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WO2012127880A1 - Dispositif d'observation et procédé d'observation - Google Patents

Dispositif d'observation et procédé d'observation Download PDF

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Publication number
WO2012127880A1
WO2012127880A1 PCT/JP2012/002048 JP2012002048W WO2012127880A1 WO 2012127880 A1 WO2012127880 A1 WO 2012127880A1 JP 2012002048 W JP2012002048 W JP 2012002048W WO 2012127880 A1 WO2012127880 A1 WO 2012127880A1
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Prior art keywords
light
observation
optical system
observed
measurement light
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Ceased
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PCT/JP2012/002048
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English (en)
Japanese (ja)
Inventor
久美子 西村
中山　繁
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Nikon Corp
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Nikon Corp
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Priority to JP2013505826A priority Critical patent/JP5610063B2/ja
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
    • G01B9/00—Measuring instruments characterised by the use of optical techniques
    • G01B9/02—Interferometers
    • G01B9/0209—Low-coherence interferometers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
    • G01B9/00—Measuring instruments characterised by the use of optical techniques
    • G01B9/02—Interferometers
    • G01B9/02034—Interferometers characterised by particularly shaped beams or wavefronts
    • G01B9/02035—Shaping the focal point, e.g. elongated focus
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/18—Arrangements with more than one light path, e.g. for comparing two specimens
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
    • G01B2290/00—Aspects of interferometers not specifically covered by any group under G01B9/02
    • G01B2290/70—Using polarization in the interferometer

Definitions

  • the present invention relates to an observation apparatus and an observation method.
  • OCT optical coherence tomography
  • Non-Patent Document 1 There is optical coherence tomography (OCT) as one of the non-destructive tomographic measurement technologies (see Non-Patent Document 1, etc.).
  • OCT can measure the refractive index distribution, spectral information, polarization information, and the like of an object to be observed by using light having a wide wavelength range as a probe.
  • OCT is suitable for in vivo cells.
  • the present invention has been made in view of the above problems, and an object thereof is to provide an observation apparatus and an observation method capable of detection based on light transmitted through an object to be observed.
  • One aspect of the observation apparatus of the present invention includes a branching unit that branches incident light into reference light and measurement light, an illumination optical system that irradiates the object to be observed along the predetermined direction, and the observation object.
  • An observation optical system that is disposed on a different side from the illumination optical system with respect to an object and receives observation light that travels in a direction different from the predetermined direction among the measurement light via the object to be observed; and the reference light
  • a reference optical system that receives the reference light and the observation light synthesized by the synthesis optical system, and combines the reference light and the observation light.
  • a detection device that detects interference intensity with the observation light.
  • the branching unit branches light in a predetermined wavelength range into the reference light and the measurement light, and the detection device splits the reference light and the observation light synthesized by the synthesis optical system.
  • An element and a detection element that detects the interference intensity for each of a plurality of different wavelength ranges among the light dispersed by the spectroscopic element may be included.
  • the detection device may include position information acquisition means for acquiring information related to the position of the observation object along a direction different from the predetermined direction based on the interference intensity for each of the plurality of wavelength ranges.
  • the detection device may include position information acquisition means for acquiring information related to the position of the observation object along a direction different from the predetermined direction based on the interference intensity.
  • the observation optical system has an optical axis along a direction different from the predetermined direction, and the illumination optical system transmits the measurement light from a direction inclined by a predetermined angle with respect to the optical axis of the observation optical system.
  • the object to be observed may be irradiated.
  • the observation optical system has an optical axis along a direction different from the predetermined direction, and the illumination optical system transmits the measurement light from a direction inclined by a predetermined angle with respect to the optical axis of the observation optical system.
  • the position information acquisition means may acquire information on the position of the object to be observed along the optical axis of the observation optical system based on the interference intensity for each of the plurality of wavelength ranges. good.
  • the illumination optical system may include shape control means arranged in the optical path of the illumination optical system in order to control the predetermined direction of the measurement light irradiated on the object to be observed.
  • the illumination optical system may include an oblique incident means that forms an intensity distribution of incident light in a ring shape in a direction along a cross section of the light beam.
  • the oblique incident means may include an optical element that deflects light incident along the optical axis of the illumination optical system or the observation optical system.
  • the oblique incidence means may include a light shielding means for shielding a part of the incident light.
  • the light shielding means may have at least one opening.
  • the light shielding means may have a plurality of openings dispersed along a substantially circular shape.
  • the light shielding means may have an opening that transmits light incident along a substantially circular shape.
  • scanning means for relatively moving the position of the illumination optical system and the object to be observed may be provided.
  • a half-wave plate or a quarter-wave plate disposed in the reference light or the measurement light; a first polarizing element for entering light of a predetermined polarization into the branching unit; A second polarizing element that transmits part of the interference light including a polarization direction that is determined based on predetermined polarization; a polarization direction that is transmitted by the first polarizing element; and a polarization that is transmitted by the second polarizing element.
  • Control means for changing the direction at the same time may be provided.
  • the numerical aperture NA of the measurement light illuminated obliquely with respect to the optical axis of the observation optical system may be 0.4 ⁇ NA ⁇ 0.8.
  • the numerical aperture NA of the measurement light illuminated obliquely with respect to the optical axis of the observation optical system may be 0.8.
  • the optical path length difference between the measurement light and the reference light may be shorter than 400 ⁇ m.
  • One aspect of the observation method of the present invention includes branching incident light into reference light and measurement light, irradiating the object to be observed along the predetermined direction with respect to the object to be observed, Receiving observation light that travels (advances) in a direction different from the predetermined direction out of the measurement light via the object to be observed from a side different from the side irradiated with the illumination light, and the reference light and the observation Combining (superimposing) light with each other, and receiving the reference light and the observation light combined with each other, and detecting interference intensity between the reference light and the observation light.
  • the branching branches light in a predetermined wavelength range into the reference light and the measurement light, and the detection includes splitting the reference light and the observation light combined with each other. , Detecting the interference intensity for each of a plurality of different wavelength ranges among the reference light and the observation light that have been dispersed.
  • the detecting may include acquiring information related to a position of the observation object along a direction different from the predetermined direction based on the interference intensity for each of the plurality of wavelength ranges.
  • FIG. 1 is a configuration diagram of the OCT apparatus according to the first embodiment.
  • the OCT apparatus includes a light source 1, a collimating lens 2, beam splitters 3 and 4, an axicon lens 5, a relay lens 6, total reflection mirrors 7 and 8, objective lenses 9 and 10, a sample 11, and a sample stage. 12, an aperture 13, a dispersion correction optical member 14, a cylindrical lens 15, a spectrum detector 16, a control device 17, an arithmetic device 18, and the like are disposed.
  • the control device 17 controls each part of the light source 1, the sample stage 12, and the spectrum detector 16 and sends the spectrum signal acquired by the spectrum detector 16 to the arithmetic device 18.
  • the observation optical system such as the objective lens 10 is different from the illumination optical system such as the objective lens 9 from the light source 1 with respect to the sample 11 that is the observation object (that is, the observation object is placed on the illumination optical system). To be sandwiched). Then, the observation optical system such as the objective lens 10 receives the observation light that travels (advances) in a direction different from the irradiation direction of the illumination light by the illumination optical system among the measurement light that has passed through the sample 11 that is the object to be observed.
  • cultured cells or the like to be observed are cultured in a container (not shown).
  • a container mentioned above various things, such as a petri dish, a flask, a well plate, a microplate, can be used.
  • a slide glass can be used instead of the container.
  • the sample 11 is set on the sample stage 12.
  • the sample stage 12 is movable in a plane (xy plane) perpendicular to the optical axis direction of the objective lenses 9 and 10, and performs scanning in the xy direction when detected by the OCT apparatus (details will be described later).
  • the light source 1 emits light having a short time coherence.
  • the light source 1 for example, a super luminescence diode (SLD), a titanium sapphire laser, a white LED, or the like is applied.
  • SLD super luminescence diode
  • titanium sapphire laser titanium sapphire laser
  • white LED white LED
  • the resolution in the z direction of the OCT apparatus depends on the coherent length of the light source 1. Further, the resolution in the xy direction of the OCT apparatus depends on the size of a condensing point to be described later, and the size of the condensing point depends on the performance of the objective lens 9.
  • the illumination light L0 emitted from the light source 1 is collimated to a predetermined beam diameter by the collimator lens 2 and enters the beam splitter 3.
  • the illumination light L0 that has entered the beam splitter 3 is branched into a reference light Lr that travels toward the dispersion correction optical member 14 and a measurement light Lm that travels toward the sample 11.
  • the reference light Lr enters the beam splitter 4 through the dispersion correction optical member 14 and the total reflection mirror 8.
  • the dispersion correction optical member 14 is an element mainly for maintaining the balance of the interferometer in the OCT apparatus.
  • the balance of the interferometer here refers to the balance of chromatic dispersion of the sample arm and the reference arm of the interferometer. This balance may be lost due to an optical material such as an objective lens and a medium (a culture solution or the like) included in the sample.
  • the dispersion correcting optical member 14 includes a member for correcting the balance loss caused by the optical material such as an objective lens, and a member for correcting the balance loss caused by the medium included in the sample. .
  • the member for correcting the balance loss caused by the optical material described above has a correction amount corresponding to each optical member arranged on the sample arm.
  • the member for correcting the balance loss caused by the medium described above is preferably a member whose correction amount is variable in accordance with the type and amount of the medium.
  • a glass block or the like is used as a member for correcting the balance loss caused by the optical material.
  • an AOPDF Acoustic-Optic programmable dispersive filter
  • a prism pair that can be inserted and changed, and the like are used.
  • the dispersion correction optical member 14 may be substituted by calculation processing.
  • the reference light Lr is adjusted by the dispersion correcting optical member 14 so as to have a dispersion balanced with the sample arm.
  • the reference light Lr that has passed through the dispersion correction optical member 14 is hereinafter referred to as reference light Lr ′.
  • the interference pattern in the spectral direction
  • the finer the interference fringes the more difficult it is to spectrally resolve and detect with a spectroscopic system. That is, the upper limit of the optical path length difference is determined by the resolution of the spectroscopic system (the spectroscope portion).
  • the measurement light Lm is incident on the axicon lens 5, bent at an angle corresponding to the apex angle of the axicon lens 5, and then guided to the relay lens 6.
  • the axicon lens 5 is a conical lens also called a conical lens, and is used for forming an annular beam (donut shape) or a Bessel beam (non-diffractive-Bessel beam).
  • annular illumination is often introduced for use in extending the depth of focus using a Bessel beam or the like.
  • FIG. 1 illustrates a convex axicon lens
  • a concave axicon lens may be applied.
  • any member may be used as long as the measurement light Lm can be changed into an annular shape in the light speed cross section.
  • at least a part of the measurement light Lm may be changed into an annular shape by combining a plurality of mirrors.
  • the annular illumination is for obliquely incident measurement light on the sample 11 (details will be described later).
  • Measurement light (hereinafter referred to as measurement light Lm ′) formed in an annular shape by the axicon lens 5 is guided to the objective lens 9 via the relay lens 6 and the total reflection mirror 7.
  • annular pattern when the light that is formed in an annular shape by the axicon lens 5 and can be brought to the pupil position by the objective lens 9 is referred to as “annular pattern”, numerical examples of the annular pattern are as follows.
  • ⁇ 54 °.
  • the Z resolution is ⁇ 10 ⁇ m.
  • the depth of focus is about 200 / 0.8 mm to 250 ⁇ m.
  • the Z resolution (10 ⁇ m) and the depth of focus (250 ⁇ m) are balanced.
  • the observation target is a living body, This is particularly effective because it is suitable for structural observation.
  • the measurement light Lm ′ from the total reflection mirror 7 is condensed by the objective lens 9 and irradiated toward one point (condensing point) in the deep part of the sample 11.
  • the relative position in the z direction between the sample 11 and the objective lens 9 is adjusted in advance so that the focal plane of the objective lens 11 covers the region where the object to be observed (such as cultured cells) in the sample 11 is present.
  • at least one pupil (Fourier transform plane) of the illumination surface in the sample 11 exists between the relay lens 6 and the sample 11.
  • the pupil position is indicated by Fa in FIG. This pupil position is also the focal plane of the objective lens 9.
  • the measurement light Lm ′ is condensed on the focal plane to form a point light source (secondary light source), so that the sample 11 is illuminated with a parallel wave (collimated light).
  • irradiation spot In the sample 11 in the irradiation region of the measurement light Lm ′ (hereinafter referred to as “irradiation spot”), diffracted light with various angles may be generated. Of these diffracted lights, the light traveling in the same direction as the measurement light Lm ′ toward the condensing point is captured by the objective lens 10 through the aperture 13.
  • the specification of the objective lens 10 is the same as the specification of the objective lens 9 described above, and the arrangement destination of the objective lens 10 is a position facing the objective lens 9 with the sample 11 interposed therebetween.
  • the focal plane of the objective lens 10 coincides with the focal plane of the objective lens 9, and the focal point of the objective lens 10 coincides with the focal point of the objective lens 9.
  • the measurement light Lm the light captured by the objective lens 10 is referred to as “measurement light Lm”.
  • the measurement light Lm ′′ passes through the objective lens 10 and then enters the beam splitter 4.
  • the illumination light L0 emitted from the light source 1 is a collimating lens. 2 is collimated to a predetermined beam diameter by 2. Then, the measurement light Lm is formed in an annular shape by the axicon lens 5. Further, the measurement light Lm ′ having passed through the relay lens 6 is irradiated onto the sample 11 by the objective lens 9. At this time, the measurement light Lm ′ is obliquely incident on the sample 11. In the sample 11, diffracted light is generated according to the contents of the observation object of the sample 11. This diffracted light is used as the objective. It is captured by the lens 10.
  • the aperture 13 blocks the 0th-order diffracted light component, as shown in FIG. It is a stop for mainly taking out the scattered component by the sample 11 among the measurement light which has been arrange
  • the aperture 13 is the aperture 13 between the objective lens 10 and the beam splitter 4.
  • the aperture 13 is disposed on the light condensing surface (Fourier transform surface) of the objective lens 10 on which the 0th-order diffracted light component transmitted through the sample 11 is condensed. .
  • FIG. 3 shows an enlarged view of the vicinity of the aperture 13.
  • the aperture 13 has a predetermined aperture, and transmits only the diffracted light near the center (shaded portion in FIG. 3) among the diffracted light by the sample 11, and blocks the other light. .
  • the measurement light Lm ′′ incident on the beam splitter 4 is integrated with the reference light Lr ′ incident on the beam splitter 4 from the reference arm side and travels toward the cylindrical lens 15.
  • the integrated reference light Lr. 'And measurement light Lm' are collectively referred to as "interference light”.
  • the interference light directed toward the cylindrical lens 15 is guided by the cylindrical lens 15 to the entrance slit of the spectrum detector 16.
  • the interference light guided to the entrance slit of the spectrum detector 16 includes various light different from the light illuminated by the illumination optical system such as the light source 1 and the objective lens 9 (transmitted through the sample 11). Light, light reflected by the sample 11, and light diffracted by the sample 11.
  • the spectrum detector 16 includes a slit plate 16a having a slit opening at a condensing point of the interference light, a collimator mirror 16b for converting the interference light that has passed through the slit plate 16a into parallel light, A reflective diffraction grating 16c that separates the interference light that has become parallel light into a plurality of wavelength components, a condensing mirror 16d that condenses the wavelength components at positions shifted from each other, and each of the light that is condensed at positions shifted from each other. And a line sensor 16e for individually detecting the intensity of the wavelength component. With this configuration, the spectrum detector 16 generates an intensity signal (that is, a spectrum signal) for each wavelength component of the interference light. This spectrum signal is sent to the control device 17.
  • an intensity signal that is, a spectrum signal
  • the sample stage 12 described above can displace the sample 11 in the xy direction under the control of the control device 17. Therefore, when the sample stage 12 is driven, the irradiation spot on the sample 11 moves in the xy direction.
  • the form of spectral spectroscopy and detection of interference light is not limited to FIG.
  • control device 17 drives the sample stage 12 to perform two-dimensional scanning on the sample 11 in the xy direction with the irradiation spot, and when the irradiation spot is at each xy position, drives the line sensor 16e to obtain the spectrum signal. By taking in, the spectrum signal of each xy position is acquired. These spectrum signals are sent to the arithmetic unit 18.
  • the arithmetic unit 18 obtains structural information in the z direction at each xy position by individually Fourier transforming the spectrum signal at each xy position. Thereby, the three-dimensional image information in the xyz direction becomes known (details will be described later).
  • the arithmetic unit 18 displays the structural information of the sample 11 that has become known on a monitor (not shown).
  • the measurement light guided from the branching unit is obliquely incident on the object to be observed by using the axicon lens, and the reference light transmitted through the reference object and the object to be measured.
  • the measurement light transmitted through the observation object is combined, and the intensity of the combined light composed of the combined reference light and measurement light is detected. Therefore, it is possible to detect the internal structure of the observation object based on the light transmitted through the observation object.
  • the object to be observed is a transparent tissue or when the object to be observed is held in a highly reflective container, it is possible to perform suitable observation by OCT. it can.
  • FIG. 4 is a configuration diagram of the OCT apparatus according to the second embodiment. 4, the same elements as those shown in FIG. 1 are denoted by the same reference numerals. As shown in FIG. 4, in the OCT apparatus of this embodiment, the axicon lens 5 and the relay lens 6 shown in FIG. 1 are omitted, and a beam expander 21 and an annular mask 22 are arranged instead.
  • the beam expander 21 is disposed between the beam splitter 3 and the total reflection mirror 7 and converts the beam diameter. If the beam diameter of the light emitted from the light source 1 is sufficiently wide, the beam expander 21 need not be used.
  • the annular mask 22 is arranged at the pupil position of the objective lens 9 (the pupil position on the side opposite to the sample 11), guided from the beam splitter 3, and the measurement light Lm whose beam diameter is converted by the beam expander 21. Shield part of the light.
  • the annular zone mask 22 is a mask having an annular opening, and the annular zone diameter is d.
  • An annular pattern having a bandwidth ⁇ is formed at the pupil position of the objective lens 9 due to the balance between the aperture band width and the distance from the annular mask 22 to the pupil position of the objective lens 9. At this time, the relationship of the following equation holds for the depth of focus.
  • f represents the focal length of the objective lens 9
  • NA represents the effective NA of the objective lens.
  • annular pattern by the annular mask 22 are as follows.
  • ⁇ 54 °.
  • the Z resolution is ⁇ 10 ⁇ m.
  • the band width ⁇ of the above-described annular pattern is 200 ⁇ m
  • the depth of focus is about 200 / 0.8 mm to 250 ⁇ m.
  • the illumination light L0 emitted from the light source 1 is a collimating lens.
  • the measurement light Lm is converted to a predetermined beam diameter by the beam expander 21. Further, the measurement light Lm that has passed through the total reflection mirror 7 is applied to the annular mask 22.
  • the measurement light Lm is formed in an annular shape by the annular mask 22, and the measurement light Lm ′ formed in an annular shape is irradiated onto the sample 11 by the objective lens 9. At this time, the measurement light Lm ′ is In the same manner as in the first embodiment, the incident light is obliquely incident on the sample 11.
  • the aperture 13 blocks part of the transmitted light (0th-order diffracted light component). This is the same as in the first embodiment.
  • the measurement light Lm ′′ incident on the beam splitter 4 is integrated with the reference light Lr ′ incident on the beam splitter 4 from the reference arm side toward the cylindrical lens 15.
  • the subsequent cylindrical lens 15 and spectrum detection are performed.
  • the configurations and operations of the device 16, the control device 17, and the calculation device 18 are the same as those in the first embodiment.
  • the measurement light guided from the branching unit is obliquely incident on the object to be observed by using the annular mask, and the reference light transmitted through the reference object and the object to be observed.
  • the measurement light transmitted through the observation object is combined, and the intensity of the interference component contained in the combined light composed of the combined reference light and measurement light is detected. Therefore, the same effect as that of the first embodiment can be obtained.
  • a mask having a single opening may be used instead of the annular zone mask 22.
  • FIG. 7A in the case of a mask having a pinhole at one point corresponding to the x-axis shape, it has resolution in the x and z directions.
  • This method can reduce dark noise and make it easier to obtain an S / N ratio when a low-intensity light source is used.
  • a mask having a plurality of discrete or continuous openings on a substantially constant circle may be used instead of the annular mask 22.
  • a mask having pinholes at one point corresponding to the x-axis and one point corresponding to the y-axis has a three-dimensional resolution.
  • the mask in the case of a mask having pinholes at two symmetrical points corresponding to the x-axis and two symmetrical points corresponding to the y-axis, the mask has three-dimensional resolution. Furthermore, distortion of PSF can be suppressed and the depth of focus can be increased.
  • Openings are formed in an annular shape, and the larger N is set, the more suitable the zonal mask can be made.
  • the openings do not necessarily have to be arranged symmetrically. Further, the shapes of the plurality of openings are not necessarily the same.
  • the control device 17 controls the objective lens 9 and the annular mask 22 in conjunction with each other according to the sample 11. By such control, it becomes possible to irradiate measurement light from various directions to the object to be observed included in the sample 11, and to capture the three-dimensional structure evenly.
  • the numerical aperture is variable
  • the objective lens that condenses the measurement light guided from the branching unit on the object to be observed is provided, and the numerical aperture of the objective lens is determined according to the content of detection by the detecting unit. To control. Therefore, suitable detection according to the object to be observed can be made possible.
  • the resolution and depth of focus required for the OCT apparatus differ depending on what structure of what observation object is desired to be detected. For example, when an object to be observed is an observation object of a relatively large scale such as a cell spheroid, and it is desired to observe an overview of the object, a resolution and a depth of focus on the order of the spheroid size are required. In addition, when the object to be observed is a minute object such as a cell nucleus and it is desired to observe its detailed structure, high resolution is required even at the expense of the depth of focus. According to the processing as described above, the resolution can be appropriately changed according to the scale of the object to be observed.
  • annular pattern the light that is formed in an annular shape by an axicon lens or an annular mask and can be brought to the pupil position by the objective lens 9 is called an “annular pattern”. numerical examples of the annular pattern are as follows. .
  • the coherence length of the light source 1 is 2 ⁇ m
  • NA is 0.4 ⁇ NA ⁇ 0.8 mm
  • the band width of the annular pattern is 200 ⁇ m
  • NA sin ⁇
  • the range of ⁇ is 6 ° ⁇ ⁇ 55 ° It becomes.
  • the Z resolution at this time is 10 mm ( ⁇ m) ⁇ Z resolution ⁇ 50 mm ( ⁇ m)
  • the focal depth is 250 mm ( ⁇ m) ⁇ focus depth ⁇ 500 mm ( ⁇ m).
  • the Z resolution is 10 ( ⁇ m) to 50 mm ( ⁇ m) and the depth of focus is 250 ( ⁇ m) to 500 mm ( ⁇ m).
  • oblique incidence illumination may be applied to a reflection type OCT assumption. Specific examples are shown below.
  • FIG. 8 is a configuration diagram of an OCT apparatus according to this modification.
  • the OCT apparatus includes a light source 1, a beam splitter 3, an objective lens 9, a sample 11, a cylindrical lens 15, a spectrum detector 16, a control device 17, a calculation device 18 and the like similar to those in the first embodiment.
  • a plane mirror (reference mirror) 41 and an optical scanner 42 are arranged.
  • the illumination light L0 emitted from the light source 1 enters the beam splitter 3 and is branched into a reference light Lr traveling toward the reference mirror 41 and a measurement light Lm traveling toward the sample 11.
  • the reference light Lr When the reference light Lr is incident on the reference mirror 41 from the front, the reference light Lr is reflected by the reference mirror 41 and returns to the beam splitter 3.
  • the measurement light Lm when the measurement light Lm is incident on the objective lens 9 via the optical scanner 42, the measurement light Lm receives the light condensing action of the objective lens 9 and is condensed toward one point (condensing point) in the deep part of the sample 11.
  • the relative position in the z direction between the sample 11 and the objective lens 9 is adjusted in advance so that the focal plane of the objective lens 9 is applied to a region where an object to be observed (such as cultured cells) in the sample 11 is present.
  • irradiation spot In the irradiation area of the measurement light Lm in the sample 11 (hereinafter referred to as “irradiation spot”), reflected light of various angles may be generated. Of the reflected light, the light that follows the optical path of the measurement light Lm toward the condensing point in the opposite direction is captured by the objective lens 9. Hereinafter, the light captured by the objective lens 9 out of the reflected light from the irradiation spot toward the objective lens 9 will be referred to as “measurement light Lm ′”. The measurement light Lm ′ follows the optical path of the measurement light Lm in the reverse direction and enters the beam splitter 3 via the optical scanner 42.
  • the measurement light Lm ′ incident on the beam splitter 3 is integrated with the reference light Lr ′ returned to the beam splitter 3 and the optical path, and travels toward the cylindrical lens 15.
  • the optical path length of the single optical path of the reference light optical path length of the reference arm
  • the optical path length of the single optical path of the measurement light optical path length of the measurement arm
  • the integrated reference light Lr ′ and measurement light Lm ′′ are collectively referred to as “interference light”.
  • the interference light directed toward the cylindrical lens 15 is guided by the cylindrical lens 15 to the entrance slit of the spectrum detector 16.
  • the configuration and operation of the spectrum detector 16 are the same as those in the first embodiment.
  • the above-described condensing point moves in the field of view of the objective lens 9, so that the irradiation spot on the sample 11 moves in the xy direction.
  • control device 17 drives the optical scanner 42 to perform two-dimensional scanning on the sample 11 in the xy direction with the irradiation spot, and when the irradiation spot is at each xy position, drives the line sensor 16e to output the spectrum signal. By taking in, the spectrum signal of each xy position is acquired. These spectrum signals are sent to the arithmetic unit 18.
  • the arithmetic unit 18 obtains structural information in the z direction at each xy position by individually Fourier transforming the spectrum signal at each xy position. As a result, the cell distribution in the xyz direction becomes known. The arithmetic unit 18 displays the distribution of the structural information in the sample 11 that has become known on a monitor (not shown).
  • the control device 17 controls the numerical aperture NA of the objective lens 9 according to the sample 11. By such control, it becomes possible to irradiate measurement light from various directions to the object to be observed included in the sample 11, and to capture the three-dimensional structure evenly.
  • FIG. 9 is a configuration diagram of the OCT apparatus of the third embodiment.
  • the same elements as those shown in FIG. in the OCT apparatus of this embodiment, the axicon lens 5 and the relay lens 6 shown in FIG. 1 are omitted, and an optical scanner 31 is arranged instead.
  • the optical scanner 31 includes two total reflection mirrors 31a and 31b, and changes the incident angle of the measurement light Lm to the objective lens 9.
  • the optical scanner 31 is controlled by the control device 17.
  • the controller 17 controls the optical scanner 31, the total reflection mirror 7, and the total reflection mirror 8 in conjunction with each other so that the measurement light Lm ′ is obliquely incident on the sample 11.
  • the controller 17 controls the optical scanner 31, the total reflection mirror 7, and the total reflection mirror 8 in conjunction with each other so that the measurement light Lm ′ is obliquely incident on the sample 11.
  • the irradiation spot that is obliquely incident is rotated and the irradiation is performed in a ring shape, substantially the same detection as in the first embodiment and the second embodiment described above can be performed.
  • Such a method can be expected to reduce dark noise and easily obtain an SN ratio when a low-intensity light source is used as the light source.
  • the measurement light Lm ′ is obliquely incident on the sample 11 by the above-described optical scanner 31 as in the first embodiment.
  • the configuration and operation of the subsequent aperture 13, beam splitter 4, cylindrical lens 15, spectrum detector 16, control device 17, and calculation device 18 are the same as those in the first embodiment.
  • the measurement light guided from the branching unit is obliquely incident on the object to be observed by using the optical scan including the annular mirror, and is transmitted through the reference object.
  • the reference light and the measurement light transmitted through the object to be observed are combined, and the intensity of the combined light composed of the combined reference light and measurement light is detected. Therefore, the same effect as that of the first embodiment can be obtained.
  • oblique incidence may be realized by electrical control using a micro device mirror, a DOE (high precision diffractive optical element), a spatial modulation element such as liquid crystal, and the like.
  • the measurement light may be formed in an annular shape using the spatial modulation element described above.
  • the OCT apparatus of each embodiment described above employs a method of displacing the sample 11 side by moving the sample stage 12 (stage scan type), but a method of displacing the irradiation spot side (beam scan type). May be adopted. Two methods may be selectively used or may be used in combination.
  • FIG. 10 is a configuration diagram of a part of a beam scan type OCT apparatus.
  • the part corresponding to between the total reflection mirror 7 and the beam splitter 4 in FIG. 4 is illustrated.
  • the sample stage 12 is omitted, and each part of the total reflection mirror 51, the mirror pairs 52a and 52b, the mirror pairs 53a and 53b, and the total reflection mirror 54 is provided.
  • the objective lens 9, the objective lens 10, and the aperture 13 are fixed.
  • the mirror pair 52a and 52b are arranged so that the center of the mirror pair 52a and 52b is the pupil position of the objective lens 9 (Fa in FIG. 10), and the center of the mirror pair 53a and 53b is the pupil position of the objective lens 10 (Fb in FIG.
  • the control apparatus 17 scans by controlling each part of mirror pair 52a and 52b and mirror pair 53a and 53b synchronously.
  • the mirror pair 52a and 52b has a three-dimensional structure as shown in FIG. 11, and emits light with an incident light velocity bent by 90 degrees. The same applies to the mirror pairs 53a and 53b.
  • the OCT apparatus of each embodiment mentioned above employ
  • a method of detecting light in a time division manner may be employed.
  • the interference light intensity at each position in the xy direction on the sample 11 can be collectively displayed. It may be detected.
  • the OCT apparatus of each embodiment mentioned above employ
  • the time domain type when the time domain type is adopted, it is not necessary to perform spectral detection, so by using an image sensor instead of the spectrum detector 18, the interference light intensity at each position in the xy direction on the sample 11 is collectively displayed. It may be detected.
  • a polarization component may be used. For example, a difference is provided between the polarization direction of the reference light toward the detection means and the polarization direction of the measurement light toward the object to be observed, and interference light is included from the combined light of the reference light and measurement light toward the detection means. Detection using the polarization component can be performed by transmitting the polarization component in the direction and removing the polarization component in the direction not including the interference light.
  • SYMBOLS 1 Light source, 3, 4 beam splitter, 5 ... Axicon lens, 9, 10 ... Objective lens, 11 ... Sample, 13 ... Aperture, 14 ... Optical member for dispersion correction, 16 ... Spectrum detector, 17 ... Control apparatus, 18 ... arithmetic unit, 22 ... annular mask, 31 ... optical scanner

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  • General Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
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  • Optics & Photonics (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)

Abstract

L'invention concerne un dispositif d'observation comprenant : des moyens de répartition pour répartir la lumière incidente en une lumière de référence et une lumière de mesure ; un système optique d'éclairage qui illumine un sujet à observer avec la lumière de mesure le long de la direction prédéterminée ; un système optique d'observation qui est disposé sur un côté différent du système optique d'éclairage par rapport au sujet à observer et reçoit la lumière d'observation voyageant dans une direction différente de la direction prédéterminée parmi la lumière de mesure passant à travers le sujet à observer ; un système optique de combinaison qui guide la lumière de référence vers le système optique d'observation pour combiner la lumière de référence et la lumière d'observation ; et un dispositif de détection qui reçoit la lumière de référence et la lumière d'observation, lesquelles sont combinées par le système optique de combinaison, pour détecter l'intensité parasite de la lumière de référence et de la lumière d'observation, permettant ainsi la détection en se basant sur la transmission de la lumière à travers le sujet à observer.
PCT/JP2012/002048 2011-03-24 2012-03-23 Dispositif d'observation et procédé d'observation Ceased WO2012127880A1 (fr)

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Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE202012008196U1 (de) 2012-08-28 2013-03-08 Martin Eberlein Mit gleichen Wagen stapelbarer Einkaufswagen
JP2015504177A (ja) * 2011-12-20 2015-02-05 エコール ポリテクニク 成形されたビームを用いた定量用非線形光学顕微鏡
JP2015049204A (ja) * 2013-09-04 2015-03-16 株式会社日立エルジーデータストレージ 光計測装置及び光断層観察方法
JP2015072152A (ja) * 2013-10-02 2015-04-16 株式会社日立エルジーデータストレージ 光計測装置
JP2017064378A (ja) * 2015-09-30 2017-04-06 キヤノン株式会社 光干渉断層撮影装置、その制御方法および光干渉断層撮影システム
JP2019536997A (ja) * 2016-10-11 2019-12-19 マルバーン パナリティカル リミテッド 粒子特性測定装置
CN111220625A (zh) * 2020-01-18 2020-06-02 哈尔滨工业大学 表面及亚表面一体化共焦显微测量装置和方法
CN111239154A (zh) * 2020-01-18 2020-06-05 哈尔滨工业大学 一种横向差动暗场共焦显微测量装置及其方法
CN111239155A (zh) * 2020-01-18 2020-06-05 哈尔滨工业大学 一种轴向差动暗场共焦显微测量装置及其方法
US20200376603A1 (en) * 2018-02-15 2020-12-03 Schott Ag Methods and devices for introducing separation lines into transparent brittle fracturing materials
JP2022179503A (ja) * 2018-11-27 2022-12-02 株式会社リコー 面発光レーザ、光源装置及び眼球の傾き位置検出装置
US12392512B2 (en) 2020-02-03 2025-08-19 Guangzhou T.K Medical Instrument Co., Ltd. Air sterilization device with heating apparatus

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH075101A (ja) * 1993-06-18 1995-01-10 Fuji Photo Film Co Ltd 光散乱媒体の吸光計測装置
JP2001066247A (ja) * 1999-08-26 2001-03-16 Japan Science & Technology Corp 光計測装置
JP2001330558A (ja) * 2000-05-25 2001-11-30 Japan Science & Technology Corp ２次元光ヘテロダイン検出法を用いた光画像計測装置
JP2005530128A (ja) * 2002-01-11 2005-10-06 ザ・ジェネラル・ホスピタル・コーポレイション 解像度と深さ領域を改善するための軸方向線焦点を用いたｏｃｔ撮像用装置
JP2005338567A (ja) * 2004-05-28 2005-12-08 Nikon Corp 光学顕微鏡および顕微鏡システム
JP2006280449A (ja) * 2005-03-31 2006-10-19 Fujinon Corp 画像診断装置
JP2007510143A (ja) * 2003-10-27 2007-04-19 ザ・ジェネラル・ホスピタル・コーポレイション 周波数ドメイン干渉測定を利用して光学撮像を実行する方法および装置

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2918992B2 (ja) * 1990-05-28 1999-07-12 科学技術振興事業団 ヘテロダイン検波受光系を用いた振幅像の検出装置
JP3076016B2 (ja) * 1997-11-13 2000-08-14 株式会社生体光情報研究所 光計測装置
SMT202000624T1 (it) * 2010-03-05 2021-01-05 Massachusetts Gen Hospital Apparecchio per fornire radiazione elettromagnetica a un campione

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH075101A (ja) * 1993-06-18 1995-01-10 Fuji Photo Film Co Ltd 光散乱媒体の吸光計測装置
JP2001066247A (ja) * 1999-08-26 2001-03-16 Japan Science & Technology Corp 光計測装置
JP2001330558A (ja) * 2000-05-25 2001-11-30 Japan Science & Technology Corp ２次元光ヘテロダイン検出法を用いた光画像計測装置
JP2005530128A (ja) * 2002-01-11 2005-10-06 ザ・ジェネラル・ホスピタル・コーポレイション 解像度と深さ領域を改善するための軸方向線焦点を用いたｏｃｔ撮像用装置
JP2007510143A (ja) * 2003-10-27 2007-04-19 ザ・ジェネラル・ホスピタル・コーポレイション 周波数ドメイン干渉測定を利用して光学撮像を実行する方法および装置
JP2005338567A (ja) * 2004-05-28 2005-12-08 Nikon Corp 光学顕微鏡および顕微鏡システム
JP2006280449A (ja) * 2005-03-31 2006-10-19 Fujinon Corp 画像診断装置

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
DING Z ET AL.: "High-resolution optical coherence tomography over a large depth range with an axicon lens", OPTICS LETTERS, vol. 27, no. 4, 15 February 2002 (2002-02-15), pages 243 - 245, XP007906548, DOI: doi:10.1364/OL.27.000243 *

Cited By (16)

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JP2015504177A (ja) * 2011-12-20 2015-02-05 エコール ポリテクニク 成形されたビームを用いた定量用非線形光学顕微鏡
US9791682B2 (en) 2011-12-20 2017-10-17 Ecole Polytechnique Quantitative nonlinear optical microscopy using a shaped beam
DE202012008196U1 (de) 2012-08-28 2013-03-08 Martin Eberlein Mit gleichen Wagen stapelbarer Einkaufswagen
JP2015049204A (ja) * 2013-09-04 2015-03-16 株式会社日立エルジーデータストレージ 光計測装置及び光断層観察方法
JP2015072152A (ja) * 2013-10-02 2015-04-16 株式会社日立エルジーデータストレージ 光計測装置
JP2017064378A (ja) * 2015-09-30 2017-04-06 キヤノン株式会社 光干渉断層撮影装置、その制御方法および光干渉断層撮影システム
JP2019536997A (ja) * 2016-10-11 2019-12-19 マルバーン パナリティカル リミテッド 粒子特性測定装置
JP7114580B2 (ja) 2016-10-11 2022-08-08 マルバーン パナリティカル リミテッド 粒子特性測定装置
US20200376603A1 (en) * 2018-02-15 2020-12-03 Schott Ag Methods and devices for introducing separation lines into transparent brittle fracturing materials
US12311469B2 (en) * 2018-02-15 2025-05-27 Schott Ag Methods and devices for introducing separation lines into transparent brittle fracturing materials
JP2022179503A (ja) * 2018-11-27 2022-12-02 株式会社リコー 面発光レーザ、光源装置及び眼球の傾き位置検出装置
JP7388507B2 (ja) 2018-11-27 2023-11-29 株式会社リコー 瞳孔又は角膜の位置検知装置用面発光レーザ、光源装置及び眼球の傾き位置検出装置
CN111239155A (zh) * 2020-01-18 2020-06-05 哈尔滨工业大学 一种轴向差动暗场共焦显微测量装置及其方法
CN111239154A (zh) * 2020-01-18 2020-06-05 哈尔滨工业大学 一种横向差动暗场共焦显微测量装置及其方法
CN111220625A (zh) * 2020-01-18 2020-06-02 哈尔滨工业大学 表面及亚表面一体化共焦显微测量装置和方法
US12392512B2 (en) 2020-02-03 2025-08-19 Guangzhou T.K Medical Instrument Co., Ltd. Air sterilization device with heating apparatus

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