JP4851871B2 - Optical microscope apparatus and microscope observation method - Google Patents

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 a 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 the degree of orientation in the tissue structure as described above, which could not be observed with the Koehler illumination method, could be observed with a conventional convergent light microscope. There is a demand for a compact microscope.

  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 of the present invention includes an inspection object mounting table on which an inspection object is mounted, an illuminating unit that outputs illumination light to irradiate the inspection object, and the same side as the illuminating unit with respect to the inspection object mounting table. Reflecting means for reflecting the illumination light output from the illuminating means toward the inspected object and allowing the light reflected from the inspected object to pass through, and the inspected object mounting table with the reflecting means interposed therebetween The illumination light output from the illumination means and reflected by the reflection means is applied to the inspection object, and the light reflected by the inspection object is between the objective lens and the reflection means. It is characterized by being incident on the objective lens after converging to one point in space.

  In this configuration, the illumination light is irradiated onto the inspection object from the same side as the objective lens, and the light reflected from the inspection object is incident on the objective lens. Therefore, the illumination means, the reflection means, and the objective lens are connected to the inspection object. It can be arranged on the same side with respect to the mounting table. As a result, the optical microscope can be made compact.

  Further, a Fourier transform image, that is, a diffraction image of the inspection object is formed on a surface including one point on the space where the light reflected from the inspection object converges (hereinafter referred to as “convergence point”). According to the above optical microscope apparatus, this diffraction image can be formed between the objective lens and the reflecting means, so that the diffraction image itself is observed or a desired process is applied to the diffraction image. Can be easily done. In addition, since a diffraction image is formed between the reflecting means and the objective lens, the object is illuminated even when the focus of the objective lens is adjusted to observe the inspection object and the diffraction image. There is no effect on light irradiation conditions. Therefore, the inspection object and the diffraction image can be easily observed under the same conditions. Further, the structure 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.

  Further, in the optical microscope apparatus according to the present invention, a convergence angle changing means for arbitrarily changing the convergence angle at a convergence point, which is one point on the space where the light reflected by the object to be converged, is in the range of 0 to 90 °. It is preferable to further provide. By changing the convergence angle by the convergence angle changing means, the effect of the high contrast and the depth of focus can be changed.

  Furthermore, the optical microscope apparatus according to the present invention includes a convergence point that is one point on the space where the light reflected by the inspection object is converged, and includes both a diffraction image plane orthogonal to the optical axis of the objective lens and the inspection object. It is preferable to provide focusing adjustment means for adjusting the focusing of the objective lens. 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 relationship between the diffracted image plane perpendicular to the optical axis of the objective lens and the object to be inspected includes a convergence point that is one point on the space where the light reflected from the object to be inspected converges. It is preferable to further include an adjustment mechanism for changing the general position.

  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 diffraction image can be enlarged by changing the relative position. can do. 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 described above, the inspection object and the diffraction image can be observed while the effects of searching for the high contrast and the depth of focus are kept constant.

  Further, the optical microscope apparatus according to the present invention is arranged at a position of a diffraction image plane that includes a convergence point that is one point on a space where light reflected by an object to be converged is orthogonal to the optical axis of the objective lens, It is preferable to further include a spatial aperture that selectively allows part of the light reflected by the object to be inspected.

  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.

  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.

  Furthermore, in the optical microscope apparatus according to the present invention, monochromatic light may be used as illumination light. 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, in the optical microscope apparatus of the present invention, the illumination light is irradiated onto the inspection object from the same side as the objective lens, and the light reflected from the inspection object is on the space between the reflecting means and the objective lens. After converging to one point, it enters the objective lens. In this case, the illumination light is applied to the inspection object as convergent light. As a result, an observation image having a very high contrast and a deep focal depth can be obtained.

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

  In this case, if the relationship between the diffraction image and the tissue structure of the inspection object is acquired in advance, the tissue structure of the inspection object can be known from the characteristics of the pattern of the diffraction image by directly observing the diffraction image. .

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

  In this case, by focusing the objective lens on the object to be inspected and the diffraction image plane, and observing the optical image and the diffraction image of the object to be inspected, the entire tissue structure that was difficult to understand only by observing the optical image. It is possible to grasp the details of the structure of the object to be inspected that contributes to the formation of the diffraction image, which is difficult to understand only by observing the characteristic features and observing the diffraction image. Further, since the convergence point is located between the reflecting means and the objective lens, the diffraction image is formed between the reflecting means and the objective lens. As a result, even if the focusing of the objective lens is adjusted in order to observe the inspection object and the diffraction image, there is no influence on the irradiation condition of the illumination light to the inspection object. Therefore, the inspection object and the diffraction image can be easily observed under the same conditions.

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

  In this microscopic observation method, since desired diffracted light is selected by passing desired light on the diffracted image plane using a spatial aperture, 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 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.

  Another microscopic observation method using the optical microscope of the present invention observes a diffraction image by illumination light of an object to be inspected formed on the diffraction image plane by matching the focus of the objective lens to the diffraction image plane. After the spatial aperture is adjusted so that light in a desired region of the diffraction image is allowed to pass, the object is observed by the light passing through the spatial aperture by adjusting the focus of the objective lens to the inspection target. Features.

  Since the diffracted light used for observing the optical image is selected based on the diffracted image, it is possible to know the diffracted light based optical image. Thereby, the tissue structure of the object to be inspected can be known in more detail. Further, as described above, since the diffraction image is formed between the reflecting means and the objective lens, even if the focusing of the objective lens is adjusted in order to observe the inspection object and the diffraction image, the diffraction image is obtained. There is no effect on the irradiation condition of the illumination light. As a result, the inspection object and the diffraction image can be easily observed under the same conditions.

  In the microscope observation method of the present invention, it is preferable to observe the inspection object by adjusting the position of the diffraction image plane with respect to the inspection object. Since the surface that includes the convergence point of the illumination light and is orthogonal to the optical axis of the objective lens is the diffraction image surface, adjusting the position of the diffraction image surface with respect to the inspection object determines the position of the convergence point with respect to the inspection object. Will be adjusted. Therefore, by adjusting and rubbing the position of the diffraction image surface, the amount of light incident on the objective lens can be adjusted, 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 diaphragm or the position on the diffraction image plane, or by changing the angle of the optical axis of the illumination light 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.

  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 the convergence point of the illumination light 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 not to change the convergence angle at the convergence point even if the position of the convergence point 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, illumination light 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. Further, according to the microscope observation method of the present invention, it is possible to observe the inspection object using a compact optical microscope apparatus.

  DESCRIPTION OF EMBODIMENTS Hereinafter, preferred embodiments of an optical microscope apparatus according to the present invention and a microscope observation method using the optical microscope apparatus will be described in detail with reference to the drawings. In the following description, the same elements are denoted by the same reference numerals, and redundant descriptions are omitted. In addition, a word indicating a direction such as “up” in the present specification is a convenient word based on the state shown in the drawings.

  FIG. 1 is a schematic configuration diagram showing a basic configuration of an embodiment of an optical microscope apparatus according to the present invention. The optical microscope apparatus 1 has a stage 10 as an object mounting table. The optical microscope apparatus 1 is a reflection type that irradiates an inspection object (specimen) 15 placed on a stage 10 with illumination light and performs various observations using light reflected from the inspection object 15. This is an optical microscope apparatus.

  Examples of the inspection object 15 to be observed with the optical microscope apparatus 1 include a polymer material (for example, a polymer film such as polyethylene or an injection-molded product of a polypropylene-based composite material), a biomaterial, ceramics, metal, and the like. . Note that an injection molded product of a polypropylene-based composite material is the most typical target material in that the tissue structure can be observed and the light transmittance is small.

  The optical microscope apparatus 1 includes a light irradiation unit 20 for irradiating the inspection object 15 with illumination light, and a light detection unit 30 for detecting light reflected from the inspection object 15 irradiated with the illumination light. The light irradiation unit 20 and the light detection unit 30 are disposed above the inspection object 15. In the following description, the optical axis L1 direction of the light detection unit 30 is defined as the Z-axis direction, and the directions orthogonal to the Z-axis direction as illustrated in FIG. 1 are defined as the X-axis direction and the Y-axis direction. In FIG. 1, it is assumed that the Z-axis direction corresponds to the vertical direction.

  FIG. 2 is a schematic configuration diagram illustrating a basic configuration of the light irradiation unit. 2, the vicinity of the light irradiation unit 20 in the optical microscope apparatus 1 shown in FIG. The light irradiation unit 20 includes a light output unit (illumination unit) 21, a reflection unit 22, and a housing 23.

  The light output unit 21 outputs illumination light for irradiating the inspection object 15. The light output unit 21 is accommodated in the housing 23, and includes a light source 21A, a diaphragm 21B, and a condenser lens 21C arranged along the X-axis direction. The light source 21A is, for example, a point light source, and the light output from the light source 21A may be white light or monochromatic light. The light source 21A can be moved by a light source moving unit 24 in a direction parallel to the optical axis L2 of the light output unit 21 with respect to the condenser lens 21C. The light source moving means 24 may be one conventionally used for position adjustment of the light source 21A, but a stage that can move in at least one axial direction using a motor, a piezo, or the like is exemplified.

  In the light output unit 21, the light that has passed through the diaphragm 21B among the light output from the light source 21A is condensed by the condenser lens 21C and output as illumination light. The illumination light output from the light output unit 21 is convergent light. In the light output unit 21 having the above configuration, the amount of light from the light source 21A can be effectively used by adjusting the opening of the diaphragm 21B (the size of the opening for allowing light to pass through). Yes.

  The reflecting means 22 is disposed on the optical axis L <b> 2 of the light output unit 21, and is held by the housing 23 by being attached to one side wall 23 a of the housing 23. The reflection means 22 is, for example, a half mirror, reflects the illumination light output from the light output unit 21 toward the inspection object 15 and irradiates the inspection object 15, and also reflects the light reflected from the inspection object 15. Let it pass. The reflecting means 22 may be accommodated in the housing 23. In this case, the light reflected by the inspection object 15 above the reflecting means 22 in FIGS. 1 and 2 and passed through the reflecting means 22. A window for passing through is formed in the side wall 23a.

  The housing 23 accommodates the light output unit 21 and holds the reflection means 22. On the side wall 23b of the housing 23 on the inspection object 15 side, there is formed a window 25 through which the illumination light reflected by the reflecting means 22 and the light reflected from the inspection object 15 irradiated with the illumination light are transmitted. Has been. The window 25 may be a simple opening, or a transparent plate such as a quartz plate may be attached to the opening. The window 25 may be formed at an appropriate position and has a sufficient size so as not to block part or all of the illumination light reflected by the reflecting means 22 and the light from the inspection object 15. The housing 23 is preferably configured so that the distance between the housing 23 and the object 15 to be inspected can be adjusted. Thereby, the position of the light irradiation part 20 can be arrange | positioned in the free position between the to-be-inspected object 15 and the diffraction image surface P1 mentioned later.

  In the optical microscope apparatus 1, the light output unit 21 and the reflection unit 22 are formed by the reflection unit 22 positioned on the optical axes L 1 and L 2 of the light detection unit 30 and the light output unit 21. The angle θi, the optical axis L1, and the angle θo formed by the reflecting means 22 are arranged to be equal. Hereinafter, it is assumed that the optical axes L1 and L2 are orthogonal unless otherwise specified.

  In the optical microscope apparatus 1, the illumination light is inspected so that the light reflected by the inspection object 15 converges at a convergence point 40 that is one point on the space above the reflecting means 22 (on the light detection unit 30 side). The object 15 is irradiated. That is, the light output from the light output unit 21 is irradiated onto the inspection object 15 as convergent light having a convergence point 40 above the reflecting means 22. On a plane P1 that passes through the convergence point 40 and is orthogonal to the optical axis L1, a Fourier transform image, that is, a diffraction image of the inspection object 15 is formed by the illumination light. Here, this plane P1 is called a diffraction image plane.

  The convergence angle θ at the convergence point 40 varies in the range of 0 to 90 ° by limiting the size of the illumination light using the stop 21B. Therefore, the diaphragm 21B also functions as a convergence angle changing unit.

  Further, by moving the position of the light source 21A by the light source moving means 24, the distance (distance d1 in FIG. 1) as the relative position between the diffraction image plane P1 and the inspection object 15 can be changed. It has become. That is, when the light source 21A moves in the direction of the optical axis L2, the distance of the convergence point 40 from the inspection object 15 changes, and as a result, the relative position between the diffraction image plane P1 and the inspection object 15 changes. Will do.

  At this time, as shown in FIGS. 1 and 2, even if the light source 21A is moved in the direction parallel to the optical axis L2 by arranging the stop 21B on the rear focal plane P2 of the condenser lens 21C, the convergence point 40 The convergence angle θ of the illumination light at can be kept constant. This is because the outermost surface P3 which is the outermost surface of the light beam incident on the condenser lens 21C before and after the movement of the light source 21A passes through the same portion on the focal plane P2, and thereby the light beam emitted from the condenser lens 21C. This is because the angle of the outermost surface P4 with respect to the optical axis L2 is constant. Accordingly, the distance between the inspection object 15 and the diffraction image plane P1 can be changed while the convergence angle θ of the illumination light at the convergence point 40 is kept constant.

  As shown in FIGS. 1 and 2, a space stop 50 constituting a part of the optical microscope apparatus 1 is provided substantially parallel to the diffraction image plane P1 at a position on or near the diffraction image plane P1. The space stop 50 can be easily detached even during observation.

  FIG. 3 is a plan view of the space stop 50. The space stop 50 is formed by forming a circular opening 52 having a diameter of, for example, several hundred microns at the center of the light shielding plate 51. The opening shape formed in the light shielding plate 51, 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 space stop 50 selectively passes a part of the light reflected from the inspection object 15 by the circular opening 52. As shown in FIG. It can move in the direction perpendicular to the direction of the axis L1 and the optical axis L1. The diaphragm moving means 53 is, for example, a three-axis stage. The observation field of the diffraction image formed on the diffraction image plane P1 can be selected by moving the spatial stop 50 in the direction substantially orthogonal to the optical axis L1, that is, in the XY plane, by the stop moving means 53. Further, since the spatial diaphragm 50 can be moved in the optical axis L1 direction (Z-axis direction) by the diaphragm moving means 53, even if the position of the diffraction image plane P1 is adjusted in the optical axis L1 direction with respect to the inspection object 15. The position of the spatial aperture 50 can be adjusted accordingly.

  Since the space stop 50 is located closer to the objective lens 31 than the reflecting means 22, even when the space stop 50 is attached or removed or the observation field of view is selected, the space stop 50 is moved to the inspection object 15. The lighting conditions can be made constant.

  As shown in FIG. 1, in the optical microscope apparatus 1, the light detection unit 30 is disposed above the spatial diaphragm 50. The light detection unit 30 includes an objective lens 31, an imaging lens 32, an eyepiece lens 33, and a lens barrel 34 that accommodates them, and the objective lens 31, the imaging lens 32, and the eyepiece lens 33 are sequentially arranged from the stage 10 side. Is arranged. The internal structure itself of the lens barrel 34 is a conventional one. In this embodiment, it is assumed that the optical axis L1 of the light detection unit 30 coincides with the optical axis of the objective lens 31, and unless otherwise specified, the light detection unit 30 has the optical axis L1 of the inspection object 15 of the stage 10. It is assumed that it is arranged with respect to the stage 10 so as to be substantially orthogonal to the surface on which is mounted.

  The objective lens 31 has a focal length that allows the objective lens 31 to be positioned above the light irradiation unit 20 and the space stop 50 when the object 15 is focused. Thereby, in the focusing operation, the light irradiation unit 20 and the space stop 50 do not get in the way. The imaging lens 32 is disposed so as to form an image captured by the objective lens 31 at an intermediate image imaging position 35 behind the imaging lens 32 (on the eyepiece lens 33 side). The focusing is adjusted so that the image can be observed.

  The lens barrel 34 is preferably recessed toward the objective lens 31 (in other words, tapered). Thereby, even if the light irradiation part 20 is brought close to the objective lens 31, it does not collide, and even if the distance d1 between the diffraction image plane P1 and the inspection object 15 is shortened, the reflected light from the inspection object 15 is reflected by the objective lens 31. It can be incident on.

  In the lens barrel 34, the distance between the lens barrel 34 and the stage 10 can be changed by the focusing adjustment means 36. Accordingly, the relative position between the objective lens 31 and the inspection object 15, that is, the distance between the objective lens 31 and the inspection object 15 (distance d2 in FIG. 1) can be changed. As a result, the objective lens 31 is changed. It is possible to perform the focusing.

  The focusing adjustment means 36 uses, for example, a rack and pinion, and may be the same as that attached to the lens barrel of a conventional optical microscope, but can be changed for focusing. The range of relative positions that can be made 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 15 and the diffraction image plane P1.

  In the optical microscope apparatus 1 having the above configuration, the optical image of the inspection object 15 can be observed by aligning the focusing of the objective lens 31 with the inspection object 15, and the focusing of the objective lens 31 on the diffraction image plane P1. A diffraction image can be observed by combining. In the diffraction image, the structural information of the object to be inspected 15 is collected, so if the relationship between the tissue structure and the diffraction image is known in advance, it is possible to know the tissue structure of the object to be inspected from the diffraction image. It is. As described above, if both the optical image and the diffraction image of the inspection object 15 can be observed, more structural information of the inspection object 15 can be acquired.

  In addition, since the optical microscope apparatus 1 uses light reflected from the inspection object 15, it is possible to suitably know the surface texture structure of the inspection object 15 having a low transmittance as the inspection object 15. Furthermore, since the objective lens 31, the reflecting means 22, and the light output unit 21 can be arranged on the same side with respect to the stage 10, the optical microscope apparatus 1 can be made compact. In addition, since the illumination light from the light output unit 21 is applied to the inspection object 15 as convergent light, the contrast, depth of focus, and the like are selected by changing the convergence angle θ using the diaphragm 21B. As a result, observation is possible with a desired contrast, depth of focus, and the like.

  As shown in FIG. 4, when the position of the diffraction image plane P1 is adjusted so that the objective lens 31 and the space stop 50 are closest to each other when the object 15 is focused, the brightest image is obtained. be able to. The diffraction image plane P1 may be adjusted by changing the position of the light source 21A from the position of the two-dot chain line (the position of the light source shown in FIG. 1) to the position of the solid line using the light source moving unit 24. In this state, the distance d1 between the inspection object 15 and the diffraction image plane P1 is substantially equal to the distance d2 between the inspection object 15 and the objective lens 31 as shown in FIG.

  Next, one of the microscope observation methods using the optical microscope apparatus 1 will be described with reference to FIGS. 1 and 5. FIG. 1 shows a state of the optical microscope apparatus 1 when the object lens 31 is focused on the object 15 to be inspected. FIG. 5 shows a state of the optical microscope apparatus 1 when the objective lens 31 is focused on the diffraction image plane P1 from the state of FIG. In the following description, it is assumed that the stop 21B is disposed on the focal plane P2.

  First, the inspection object 15 is placed on the stage 10, and the illumination light is output from the light output unit 21 to output the illumination light to the inspection object 15. Next, with the spatial aperture 50 removed, as shown in FIG. 5, the focusing of the objective lens 31 is adjusted to the diffraction image plane P1, and the diffraction image is observed. Then, the size of the diffraction image is adjusted to a desired size by moving the position of the light source 21 </ b> A in the X-axis direction using the light source moving means 24. Thus, even if the size of the diffraction image is adjusted, the convergence angle θ at the convergence point 40 is constant because the diaphragm 21B is disposed on the focal plane P2.

  Subsequently, as shown in FIG. 5, a spatial diaphragm 50 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 by the inspection object 15. That is, a desired diffracted light is selected from among the diffracted lights that form a diffraction image by the space stop 50. The diffracted light is meant to include zero-order diffracted light (that is, direct light). Since the space stop 50 is disposed on the objective lens 31 side with respect to the reflecting means 22, the illumination condition of the inspection object 15 is not affected even if the space stop 50 is attached.

  After the desired diffracted light is selected by the spatial aperture 50, the focusing of the objective lens 31 is adjusted to the inspection object 15 as shown in FIG. At this time, since the opening of the aperture 21B is kept constant, the convergence angle θ is also kept constant. Therefore, as shown in FIG. 1, even when the focus of the objective lens 31 is adjusted to the inspection object 15, the desired diffracted light selected by the spatial diaphragm 50 is used under the same illumination conditions as when the diffraction image is observed. Thus, an optical image of the inspection object 15 can be observed.

  As described above, since the diffracted light to be imaged can be freely selected by the spatial diaphragm 50, a higher-order diffracted light is selected by the spatial diaphragm 50, and an image is obtained using the higher-order diffracted light. It can also be formed. As described above, when forming an image with high-order diffracted light from the same inspection object 15, the inclination of the lens barrel 34 is adjusted by, for example, the inclination adjusting means 37 included in the optical microscope apparatus 1. Thus, it is preferable to make the selected diffracted light as close to the optical axis L1 as possible. This is because a preferable result (image) with reduced distortion can be obtained.

  Further, when the condenser lens 21C to the reflecting means 22 in the optical axis L2 of the light output unit 21 are particularly used as the optical axis of the illumination light, in this embodiment, the optical axis L1 of the objective lens 31 and the optical axis of the illumination light However, it is also preferable that the angle of the optical axis of the illumination light with respect to the optical axis L1 of the objective lens 31 can be changed. As described above, by changing the angle of the optical axis of the illumination light with respect to the optical axis L1, the diffracted light applied to the observation can be changed, and as a result, image information for knowing the tissue structure and orientation can be increased. it can. Note that changing the angle of the optical axis of the illumination light may be, for example, changing the inclination angle of the condenser lens 21C with respect to the Z-axis direction or changing the inclination angle of the entire light output unit 21 with respect to the Z-axis direction.

  In the description of the observation method, the case where the diffraction image plane P1 and the objective lens 31 are separated when focusing on the inspection object 15 has been described. However, as shown in FIGS. When focusing on the inspection object 15, the optical image and the diffraction image of the inspection object 15 may be observed so that the diffraction image plane P1 and the objective lens 31 are closest to each other. FIG. 6 is a diagram showing a state of the optical microscope apparatus 1 when the objective lens 31 is focused on the diffraction image plane P1 from the state of FIG.

  Also in this case, first, as shown in FIG. 6, the diffraction image is observed by focusing the objective lens 31 on the diffraction image plane P1. And after selecting a desired area | region with the space aperture 50, as shown in FIG. 4, focusing on the to-be-inspected object 15, the optical image of the to-be-inspected object 15 is observed. Since the light reflected from the inspection object 15 converges at the convergence point 40, the light reflected from the inspection object 15 enters the objective lens 31 without loss by bringing the diffraction image plane P1 and the objective lens 31 close to each other. As a result, a bright image can be acquired.

  By the way, as a reflection type optical microscope apparatus, the structure which arrange | positions a half mirror above an objective lens can also be considered, for example. In this configuration, the illumination light is irradiated onto the inspection object through the objective lens, and the image is acquired by causing the light from the inspection object to enter the objective lens again. However, with this configuration, as in the present embodiment, observe the diffraction image formed at the position of the convergence point by irradiating the inspection object with illumination light as convergent light and the optical image of the inspection object. Then, the objective lens contributes to the formation of convergent light, and a spatial aperture is disposed between the half mirror and the inspection object. As a result, the illumination conditions for the inspection object may change depending on the arrangement of the spatial aperture between the inspection object and the focusing of the objective lens.

  On the other hand, in the optical microscope apparatus 1, the reflecting means 22 is disposed between the objective lens 31 and the inspection object 15, and the light output from the light source 21A is collected using the condenser lens 21C. As a result, convergent light as illumination light is formed. Further, since the convergence point of the convergent light is located between the reflecting means 22 and the objective lens 31, the diffraction image plane P1 is also located between the reflecting means 22 and the objective lens 31.

  In this case, the objective lens 31 does not contribute to the formation of the convergent light, and the spatial aperture 50 is also located between the reflecting means 22 and the objective lens 31, so the focusing of the objective lens 31 is adjusted. Even in this case, the irradiation condition of the inspection object 15, the distance d1 between the diffraction image plane Pl and the inspection object 15, and the convergence angle θ do not change.

  Therefore, by focusing the objective lens 31 to the inspection object 15 and the diffraction image plane P1, respectively, the optical image and the diffraction image of the inspection object 15 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 an optical image or a diffraction image of the inspection object 15 while maintaining effects such as the high contrast and the depth of focus.

  In addition, by appropriately inserting or moving the space stop 50, an optical image and a diffraction image of the inspection object 15 by a desired diffracted light can be obtained. Therefore, 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 50, various optical images corresponding to the diffracted light can be observed for the same inspection object 15. As a result, it is possible to know the tissue structure of the inspection object 15 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.

  In the above embodiment, by adjusting the position of the light source 21A using the light source moving means 24, the distance d1 is changed while maintaining the convergence angle θ substantially constant. It may be moved integrally in the X-axis direction. Convergence by changing the position of the light output unit 21 as a whole, that is, by moving the light source 21A, the diaphragm 21B and the condenser lens 21C while keeping their positional relationship and the aperture of the diaphragm 21B constant. The angle θ can be kept constant. This is by changing the position of the light output unit 21 as a whole, so that the outermost surface P4 of the light beam emitted from the condenser lens 21C even when the light output unit 21 is moved in the direction parallel to the optical axis L2 (FIG. 2) with respect to the optical axis L2.

  As a method of changing the position of the entire light output unit 21, the light output unit moving means 27 for moving the light output unit 21 may be further provided as shown in FIG. Since the distance d1 between the diffraction image plane P1 and the inspection object 15 can also be changed by the light output section moving means 27, the light output section moving means 27 adjusts the relative position of the diffraction image plane P1 and the inspection object 15. It is functioning as a mechanism. As the light output unit moving means 27, for example, a light source 21A, a diaphragm 21B, a condenser lens 21C, and a position adjusting means thereof can be mounted on one stage and can be moved at least in the X-axis direction. . The position adjusting means of the light source 21A corresponds to the light source moving means 24 described above.

  Further, changing the position of the light output unit 21 as a whole means that the positional relationship between the light source 21A, the diaphragm 21B, and the condenser lens 21C is fixed by the respective position adjusting means. You may move those positions, keeping it.

  When the distance d1 between the convergence point 40 and the inspection object 15 is changed by changing the position of the entire light output unit 21, the positions of the light source 21A, the diaphragm 21B, and the condenser lens 21C are mutually changed. For example, the position of the light source 21 </ b> A may be further adjusted using the light source moving unit 24 as long as the relationship is kept constant. Thereby, the range of the observation method of the image of the inspection object 15 itself and the diffraction image can be widened. This will be described more specifically with reference to FIG.

  FIG. 8 is a diagram showing the optical microscope in a state in which the position of the light output unit 21 is moved in the direction of the optical axis L2 using the light output unit moving unit 27 from the state of FIG.

  In the state of the optical microscope apparatus 1 shown in FIG. 8, the position of the light output unit 21 is farther from the reflecting means 22 than in the state shown in FIG. 1, and further, the condenser lens 21C and the light source 21A in the light output unit 21 In the positional relationship, the light source 21A has moved to the condenser lens 21C side from the state shown in FIG. 1 (the position of the two-dot chain line in FIG. 8) with respect to the condenser lens 21C. The distance d1 between the convergence point 40 and the inspection object 15 in FIG. 8 is the same as the distance d1 between the convergence point 40 and the inspection object 15 in FIG. 1, and the object lens 31 is focused on the inspection object 15. ing. Thus, the optical image of the inspection object 15 can be observed in a state where the distance between the diffraction image plane P1 and the inspection object 15 is reduced as in the case of FIG. When observing the diffraction image, the aim of the objective lens 31 may be aligned with the diffraction image plane P1.

  Further, in the embodiment, the lens barrel 34 is moved in order to adjust the focusing of the objective lens 31 to the object 15 to be inspected. However, the stage (inspection object mounting table) 3 and the light irradiation unit 20 are moved together. You may let them. Alternatively, both the lens barrel 34 and the stage 10 and the light irradiation unit 20 may be moved together.

  In the above embodiment, the optical image is observed by focusing on the inspection object 15 after focusing the diffraction image plane P1 and observing the diffraction image. However, the diffraction is performed after observing the optical image. An image may be observed. Further, either one of the optical image and the diffraction image of the inspection object 15 may be observed. Furthermore, it is not always necessary to select a desired diffracted light with the space stop 50. Furthermore, although the light output unit 21 is provided with the stop 21B, the light output unit 21 is not necessarily provided with the stop 21B as long as it can output illumination light as convergent light.

It is the schematic which shows the structure of one Embodiment of the optical microscope apparatus of this invention. It is a schematic block diagram which shows the basic composition of a light irradiation part. It is a top view which shows-example of a space stop. It is a figure which shows the state of an optical microscope apparatus when observing an optical image in the state which made the convergence point adjoin to the objective lens in FIG. It is a figure which shows the state of an optical microscope apparatus when the focus of an objective lens is matched with the diffraction image surface from the state of FIG. FIG. 5 is a diagram showing a state of the optical microscope apparatus when the objective lens is focused on the diffraction image plane from the state of FIG. 4. It is a schematic block diagram which shows the basic composition of the light irradiation part in the case of having a light output part moving means. It is a figure which shows the state of an optical microscope when the position of the light output part is separated from the reflection means from the state shown in FIG.

Explanation of symbols

  DESCRIPTION OF SYMBOLS 1 ... Optical microscope apparatus, 10 ... Stage (inspection object mounting base), 15 ... Inspection object, 21 ... Light output part, 21C ... Diaphragm (convergence angle changing means), 22 ... Reflection means, 24 ... Light source moving means ( (Adjustment mechanism for relative position between diffraction image plane and object to be inspected) 31... Objective lens 36. Focus adjustment means 37. Inclination adjustment means 40. Convergence point 50. Means, d1... Distance between the object to be inspected and the diffraction image plane, L1... Optical axis of optical detector (optical axis of objective lens), P1.

Claims (18)

An inspection object mounting table for mounting the inspection object;
Illuminating means for outputting illumination light to irradiate the inspection object;
Provided on the same side as the illumination means with respect to the inspection object mounting table, and reflects the illumination light output from the illumination means toward the inspection object and is reflected from the inspection object. Reflection means for passing the light,
An objective lens provided on the opposite side of the inspection object mounting table across the reflecting means,
The illumination light output from the illumination means and reflected by the reflection means is applied to the object to be inspected,
The optical microscope apparatus characterized in that the light reflected by the object to be inspected is incident on the objective lens after converging to one point on the space between the objective lens and the reflecting means.   2. The apparatus according to claim 1, further comprising a convergence angle changing unit that arbitrarily changes a convergence angle at a convergence point that is one point on a space where light reflected by the inspection object converges. The optical microscope apparatus described.   The objective lens is focused on both the diffracted image plane orthogonal to the optical axis of the objective lens and the object to be inspected, which includes a convergence point that is one point in the space where the light reflected by the object is converged. An optical microscope apparatus according to claim 1 or 2, further comprising a focusing adjustment means for the purpose.   Adjustment for changing the relative position of the diffracted image plane perpendicular to the optical axis of the objective lens including a convergence point that is one point in the space where the light reflected by the inspection object converges The optical microscope apparatus according to claim 1, further comprising a mechanism.   The light reflected from the inspection object is disposed at a position of a diffraction image plane that includes a convergence point that is one point on a converged space and orthogonal to the optical axis of the objective lens. The optical microscope apparatus according to any one of claims 1 to 4, further comprising a spatial aperture that selectively allows a portion to pass therethrough.   The optical microscope apparatus according to claim 5, further comprising an adjustment mechanism that substantially matches a direction of light that has passed through the spatial aperture and an optical axis of the objective lens.   The optical microscope apparatus according to claim 1, wherein the illumination light 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,
By focusing the objective lens on a diffractive image plane that includes a convergence point that is one point on a space where light reflected by the inspection object is converged, and orthogonal to the optical axis of the objective lens, A microscope observation method characterized by observing a diffraction image of the object to be inspected by the illumination light. 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,
Passing light in a desired region of the diffraction image plane using the spatial diaphragm, aligning the focus of the objective lens with the inspection object, and observing the inspection object with the light passing through the spatial diaphragm A microscope observation method characterized by the above. In the microscope observation method using the optical microscope apparatus according to claim 5,
By aligning the focus of the objective lens with the diffraction image plane, a diffraction image by the illumination light of the inspection object formed on the diffraction image plane is observed, and light in a desired region of the diffraction image is observed. A microscope observation characterized by observing the object to be inspected by light that has passed through the space diaphragm by adjusting the spatial aperture so as to pass through, and adjusting the focus of the objective lens to the object to be inspected Method.   The microscope observation method according to claim 11 or 12, wherein the inspection object is observed by adjusting a relative position between the diffraction image plane and the inspection object.   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 illumination light with respect to the optical axis of the objective lens, The microscope observation method according to any one of claims 11 to 13, wherein the inspection object is observed by selecting diffracted light that participates in optical image formation.   The microscope observation method according to any one of claims 11 to 14, wherein the object to be inspected is observed with the direction of light passing through the spatial aperture substantially matched with the optical axis of the objective lens. .   The microscope observation according to any one of claims 9 to 15, wherein the size of the diffraction image is adjusted by changing a position of a convergence point of the illumination light in an optical axis direction of the objective lens. Method.   The microscope observation method according to any one of claims 8 to 16, wherein the illumination light is monochromatic light.   The microscope observation method according to any one of claims 8 to 17, wherein the inspection object is a polymer material.

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