefront recovery device 6 is arranged spaced a distance b F to the image I side from the field lens 4 Yes.
- symbol f O is the focal length of the imaging lens 2
- symbol f I is the focal length of the imaging lens 3
- symbols F O and F O ′ are focal positions of the imaging lens 2
- 'I is the focal position of the imaging lens 3
- symbols II 0 , II A , and II B are intermediate images.
- the wavefront confusion element 5 need not be necessarily disposed at the pupil position PP O vicinity of the imaging lens 2, the wave front recovery device 6 also necessarily have to be arranged near the pupil position PP I of the imaging lens 3 There is no. However, the wavefront confusion element 5 and the wavefront recovery element 6 need to be arranged in a positional relationship conjugated with each other as shown in Expression (1) with respect to the image formation by the field lens 4.
- f F 1 / a F + 1 / b F (1)
- f F is the focal length of the field lens 4.
- Figure 3 is a diagram showing in detail from the pupil position PP O of the object O side of Fig. 2 to the wavefront recovery device 6.
- ⁇ L is a phase advance amount based on a light beam transmitted through a specific position (that is, a light beam height), which is given by the light passing through the optical element.
- ⁇ L O (x O ) and ⁇ L I (x I ) satisfy the following expression (2).
- ⁇ F is a lateral magnification in the conjugate relationship between the wavefront confusion element 5 and the wavefront recovery element 6 by the field lens 4 and is represented by the following expression (3).
- ⁇ F ⁇ b F / a F (3)
- one light beam R enters such an imaging optical system 1 and passes through the position x O on the wavefront confusion element 5, it undergoes phase modulation of ⁇ L O (x O ), and is refracted, diffracted and scattered.
- a confusion ray Rc due to the above is generated.
- the wavefront confusion element 5 and the wavefront recovery element 6 are in a conjugate positional relationship and have the characteristic of equation (2), the light beam that has undergone phase modulation via one position on the wavefront confusion element 5 is It always passes through a specific position of the wavefront recovery element 6 that has a one-to-one correspondence with the position and applies phase modulation that cancels the phase modulation received from the wavefront confusion element 5.
- the optical system shown in FIGS. 2 and 3 acts on the light ray R as described above regardless of the incident position x O and the incident angle in the wavefront confusion element 5. That is, the intermediate image II can be made unclear and the final image I can be clearly formed with respect to all the light rays R.
- FIG. 4 shows a conventional imaging optical system.
- this imaging optical system the light condensed by the imaging lens 2 on the object O side forms a clear intermediate image II in the field lens 4 arranged on the intermediate imaging surface, and then the image I side. It is condensed by the imaging lens 3 to form a clear final image I.
- the imaging optical system 1 according to the present embodiment, the intermediate image II blurred by the wavefront confusion element 5 is formed on the intermediate imaging surface arranged at a position coincident with the field lens 4.
- the foreign object image superimposed on the intermediate image II is blurred by the same phase modulation when the wavefront recovery element 6 undergoes phase modulation to sharpen the blurred intermediate image II. Therefore, it is possible to prevent the image of the foreign matter on the intermediate image plane from overlapping the clear final image I.
- the two imaging lenses 2 and 3 are described as being telecentric.
- the present invention is not limited to this, and the same effect is obtained even in a non-telecentric system.
- the phase advance amount function is a one-dimensional function, it can be similarly operated as a two-dimensional function instead.
- the space between the imaging lens 2, the wavefront confusion element 5, and the field lens 4 and the space between the field lens 4, the wavefront recovery element 6, and the imaging lens 3 are not necessarily required. May be optically bonded.
- each lens constituting the imaging optical system 1 that is, each of the imaging lenses 2 and 3 and the field lens 4 is configured to clearly share the functions of imaging and pupil relay.
- US Pat. No. 5,637 a configuration in which one lens has both functions of image formation and pupil relay is also used. Even in such a case, if the above condition is satisfied, the wavefront confusion element 5 imparts a disturbance to the wavefront to blur the intermediate image II, and the wavefront recovery element 6 cancels the wavefront disturbance and obtains the final image I. It can be sharpened.
- the observation apparatus 10 includes a light source 11 that generates non-coherent illumination light, an illumination optical system 12 that irradiates the observation object A with illumination light from the light source 11, and An imaging optical system 13 that condenses the light from the observation object A, and an imaging element (photodetector) 14 that captures the light collected by the imaging optical system 13 and obtains an image are provided. Yes.
- the illumination optical system 12 includes condenser lenses 15a and 15b that collect the illumination light from the light source 11, and an objective lens 16 that irradiates the observation object A with the illumination light collected by the condenser lenses 15a and 15b. It has.
- the illumination optical system 12 is so-called Koehler illumination, and the condenser lenses 15a and 15b are arranged so that the light emitting surface of the light source 11 and the pupil surface of the objective lens 16 are conjugate with each other.
- the imaging optical system 13 includes the objective lens (imaging lens) 16 that collects the observation light (for example, reflected light) emitted from the observation object A arranged on the object side, and the objective lens 16 collects the observation light.
- a wavefront confusion element 17 that gives disturbance to the wavefront of the illuminated observation light
- a first beam splitter 18 that branches the light given disturbance to the wavefront from the illumination optical path from the light source 11, and an interval in the optical axis direction.
- Optical path length varying means 22 and second bee A wavefront recovery element 23 disposed between the splitter 20 and the second intermediate imaging lens 21 and the light transmitted through the wavefront recovery element 23 and the second beam splitter 20 are condensed to form a final image.
- the imaging lens 24 is provided.
- the imaging device 14 is a two-dimensional image sensor such as a CCD or a CMOS, for example, and includes an imaging surface 14a arranged at the imaging position of the final image by the imaging lens 24, and images incident light.
- the wavefront confusion element 17 is disposed in the vicinity of the pupil position of the objective lens 16.
- the wavefront confusion element 17 is made of an optically transparent material that can transmit light. When the light is transmitted, the wavefront confusion element 17 imparts phase modulation to the light wavefront according to the uneven shape of the surface. In the present embodiment, the necessary wavefront disturbance is imparted by transmitting the observation light from the observation object A once.
- the wavefront recovery element 23 is disposed in the vicinity of the pupil position of the second intermediate imaging lens 21.
- the wavefront recovery element 23 is also made of an optically transparent material that can transmit light, and when the light is transmitted, phase modulation according to the uneven shape of the surface is applied to the wavefront of the light.
- the wavefront recovery element 23 transmits the observation light deflected by the beam splitter 20 and the observation light reflected so as to be folded back by the optical path length varying unit 22 twice, so that the wavefront confusion element 17 is reciprocated twice. Is applied to the wavefront of the light so as to cancel the disturbance of the wavefront imparted by.
- the optical path length varying means 22 as an optical axis (Z-axis) scanning system includes a plane mirror 22a disposed orthogonal to the optical axis, and an actuator 22b that displaces the plane mirror 22a in the optical axis direction.
- the optical path length between the second intermediate imaging lens 21 and the plane mirror 22a is changed.
- the position of the object A conjugate with the imaging surface 14a, that is, the in-focus position in front of the objective lens 16 can be changed in the optical axis direction.
- the illumination optical system 12 irradiates the observation object A with illumination light from the light source 11.
- Observation light composed of fluorescence, reflected light, scattered light, and the like emitted from the observation object A is collected by the objective lens 16 and transmitted once through the wavefront confusion element 17 to form the first beam splitter 18 and the intermediate image.
- the light passes through the optical system 19, is deflected by 90 ° at the second beam splitter 20, and passes through the wavefront recovery element 23.
- the observation light is reflected so as to be folded back by the plane mirror 22 a of the optical path length varying means 22, passes through the wavefront recovery element 23 again, and passes through the beam splitter 20.
- the final image formed by the imaging lens 24 is photographed by the image sensor 14.
- the optical path length between the second intermediate imaging lens 21 and the plane mirror 22a can be changed by operating the actuator 22b of the optical path length varying means 22 and moving the plane mirror 22a in the optical axis direction. Accordingly, the focal position in front of the objective lens 16 can be moved in the optical axis direction for scanning.
- a plurality of images focused on different positions in the depth direction of the observation object A can be acquired by photographing the observation light at different focal positions.
- an image with a deep depth of field can be acquired by performing high-frequency emphasis processing after combining these images by addition averaging.
- an intermediate image is formed by the second intermediate imaging lens 21 in the vicinity of the plane mirror 22 a of the optical path length varying means 22, and this intermediate image is given by passing through the wavefront confusion element 17.
- the wavefront disturbance is smeared by the wavefront disturbance left partially canceled by passing through the wavefront recovery element 23 once.
- the light after forming the blurred intermediate image is condensed by the second intermediate imaging lens 21 and then passed again through the wavefront recovery element 23, so that the wavefront disturbance is completely eliminated. Be countered.
- the observation apparatus 10 even if foreign matter such as scratches and dust is present on the surface of the plane mirror 22a, the foreign matter image is prevented from being captured on the final image.
- a clear image of the observation object A can be obtained.
- the intermediate image formed by the first intermediate imaging lens pair 19 also varies greatly in the optical axis direction.
- the intermediate image is blurred. Therefore, it is possible to prevent the image of the foreign object from being captured on the final image.
- the above-described scanning system is mounted, no noise image is generated even if light moves on the Z axis on any optical element arranged in the imaging optical system.
- an observation apparatus 30 according to a second embodiment of the present invention will be described below with reference to the drawings.
- portions having the same configuration as those of the observation apparatus 10 according to the first embodiment described above are denoted by the same reference numerals and description thereof is omitted.
- the observation device 30 condenses the laser light source 31 and the laser light from the laser light source 31 onto the observation object A, while the light from the observation object A is condensed.
- An imaging optical system 32 that condenses, an image sensor (photodetector) 33 that captures the light collected by the imaging optical system 32, and between the light source 31, the image sensor 33, and the imaging optical system 32.
- the Niipou disc type confocal optical system 34 is provided.
- the Nipkow disc type confocal optical system 34 includes two discs 34a and 34b arranged at a parallel interval, and an actuator 34c that simultaneously rotates the discs 34a and 34b.
- a number of microlenses (not shown) are arranged on the disk 34a on the laser light source 31 side, and a number of pinholes (not shown) are provided on the object side disk 34b at positions corresponding to the respective microlenses.
- a dichroic mirror 34d that divides the light that has passed through the pinhole is fixed in the space between the two disks 34a and 34b.
- the light branched by the dichroic mirror 34d is condensed by the condenser lens 35. Then, the final image is formed on the imaging surface 33a of the imaging device 33, and the image is acquired.
- the first beam splitter 18 and the second beam splitter 20 in the first embodiment are shared to form a single beam splitter 36, and the pinhole of the Niipou disc type confocal optical system 34 is formed.
- the optical path for irradiating the observation object A with the passed light and the optical path generated in the observation object A and entering the pinhole of the Niipou disc type confocal optical system 34 are completely made common.
- the observation device 30 configured as described above will be described below.
- the light that has entered the imaging optical system 32 from the pinhole of the Niipou disc type confocal optical system 34 passes through the beam splitter 36 and the phase modulation element 23, and then the second The light is condensed by the intermediate imaging lens 21 and reflected so as to be folded back by the plane mirror 22a of the optical path length varying means 22.
- the second intermediate imaging lens 21 After passing through the second intermediate imaging lens 21, the light passes through the phase modulation element 23 again, is deflected by 90 ° by the beam splitter 36, and passes through the first intermediate imaging lens pair 19 and the phase modulation element 17. Then, the light is condensed on the observation object A by the objective lens 16.
- the phase modulation element 23 through which the laser light is initially transmitted twice functions as a wavefront confusion element that imparts a disturbance to the wavefront of the laser light, and the phase modulation element 17 that is transmitted once thereafter has the phase It functions as a wavefront recovery element that applies phase modulation that cancels the disturbance of the wavefront applied by the modulation element 23.
- the image of the light source formed in a number of point light sources by the Niipou disc type confocal optical system 34 is formed as an intermediate image on the plane mirror 22a by the second intermediate imaging lens 21, but the second intermediate connection is formed. Since the intermediate image formed by the image lens 21 is blurred by passing through the phase modulation element 23 once, the inconvenience that the image of the foreign matter existing on the intermediate imaging surface overlaps the final image is prevented. it can.
- the disturbance imparted to the wavefront by passing through the phase modulation element 23 twice is canceled by passing through the phase modulation element 17 once, so that a clear image of many point light sources is displayed on the observation object A.
- An image can be formed.
- light for example, fluorescence
- the objective lens 16 is collected by the objective lens 16 and transmitted through the phase modulation element 17 and the first intermediate imaging lens pair 19. Thereafter, it is deflected by 90 ° by the beam splitter 36, passes through the phase modulation element 23, is collected by the second intermediate imaging lens 21, and is reflected so as to be folded by the plane mirror 22 a. Thereafter, the fluorescence is again condensed by the second intermediate imaging lens 21, transmitted through the phase modulation element 23 and the beam splitter 36, and condensed by the imaging lens 24. An image is formed at the pinhole position.
- the light that has passed through the pinhole is branched from the optical path from the laser light source by the dichroic mirror, condensed by the condenser lens, and formed as a final image on the imaging surface of the imaging device.
- the phase modulation element 17 through which the fluorescence generated in a large number of dots in the observation object passes functions as a wavefront confusion element as in the first embodiment, and the phase modulation element 23 functions as a wavefront recovery element. .
- the fluorescent light whose disturbance is given to the wavefront by passing through the phase modulation element 17 is in a state where the disturbance is partially canceled by passing through the phase modulation element 23 once, but is connected to the plane mirror 22a.
- the intermediate image to be imaged is blurred.
- the fluorescence whose wavefront disturbance has been completely cancelled forms an image on the pinhole of the Niipou disc type confocal optical system 34 and passes through the pinhole.
- the light is branched by the dichroic mirror 34d, condensed by the condenser lens 35, and a clear final image is formed on the imaging surface 33a of the imaging device 33.
- the intermediate image is blurred, both as an illumination device for irradiating the observation target A with laser light and as an observation device for photographing the fluorescence generated in the observation target A.
- the intermediate image is blurred, both as an illumination device for irradiating the observation target A with laser light and as an observation device for photographing the fluorescence generated in the observation target A.
- an observation apparatus 40 according to a third embodiment of the present invention will be described below with reference to the drawings.
- portions having the same configuration as those of the observation apparatus 30 according to the second embodiment described above are denoted by the same reference numerals and description thereof is omitted.
- the observation device 40 is a laser scanning confocal observation device as shown in FIG.
- the observation device 40 includes a laser light source 41, an imaging optical system 42 that condenses the laser light from the laser light source 41 on the observation object A, and condenses the light from the observation object A, and the connection.
- a confocal pinhole 43 that allows the fluorescence condensed by the image optical system 42 to pass therethrough and a photodetector 44 that detects the fluorescence that has passed through the confocal pinhole 43 are provided.
- the imaging optical system 42 is disposed in the vicinity of a position conjugate with the pupil of the objective lens 16, a beam expander 45 that expands the beam diameter of the laser light, a dichroic mirror 46 that deflects the laser light and transmits fluorescence.
- the galvanometer mirror 47 and the third intermediate imaging lens pair 48 are provided as different configurations from the observation device 30 according to the second embodiment.
- a phase modulation element 23 that imparts disturbance to the wavefront of the laser light is disposed in the vicinity of the galvanometer mirror 47.
- reference numeral 49 denotes a mirror.
- the laser light emitted from the laser light source 41 is enlarged by the beam expander 45, deflected by the dichroic mirror 46, and scanned two-dimensionally by the galvano mirror 47. After that, the light passes through the phase modulation element 23 and the third intermediate imaging lens pair 48 and enters the beam splitter 36.
- the laser light incident on the beam splitter 36 is incident on the plane mirror 22a of the optical path length varying means 22 to form an intermediate image.
- the wave front is disturbed by the phase modulation element 23 and the intermediate image is unclear. Therefore, it is possible to prevent the images of foreign matters existing on the intermediate image plane from overlapping.
- the wavefront disturbance is canceled out by the phase modulation element 17 disposed at the pupil position of the objective lens 16, a sharpened final image can be formed on the observation object A. Further, the imaging depth of the final image can be arbitrarily adjusted by the optical path length varying means 22.
- the fluorescence generated at the imaging position of the final image of the laser light on the observation object A is collected by the objective lens 16 and passes through the phase modulation element 17, and then follows an optical path opposite to the laser light. It is deflected by the beam splitter 36. Then, after passing through the third intermediate imaging lens pair 48, the phase modulation element 23, the galvano mirror 47, and the dichroic mirror 46, the fluorescence is condensed on the confocal pinhole 43 by the imaging lens 24, Only the fluorescence that has passed through the hole 43 is detected by the photodetector 44.
- the fluorescence condensed by the objective lens 16 forms an intermediate image after the wave front is disturbed by the phase modulation element 17, the intermediate image is blurred and exists on the intermediate image plane. It can prevent that the image of the foreign material to overlap. Since the wavefront disturbance is canceled by transmitting through the phase modulation element 23, a sharpened image can be formed on the confocal pinhole 43, and the final image of the laser beam is observed on the observation object A. The fluorescence generated at the image position can be detected efficiently. As a result, there is an advantage that a bright high-resolution confocal image can be acquired. In the present embodiment, when the above-described scanning system is mounted, no noise image is generated even if light moves on the Z axis on any optical element arranged in the imaging optical system.
- the laser scanning confocal observation device is illustrated, but instead, it may be applied to a laser scanning multiphoton excitation observation device as shown in FIG.
- an ultrashort pulse laser light source may be employed as the laser light source 41
- the dichroic mirror 46 may be eliminated, and the dichroic mirror 46 may be employed in place of the mirror 49.
- the intermediate image can be made unclear and the final image can be made clear by the function of the illumination device that irradiates the observation object A with the ultrashort pulse laser beam.
- the fluorescence generated in the observation object A is collected by the objective lens 16, and after being transmitted through the phase modulation element 17 and the dichroic mirror 46, is collected by the condenser lens 51 without forming an intermediate image.
- the light detector 44 detects the light as it is.
- the focal point position in front of the objective lens is changed in the optical axis direction by the optical path length varying means 22 that changes the optical path length by moving the plane mirror that turns the optical path.
- the optical path length varying means as shown in FIG. 9, one of the lenses 61a and 61b constituting the intermediate imaging optical system 61 is moved in the optical axis direction by an actuator 62.
- an observation device 60 that employs a device that changes the optical path length may be configured.
- reference numeral 63 denotes another intermediate imaging optical system.
- another intermediate imaging optical system 80 is disposed between two galvanometer mirrors 47 constituting a two-dimensional optical scanner, and the two galvanometer mirrors 47 are phase modulation elements 17. , 23 and the aperture stop 81 arranged in the pupil of the objective lens 16 may be arranged in an optically conjugate positional relationship with high accuracy.
- a spatial light modulation element (SLM) 64 such as a reflective LCOS may be employed as the optical path length varying means.
- SLM spatial light modulation element
- phase modulation applied to the wavefront can be changed at high speed by controlling the LCOS liquid crystal, and the focal position in front of the objective lens 16 can be changed at high speed in the optical axis direction.
- reference numeral 65 denotes a mirror.
- a spatial light modulator 66 such as the transmissive LCOS may be employed as shown in FIG. Compared with the reflective LCOS, the mirror 65 is not required, so that the configuration can be simplified.
- Means for moving the in-focus position in the observation object A in the optical axis direction are those shown in the above embodiments (optical path length varying means 22, intermediate imaging optical system 61 and actuator 62, or reflective spatial light).
- various power variable optical elements known as active optical elements can be used.
- a variable optical element having a mechanically movable portion there are a deformable mirror (DFM: Deformable Mirror) and a variable shape lens using liquid or gel.
- DFM Deformable Mirror
- variable optical element As a similar variable optical element having no mechanical movable part, a liquid crystal lens, a potassium tantalate niobate (KTN: KTa 1-x Nb x O 3 ) crystal lens that controls the refractive index of the medium by an electric field, Acousto-optic deflector (AOD / Acousto-Optical) There is a lens applying the cylindrical lens effect in Defect).
- KTN potassium tantalate niobate
- AOD / Acousto-Optical Acousto-optic deflector
- each of the embodiments of the microscope of the present invention has some means for moving the in-focus position on the observation object A in the optical axis direction. Further, these in-focus position optical axis direction moving means are compared with the means in the conventional microscope for the same purpose (which moves either the objective lens or the observation object in the optical axis direction).
- the operating speed can be greatly increased because of the use of a physical phenomenon with a small mass or a fast response speed. This has the advantage that a faster phenomenon can be detected in the observation object (for example, a living biological tissue specimen).
- the spatial light modulators 64 and 66 such as transmissive or reflective LCOS are employed, the spatial light modulators 64 and 66 can have the function of the phase modulator 23.
- the phase modulation element 23 as a wavefront confusion element can be abbreviate
- the phase modulation element 23 is omitted in the combination of the spatial light modulation element and the laser scanning type multiphoton excitation observation apparatus.
- the spatial light modulation element and the laser scanning type common apparatus are omitted.
- the phase modulation element 23 can be omitted. That is, in FIGS. 11 and 12, a mirror 49 is employed instead of the dichroic prism 36, a dichroic mirror 46 is employed between the beam expander 45 and the spatial light modulators 64 and 66, and a branched optical path is formed.
- the spatial light modulators 64 and 66 can have the function of the phase modulation element 23 after employing the imaging lens 24, the confocal pinhole 43, and the photodetector 44.
- the spatial light modulators 64 and 66 impart a disturbance to the wavefront as a wavefront confusion element for the laser light from the laser light source 41, while phase is applied to the fluorescence from the observation object A. It acts as a wavefront recovery element that cancels the disturbance of the wavefront imparted by the modulation element 17.
- phase modulation element for example, cylindrical lenses 17 and 23 as shown in FIG. 13 may be adopted.
- the intermediate image since the intermediate image is linearly extended by the cylindrical lens 17 due to astigmatism, the intermediate image can be blurred by this action, and the cylindrical lens 23 having a shape complementary thereto.
- the final image can be sharpened.
- either a convex lens or a concave lens may be used as the wavefront confusion element or a wavefront recovery element.
- FIG. 14 shows an example in which cylindrical lenses 5 and 6 are used as the phase modulation elements in FIGS. 2 and 3.
- a cylindrical lens having a power ⁇ O x in the x direction is used as the phase modulation element (wavefront confusion element) 5 on the object O side.
- a cylindrical lens having power ⁇ I x in the x direction is used as the phase modulation element (wavefront recovery element) 6 on the image I side.
- C position in the cylindrical lens 5 of the axial ray R x of the xz plane (ray height) and x O.
- D position in the cylindrical lens 6 in the axial ray R x of the xz plane (ray height) to x I.
- symbols II 0X and II 0Y are intermediate images.
- the optical path length difference L (x) ⁇ L (0) has the same absolute value as the phase advance amount of the emitted light at the height x with respect to the emitted light at the height 0, but the opposite sign. Therefore, the phase advance amount is expressed by the following equation (6) in which the sign of equation (5) is inverted.
- L (0) -L (x) (x 2/2) (n-1) (1 / r 1 -1 / r 2) ⁇ (6)
- the optical power ⁇ of the thin lens is expressed by the following equation (7).
- L Oc (x O ) is a function of the optical path length from the incident side tangent plane to the exit side tangent plane along the light beam having the height x O in the cylindrical lens 5.
- phase lead amount ⁇ L Ic for the axial principal ray that is, the ray RA along the optical axis, received by the cylindrical lens 6 by the axial ray Rx on the xz plane is expressed by the following equation (10).
- L Ic (x I ) is a function of the optical path length from the incident side tangent plane to the exit side tangent plane along the light beam of height x I in the cylindrical lens 6.
- the values of ⁇ OX and ⁇ IX have opposite signs, and the ratio of their absolute values needs to be proportional to the square of the lateral magnification of the field lens 4.
- the description has been made based on the on-axis light beam.
- the cylindrical lenses 5 and 6 similarly perform the function of wavefront confusion and wavefront recovery for the off-axis light beam.
- phase modulation elements 5, 6, 17, and 23 shown as phase modulation elements 5 and 6 in the figure
- a one-dimensional binary diffraction grating as shown in FIG. 16 is used instead of the cylindrical lens. Adopting a one-dimensional sinusoidal diffraction grating as shown in FIG. 17, a free-form surface lens as shown in FIG. 18, a cone lens as shown in FIG. 19, and a concentric binary diffraction grating as shown in FIG. Also good.
- the concentric diffraction grating is not limited to the binary type, and any form such as a blazed type or a sine wave type can be adopted.
- the diffraction gratings 5 and 6 are used as the wavefront modulation element.
- the intermediate image II in this case, one point image is separated into a plurality of point images by diffraction. By this action, it is possible to prevent the intermediate image II from being blurred, and the foreign object image on the intermediate imaging surface from appearing overlapping the final image.
- FIG. 21 shows an example of a preferable path of the axial principal ray, that is, the light beam RA along the optical axis when the diffraction gratings 5 and 6 are used as the phase modulation element, and a preferable path of the axial light beam R X.
- a preferable path of the axial principal ray that is, the light beam RA along the optical axis when the diffraction gratings 5 and 6 are used as the phase modulation element
- R X An example of each is shown in FIG. In these drawings, the light rays R A and R X are separated into a plurality of diffracted lights through the diffraction grating 5, but are converted into a single original light beam through the diffraction grating 6.
- equation (2) is the sum of the phase modulation received by axial rays R X of "one diffraction grating 5 and 6, the axial principal ray R A diffraction grating 5 In other words, it is always equal to the sum of the phase modulations received at 6.
- FIG. 23 is a detailed view of the diffraction grating 5, and FIG.
- the conditions for the diffraction gratings 5 and 6 to satisfy Expression (2) are as follows. That is, the modulation period p I in the diffraction grating 6 is equal to the modulation period p O by the diffraction grating 5 projected by the field lens 4, and the modulation phase by the diffraction grating 6 is due to the diffraction grating 5 projected by the field lens 4. The phase of the modulation is inverted, and the magnitude of the phase modulation by the diffraction grating 6 and the magnitude of the phase modulation by the diffraction grating 6 must be equal in absolute value.
- the diffraction grating 5 is The center of one of the mountain regions may be arranged so as to coincide with the optical axis, and the diffraction grating 6 may be arranged so that one of the centers of its valley regions may coincide with the optical axis.
- FIG. 23 and FIG. 24 are nothing but examples.
- phase advance amount ⁇ L Idt for the light ray RA along the optical axis (transmitting through the valley region) is expressed by the following equation (14).
- the diffraction grating 5 functions as a wavefront scattering and the diffraction grating 6 functions as a wavefront recovery for off-axis light beams as long as the above condition is satisfied.
- the sectional shape of the diffraction gratings 5 and 6 has been described as a trapezoid here, it is needless to say that other shapes can perform the same function.
- phase modulation elements 5 and 6 a spherical aberration element as shown in FIG. 25, an irregularly shaped element as shown in FIG. 26, a transmissive spatial light modulation element 64 as shown in FIG. A reflection type wavefront modulation element by a combination of the above and a gradient index element as shown in FIG. 28 may be adopted.
- phase modulation elements 5 and 6 a fly-eye lens or a micro lens array in which a large number of minute lenses are arranged, or a micro prism array in which a large number of minute prisms are arranged may be employed.
- the phase confusion element 5 is disposed inside the objective lens (imaging lens) 70
- the phase recovery element 6 may be disposed in the vicinity of the eyepiece lens 73 disposed on the opposite side of the objective lens 70 with the relay optical system 72 including the plurality of field lenses 4 and the condenser lens 71 interposed therebetween.
- the wavefront confusion element 5 is provided in an endoscope-type thin objective lens 74 with an inner focus function that drives a lens 61 a by an actuator 62, and a tube lens provided in a microscope main body 75.
- the wavefront recovery element 6 may be disposed near the pupil position of the (imaging lens) 76.
- the actuator itself may be a known lens driving unit (for example, a piezoelectric element), but the spatial modulation of the intermediate image can be performed from the same viewpoint as the above-described embodiment in terms of the movement of the intermediate image in the Z-axis direction. It is important that the arrangement is as follows.
- the wavefront confusion elements 5 and 23 and the wavefront recovery elements 6 and 17 are disposed in a conjugate relationship with each other.
- the wavefront confusion elements 5 and 23 and the wavefront recovery elements 6 and 17 are arranged. May be arranged in a non-conjugated positional relationship. In this case, it is desirable to employ cylindrical lenses as the wavefront confusion elements 5 and 23 and the wavefront recovery elements 6 and 17.
- the wavefront confusion element 5 and the wavefront recovery element 6 are illustrated in the case where the wavefront confusion elements 5 and 23 and the wavefront recovery elements 6 and 17 are arranged in a conjugate relationship with each other.
- the focal length f PMO 2l of the wavefront confusion element 5
- the focal length f PMI -2l of the wavefront recovery element 6
- ⁇ OX ⁇ IX
- ⁇ OY ⁇ IY
- the imaging lateral magnification from the object O to the image point I is equal to 1 in both the X direction ( ⁇ X ) and the Y direction ( ⁇ Y ).
- the pupil imaging magnification from the wavefront confusion element 5 arranged on the pupil plane to the wavefront restoration element 6 arranged on the pupil conjugate plane is equal to -1.
- An intermediate image II X ′ in the X direction which is a virtual image formed by, for example, a marginal ray R (O) as an outgoing ray from the wavefront recovery element 6 is generated on the field lens 4.
- the light emitted from the field lens 4 is parallel in the X direction.
- each wavefront recovery element 6 in these embodiments, each arrangement, and the above condition that the incident light to each wavefront recovery element 6 is parallel light in the X direction,
- a marginal ray R ( ⁇ ) as an outgoing ray from the wavefront recovery element 6 causes the wavefront confusion element 5 and the wavefront recovery element 6 to pass through. It is necessary to diverge more broadly than the marginal ray R (O) from the wavefront recovery element 6 when arranged in a conjugate relationship with each other. That is, the wavefront recovery element 6 needs to have a stronger negative power as a cylindrical lens. Specifically, regarding the distance m ( ⁇ 2l) from the field lens 4 to the wavefront recovery element 6, the focal length f PMI of the wavefront recovery element 5 must be ⁇ m.
- the image I is generated by the wavefront recovery element 6 without causing astigmatism.
- the marginal ray R ( ⁇ ) from the wavefront recovery element 6 diverges more widely than the marginal ray R (O) when the wavefront confusion element 5 and the wavefront recovery element 6 are arranged in a conjugate relationship with each other.
- This means that there is a difference in imaging lateral magnification ⁇ between the X direction and the Y direction, the Y direction remains the same magnification ( ⁇ Y 1), but the X direction is reduced ( ⁇ X ⁇ 1). .
- FIGS. 33A and 33B show a case where the wavefront recovery element 6 and the wavefront recovery element 6 are arranged closer to the image I side than when the wavefront recovery element 6 and the wavefront recovery element 6 are arranged in a conjugate relationship with each other. .
- the marginal ray R (+) as the light emitted from the wavefront recovery element 6 causes the wavefront confusion element 5 and the wavefront recovery element 6 to pass through. It is necessary to diverge narrower than the marginal ray R (O) when arranged in a conjugate positional relationship. That is, the wavefront recovery element 6 needs to have a weaker negative power as a cylindrical lens.
- the focal length f PMI of the wavefront recovery element 6 must be ⁇ n. Thereby, the intermediate image II X ′ formed by the marginal ray R (+) can be generated on the field lens 4.
- the image I is generated by the wavefront recovery element 6 without causing astigmatism.
- the marginal ray R (+) from the wavefront recovery element 6 diverges narrower than the marginal ray R (O) when the wavefront confusion element 5 and the wavefront recovery element 6 are arranged in a conjugate relationship with each other.
- the wavefront confusion element 5 and the wavefront recovery element 6 are not arranged in a conjugate positional relationship, by appropriately selecting the power of the cylindrical lens as the wavefront confusion element 5 and the wavefront recovery element 6, respectively.
- the image I can be formed without causing astigmatism. That is, the wavefront disturbance generated by the wavefront confusion element 5 can be canceled by the wavefront recovery element 6.
- an aspect ratio conversion optical system 121 configured by a cylindrical lens or a toroidal lens may be employed.
- the aspect ratio conversion optical system 121 includes a convex cylindrical lens 123A and a concave cylindrical lens 123B, and is disposed, for example, in front of the image sensor 33 (see FIG. 6). Yes.
- the aspect ratio conversion optical system 121 has the same magnification in the X direction and expands the magnification in the Y direction, and the focal position is matched in the X direction and the Y direction. That is, the aspect ratio conversion optical system 121 is configured such that the magnification changes in the X direction and the Y direction, but the focal position does not change.
- the solid line indicates the light beam in the Y direction
- the broken line indicates the light beam in the X direction.
- the aspect ratio conversion mechanism 125 capable of converting the aspect ratio of the image may be employed by changing the ratio of the X-scanning and Y-scanning amplitude with respect to a predetermined number of samplings.
- the aspect ratio conversion mechanism 125 includes an X-direction signal source 127A, a Y-direction signal source 127B, variable resistors 129A and 129B, and drive amplifiers 131A and 131B.
- the X-direction signal source 127A and the Y-direction signal source 127B each output a sawtooth signal.
- the voltage of each signal is relatively adjusted via the variable resistors 129A and 129B.
- the shake width in the X direction and the shake width in the Y direction of the galvanometer mirror 47 can be changed.
- the image information acquired by the observation apparatus 10 is subjected to an aspect ratio correction process to thereby obtain an image.
- An aspect ratio conversion circuit 133 or an aspect ratio conversion program that can convert the aspect ratio may be employed. As shown in FIG. 37, for example, when the observation object A is circular, the aspect ratio conversion circuit 133 can correct an image image acquired in an elliptical shape into a circular image image.
- FIGS. 32A and 32B The above-described properties for the case where the wavefront confusion element 5 and the wavefront recovery element 6 which are a combination of a phase modulation element and a phase demodulation element made of a cylindrical lens are arranged in an optically non-conjugated positional relationship are shown in FIGS. 32A and 32B.
- the above description is not limited to the configurations of FIGS. 33A and 33B, and includes the case where the basic arrangement is a so-called 4f optical system and the case where a lens having any power and a cylindrical lens having any power are combined. It is common to all those on the extension line.
- the wavefront confusion element 5 and the wavefront recovery element 6 according to this modification can be applied to the observation devices 10, 30, 40, 50, 60 as the microscopes of the above embodiments. Further, the wavefront confusion element 5 and the wavefront recovery element 6 according to this modification may be combined with various other microscopes.
- Each of the embodiments in which the wavefront confusion element 5 and the wavefront recovery element 6 are arranged in a conjugate relationship with each other can be applied to the observation apparatuses 10, 30, 40, 50, and 60 as a microscope as described above. Needless to say, it can be combined with various other microscopes.
- the optical system of FIG. 38 may be combined with the observation apparatus 10, 30, 40, 50, 60 as a microscope, and the wavefront confusion element 5 and the wavefront recovery element 6 may be arranged in a conjugate manner. Alternatively, it may be applied in a non-conjugated manner.
- the parallel plate 135 is formed of a glass member having a stepped shape with different thicknesses, and is disposed in the vicinity of the focal position of the lenses 139A and 139B facing each other.
- the parallel plate 135 is rotated around the axis by the motor 137 so that the thickness of the parallel plate 135 disposed in the vicinity of the focal position of the lenses 139A and 139B can be changed.
- the optical path length can be changed at high speed.
- the illumination device, the X-axis scanning device, and the observation light detection device in the line-scan microscope are replaced with the Niipou disc type confocal optical system 34 in the observation device 30, or the observation device 40.
- the wavefront confusion element 5 and the wavefront recovery element 6 may be applied in a conjugate manner to the observation device replaced with the laser light source 41, the imaging optical system 42, the confocal pinhole 43, and the photodetector 44 in FIG. Alternatively, it may be applied in a non-conjugated manner.
- a disk-type microscope with a slit pattern as described in Japanese Patent No. 4334801, and the non-patent literature “Ultrafast superresolution fluorescence imaging with spinning confocal microscopical.” 26, p. 1743-1751, May 1, 2015 may be combined with a disk-type super-resolution microscope with a slit pattern.
- the wavefront confusion element 5 and the observation device in which the illumination device, the rotary scanning device, and the observation light detection device in the disk-type microscope with a slit pattern are replaced with the Niipou disc type confocal optical system 34 in the observation device 30.
- the wavefront recovery element 6 may be applied in a conjugate manner or may be applied in a non-conjugated manner.
- non-patent literature “Breaking the diffraction resolution limit by stimulated emission: stimulated-mission-depletion fluorescence microscopy, Optics Letters. 19, p. It may be combined with a STED (Stimulated Emission Depletion) microscope as described in 780-782, 1994.
- the wavefront confusion element 5 and the wavefront recovery element 6 may be conjugated and applied to an observation apparatus in which the illumination apparatus in the STED microscope is replaced with the laser light source 41 in the observation apparatuses 40, 50, 60. However, they may be applied in a non-conjugated manner.
- the embodiment described above discusses a method of applying the smearing of the intermediate image by phase modulation to the imaging optical system of the observation apparatus from the viewpoint of moving the intermediate image and the final image in the Z-axis direction. .
- the movement of the intermediate image and the final image in the XY axis direction (or on the image plane), which is another viewpoint in the imaging optical system, will be discussed below. Therefore, the present invention includes not only optical scanning in the Z-axis direction but also optical scanning in the XY-axis direction.
- the present invention can also be applied to three-dimensional observation that combines the movement of the intermediate image and the final image in both directions in the Z-axis direction and the XY-axis direction.
- the moving means for executing only the movement of the intermediate image in the Z-axis direction the moving means for executing only the movement of the intermediate image and the final image in the XY-axis direction is referred to as a scanner. .
- One aspect of the present invention includes a plurality of imaging lenses that form a final image and at least one intermediate image, and are disposed closer to the object side than any of the intermediate images formed by the imaging lens.
- a first phase modulation element that imparts spatial disturbance to the wavefront of the light, and at least one intermediate image interposed between the first phase modulation element and the first phase modulation element
- An imaging optical system comprising: a second phase modulation element that cancels a spatial disturbance imparted to the wavefront of light from the object, and the imaging optical system disposed on the object side of the imaging optical system;
- a light detector for detecting, and the first phase modulation element and the second phase modulation element are disposed at a position optically conjugate with the first scanner
- the final image is formed by being condensed by the imaging lens.
- a spatial disturbance is given to the wavefront of the illumination light by passing through the first phase modulation element arranged on the object side of one of the intermediate images, and the formed intermediate image is blurred.
- the illumination light that forms the intermediate image passes through the second phase modulation element, thereby canceling the spatial disturbance of the wavefront imparted by the first phase modulation element.
- a clear image can be obtained in the final image formed after the second phase modulation element.
- the illumination light from the light source is scanned two-dimensionally by the first scanner and the second scanner, so that the final image formed on the observation object can be scanned two-dimensionally.
- the first scanner when the first scanner is operated, the luminous flux of the illumination light moves in a one-dimensional linear direction, but the first scanner and the second phase modulation element are arranged at optically conjugate positions. For this reason, the position of the light beam passing through the second phase modulation element does not fluctuate.
- the second scanner which is spaced from the first scanner in the optical axis direction, is not disposed in an optically conjugate positional relationship with the second phase modulation element, and thus activates the second scanner. Then, the luminous flux of the illumination light moves so as to change the passing position of the second phase modulation element. Since the direction in which the phase distribution characteristic of the second phase modulation element changes coincides with the scanning direction of the illumination light by the first scanner, the phase distribution in the direction orthogonal to this, that is, the scanning direction of the illumination by the second scanner The characteristic does not change, and the phase modulation applied to the illumination light does not change even if the passage position of the illumination light beam changes.
- the second phase is not affected by the scanning state of the illumination light.
- a constant state can be maintained without changing the phase modulation by the modulation element, and the spatial disturbance of the wavefront provided by the first phase modulation element can be completely canceled.
- the first phase modulation element and the second phase modulation element may be lenticular elements.
- the first phase modulation element and the second phase modulation element may be a prism array.
- the first phase modulation element and the second phase modulation element may be diffraction gratings.
- the first phase modulation element and the second phase modulation element may be cylindrical lenses.
- the observation apparatus 101 is, for example, a multiphoton excitation microscope.
- the observation apparatus 101 includes an illumination apparatus 102 that irradiates the observation target A with an ultrashort pulse laser beam (hereinafter simply referred to as laser light (illumination light)), and a laser by the illumination apparatus 102.
- a detector optical system 104 that guides the fluorescence generated in the observation object A due to light irradiation to the photodetector 105 and a photodetector 105 that detects the fluorescence guided by the detector optical system 104 are provided.
- the illumination device 102 includes a light source 106 that generates laser light and an imaging optical system 103 that irradiates the observation object A with the laser light from the light source 106.
- the imaging optical system 103 condenses the beam expander 107 that expands the beam diameter of the laser light from the light source 106 and the laser light that has passed through the beam expander 107 to form an intermediate image, and its imaging position. Are moved in a direction along the optical axis S, and a collimating lens 109 for converting the laser light that has passed through the Z scanning unit 108 and formed an intermediate image into substantially parallel light.
- the imaging optical system 103 is formed by a wavefront confusion element (first phase modulation element) 110 disposed at a position where laser light that has been substantially collimated by the collimator lens 109 passes, and a Z scanning unit 108.
- the laser beam that has passed through the element (second phase modulation element) 114 and the wavefront recovery element 114 is condensed and irradiated onto the observation object A, while the observation object A
- the Z scanning unit 108 includes a condensing lens 108a that condenses the laser light whose beam diameter has been expanded by the beam expander 107, and an actuator 108b that moves the condensing lens 108a in a direction along the optical axis S. ing.
- the focusing position 108a can be moved in the direction along the optical axis S by moving the condenser lens 108a in the direction along the optical axis S by the actuator 108b.
- the wavefront confusion element 110 is a lenticular element made of an optically transparent material that can transmit light.
- the wavefront confusion element 110 imparts phase modulation that changes in a one-dimensional direction perpendicular to the optical axis S to the wavefront of the laser light according to the shape of the surface 116 when the laser light is transmitted.
- the necessary wavefront disturbance is imparted by transmitting the laser light from the light source 106 once.
- the relay lens pair 111 condenses the laser light, which has become substantially parallel light by the collimator lens 109, by one lens 111a to form an intermediate image II, and then condenses the diffusing laser light again by the other lens 111b. So that it returns to almost parallel light.
- the two relay lens pairs 111 and 112 are arranged at an interval so as to sandwich the XY scanning unit 113 in the direction along the optical axis S.
- the galvanometer mirrors 113a and 113b are provided so as to be swingable around an axis perpendicular to the optical axis S and in a twisted relationship with each other. These galvano mirrors 113a, 113b, by which is oscillated, is changed to the two-dimensional direction perpendicular to the inclination angle of the laser beam to the optical axis S, the position of the final image I F by the objective lens 115 in the optical axis S It is possible to scan in the intersecting two-dimensional direction.
- the wavefront recovery element 114 is a lenticular element made of an optically transparent material capable of transmitting light and having a phase distribution characteristic opposite to that of the wavefront confusion element 110.
- the wavefront recovery element 114 imparts phase modulation that changes only in a one-dimensional direction orthogonal to the optical axis S according to the shape of the surface 117 to the wavefront of the light when the laser light is transmitted, and is imparted by the wavefront confusion element 110. It is designed to cancel the wave front disturbance.
- the two galvanometer mirrors 113a and 113b are arranged with a gap in the direction along the optical axis S, and their intermediate position 113c is optically substantially the same as the pupil position POB of the objective lens 115. It arrange
- the galvanometer mirror 113a on the light source 106 side is disposed at a position optically conjugate with the wavefront confusion element 110 and the wavefront recovery element 114.
- the central ray Ra of the light beam P of the laser beam is reflected by the wavefront recovery element.
- 114 intersects the optical axis S on the surface 117 of the surface. That is, the laser beam P can pass through the same region without changing the passage position in the wavefront recovery element 114.
- this galvanometer mirror 113a is arrange
- the light beam P of the laser beam passes through the same region of the wavefront recovery element 114 regardless of the oscillation of the galvanometer mirror 113a, so that the phase modulation applied to the laser beam changes even if the galvanometer mirror 113a oscillates. You do n’t have to.
- the galvanometer mirror 113b on the observation object A side is disposed at a position optically unconjugated to the wavefront recovery element 114.
- the central ray Rb of the light beam P of the laser beam is restored to the wavefront.
- the surface of the element 114 is separated from the optical axis S.
- the galvano mirror 113b matches the swing direction (the direction of arrow Y in FIG. 41) with the direction orthogonal to the direction in which the phase distribution characteristic of the wavefront recovery element 114 changes (the direction in which the phase distribution characteristic does not change). Are arranged.
- the laser beam is caused by the oscillation of the galvanometer mirrors 113a and 113b.
- 43 moves in the two-dimensional directions of arrows X and Y as shown in FIG. 43 at the pupil position POB of the objective lens 115.
- the movement range is limited to the movement of a minute range that can pass without being kicked by the opening 118a of the aperture stop 118 disposed at the pupil position POB of the objective lens 115.
- the detector optical system 104 includes a dichroic mirror 119 that branches the fluorescence collected by the objective lens 115 from the optical path of the laser beam, and two condenser lenses 104a and 104b that collect the fluorescence branched by the dichroic mirror 119. And.
- the photodetector 105 is, for example, a photomultiplier tube, and detects the intensity of incident fluorescence.
- the image formation optical system 103 irradiates the observation object A with the laser light emitted from the light source 106.
- the beam diameter of the laser beam is expanded by the beam expander 107 and passed through the Z scanning unit 108, the collimating lens 109, and the wavefront confusion element 110.
- Laser light is condensed by the condensing lens 108a of the Z scanning unit 108, and the condensing position can be adjusted in the direction along the optical axis S by the operation of the actuator 108b. Further, the laser light is allowed to pass through the wavefront confusion element 110, so that spatial disturbance is imparted to the wavefront.
- the laser light is passed through the two relay lens pairs 111 and 112 and the XY scanning unit 113, whereby the inclination angle of the light beam P is changed while forming the intermediate image II, and passes through the dichroic mirror 119. . Then, the laser light that has passed through the dichroic mirror 119 passes through the wavefront recovery element 114, cancels the spatial disturbance imparted by the wavefront confusion element 110, and is condensed by the objective lens 115, and the final image IF is observed. An image is formed on the object A.
- Focus position of the imaging optical system 103 is the position of the final image I F imaged laser beam, the operation of the actuator 108b by moving the condenser lens 108a, is moved in the direction along the optical axis S It is done. Thereby, the observation depth of the observation object A can be adjusted. Further, the focus position of the laser beam on the observation object A can be two-dimensionally scanned in the direction orthogonal to the optical axis S by swinging the galvanometer mirrors 113a and 113b.
- the laser light to which the wavefront confusion element 110 imparts a spatial disturbance of the wavefront is used for the lenticular element that forms the wavefront confusion element 110, that is, the cylindrical lens array.
- astigmatism is given after one light beam P is divided into a large number of small light beams.
- a point image that is originally one is blurred and formed as a collection of a large number of circular images, elliptical images, or linear images arranged in a straight line.
- the laser light passes through the wavefront recovery element 114, since the spatial disturbance of the wavefront applied by the wavefront confusion element 110 is canceled, the final image I F to be imaged in the wavefront recovery device 114 later It becomes clear.
- the intermediate image II is located in the vicinity of an optical element in which scratches, foreign matter, or defects are present on the surface or inside because the intermediate image II is blurred and blurred, the scratches, foreign matter, or defects etc. is superimposed over the intermediate image II, the final image I F which is formed on the observation object a can be prevented from becoming unclear. As a result, a very small spot as the final image I F can be imaged.
- the light beam P of the laser light moves in a one-dimensional linear direction, but is in a positional relationship optically conjugate with the galvano mirror 113a.
- the light flux P in the wavefront recovery element 114 passes through the same region in the direction of the arrow X. Therefore, it is not necessary to change the phase modulation applied to the laser beam by the wavefront recovery element 114 regardless of the oscillation of the galvanometer mirror 113a.
- the galvanometer mirror 113b on the observation object A side is swung, the tilt of the light beam P of the laser light is changed by the swing of the galvanometer mirror 113b, and the passing position of the light beam P in the wavefront recovery element 114 is indicated by an arrow.
- Move in the Y direction Since the direction of the arrow Y coincides with the direction in which the phase distribution characteristics of the wavefront recovery element 114 do not change, it is given even if the wavefront recovery element 114 passes through a different region in the direction of the arrow Y due to the movement of the passage position of the light beam P.
- the phase modulation does not change. Therefore, even if the galvano mirror 113b is swung, it is not necessary to change the phase modulation applied to the laser light by the wavefront recovery element 114.
- the positional relationship for the elements 114 to be complementary is destroyed, and as a result, the wavefront disturbance applied by the wavefront confusion element 110 cannot be canceled by the wavefront recovery element 114.
- the wavefront confusion element 110 and the wavefront recovery element 114 are complementary even if the galvanometer mirrors 113a and 113b are swung.
- the wavefront disturbance imparted by the wavefront confusion element 110 can always be completely canceled out by the wavefront recovery element 114.
- fluorescence can be generated by increasing the photon density in an extremely small region, and the generated fluorescence is condensed by the objective lens 115 and is then dichroic mirror 119. Can be detected by directing fluorescence to the photodetector 105 by the detector optical system 104.
- the fluorescence intensity detected by the photodetector 105 corresponds to the scanning position of the three-dimensional laser beam by the position in the directions of arrows X and Y by the galvanometer mirrors 113a and 113b and the position in the direction along the optical axis S by the actuator 108b.
- the fluorescence image of the observation object A is acquired by adding and storing. That is, according to the observation apparatus 101 according to the present embodiment, since fluorescence is generated in an extremely small spot area at each scanning position, there is an advantage that a fluorescence image with high spatial resolution can be acquired.
- the observation apparatus 101 since the observation apparatus 101 according to the present embodiment does not need to arrange a relay lens pair between the two galvanometer mirrors 113a and 113b, the number of parts of the apparatus can be reduced. Further, by adopting a configuration in which the galvanometer mirrors 113a and 113b are arranged close to each other without arranging the relay lens pair, the apparatus can be reduced in size.
- lenticular elements are illustrated as the wavefront confusion element 110 and the wavefront recovery element 114, but instead, elements having a one-dimensional phase distribution characteristic may be employed.
- elements having a one-dimensional phase distribution characteristic may be employed.
- a prism array, a diffraction grating, or a cylindrical lens may be employed.
- the galvanometer mirrors 113a and 113b are exemplified as the first scanner and the second scanner which are moving means of the intermediate image on the XY axes.
- Different types of scanners may be used instead.
- a polygon mirror, AOD (acousto-optic element), KTN (potassium tantalate niobate) crystal, or the like may be employed.
- the observation apparatus 101 which concerns on this embodiment illustrated the multiphoton excitation microscope, it may replace with this and may apply to a confocal microscope.
- a very small spot is imaged on the observation object A as the final image I F which is sharpened, it is possible to generate fluorescence to increase the photon density in a very small area, a confocal pin A bright confocal image can be acquired by increasing the fluorescence passing through the hole.
- FIG. A specific example of the optical condition in the illumination device 102 of the present embodiment shown in FIG. 39 is a wavefront confusion element at a position optically conjugate with the galvano mirror 113a between the galvano mirror 113a on the light source 106 side and the light source 106. 110 is disposed, and the wavefront recovery element 114 is disposed at a position optically conjugate with the galvano mirror 113 a on the light source 106 side behind the objectives 115.
- the wavefront recovery element 114 is arranged so that the phase distribution characteristic thereof coincides with the scanning direction of the laser light (the direction of the arrow X) by the galvanometer mirror 113a.
- the wavefront recovery element 114 can always cancel the spatial disturbance of the wavefront imparted by the wavefront confusion element 110 regardless of the swing angle of the galvanometer mirrors 113a and 113b. Therefore, it is possible to image the objects blocking the intermediate image II is blurred intermediate image II imaging position is prevented from overlapping the intermediate image II, and always sharpen the final image I F.
- b b (f TL / f PL ) 2 (16)
- b is the distance to the two galvanometer mirrors 113a, a substantially conjugate position 113c to the pupil position POB of the object lens 115 positioned sandwiched 113b of the light source 106 side galvanomirror 113a, f PL relay lens pair 112
- the focal length of the lens 112 a on the light source 106 side, f TL indicates the focal length of the lens 112 b on the observation object A side of the relay lens pair 112.
- the distance c from the rear end of the mounting screw of the objective lens 115 to the wavefront recovery element 114 satisfies the condition of Expression (17).
- d the protrusion amount of the mounting screw of the objective lens 115
- e the distance from the body surface of the objective lens 115 to the pupil position POB of the objective lens 115.
- the wavefront recovery element 114 is disposed at a position optically conjugate with the galvano mirror 113a on the light source 106 side behind the objective lens 115 without contacting the rear end of the outer frame of the objective lens 115, that is, the mounting screw.
- the present invention is more useful for the microscope observation in combination with the above aspect relating to the movement of the intermediate image and the final image in the Z axis direction. It shall be Therefore, the present invention is arranged in a conjugate manner with respect to the intermediate image moving in the Z-axis direction as referred to in FIGS. 1 to 38, as illustrated in FIGS. 39 to 44. From the viewpoint of maintaining the complementarity of the wavefront confusion element and the wavefront recovery element with respect to scanning in the XY-axis direction by a pair of galvanometer mirrors, the following additional items are also included.
- a plurality of imaging lenses that form a final image and at least one intermediate image, and light from the object that is disposed closer to the object side than any of the intermediate images formed by the imaging lens
- a first phase modulation element that imparts spatial disturbance to the wavefront of the first phase modulation element, and at least one intermediate image sandwiched between the first phase modulation element and the first phase modulation element
- An imaging optical system comprising: a second phase modulation element that cancels a spatial disturbance applied to a wavefront of light from the object; A light source that is disposed on the object side of the imaging optical system and generates illumination light incident on the imaging optical system, and a light source that is disposed at an interval in the optical axis direction and that scans illumination light from the light source.
- a second detector, and a photodetector for detecting light emitted from the observation object disposed at the final image position of the imaging optical system, and the first phase modulation element and the first scanner The two phase modulation elements are arranged at a position optically conjugate with the first scanner arranged on the light source side, and change in a direction that coincides with the scanning direction of illumination light by the first scanner.
- An observation apparatus applied to an optical axis direction scanning microscope apparatus having a one-dimensional phase distribution characteristic.
- the first phase modulation element and the second phase modulation element are arranged at a position optically conjugate with the second scanner arranged on the object side, and illumination by the second scanner It has a one-dimensional phase distribution characteristic that changes in a direction that coincides with the scanning direction of light, and the other configuration is applied to an optical axis direction scanning microscope apparatus that conforms to the observation apparatus according to appendix 1.
- Observation device (Additional Item 3) The observation apparatus according to Additional Item 1, wherein the first phase modulation element and the second phase modulation element are lenticular elements.
- the observation apparatus according to Additional Item 1, wherein the first phase modulation element and the second phase modulation element are prism arrays.
- the said aspect can also be summarized as follows. That is, in the above supplementary item, even if the intermediate image is formed at a position that coincides with the optical element, the intermediate image is prevented from overlapping with scratches, foreign matters, defects, or the like of the optical element to obtain a clear final image. This is a technical issue.
- an imaging lens 111,112,115 to form the final image I F and the intermediate image II, one of the intermediate image to solve the technical problem by the appended claim
- the first phase modulation element 110 disposed on the object side from II and imparting spatial disturbance to the wavefront of light, and the space disposed on the final image IF side from one or more intermediate images II and imparted to the wavefront of light
- the imaging optical system 103 including the second phase modulation element 114 that cancels the general disturbance, the light source 106 disposed on the object side, and the first and second light sources disposed at intervals in the optical axis S direction.
- An XY scanning unit 113 including scanners 113a and 113b and a photodetector 105 for detecting light are provided, and two phase modulation elements 110 and 114 are optically conjugate with the first scanner 113a disposed on the light source 106 side. Arranged at various positions Is, having a one-dimensional phase distribution characteristic that varies in a direction corresponding to the scanning direction of the illumination light by the first scanner 113a, to provide a viewing device 101.
- the wavefront confusion element 110 and the wavefront recovery element 114 may be arranged in a non-conjugated positional relationship.
- a cylindrical lens may be employed as the wavefront confusion element 110 and the wavefront recovery element 114.
- the first scanner 113a and the wavefront recovery element 114 may be arranged in a conjugate manner, and the first scanner 113a and the wavefront confusion element 110 may be arranged in a nonconjugated manner.
- a means for eliminating the difference between the imaging magnification in the X direction and the imaging magnification in the Y direction such as the aspect ratio conversion optical system 121, the amplitude ratio changing mechanism 125, and the aspect ratio correction circuit 133, may be adopted. That's fine.
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Abstract
Description
本発明の一態様は、最終像および少なくとも1つの中間像を形成する複数の結像レンズと、該結像レンズにより形成されるいずれかの前記中間像よりも物体側に配置され、前記物体からの光の波面に空間的な乱れを付与する第1の位相変調素子と、該第1の位相変調素子との間に少なくとも1つの中間像を挟む位置に配置され、前記第1の位相変調素子により前記物体からの光の波面に付与された空間的な乱れを打ち消す第2の位相変調素子とを備える結像光学系と、前記物体からの波面が前記結像光学系を通過することにより結像される像を光軸方向に走査するための走査系とを備える光軸方向走査型顕微鏡装置である。
まず「鮮明な像」とは、物体から発した光の波面に、空間的な乱れが付与されていない状態で、あるいは一旦付与された乱れが打ち消され解消された状態で、結像レンズを介して生成された像であり、光の波長と結像レンズの開口数とで決まる空間周波数帯域、あるいはそれに準ずる空間周波数帯域、あるいは目的に応じた所望の空間周波数帯域を有するものを意味する。
上記態様においては、前記第1の位相変調素子および前記第2の位相変調素子が、光学的に共役な位置に配置されていることとしてもよい。
このようにすることで、光束の変動しない瞳位置近傍に配置して第1の位相変調素子および第2の位相変調素子を小型化することができる。
このようにすることで、光路長可変手段の作動により、2つの結像レンズ間の光路長を変更することにより、最終像の結像位置を光軸方向に容易に変更することができる。
このようにすることで、物体側の結像レンズにより集光された物体側からの光が平面鏡によって反射されて折り返された後、ビームスプリッタによって分岐されて像側の結像レンズに入射される。この場合において、アクチュエータを作動させて平面鏡を光軸方向に移動させることにより、2つの結像レンズ間の光路長を容易に変更することができ、最終像の結像位置を光軸方向に容易に変更することができる。
このようにすることで、可変空間位相変調素子によって最終像位置を光軸方向に変化させるような空間的な位相変調を光の波面に付与することができる。付与する位相変調を調節することにより、最終像の結像位置を光軸方向に容易に変更することができる。
このようにすることで、可変空間位相変調素子に最終像位置を光軸方向に変化させるような空間的な位相変調と、中間像をぼやけさせるような位相変調あるいは中間像のぼやけを打ち消すような位相変調との両方を受け持たせることができる。これにより、構成部品を少なくして簡易な結像光学系を構成することができる。
このようにすることで、第1の位相変調素子により光軸に直交する1次元方向に変化する位相変調を光の波面に付与して、中間像をぼやけさせることができる。そして、中間像位置に何らかの光学素子が配置されて、該光学素子の表面や内部に傷、異物あるいは欠陥等が存在していても、それら光学素子の傷、異物あるいは欠陥等が中間像に重なって、最終的に最終像の一部として形成されてしまう不都合の発生を防止することができる。また、1次元方向に変化した位相変調を打ち消すような位相変調を第2の位相変調素子により光の波面に付与して、ぼやけない鮮明な最終像を結像させることができる。
このようにすることで、第1の位相変調素子により光軸に直交する2次元方向に変化する位相変調を光の波面に付与して、中間像をより確実にぼやけさせることができる。また、2次元方向に変化した位相変調を打ち消すような位相変調を第2の位相変調素子により光の波面に付与して、より鮮明な最終像を結像させることができる。
また、上記態様においては、前記第1の位相変調素子および前記第2の位相変調素子が、光を反射させる際に波面に位相変調を付与する反射型素子であってもよい。
このようにすることで、中間像をぼやけさせる空間的な乱れを波面に付与する第1の位相変調素子と、波面に付与された空間的な乱れを打ち消すような位相変調を付与する第2の位相変調素子とを簡易に構成することができる。
このようにすることで、第1の位相変調素子を光が透過する際に屈折率分布に従う波面の乱れを生じさせ、第2の位相変調素子を光が透過する際に屈折率分布によって波面の乱れを打ち消すような位相変調を光の波面に付与することができる。
本態様によれば、物体側に配置された光源から発せられた照明光が結像光学系に入射されることにより、最終像側に配置された照明対象物に照明光を照射することができる。この場合に、第1の位相変調素子によって、結像光学系により形成される中間像がぼやけさせられるので、中間像位置に何らかの光学素子が配置されて、該光学素子の表面や内部に傷、異物あるいは欠陥等が存在していても、それら光学素子の傷、異物あるいは欠陥等が中間像に重なって、最終的に最終像の一部として形成されてしまう不都合の発生を防止することができる。
本態様によれば、結像光学系により、光学素子の表面や内部に傷、異物あるいは欠陥等の像が中間像に重なることが防止されることによって形成された鮮明な最終像を光検出器によって検出することができる。
このようにすることで、結像光学系の最終像位置に配置された撮像素子により、鮮明な最終像を撮影して、精度の高い観察を行うことができる。
このようにすることで、観察対象物に多点のスポット光を走査させて観察対象物の鮮明な画像を高速に取得することができる。
このようにすることで、中間像位置における傷や異物や欠陥等の像の写り込みのない、鮮明な共焦点画像による観察対象物の観察を行うことができる。
このようにすることで、中間像位置における傷や異物や欠陥等の像の写り込みのない、鮮明な多光子励起画像による観察対象物の観察を行うことができる。
上記態様においては、光スキャナを備え、該光スキャナが、前記第1の位相変調素子、前記第2の位相変調素子および前記結像レンズの瞳に対して光学的に共役な位置に配置されていることとしてもよい。
すなわち、適切なパワーのシリンドリカルレンズを適切な場所に配置することによって、第1の位相変調素子と第2の位相変調素子が光学的に非共役であっても、第1の位相変調素子により生じた光の波面の乱れを第2の位相変調素子により打ち消して、非点収差を生じることなく結像させることができる。これにより、たとえ空間的な制約等によって、第1の位相変調素子と第2の位相変調素子を光学的に共役に配置することが出来ない光学系であっても、中間像をぼやけさせることにより、中間像位置に配置された光学素子の表面や内部に存在する傷、異物あるいは欠陥等が中間像に重なって、最終的に最終像の一部として形成されてしまう不都合の発生を防止することができる。
上記態様においては、いずれかの前記中間像を挟む位置に配置される2つの前記結像レンズ間の光路長を変更可能な光路長可変手段を備えていてもよい。
上記態様においては、前記第1の位相変調素子または前記第2の位相変調素子の少なくとも一方の機能が、前記可変空間位相変調素子によって担われていてもよい。
上記態様においては、前記第1の位相変調素子および前記第2の位相変調素子が、光を反射させる際に波面に位相変調を付与する反射型素子であってもよい。
上記態様においては、前記第1の位相変調素子および前記第2の位相変調素子が、透明材料の屈折率分布によって波面に位相変調を付与してもよい。
上記態様においては、前記結像光学系の最終像側に配置され、観察対象物から発せられた光を検出する光検出器をさらに備えていてもよい。
上記態様においては、前記結像光学系の物体側に配置され、該結像光学系に入射させる照明光を発生する光源と、前記結像光学系の最終像側に配置され、観察対象物から発せられた光を検出する光検出器とをさらに備えていてもよい。
上記態様においては、前記光源がレーザ光源であり、前記光検出器が共焦点ピンホールおよび光電変換素子を備えていてもよい。
上記態様においては、光スキャナを備え、該光スキャナが、前記第1の位相変調素子、前記第2の位相変調素子および前記結像レンズの瞳に対して光学的に共役な位置に配置されていることとしてもよい。
本実施形態に係る結像光学系1は、図1に示されるように、間隔をあけて配置された2つ1組の結像レンズ2,3と、これらの結像レンズ2,3の中間結像面に配置されたフィールドレンズ4と、物体O側の結像レンズ2の瞳位置PPO近傍に配置された波面錯乱素子(第1の位相変調素子)5と、像I側の結像レンズ3の瞳位置PPI近傍に配置された波面回復素子(第2の位相変調素子)6とを備えている。図中、符号7は開口絞りである。
図2に示される例では、結像光学系1は、物体O側および像I側に関してテレセントリックな配置になっている。また、波面錯乱素子5はフィールドレンズ4から物体O側に距離aFだけ離れた位置に配置され、波面回復素子6はフィールドレンズ4から像I側に距離bFだけ離れた位置に配置されている。
ただし、波面錯乱素子5と波面回復素子6は、フィールドレンズ4による結像に関して、式(1)に示されるように、互いに共役な位置関係に配置されている必要がある。
ここで、fFはフィールドレンズ4の焦点距離である。
ここで、ΔLは、光が光学素子を透過することによって付与される、特定の位置(すなわち光線高さ)を透過する光線を基準とした、位相の進み量である。
さらに、ΔLI(xI)は、光が波面回復素子6の光軸上(x=0)を通過する場合を基準とした、波面回復素子6の任意の光線高さxIを通過する場合の位相の進み量を与える関数である。
ΔLO(xO)+ΔLI(xI)=ΔLO(xO)+ΔLI(βF・xO)=0・・・(2)
ここで、βFは、フィールドレンズ4による波面錯乱素子5と波面回復素子6の共役関係における横倍率であり、下式(3)により表される。
βF=-bF/aF・・・(3)
また、位相進み量の関数を1次元的な関数としたが、これに代えて、2次元的な関数としても同様に作用し得る。
本実施形態に係る観察装置10は、図5に示されるように、非コヒーレントな照明光を発生する光源11と、光源11からの照明光を観察対象物Aに照射する照明光学系12と、観察対象物Aからの光を集光する結像光学系13と、該結像光学系13により集光された光を撮影して画像を取得する撮像素子(光検出器)14とを備えている。
また、この照明光学系12は、いわゆるケーラー照明であり、集光レンズ15a,15bは、光源11の発光面と対物レンズ16の瞳面とが互いに共役になるように配置されている。
波面錯乱素子17は、対物レンズ16の瞳位置近傍に配置されている。波面錯乱素子17は、光を透過可能な光学的に透明な材料により構成され、光が透過する際に、表面の凹凸形状に従う位相変調を光の波面に付与するようになっている。本実施形態においては、観察対象物Aからの観察光を1回透過させることにより、必要な波面の乱れを付与するようになっている。
また、同様にして、観察対象物Aにおける合焦点位置を光軸方向に移動させると、第1の中間結像レンズ対19によって形成される中間像も光軸方向に大きく変動するが、その変動の結果、中間像が第1の中間結像レンズ対19の位置に重なったとしても、あるいはまた、その変動範囲内に何らかの他の光学素子が存在する場合であっても、中間像が不鮮明化されているので、異物の像が最終像に重なって撮影されてしまうことを防止することができる。本実施形態において、上述したような走査系を搭載した場合には、結像光学系に配置されるあらゆる光学素子上で、光がZ軸移動してもノイズ画像を生じない。
本実施形態の説明において、上述した第1の実施形態に係る観察装置10と構成を共通とする箇所には同一符号を付して説明を省略する。
本実施形態に係る観察装置30によれば、ニポウディスク型コンフォーカル光学系34のピンホールから結像光学系32に入射した光は、ビームススプリッタ36および位相変調素子23を透過した後に、第2の中間結像レンズ21によって集光され、光路長可変手段22の平面鏡22aによって折り返されるように反射される。そして、第2の中間結像レンズ21を通過した後に、位相変調素子23を再度透過し、ビームスプリッタ36によって90°偏向され、第1の中間結像レンズ対19および位相変調素子17を透過して対物レンズ16により観察対象物Aに集光される。
この場合において、観察対象物において多数の点状に発生した蛍光が透過する位相変調素子17は第1の実施形態と同様に波面錯乱素子として機能し、位相変調素子23は波面回復素子として機能する。
本実施形態の説明において、上述した第2の実施形態に係る観察装置30と構成を共通とする箇所には同一符号を付して説明を省略する。
この観察装置40は、レーザ光源41と、該レーザ光源41からのレーザ光を観察対象物Aに集光させる一方、観察対象物Aからの光を集光する結像光学系42と、該結像光学系42により集光された蛍光を通過させる共焦点ピンホール43と、該共焦点ピンホール43を通過した蛍光を検出する光検出器44とを備えている。
本実施形態に係る観察装置40によれば、レーザ光源41から発せられたレーザ光は、ビームエキスパンダ45によってビーム径が拡大されてダイクロイックミラー46により偏向され、ガルバノミラー47によって2次元的に走査された後、位相変調素子23および第3の中間結像レンズ対48を通過してビームスプリッタ36に入射する。
この場合、レーザ光源41として、極短パルスレーザ光源を採用し、ダイクロイックミラー46を無くし、ミラー49に代えて、ダイクロイックミラー46を採用すればよい。
Deflector)におけるシリンドリカルレンズ効果を応用したレンズ、等がある。
このことには、観察対象物(例えば、生きた生体組織標本)における、より高速な現象を検出し得る、という利点がある。
この場合には、シリンドリカルレンズ17によって中間像は非点収差によって点像が線状に伸ばされるので、この作用により、中間像を不鮮明化することができ、これと相補的な形状のシリンドリカルレンズ23により、最終像を鮮明化することができる。
図13の場合、凸レンズまたは凹レンズのいずれを波面錯乱素子として使用してもよいし、波面回復素子として使用してもよい。
(a)物体O側の位相変調素子(波面錯乱素子)5として、x方向にパワーψOxを有するシリンドリカルレンズを用いる。
(b)像I側の位相変調素子(波面回復素子)6として、x方向にパワーψIxを有するシリンドリカルレンズを用いる。
(c)xz平面上の軸上光線Rxのシリンドリカルレンズ5における位置(光線高さ)をxOとする。
(d)xz平面上の軸上光線Rxのシリンドリカルレンズ6における位置(光線高さ)をxIとする。
図14において、符号II0X,II0Yは中間像である。
図15において、高さ(光軸からの距離)xでのレンズの厚さをd(x)、高さ0(光軸上)でのレンズの厚さをd0とすると、高さxの光線に沿った入射側接平面から射出側接平面までの光路長L(x)は次式(4)で表される。
L(x)=(d0-d(x))+n・d(x)・・・(4)
L(x)-L(0)=(-x2/2)(n-1)(1/r1-1/r2)・・・(5)
L(0)-L(x)=(x2/2)(n-1)(1/r1-1/r2)・・・(6)
ψ=1/f=(n-1)(1/r1-1/r2)・・・(7)
L(0)-L(x)=ψ・x2/2・・・(8)
xz面上の軸上光線Rxがシリンドリカルレンズ5において受ける軸上主光線すなわち光軸に沿った光線RAに対する位相進み量ΔLOcは、式(8)に基づいて次式(9)で表される。
ΔLOc(xO)=LOc(0)-LOc(xO)=ψOx・xO 2/2・・・(9)
ここで、LOc(xO)はシリンドリカルレンズ5における高さxOの光線に沿った、入射側接平面から射出側接平面までの光路長の関数である。
ΔLIc(xI)=LIc(0)-LIc(xI)=ψIx・xI 2/2・・・(10)
ここで、LIc(xI)はシリンドリカルレンズ6における高さxIの光線に沿った、入射側接平面から射出側接平面までの光路長の関数である。
ψOX/ψIX=-βF 2・・・(11)
なお、ここでは軸上光線に基づいて説明したが、上記条件を満たすならば、シリンドリカルレンズ5,6は軸外光線に対しても同様に波面錯乱と波面回復の機能を果たす。
この場合の中間像IIにおいては回折によって1つの点像が複数の点像に分離される。
この作用によって、中間像IIが不鮮明化され、中間結像面の異物の像が最終像に重なって表れることを防止することができる。
ここで、図21および図22に準じて、式(2)は「1本の軸上光線RXが回折格子5,6で受ける位相変調の和は、軸上主光線RAが回折格子5,6で受ける位相変調の和と常に等しい。」と言い換えることができる。
そこで、回折格子5,6の中央部、すなわち、光軸近傍領域に着目して説明する。図23は回折格子5の、図24は回折格子6の、それぞれ中央部の詳細図である。
すなわち、回折格子6における変調の周期pIがフィールドレンズ4によって投影された回折格子5による変調の周期pOと等しく、回折格子6による変調の位相がフィールドレンズ4によって投影された回折格子5による変調の位相に対して反転しており、かつ、回折格子6による位相変調の大きさと回折格子6による位相変調の大きさとが絶対値で等しくなければならない。
pI=|βF|・pO・・・(12)
回折格子5の光学的なパラメータ(山領域厚さtOC、谷領域厚さtOt、屈折率nO)より、回折格子5の谷領域を透過する軸上光線RXに付与される、光軸に沿った(山領域を透過する)光線RAに対する位相進み量ΔLOdtは、次式(13)で表される。
ΔLIdt=(nI・tIt+(tIc-tIt))-nI・tIc=-(nI-1)(tIc-tIt)・・・(14)
ΔLOdt+ΔLIdt=(nO-1)(tOc-tOt)-(nI-1)(tIc-tIc)=0・・・(15)
また、ここでは回折格子5,6の断面形状を台形として説明したが、他の形状でも同様の機能を果たし得ることは言うまでもない。
次に、上記各実施形態の観察装置に用いられる結像光学系の変形例について図を参照して説明する。
上記実施形態においては、波面錯乱素子5,23と波面回復素子6,17とが互いに共役な位置関係に配置されていることとしたが、これら波面錯乱素子5,23と波面回復素子6,17とを非共役な位置関係に配置することとしてもよい。この場合、波面錯乱素子5,23および波面回復素子6,17として、シリンドリカルレンズを採用することが望ましい。
図31Aおよび図31Bにおいて、焦点距離f0=fF=fI=l、波面錯乱素子5の焦点距離fPMO=2l、波面回復素子6の焦点距離fPMI=-2l、ΘOX=ΘIX、ΘOY=ΘIY、βX=βY=1とする。
なお、波面錯乱素子5と波面回復素子6とが互いに共役な位置関係に配置される前記各実施形態は、既述の通り顕微鏡としての観察装置10,30,40,50,60に適用し得るのみならず、その他の顕微鏡各種と組み合わせることも可能であるのは、言うまでもない。
集光点(最終像IF)において発生した蛍光を集光する対物レンズ(結像レンズ)115とを備えている。
上述したようにガルバノミラー113aの揺動に関わらず、レーザ光の光束Pが波面回復素子114の同一領域を通過するので、ガルバノミラー113aが揺動してもレーザ光に付与する位相変調を変化させずに済むようになっている。
光検出器105は、例えば、光電子増倍管であり、入射された蛍光の強度を検出するようになっている。
本実施例のように、中間にリレーレンズ対を配置せずにガルバノミラー113a,113bを近接させて配置する構成の場合、ガルバノミラー113a,113bの両方に対して光学的に共役な位置は存在しない。すなわち、波面錯乱素子110と波面回復素子114をたとえ共役に配置しても、ガルバノミラー113a,113bの揺動による光の二次元方向の走査に伴って、通常ならば波面錯乱素子110と波面回復素子114が相補的になるための位置関係が崩れ、その結果として波面錯乱素子110によって付与された波面の乱れが波面回復素子114によって打ち消すことが出来なくなる。しかしながら本実施例では、波面錯乱素子110と波面回復素子114の形状と配置を工夫することによって、ガルバノミラー113a,113bが揺動しても、波面錯乱素子110と波面回復素子114が相補的になる位置関係が実質的には保たれ、その結果として波面錯乱素子110によって付与された波面の乱れを波面回復素子114によって常に完全に打ち消すようにできるのである。
これによれば、鮮明化された最終像IFとして観察対象物Aに極めて小さいスポットが結像されることにより、極めて小さい領域において光子密度を高めて蛍光を発生させることができ、共焦点ピンホールを通過する蛍光を増加させて明るい共焦点画像を取得することができる。
図39に示される、本実施形態の照明装置102における光学的条件の具体例は、光源106側のガルバノミラー113aと光源106との間のガルバノミラー113aと光学的に共役な位置に波面錯乱素子110を配置し、対物ンズ115の後ろ側の光源106側のガルバノミラー113aと光学的に共役な位置に、波面回復素子114を配置する。波面回復素子114は、その位相分布特性が、ガルバノミラー113aによるレーザ光の走査方向(矢印Xの方向)に一致するように配置する。
図4における、対物レンズ115の瞳位置POBから波面回復素子114までの距離aは、式(16)の条件を満足する。
ここで、bは2つのガルバノミラー113a,113bに挟まれて位置する対物レンズ115の瞳位置POBに略共役な位置113cから光源106側のガルバノミラー113aまでの距離、fPLはリレーレンズ対112の光源106側のレンズ112aの焦点距離、fTLはリレーレンズ対112の観察対象物A側のレンズ112bの焦点距離を示している。また、対物レンズ115の取り付けねじ後端から波面回復素子114までの距離cは、式(17)の条件を満足する。
c=a-(d+e)・・・(17)
ここで、dは対物レンズ115の取り付けねじの突出量、eは対物レンズ115の胴付面から対物レンズ115の瞳位置POBまでの距離を示している。
b=2.7(mm)
fPL=52(mm)
fTL=200(mm)
d=5(mm)
e=28(mm)
となる。
該結像光学系の物体側に配置され、該結像光学系に入射させる照明光を発生する光源と、光軸方向に間隔をあけて配置され、前記光源からの照明光を走査する第1のスキャナおよび第2のスキャナと、前記結像光学系の最終像位置に配置された観察対象物から発せられた光を検出する光検出器とを備え、前記第1の位相変調素子および前記第2の位相変調素子が、前記光源側に配置された前記第1のスキャナと光学的に共役な位置に配置されるとともに、前記第1のスキャナによる照明光の走査方向に一致する方向に変化する一次元的な位相分布特性を有する、光軸方向走査型顕微鏡装置に適用される観察装置。
(付記項2) 第1の位相変調素子および第2の位相変調素子が、物体側に配置された第2のスキャナと光学的に共役な位置に配置されるとともに、該第2のスキャナによる照明光の走査方向に一致する方向に変化する一次元的な位相分布特性を有し、それ以外の構成は、付記項1に記載の観察装置に準じる、光軸方向走査型顕微鏡装置に適用される観察装置。
(付記項3) 前記第1の位相変調素子および前記第2の位相変調素子がレンチキュラー素子である付記項1に記載の観察装置。
(付記項4) 前記第1の位相変調素子および前記第2の位相変調素子がプリズムアレイである付記項1に記載の観察装置。
(付記項5) 前記第1の位相変調素子および前記第2の位相変調素子が回折格子である付記項1に記載の観察装置。
(付記項6) 前記第1の位相変調素子および前記第2の位相変調素子がシリンドリカルレンズである付記項1に記載の観察装置。
すなわち、上記付記項においては、中間像が光学素子に一致する位置で結像されても、中間像に光学素子の傷、異物および欠陥等が重なることを防止して鮮明な最終像を取得することが技術的課題であるといえる。また、上記付記項による技術課題を解決する手段は、概して図39に示されるように、最終像IFと中間像IIとを形成する結像レンズ111,112,115と、いずれかの中間像IIより物体側に配置され光の波面に空間的な乱れを付与する第1の位相変調素子110と、1以上の中間像IIより最終像IF側に配置され光の波面に付与された空間的な乱れを打ち消す第2の位相変調素子114とを備える結像光学系103と、物体側に配置される光源106と、光軸S方向に間隔をあけて配置された第1および第2のスキャナ113a,113bを備えるXY走査部113と、光を検出する光検出器105とを備え、2つの位相変調素子110,114が光源106側に配置された第1のスキャナ113aと光学的に共役な位置に配置され、前記第1のスキャナ113aによる照明光の走査方向に一致する方向に変化する一次元的な位相分布特性を有する、観察装置101を提供する。
II 中間像
O 物体
PPO,PPI 瞳位置
1,13,32,42 結像光学系
2,3 結像レンズ
5 波面錯乱素子(第1の位相変調素子)
6 波面回復素子(第2の位相変調素子)
10,30,40,50,60 観察装置
11,31,41 光源
14,33 撮像素子(光検出器)
17,23 位相変調素子
20,36 ビームスプリッタ
22 光路長可変手段
22a 平面鏡
22b アクチュエータ
34 ニポウディスク型コンフォーカル光学系
43 共焦点ピンホール
44 光検出器(光電子変換素子)
61a レンズ(光路長可変手段)
62 アクチュエータ(光路長可変手段)
64 空間光変調素子(可変空間位相変調素子)
101 観察装置
103 結像光学系
105 光検出器
106 極短パルスレーザ光(光源)
110 波面錯乱素子(第1の位相変調素子)
111,112 リレーレンズ対(結像レンズ)
113 XY走査部
113a ガルバノミラー(第1のスキャナ)
113b ガルバノミラー(第2のスキャナ)
114 波面回復素子(第2の位相変調素子)
115 対物レンズ(結像レンズ)
Claims (39)
- 最終像および少なくとも1つの中間像を形成する複数の結像レンズと、該結像レンズにより形成されるいずれかの前記中間像よりも物体側に配置され、前記物体からの光の波面に空間的な乱れを付与する第1の位相変調素子と、該第1の位相変調素子との間に少なくとも1つの中間像を挟む位置に配置され、前記第1の位相変調素子により前記物体からの光の波面に付与された空間的な乱れを打ち消す第2の位相変調素子とを備える結像光学系と、
前記物体からの波面が前記結像光学系を通過することにより結像される像を光軸方向に走査するための走査系とを備える光軸方向走査型顕微鏡装置。 - 前記第1の位相変調素子および前記第2の位相変調素子が、光学的に共役な位置に配置されている請求項1に記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子および前記第2の位相変調素子が、前記結像レンズの瞳位置近傍に配置されている請求項1または請求項2に記載の光軸方向走査型顕微鏡装置。
- いずれかの前記中間像を挟む位置に配置される2つの前記結像レンズ間の光路長を変更可能な光路長可変手段を備える請求項1から請求項3のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記光路長可変手段が、光軸に直交して配置され前記中間像を形成する光を折り返すように反射する平面鏡と、該平面鏡を光軸方向に移動させるアクチュエータと、前記平面鏡により反射された光を2方向に分岐するビームスプリッタとを備える請求項4に記載の光軸方向走査型顕微鏡装置。
- いずれかの前記結像レンズの瞳位置近傍に、光の波面に付与する空間的な位相変調を変更することにより、前記最終像位置を光軸方向に変化させる可変空間位相変調素子を備える請求項1から請求項3のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子または前記第2の位相変調素子の少なくとも一方の機能が、前記可変空間位相変調素子によって担われる請求項6に記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子および前記第2の位相変調素子が、光軸に直交する1次元方向に変化する位相変調を光束の波面に付与する請求項1から請求項7のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子および前記第2の位相変調素子が、光軸に直交する2次元方向に変化する位相変調を光束の波面に付与する請求項1から請求項7のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子および前記第2の位相変調素子が、光を透過させる際に波面に位相変調を付与する透過型素子である請求項1から請求項9のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子および前記第2の位相変調素子が、光を反射させる際に波面に位相変調を付与する反射型素子である請求項1から請求項9のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子と前記第2の位相変調素子とが、相補的な形状を有する請求項1から請求項11のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子および前記第2の位相変調素子が、透明材料の屈折率分布によって波面に位相変調を付与する請求項10に記載の光軸方向走査型顕微鏡装置。
- 請求項1から請求項13のいずれかに記載の光軸方向走査型顕微鏡装置において、前記結像光学系の物体側に配置され、該結像光学系に入射させる照明光を発生するための光源をさらに備える、光軸方向走査型顕微鏡装置。
- 請求項1から請求項13のいずれかに記載の光軸方向走査型顕微鏡装置において、前記結像光学系の最終像側に配置され、観察対象物から発せられた光を検出する光検出器をさらに備える、光軸方向走査型顕微鏡装置。
- 前記光検出器が、前記結像光学系の最終像位置に配置され、該最終像を撮影する撮像素子である請求項15に記載の光軸方向走査型顕微鏡装置。
- 請求項1から請求項13のいずれかに記載の光軸方向走査型顕微鏡装置において、前記結像光学系の物体側に配置され、該結像光学系に入射させる照明光を発生する光源と、前記結像光学系の最終像側に配置され、観察対象物から発せられた光を検出する光検出器とをさらに備える、光軸方向走査型顕微鏡装置。
- 前記光源および前記光検出器と前記結像光学系との間に配置されたニポウディスク型コンフォーカル光学系を備える請求項17に記載の光軸方向走査型顕微鏡装置。
- 前記光源がレーザ光源であり、
前記光検出器が共焦点ピンホールおよび光電変換素子を備える請求項17に記載の光軸方向走査型顕微鏡装置。 - 前記光源によって照明された観察対象物から発せられた光を検出する光検出器を備え、
前記光源がパルスレーザ光源である請求項14に記載の光軸方向走査型顕微鏡装置。 - 光スキャナを備え、
該光スキャナが、前記第1の位相変調素子、前記第2の位相変調素子および前記結像レンズの瞳に対して光学的に共役な位置に配置されている請求項19または請求項20に記載の光軸方向走査型顕微鏡装置。 - 前記第1の位相変調素子および前記第2の位相変調素子が、光学的に非共役な位置に配置されたシリンドリカルレンズの組合せである請求項1に記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子および前記第2の位相変調素子の少なくとも1つが、前記結像レンズの瞳位置近傍に配置されている請求項22に記載の光軸方向走査型顕微鏡装置。
- いずれかの前記中間像を挟む位置に配置される2つの前記結像レンズ間の光路長を変更可能な光路長可変手段を備える請求項22から請求項23のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記光路長可変手段が、光軸に直交して配置され前記中間像を形成する光を折り返すように反射する平面鏡と、該平面鏡を光軸方向に移動させるアクチュエータと、前記平面鏡により反射された光を2方向に分岐するビームスプリッタとを備える請求項24に記載の光軸方向走査型顕微鏡装置。
- いずれかの前記結像レンズの瞳位置近傍に、光の波面に付与する空間的な位相変調を変更することにより、前記最終像位置を光軸方向に変化させる可変空間位相変調素子を備える請求項22または請求項23に記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子または前記第2の位相変調素子の少なくとも一方の機能が、前記可変空間位相変調素子によって担われる請求項26に記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子および前記第2の位相変調素子が、光を透過させる際に波面に位相変調を付与する透過型素子である請求項22から請求項27のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子および前記第2の位相変調素子が、光を反射させる際に波面に位相変調を付与する反射型素子である請求項22から請求項27のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子と前記第2の位相変調素子とが、相補的な形状を有する請求項22から請求項29のいずれかに記載の光軸方向走査型顕微鏡装置。
- 前記第1の位相変調素子および前記第2の位相変調素子が、透明材料の屈折率分布によって波面に位相変調を付与する請求項28に記載の光軸方向走査型顕微鏡装置。
- 請求項22から請求項31のいずれかに記載の光軸方向走査型顕微鏡装置において、前記結像光学系の物体側に配置され、該結像光学系に入射させる照明光を発生するための光源をさらに備える、光軸方向走査型顕微鏡装置。
- 請求項22から請求項31のいずれかに記載の光軸方向走査型顕微鏡装置において、前記結像光学系の最終像側に配置され、観察対象物から発せられた光を検出する光検出器をさらに備える、光軸方向走査型顕微鏡装置。
- 前記光検出器が、前記結像光学系の最終像位置に配置され、該最終像を撮影する撮像素子である請求項33に記載の光軸方向走査型顕微鏡装置。
- 請求項22から請求項31のいずれかに記載の光軸方向走査型顕微鏡装置において、前記結像光学系の物体側に配置され、該結像光学系に入射させる照明光を発生する光源と、前記結像光学系の最終像側に配置され、観察対象物から発せられた光を検出する光検出器とをさらに備える、光軸方向走査型顕微鏡装置。
- 前記光源および前記光検出器と前記結像光学系との間に配置されたニポウディスク型コンフォーカル光学系を備える請求項35に記載の光軸方向走査型顕微鏡装置。
- 前記光源がレーザ光源であり、
前記光検出器が共焦点ピンホールおよび光電変換素子を備える請求項35に記載の光軸方向走査型顕微鏡装置。 - 前記光源によって照明された観察対象物から発せられた光を検出する光検出器を備え、
前記光源がパルスレーザ光源である請求項32に記載の光軸方向走査型顕微鏡装置。 - 光スキャナを備え、
該光スキャナが、前記第1の位相変調素子、前記第2の位相変調素子および前記結像レンズの瞳に対して光学的に共役な位置に配置されている請求項37または請求項38に記載の光軸方向走査型顕微鏡装置。
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Citations (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH06265814A (ja) * | 1992-11-26 | 1994-09-22 | Asahi Optical Co Ltd | 投影光学系の瞳共役結合装置 |
JPH10257373A (ja) * | 1997-01-10 | 1998-09-25 | Olympus Optical Co Ltd | 画像入力装置 |
JPH11109243A (ja) * | 1997-08-04 | 1999-04-23 | Canon Inc | 光学素子及びそれを用いた光学装置 |
JPH11326860A (ja) * | 1998-05-18 | 1999-11-26 | Olympus Optical Co Ltd | 波面変換素子及びそれを用いたレーザ走査装置 |
JP2002196246A (ja) * | 2000-12-26 | 2002-07-12 | Olympus Optical Co Ltd | 走査型光学顕微鏡 |
JP2008113860A (ja) * | 2006-11-06 | 2008-05-22 | Kyocera Corp | 生体認証装置 |
JP2008245157A (ja) * | 2007-03-28 | 2008-10-09 | Kyocera Corp | 撮像装置およびその方法 |
JP2010513968A (ja) * | 2006-12-22 | 2010-04-30 | アイシス イノベイシヨン リミテツド | 焦点調整装置および焦点調整方法 |
JP2010266813A (ja) * | 2009-05-18 | 2010-11-25 | Olympus Corp | 観察装置 |
JP2013083817A (ja) * | 2011-10-11 | 2013-05-09 | Ricoh Co Ltd | 画像表示装置 |
WO2014163114A1 (ja) * | 2013-04-03 | 2014-10-09 | オリンパス株式会社 | 結像光学系、照明装置および観察装置 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8064041B2 (en) * | 2004-06-10 | 2011-11-22 | Carl Zeiss Smt Gmbh | Projection objective for a microlithographic projection exposure apparatus |
JP4606831B2 (ja) * | 2004-08-31 | 2011-01-05 | 浜松ホトニクス株式会社 | 光パターン形成方法および装置、ならびに光ピンセット装置 |
CN100403087C (zh) * | 2006-09-26 | 2008-07-16 | 浙江大学 | 基于数字微镜器件的无串扰并行oct成像方法及*** |
-
2015
- 2015-10-02 CN CN201580053458.6A patent/CN107076974A/zh active Pending
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Patent Citations (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH06265814A (ja) * | 1992-11-26 | 1994-09-22 | Asahi Optical Co Ltd | 投影光学系の瞳共役結合装置 |
JPH10257373A (ja) * | 1997-01-10 | 1998-09-25 | Olympus Optical Co Ltd | 画像入力装置 |
JPH11109243A (ja) * | 1997-08-04 | 1999-04-23 | Canon Inc | 光学素子及びそれを用いた光学装置 |
JPH11326860A (ja) * | 1998-05-18 | 1999-11-26 | Olympus Optical Co Ltd | 波面変換素子及びそれを用いたレーザ走査装置 |
JP2002196246A (ja) * | 2000-12-26 | 2002-07-12 | Olympus Optical Co Ltd | 走査型光学顕微鏡 |
JP2008113860A (ja) * | 2006-11-06 | 2008-05-22 | Kyocera Corp | 生体認証装置 |
JP2010513968A (ja) * | 2006-12-22 | 2010-04-30 | アイシス イノベイシヨン リミテツド | 焦点調整装置および焦点調整方法 |
JP2008245157A (ja) * | 2007-03-28 | 2008-10-09 | Kyocera Corp | 撮像装置およびその方法 |
JP2010266813A (ja) * | 2009-05-18 | 2010-11-25 | Olympus Corp | 観察装置 |
JP2013083817A (ja) * | 2011-10-11 | 2013-05-09 | Ricoh Co Ltd | 画像表示装置 |
WO2014163114A1 (ja) * | 2013-04-03 | 2014-10-09 | オリンパス株式会社 | 結像光学系、照明装置および観察装置 |
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