JP2006284686A - Optical microscopic device and microscopic observation method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an optical microscopic device which can be made compact, and a microscopic observation method using the optical microscopic device. <P>SOLUTION: The optical microscopic device 1 includes: an illumination means 7 having a condenser lens 11 outputting light emitted from a light source 9 as converged light; a reflection means 13 reflecting the converged light output from the condenser lens to the illumination means side; an object-to-be-inspected placing table 3 which is provided between the illumination means and the reflection means and on which an object to be inspected is placed; and an objective lens 15 provided on an opposite side to the object-to-be-inspected placing table with respect to the reflection means. The converged light output from the illumination means and reflected by the reflection means is radiated to the object to be inspected. The reflection means has a light passing part 13a through which the light reflected on the object to be inspected passes, and the light reflected on the object to be inspected is made incident on the objective lens through the light passing part after it is converged on one point 23 in space between the reflection means and the object to be inspected. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to an optical microscope apparatus and a microscope observation method suitable for observing the structure of various materials. Examples of materials that can be observed in the optical microscope apparatus and microscope observation method of the present invention include polymer materials such as polyethylene film and injection-molded polypropylene composite materials, biomaterials such as plants and pathological tissues, coating liquids, and emulsions. And suspensions such as semiconductor materials.

  Since the structure of various materials is closely correlated with their physical properties, it is important to accurately evaluate and analyze the structure. For this purpose, many techniques have been developed and utilized, but among them, the optical microscope apparatus is most commonly used as a method for observing the structure of materials because of its ease of use and the diversity of information obtained. It is a method.

  In the conventional optical microscope apparatus, as an illumination method for the inspection object, a Koehler illumination method in which parallel light is incident is usually used in order to uniformly irradiate the inspection object and increase the resolution of the image. .

However, the conventional optical microscope using the Koehler illumination method in which parallel light is incident only observes an image caused by the intensity of light from the object illuminated by the illumination light. Features such as presence / absence and degree of orientation could not be observed. In order to solve such a problem, an optical microscope apparatus including an illuminating unit that irradiates convergent light that converges to one point in space as illumination light and an observation method thereof are known (for example, Patent Document 1). Here, this microscope is called a convergent light microscope.
JP 2001-264638 A

  In such a convergent light microscope, convergent light is output from the illuminating means to irradiate the inspection object. The objective lens is arranged so that light from the object to be inspected once converges at the convergence point and then enters the objective lens. In this case, since the diffraction image of the inspection object is formed on the surface including the convergence point, the diffraction image and the inspection object are aligned by adjusting the position of the convergence point and the focus of the objective lens to the inspection object, respectively. An optical image can be observed.

  As a result, features such as the presence or absence of anisotropy and orientation degree in the tissue structure as described above, which could not be observed with the Koehler illumination method, can be observed with a conventional focusing microscope. There has been a demand for compactness.

  Therefore, an object of the present invention is to provide an optical microscope apparatus that can be made compact and a microscope observation method using the optical microscope apparatus.

  The optical microscope apparatus and the microscope observation method of the present invention have been made in order to solve such problems.

  An optical microscope apparatus according to the present invention includes an illuminating unit having a condenser lens that outputs light emitted from a light source as convergent light, a reflecting unit that reflects convergent light output from the condenser lens toward the illuminating unit, and an illuminating unit. Provided between the reflecting means and an object mounting table for placing the object to be inspected, and an objective lens provided on the opposite side of the object mounting table with respect to the reflecting means. The convergent light output from the means and reflected by the reflecting means is applied to the object to be inspected, and the reflecting means has a light passing portion through which the light reflected by the object to be inspected, and the light reflected by the object to be inspected is The light beam is converged at a point on the space between the objective lens and the object to be inspected and then enters the objective lens, and enters the objective lens through the light transmitting portion.

  In this configuration, the convergent light output from the illuminating means is once reflected by the reflecting means, and then irradiated to the inspection object on the inspection object mounting table provided between the illuminating means and the reflecting means. Then, the light reflected by the inspection object illuminated by the convergent light converges at one point on the space between the objective lens and the inspection object, and passes through the light passing portion of the reflecting means to the objective lens. Incident. In this way, since the convergent light is reflected once by the reflecting means and irradiated onto the object to be inspected, the length of the objective lens in the optical axis direction can be shortened, and as a result, the optical microscope can be made compact. be able to.

  In addition, a Fourier transform image, that is, a diffraction image of the object to be inspected by the convergent light as the illumination light is formed on a surface including a point on the space where the light reflected from the object to be converged (hereinafter referred to as “convergence point”). Is done. According to the above optical microscope apparatus, this diffraction image can be formed between the objective lens and the object to be inspected, so that the diffraction image itself can be observed or a desired process can be performed on the diffraction image. Can be applied.

  In addition, structural information of the inspection object is collected in the diffraction image. In other words, a diffraction image corresponding to the tissue structure of the inspection object is formed. If the tissue structure of the inspection object is different, the diffraction image is also different. Therefore, if the relationship between the tissue structure and the diffraction image is known, the tissue structure of the object to be inspected can be known from the diffraction image.

  In the optical microscope apparatus according to the present invention, it is preferable that the optical microscope apparatus further includes a convergence angle changing unit that changes the convergence angle of the light reflected from the inspection object in a range of 0 to 90 °. In this case, the effect of the high contrast and the depth of focus can be changed by changing the convergence angle by the convergence angle changing means.

  Furthermore, the optical microscope apparatus according to the present invention includes a focus of the objective lens on both the diffraction image plane perpendicular to the optical axis of the objective lens and the object to be examined, including one point on the space where the light reflected by the object is converged. It is preferable to provide a focusing adjustment means for adjusting the level. Thereby, an optical image and a diffraction image of the inspection object can be observed, respectively, and more structural information of the inspection object can be acquired.

  Furthermore, in the optical microscope apparatus of the present invention, the relative position between the diffracted image plane perpendicular to the optical axis of the objective lens including one point on the space where the light reflected from the object is converged is determined. It is preferable to further include an adjustment mechanism for changing.

  In this case, it is possible to change the size of the diffraction image by changing the relative position between the diffraction image surface and the object to be inspected. For example, the larger the distance from the relative position, the larger the diffraction image. be able to. Further, when the convergence angle changing means is provided, the convergence angle can be made constant by the convergence angle changing means even if the relative position between the diffraction image plane and the inspection object is changed. As a result, the inspection object and the diffraction image can be observed while maintaining the effects of high contrast and depth of focus constant.

  Further, the optical microscope apparatus according to the present invention is disposed at a position of a diffraction image plane that includes one point on the space where the light reflected by the inspection object is converged and is orthogonal to the optical axis of the objective lens. It is preferable to further include a spatial aperture that selectively allows a portion of the reflected light to pass through.

  Since this spatial stop allows a part of the light reflected from the inspection object to be selected, it is possible to select a desired diffracted light from the inspection object and enter the objective lens. Here, the diffracted light means to include 0th-order diffracted light (that is, direct light). After selecting the diffracted light, if the focus of the objective lens is adjusted to the object to be inspected, an optical image of the object to be inspected by the selected desired diffracted light can be observed. Since the diffracted light can be freely selected, various optical images of the inspection object corresponding to the desired diffracted light can be observed for the same inspection object.

  Furthermore, in the optical microscope apparatus of the present invention having a spatial diaphragm, a shielding object that shields part of the convergent light so that the convergent light does not illuminate the spatial diaphragm is further provided between the object mounting table and the light source. It is preferable. In this configuration, since the converging light is not applied to the space stop by blocking a part of the converging light with the shielding object, the converging light irradiated to the inspection object is not easily affected by the space stop. Thereby, for example, even if the spatial aperture is taken in and out or the type of the spatial aperture is changed, the irradiation condition of the convergent light to the inspection object is hardly changed.

  Moreover, in the optical microscope apparatus according to the present invention including a spatial aperture, it is preferable to further include an adjustment mechanism that substantially matches the direction of light that has passed through the spatial aperture and the optical axis of the objective lens. Although the amount of light is reduced by the spatial diaphragm, a bright image with little distortion can be obtained by making the direction of light passing through the spatial diaphragm substantially coincide with the optical axis of the objective lens.

  The objective lens can be focused on the entire field of view by arranging the objective lens so that its focal plane is parallel to the object to be inspected. In this case, the “optical axis of the objective lens” refers to a direction connecting the center of the objective lens and the intersection of the optical axis of the object to be inspected and the condenser lens.

  Furthermore, in the optical microscope apparatus according to the present invention, monochromatic light may be used as the light output from the light source. By using monochromatic light, it is possible to obtain an image that is important for knowing the tissue structure, which cannot be obtained with white light.

  One of the microscope observation methods of the present invention using the optical microscope apparatus according to the present invention is characterized by observing the inspection object with the focusing of the objective lens aligned with the inspection object. As described above, the optical microscope apparatus of the present invention is compact, and the convergent light output from the condenser lens is reflected by the reflecting means and applied to the object to be inspected.

  Then, the light reflected by the object to be inspected converges to one point on the space between the object to be inspected and the objective lens, and enters the objective lens through the light passing portion. In this case, since the inspecting object is irradiated with light that is converging, an observation image with a very high contrast and a deep depth of focus can be obtained by observing the inspecting object while focusing. .

  Further, another microscope observation method of the present invention using the optical microscope apparatus according to the present invention has a diffraction image plane orthogonal to the optical axis of the objective lens including one point on the space where the light reflected from the object to be inspected converges. By focusing the objective lens, the diffraction image of the object to be inspected formed on the diffraction image surface is observed.

  As described above, the optical microscope apparatus of the present invention is compact, and the convergent light output from the condenser lens is reflected by the reflecting means and applied to the object to be inspected. The light reflected by the object to be inspected converges at one point on the space between the object to be inspected and the objective lens, and enters the objective lens through the light passing portion.

  In this case, a diffraction image of the inspection object is formed on the diffraction image surface. In the above observation method, since the diffraction image is observed by focusing on the diffraction image, if the relationship between the diffraction image and the tissue structure is acquired in advance for the object to be inspected, the characteristics of the pattern of the diffraction image are obtained. It is possible to know the tissue structure of the inspection object.

  Further, the microscope observation method of the present invention using the optical microscope apparatus according to the present invention provided with the focusing adjustment means includes a step of observing the inspection object by aligning the focusing of the objective lens with the inspection object, and the objective lens. And observing the diffraction image in accordance with the diffraction image formed on the diffraction image surface.

  As described above, the optical microscope apparatus of the present invention is compact, and the convergent light output from the condenser lens is reflected by the reflecting means and applied to the object to be inspected. The light reflected by the object to be inspected converges at one point on the space between the object to be inspected and the objective lens, and enters the objective lens through the light passing portion. In this case, a diffraction image of the inspection object is formed on the diffraction image surface.

  In the above observation method, it is difficult to understand only by observing the optical image by observing the optical image and the diffraction image of the inspection object by adjusting the focusing of the objective lens to the inspection object and the diffraction image plane. It is possible to grasp the overall characteristics of the tissue structure and to know the details of the tissue structure from which the diffraction image, which was difficult to understand simply by observing the diffraction image, can be obtained.

  In addition, the microscope observation method according to the present invention using the optical microscope apparatus according to the present invention having a spatial aperture allows the light in a desired region on the diffraction image plane to pass through using the spatial aperture, and focuses the objective lens. The object to be inspected is observed using light that has passed through a spatial aperture in accordance with the object to be inspected.

  As described above, the optical microscope apparatus of the present invention is compact, and the convergent light output from the condenser lens is reflected by the reflecting means and applied to the object to be inspected. The light reflected by the object to be inspected converges at one point on the space between the object to be inspected and the objective lens, and enters the objective lens through the light passing portion. In this case, a diffraction image of the inspection object is formed on the diffraction image surface.

  And in the said microscope observation method, desired diffracted light can be selected by allowing desired light on a diffraction image surface to pass through using a space stop. As a result, it is possible to observe an optical image of the inspection object with the selected desired diffracted light. Since the diffracted light can be freely selected, various optical images corresponding to the diffracted light can be observed for the same inspection object, and as a result, the tissue structure of the inspection object can be known in more detail.

  Furthermore, another microscope observation method of the present invention using the optical microscope apparatus according to the present invention provided with a spatial aperture is to adjust the focus of the objective lens to the diffraction image surface, thereby forming an object formed on the diffraction image surface. After observing the diffraction image of the inspection object and adjusting the spatial aperture so that the light in the desired region of the diffraction image can pass through, the focus of the objective lens is adjusted to the inspection object, so that the light passing through the spatial aperture is It is characterized by observing an object to be inspected.

  As described above, the optical microscope apparatus of the present invention is compact, and the convergent light output from the condenser lens is reflected by the reflecting means and applied to the object to be inspected. Then, the light reflected by the object to be inspected converges to one point on the space between the object to be inspected and the objective lens, and enters the objective lens through the light passing portion. In this case, a diffraction image of the inspection object is formed on the diffraction image surface. And in the said observation method, since the diffracted light used for observation of an optical image is selected based on a diffraction image, it can be known what kind of diffracted light is an optical image. Thereby, the tissue structure of the object to be inspected can be known in more detail.

  In another microscope observation method of the present invention, it is preferable to adjust the position of the diffraction image plane and observe the inspection object. Since the diffracted image plane is a position where convergent light converges, the amount of light incident on the objective lens can be adjusted by adjusting the position of the diffracted image plane, and as a result, the brightness of the image to be observed can be adjusted. . For example, by observing the inspection object by adjusting the position of the diffraction image surface so that the inspection object is focused when the objective lens is positioned in the vicinity of the diffraction image surface, the diffraction light is not lost. Since it can enter the objective lens, the image is brightest.

  In the microscope observation method of the present invention, the object to be inspected by the objective lens is changed by changing the shape of the spatial aperture or the position on the diffraction image plane, or by changing the angle of the optical axis of the condenser lens with respect to the optical axis of the objective lens. It is preferable to select the diffracted light that participates in the optical image formation of the object and observe the object to be inspected.

  In the microscope observation method of the present invention, it is preferable to observe the object to be inspected with the direction of the light passing through the space stop and the optical axis of the objective lens being substantially matched. Since the direction of the light that has passed through the spatial stop and the optical axis of the objective lens substantially coincide with each other, a bright image with little distortion can be obtained.

  The objective lens can be focused on the entire field of view by arranging the objective lens so that its focal plane is parallel to the object to be inspected. In this case, the “optical axis of the objective lens” refers to a direction connecting the center of the objective lens and the intersection of the optical axis of the object to be inspected and the condenser lens.

  In the microscope observation method of the present invention, it is preferable to adjust the size of the diffraction image by changing the position of one point in the space in the optical axis direction of the objective lens. Thereby, a diffraction image of a desired size can be observed.

  At this time, it is preferable that the convergence angle at the convergence point is not changed even if the position of one point (convergence point) in the space described above is changed in the optical axis direction of the objective lens. When the convergence angle is constant, even if the relative position between the diffraction image plane and the object to be inspected is changed, observation can be performed while maintaining the effects of high contrast and depth of focus. .

  Moreover, in the microscope observation method of this invention, the light emitted from a light source can be made into monochromatic light.

  Furthermore, a polymer material can be considered as an object to be inspected by the microscope observation method according to the present invention.

  According to the optical microscope apparatus of the present invention, the optical microscope apparatus can be made compact. In addition, according to the microscope observation method of the present invention, a microscope can be observed using a compact optical microscope.

  DESCRIPTION OF EMBODIMENTS Hereinafter, preferred embodiments of an optical microscope according to the present invention and a microscope observation method using the optical microscope will be described in detail with reference to the drawings.

  FIG. 1 is a schematic configuration diagram showing a basic configuration of an embodiment of an optical microscope according to the present invention. The optical microscope apparatus 1 has a stage 3 as an inspection object mounting table. The optical microscope apparatus 1 irradiates an inspection object (specimen) 5 on the stage 3 with illumination light from the inspection object 5. Various observations are performed using the reflected light.

  The optical microscope apparatus 1 has an illuminating means 7 that outputs illumination light that irradiates the inspection object 5, and the illuminating means 7 is below the stage 3, that is, on the back side of the stage 3 (the inspection object 5 is It is arranged on the opposite side to the surface to be placed. The illumination means 7 includes a point light source 9 and a condenser lens 11, and the light output from the point light source 9 is converged by the condenser lens 11. Thereby, the illumination light as the convergent light is output from the illumination means 7. The light output from the point light source 9 may be white light or monochromatic light.

  In order to irradiate the inspection object 5 with the illumination light output from the illumination unit 7, the optical microscope apparatus 1 has a reflection unit 13 provided on the opposite side of the stage 3 from the illumination unit 7. The reflecting means 13 reflects the illumination light so as to be folded back to the condenser lens 11 side. Although the mirror is illustrated as the reflection means 13, if it can reflect illumination light to the condenser lens 11 side, it will not be specifically limited.

  The reflecting means 13 is arranged so as to be orthogonal to the optical axis L1 of the condenser lens 11 and irradiates the inspection object 5 with the illumination light output from the illumination means 7. The reflecting means 13 has an opening 13a on the optical axis L1 as a light transmitting portion for allowing the light reflected from the inspection object 5 that has received the illumination light to pass through.

  The opening 13a is, for example, a circular opening with the optical axis L1 as the center, but is not particularly limited as long as light from the inspection object 5 is allowed to pass through. Further, the size of the opening 13a is not particularly limited as long as the area of the region for reflecting the illumination light by the reflecting means 13 is sufficiently large and the size does not block the light reflected from the inspection object 5.

  Since the stage 3 is arranged between the illumination means 7 and the reflection means 13 as described above, the stage 3 and the inspection object 5 should be as small as possible so that a large amount of illumination light is irradiated onto the inspection object 5. Is preferred.

  In order to form an image using the light passing through the opening 13a, the optical microscope apparatus 1 reflects the lens barrel 21 in which the objective lens 15, the imaging lens 17, and the eyepiece lens 19 are arranged in order from the reflecting means 13 side. Above the means 13 (upper side in the figure). The internal configuration itself of the lens barrel 21 is a conventional one, and focusing can be performed by changing the relative distance from the inspection object 5. The lens barrel 21 is arranged so that the optical axis L2 of the objective lens 15 substantially coincides with the optical axis L1 of the condenser lens 11.

  The objective lens 15 has a focal length such that the position when the object 5 is focused is behind the reflecting means 13 (upper side in the figure). Thereby, the reflecting means 13 does not get in the way in the focusing operation.

  The image captured by the objective lens 15 is formed at an intermediate image position 18 behind the imaging lens 17 (upper side in the figure), and the eyepiece 19 is adjusted in focus so that the image can be observed. Yes.

  In the above configuration, the illumination unit 7, the stage 3, and the reflection unit 13 are positioned so that the convergence point 23 of the converged light output from the illumination unit 7 is located between the object 5 to be inspected and the objective lens 15. Yes.

  More specifically, the reflecting means 13 is disposed closer to the condenser lens 11 than the position where the convergent light output from the condenser lens 11 converges at one point on the optical axis L1. Further, the stage 3 is arranged so that the illumination light reflected by the reflecting means 13 is irradiated on the inspection object 5 before the illumination light further converges on the optical axis L1. As a result, convergent light that is converging is irradiated onto the inspection object 5, so that the convergent light reflected by the inspection object 5 is on the space between the inspection object 5 and the objective lens 15. Converge to one point.

  Then, the illumination light as the convergent light passes through a convergence point 23 that is one point on the converged space after being reflected by the inspection object 5, and a plane orthogonal to the optical axis L2 of the objective lens 15 is covered with the illumination light. A Fourier transform image, that is, a diffraction image of the inspection object 5 is formed. Here, the plane P1 is referred to as a diffraction image plane P1.

  A focusing adjustment means 25 is attached to the lens barrel 21 in order to observe the diffraction image formed on the diffraction image plane P1 and the image of the inspection object 5. The focusing adjustment means 25 changes the distance between the lens barrel 21 and the stage 3. Accordingly, the relative position between the objective lens 15 and the inspection object 5, that is, the distance between the objective lens 15 and the inspection object 5 (distance d1 in FIG. 1) can be changed. Semi-alignment can be performed.

  The focusing adjustment means 25 uses, for example, a rack and pinion, and may be the same as that attached to a lens barrel or the like of a conventional optical microscope, but can be changed for focusing. The range of the relative position needs to be sufficiently longer than that of a conventional general microscope. That is, focusing can be performed at least on both the inspection object 5 and the diffraction image plane P1.

  Accordingly, the optical image of the inspection object 5 can be observed by adjusting the focusing of the objective lens 15 to the inspection object 5, and the diffraction image can be observed by adjusting the focusing of the objective lens 15 to the diffraction image plane P1. can do.

  Since the structure information of the inspection object 5 is collected in the diffraction image, if the relationship between the tissue structure and the diffraction image is known in advance, the tissue structure of the inspection object 5 can be known from the diffraction image. Is possible. As described above, if both the optical image and the diffraction image of the inspection object 5 can be observed, more structural information of the inspection object 5 can be acquired.

  In the optical microscope apparatus 1, the light source moving means 27 is provided in the point light source 9 in order to change the distance (distance d 2 in FIG. 1) that is the relative position between the diffraction image plane P 1 and the inspection object 5. It is attached. The light source moving means 27 is, for example, a motor, and moves the point light source 9 relative to the condenser lens 11 in a direction parallel to the optical axis L1 of the illumination means 7.

  As the point light source 9 moves in the direction of the optical axis L1, the distance of the convergence point 23 from the inspection object 5 changes, so that the relative position between the diffraction image plane P1 and the inspection object 5 changes. In this way, by changing the distance between the diffraction image plane P1 and the inspection object 5, the size of the diffraction image and the brightness of the image to be observed can be adjusted.

  Further, in the optical microscope apparatus 1, a diaphragm 29 as a convergence angle changing unit is disposed on the front focal plane P <b> 2 of the condenser lens 11. Thereby, even if the point light source 9 is moved in a direction parallel to the optical axis L1, the convergence angle θ of the illumination light at the convergence point 23 can be kept constant.

  This is because the outermost surface P3 that is the outermost surface of the light beam incident on the condenser lens 11 passes through the same part on the focal plane P2 before and after the movement of the point light source 9, and is emitted from the condenser lens 11. This is because the angle of the outermost surface P4 of the emitted light beam with respect to the optical axis L1 is constant. As a result, the distance between the inspection object 5 and the diffraction image plane P1 is maintained without changing the distance between the inspection object 5 and the objective lens 15, that is, while the focusing of the objective lens 15 is kept in focus. Can be changed.

  The size of the convergence angle θ may be selected by changing the convergence angle θ in the range of 0 to 90 ° by limiting the size of the illumination light by the diaphragm 29.

  The means for making the convergence angle θ constant is not limited to the means for disposing the diaphragm 29 on the focal plane P2 as described above. For example, the size of the illumination light may be limited by the diaphragm 29 so that the convergence angle θ becomes constant according to the movement of the light source 9. In this case, the diaphragm 29 may be disposed between the light source 9 and the condenser lens 11.

  Further, in the optical microscope apparatus 1, a spatial stop 35 is provided in parallel with the diffraction image plane P1 at a position on or near the diffraction image plane P1 in order to select an observation field. FIG. 2 is a plan view of the space stop 35. The space stop 35 is formed by forming a circular opening 35b having a diameter of, for example, several hundred microns in the center of the light shielding plate 35a.

  The space stop 35 is movable in a direction perpendicular to the optical axis L2, and selectively passes a part of the light reflected from the inspection object 5 through the circular opening 35b. As a result, an observation field of the diffraction image formed on the diffraction image plane P1 can be selected. The space diaphragm 35 can be easily detached even during observation. The opening shape formed on the light shielding plate 35a, that is, the observation field of view, does not necessarily have to be a circle, and a square, a semicircle, a fan, or the like can be appropriately selected according to the purpose.

  The brightest image can be obtained by adjusting the position of the diffraction image plane P1 so that the object 5 is focused when the objective lens 15 is closest to the spatial aperture 35.

  Further, in the optical microscope apparatus 1, the condenser lens 11 and the light source are arranged so that the illumination condition for the object 5 to be inspected does not change even when the spatial diaphragm 35 is attached or detached or when the observation visual field is selected. 9 has a shield 31 on the optical axis L1. The shielding object 31 is for shielding a part of the illumination light so that the illumination light does not strike the space stop 35.

  When the area surrounded by the light ray (dotted line R in the figure) passing through the edge of the space stop 35 is cut by a plane perpendicular to the optical axis L1, the size of the area of the shield 31 is the same as the size of the space stop 35. It is preferable that they lie on the same plane P5. Thereby, it is not necessary to change the size of the shielding object 31 even if the relative position between the diffraction image plane P1 and the inspection object 5 is changed. The surface on which the shielding object 31 is arranged is also referred to as a shielding object arrangement surface P5, and this shielding object arrangement surface P5 moves correspondingly when the light source 9 moves along the optical axis L1. become.

  The size and shape of the shield 31 need only be such that the spatial stop 35 is positioned within the area shielded by the shield 31, but the spatial stop 35 is moved to select the observation field of the diffraction image. Therefore, it is preferable that the illumination light is shielded including the moving range. The shape of the shield 31 can be similar to the outer shape of the space diaphragm 35. For example, a rectangular shape, a circular shape, or the like is appropriately selected according to the outer shape of the space diaphragm 35.

  Since the illumination light is not applied to the space stop 35 by having the shielding object 31, the illumination condition of the inspection object 5 is made constant even if the shape of the space stop 35 is changed or the space stop 35 is removed. be able to. Further, since the shielding object 31 is arranged on the shielding object arrangement surface P5, the size of the shielding object 31 can be increased even if the relative position between the diffraction image surface P1 and the inspection object 5 is changed. There is no need to change.

  Here, although the position of the shielding object 31 is on the shielding object placement surface P <b> 5, the shielding object 31 may be disposed between the stage 3 and the light source 9.

  As described above, when the shielding object 31 is provided on the optical axis L1, the illuminated area in the field of view becomes, for example, a donut shape, and the center of the field of view is not illuminated, but this can be achieved by changing the illumination method during optical image observation. Good. Illumination methods include a method of illuminating by replacing the point light source 9 with a light source having a size or changing the size of the point light source 9. However, in order to accurately correspond the diffraction image and the optical image, it is preferable to illuminate with the same illumination method when observing the optical image.

  In the above configuration, the illumination light as the convergent light output from the illumination unit 7 is reflected by the reflection unit 13 and applied to the inspection object 5 between the reflection unit 13 and the illumination unit 7 and reflected by the inspection object 5. The incident light is incident on the objective lens 15 after being converged to one point in the space. As a result, the length of the objective lens 15 in the direction of the optical axis L2 can be shortened compared to the case where the illumination light from the illumination means 7 is transmitted through the inspection object 5, and the optical microscope apparatus 1 can be made compact. Is planned.

  Moreover, in the said structure, since the illumination light from the illumination means 7 is irradiated to the to-be-inspected object 5 as convergent light, an observation image with very high contrast and a deep focal depth can be obtained. Then, by changing the convergence angle θ using the diaphragm 29, the contrast, the depth of focus, and the like can be selected.

  Next, one of the microscope observation methods using the optical microscope apparatus 1 will be described with reference to FIGS. 1 and 3. FIG. 3 is a diagram showing the optical microscope apparatus 1 when the objective lens 15 is focused on the diffraction image plane P1 from the state of FIG. In the following description, it is assumed that the diaphragm 29 is disposed on the focal plane P2.

  First, the inspection object 5 is placed on the stage 3, and then the illumination light is output from the illuminating means 7 to irradiate the inspection object 5 with the illumination light. Examples of the inspection object 5 to be observed with the optical microscope apparatus 1 include polymer materials (for example, polymer films such as polyethylene and injection-molded products of polypropylene composite materials), biomaterials, ceramics, metals, and the like. However, an injection-molded product of a polypropylene-based composite material is the most typical target material in that the structure can be observed and the light transmittance is small.

  Next, with the spatial aperture 35 removed, as shown in FIG. 3, the diffraction image is observed by aligning the focus of the objective lens 15 with the diffraction image plane P1. Then, by adjusting the position of the point light source 9 using the light source moving means 27, the size of the diffraction image is set to a desired size. Thus, even if the size of the diffraction image is adjusted, the convergence angle θ at the convergence point 23 is constant because the diaphragm 29 is disposed on the focal plane P2. At this time, the position of the shielding object 31 is moved according to the movement of the light source 9 so as to be positioned on the shielding object placement surface P5.

  Subsequently, as shown in FIG. 3, a spatial diaphragm 35 is attached, and the observation field of the diffraction image is selected by allowing light in a desired region to pass through the light reflected from the inspection object 5. That is, a desired diffracted light is selected from among the diffracted lights that form a diffraction image by the spatial diaphragm 35. The diffracted light is meant to include zero-order diffracted light (that is, direct light). Since the space stop 35 is disposed in a region where the illumination light is previously shielded by the shield 31, the illumination condition of the inspection object 5 is not affected even when the space stop 35 is attached.

  After the desired diffracted light is selected by the spatial diaphragm 35, the focusing of the objective lens 15 is adjusted to the inspection object 5 by the focusing adjustment means 25 as shown in FIG. As a result, the optical image of the inspection object 5 can be observed using the desired diffracted light selected by the spatial diaphragm 35 under the same illumination conditions as when the diffraction image was observed.

  Note that when an image is formed using high-order diffracted light, the light applied to the image is greatly deviated from the optical axis L2 of the objective lens 15, and the obtained image becomes an image with large distortion. Therefore, in such a case, a preferable result can be obtained by adjusting the position of the lens barrel 21 and bringing the selected diffracted light as close to the optical axis L2 as possible.

  In order to adjust the position of the lens barrel 21 in this way, the optical microscope apparatus 1 includes tilt adjusting means 33 that rotates the lens barrel 21 around a point passing through a predetermined position (for example, the focal point of the objective lens 15). .

  When the image of the inspection object 5 is observed, the inclination adjusting means 33 passes through the intersection between the inspection object 5 where the focal point of the objective lens 15 is located and the optical axis L2, and the intersection and the selected diffracted light. By rotating the lens barrel 21 around a straight line perpendicular to the plane including the optical axis L2, the selected diffracted light is made as close as possible to the optical axis L2 by tilting the lens barrel 21.

  Further, when observing the diffraction image, the inclination adjusting means 33 passes through the intersection of the diffraction image plane P1 where the focal point of the objective lens 15 is located and the optical axis L2, and the intersection, the selected diffracted light and the optical axis. By rotating the lens barrel 21 about a line perpendicular to the plane including L2, the lens barrel 21 is tilted to bring the selected diffracted light as close as possible to the optical axis L2.

  When the optical axis L2 of the objective lens 15 is tilted with respect to the optical axis L1 of the condenser lens 11 as described above, the reflecting means 13 is preferably disposed so as to be orthogonal to the optical axis L1 as described above. It is. This is because if the reflecting means 13 is tilted with respect to the optical axis L1, the irradiation area on the inspection object 5 moves. When the optical axis L2 is inclined with respect to the optical axis L1, the objective lens 15, the imaging lens 17, the eyepiece lens 19, and the reflecting means 13 are made parallel to the object to be inspected 5, so Fifteen focusing points can be adjusted. In this case, the optical axis L2 of the objective lens 15 refers to the direction connecting the center of the objective lens 15 and the intersection of the optical axis L1 of the object to be inspected 5 and the condenser lens 11.

  Note that the objective lens 15, the imaging lens 17, and the eyepiece lens 19 are parallel to the inspection object 5. The objective lens 15, the imaging lens 17, and the eyepiece lens 19 have their focal planes parallel to the inspection object 5. It is arranged so that it becomes.

  In the present embodiment, the optical axis L2 of the objective lens 15 and the optical axis L1 of the illumination means 7 are made to coincide with each other. However, the angle of the optical axis L1 of the condenser lens 11 with respect to the optical axis L2 of the objective lens 15 can be varied. You may comprise so that it can do. By changing the angle of the optical axis L2 of the illuminating means 7, diffracted light for observation can be changed, and image information for knowing the tissue structure and orientation can be increased.

  In this case, the reflection means 13 is preferably arranged so as to be orthogonal to the optical axis L 2 as described above, and the reflection means 13, the condenser lens 11, the diaphragm 29, and the shielding object 31 are located with respect to the inspection object 5. By maintaining the parallelism, the parallelism and distance among the diffraction image plane P1, the focal plane P2, and the shielding object arrangement plane P5 can be substantially maintained. To maintain the parallelism between the condenser lens 11 and the inspection object 5 means to arrange the condenser lens 11 so that its focal plane is parallel to the inspection object 5.

  In the above description of the observation method, the case where the diffraction image plane P1 and the objective lens 15 are separated when focusing on the inspection object 5 is described. However, focusing is performed on the inspection object 5. Sometimes, the optical image and the diffraction image of the inspection object 5 may be observed so that the diffraction image plane P1 and the objective lens 15 are closest to each other. Thereby, a bright image can be acquired. The diffraction image plane P1 may be brought closer to the objective lens 15 by moving the light source 9 by the light source moving means 27 as described above. At this time, the shield 31 is also moved as described above.

  By the way, as in the case of a transmission type conventional optical microscope using convergent light, the object to be inspected is irradiated with convergent light as illumination light output from the illumination means, and the light transmitted through the object is once converged. When the light is incident on the objective lens, the irradiation condition to the inspection object does not change even if a spatial aperture is arranged between the inspection object and the objective lens. However, in a reflective optical microscope, the illumination light is irradiated onto the object to be inspected from the same side as the objective lens, and the image is observed using the light reflected by the object to be inspected. There is a risk of being affected.

  On the other hand, according to the optical microscope apparatus 1 and the microscope observation method using the same, since a part of the illumination light is shielded in advance by the shield 31, it is between the object to be inspected 5 and the objective lens 15. Even if the space stop 35 is arranged on the diffraction image plane P1, the illumination light can be prevented from being newly blocked by the space stop 35. As a result, the illumination light can be used without being affected by the space stop 35.

  Further, the diaphragm 29 is arranged on the focal plane P2, and the position of the point light source 9 is adjusted by the light source moving means 27 in accordance with the focusing of the objective lens 15, so that the space between the diffraction image plane P1 and the inspection object 5 is adjusted. And the convergence angle θ can be kept constant.

  Therefore, by focusing the objective lens 15 to the inspection object 5 and the diffraction image plane P1, respectively, the optical image and the diffraction image of the inspection object 5 having a small light transmittance are selectively observed under the same illumination condition. can do. In particular, by making the convergence angle θ constant, it is possible to observe the optical image and the diffraction image of the inspection object 5 while keeping the effects such as the high contrast and the depth of focus constant.

  Further, by appropriately inserting or moving the space stop 35, an optical image and a diffraction image of the inspection object 5 by desired diffracted light can be obtained, and thus a tissue structure that cannot be obtained by a conventional optical microscope apparatus. Information and orientation information can be obtained as an optical image or a diffraction image. Furthermore, since the diffracted light can be freely selected by the space stop 35, various optical images corresponding to the diffracted light can be observed for the same inspection object 5. As a result, it is possible to know the tissue structure of the inspection object 5 in more detail.

  The preferred embodiment of the present invention has been described above, but the present invention is not limited to the above embodiment.

  For example, in the above embodiment, the focused image is focused on the diffraction image plane P1 and the diffraction image is observed, and then the focused image is focused on the inspection object 5, and the optical image is observed. However, the diffraction is performed after the optical image is observed. An image may be observed. Moreover, you may make it observe any one of the optical image of the to-be-inspected object 5, and a diffraction image. Furthermore, it is not always necessary to select a desired diffracted light by the spatial aperture 35.

  Moreover, in the said embodiment, although the distance d2 of the to-be-inspected object 5 and the diffraction image plane P1 is adjusted by moving the light source 9, it is not limited to this. For example, the condenser lens 11, the light source 9, and the diaphragm 29 may be moved together in the direction of the optical axis L2 while the positional relationship between the condenser lens 11 and the light source 9 as the illumination unit 7 and the diaphragm 29 is fixed. Furthermore, the distance d2 can also be adjusted by changing the distance between the reflecting means 13 and the stage 3 (the distance along the optical axis L1).

  Thus, even when the condenser lens 11 and the diaphragm 29 are moved together with the light source 9 or when the reflecting means 13 is moved, the angle of the light beam output from the condenser lens 11 with respect to the optical axis L2 of the outermost surface P4 is constant. Therefore, the convergence angle θ is constant. Further, when the reflecting means 13 is moved, the position of the shielding object 31 is preferably on the shielding object placement surface P5 when the objective lens 15 is closest to the reflecting means 13.

  Furthermore, although the light transmission part which the reflection means 13 has was set as the opening part 13a, what has the transparency with respect to the light from the to-be-inspected object 5 may be provided, for example. Further, in the above embodiment, the convergence point 23 is located between the inspection object 5 and the reflection means 13, but is not limited to this. For example, the convergence point 23 is located between the reflection means 13 and the objective lens 15. May be. That is, the light reflected by the inspection object 5 may be incident on the objective lens 15 through the light passing portion of the reflection means 13 after being converged once, or the light reflected by the inspection object 5 is reflected by the reflection means 13. The light may converge once after passing through the light transmitting portion, and may enter the objective lens 15.

1 is a schematic configuration diagram of an embodiment of an optical microscope apparatus according to the present invention. It is a top view which shows an example of a space stop. It is a figure which shows an optical microscope apparatus when the focusing of an objective lens is matched with the diffraction image surface from the state of FIG.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Optical microscope apparatus, 3 ... Stage (inspection object mounting base), 5 ... Inspection object, 7 ... Illuminating means, 9 ... Point light source (light source), 11 ... Condenser lens, 13 ... Reflecting means, 13a ... Opening part (Light transmitting portion), 15 ... objective lens, 21 ... lens barrel, 23 ... convergence point (one point in space), 25 ... focusing adjustment means, 27 ... light source moving means (distance between the object to be inspected and the diffraction image plane) Adjustment mechanism), 29 ... aperture (convergence angle changing means), 31 ... shielding object, 33 ... adjustment means (adjustment mechanism for matching the light direction after passing through the spatial aperture and the optical axis of the objective lens), 35 ... space. Aperture, L1... Optical axis of condenser lens, L2... Optical axis of objective lens, P1... Diffraction image plane, P2.

Claims (20)

An illumination unit having a condenser lens that outputs light emitted from a light source as convergent light;
Reflecting means for reflecting the convergent light output from the condenser lens toward the illumination means;
An inspection object mounting table provided between the illumination means and the reflection means, for mounting the inspection object;
An objective lens provided on the opposite side of the inspection object mounting table with respect to the reflecting means,
The convergent light output from the illuminating means and reflected by the reflecting means is applied to the object to be inspected,
The reflection means has a light transmission part that transmits light reflected by the inspection object,
The light reflected from the object to be inspected is incident on the objective lens after converging to a point in the space between the objective lens and the object to be inspected, and is incident on the object lens through the light passing portion. An optical microscope apparatus characterized by:   The optical microscope apparatus according to claim 1, further comprising a convergence angle changing unit that changes a convergence angle of light reflected from the inspection object in a range of 0 to 90 °.   Focusing for focusing the objective lens on both the diffraction image plane that includes one point on the space where the light reflected by the inspection object converges and is orthogonal to the optical axis of the objective lens, and the inspection object The optical microscope apparatus according to claim 1, further comprising an adjusting unit.   It further includes an adjustment mechanism for changing the relative position of the diffractive image plane that includes one point on the space where the light reflected by the inspection object converges and is orthogonal to the optical axis of the objective lens, and the inspection object. The optical microscope apparatus according to any one of claims 1 to 3, wherein   A part of the light reflected by the object to be inspected is arranged at a position of a diffraction image plane that includes one point on the converged space and orthogonal to the optical axis of the objective lens, and a part of the light reflected by the object to be inspected is selected. The optical microscope apparatus according to any one of claims 1 to 4, further comprising a spatial aperture that allows the light to pass therethrough.   The shielding object which shields a part of the convergent light so that the convergent light does not illuminate the space stop is further provided between the inspection object mounting table and the light source. Optical microscope device.   The optical microscope apparatus according to claim 5, further comprising an adjustment mechanism that substantially matches the direction of the light that has passed through the spatial aperture and the optical axis of the objective lens.   The optical microscope apparatus according to claim 1, wherein the light emitted from the light source is monochromatic light. In the microscope observation method using the optical microscope apparatus according to claim 1,
A microscope observation method, wherein the object is observed with the focusing of the objective lens adjusted to the object to be inspected. In the microscope observation method using the optical microscope apparatus according to claim 1,
It is formed on the diffraction image plane by focusing the objective lens on a diffraction image plane that includes one point on the space where the light reflected by the inspection object converges and is orthogonal to the optical axis of the objective lens A microscope observation method characterized by observing a diffraction image of the inspection object. In the microscope observation method using the optical microscope apparatus according to claim 3,
Observing the inspection object with the focusing of the objective lens aligned with the inspection object;
And a step of observing the diffraction image by aligning the focusing of the objective lens with a diffraction image formed on the diffraction image surface. In the microscope observation method using the optical microscope apparatus according to claim 5,
Using the spatial diaphragm, the light in a desired region on the diffraction image plane is allowed to pass, the focusing of the objective lens is adjusted to the object to be inspected, and the light passing through the spatial diaphragm is used for the inspection object. A microscope observation method characterized by observing the above. In the microscope observation method using the optical microscope apparatus according to claim 5,
By focusing the objective lens on the diffraction image surface, the diffraction image of the inspection object formed on the diffraction image surface is observed, and light in a desired region of the diffraction image is passed. A microscope observation method characterized by observing the object to be inspected by using the light that has passed through the space diaphragm by adjusting the focus of the objective lens to the object to be inspected after adjusting the space diaphragm .   The microscope observation method according to claim 12 or 13, wherein the inspection object is observed by adjusting a position of the diffraction image plane.   By changing the shape of the spatial aperture or the position on the diffraction image plane, or by changing the angle of the optical axis of the condenser lens with respect to the optical axis of the objective lens, the object to be inspected by the objective lens is changed. The microscope observation method according to any one of claims 12 to 14, wherein the object to be inspected is selected by selecting diffracted light that participates in optical image formation.   The microscope observation method according to any one of claims 12 to 15, wherein the inspection object is observed with a direction of light passing through the spatial aperture substantially matched with an optical axis of the objective lens. .   The microscope observation method according to any one of claims 10 to 16, wherein the size of the diffraction image is adjusted by changing the position of one point on the space in the optical axis direction of the objective lens. .   18. The microscope observation method according to claim 17, wherein even if the position of one point on the space is changed in the optical axis direction of the objective lens, the convergence angle at the one point on the space is not changed.   The microscope observation method according to claim 9, wherein the light emitted from the light source is monochromatic light.   The microscope observation method according to claim 9, wherein the inspection object is a polymer material.

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