JP2017097343A - Optical observation device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an optical observation device in which a simple structure makes an observation object unnecessary to be loaded on a specific instrument in the optical observation device applying a sample with light at two angles of incidence, processing two photograph images to be acquired, and preparing a sharp image.SOLUTION: LED light from a first LED light irradiation device 13 and second LED light irradiation device 14 are caused to be incident upon a first vertical direction light guide member 21 and second vertical direction light guide member 22 allowing LED light to be incident along an optical axis of an objective lens 10, and respective LED light are caused to be reflected upon a first reflection part 23 and second reflection unit 24 at a different angle, and are caused to be incident somewhere around a part crossing the optical axis of the objective lens 10 at a different angle of incidence in a low surface of a lateral direction light guide member 25. Due to a mechanism changing an interval between a stage 30 and the objective lens 10, the low surface of the lateral direction light guide member 25 is located in the vicinity of an inner side bottom surface of a glass plate P, and a picture is taken by an imaging unit including the objective lens 10.SELECTED DRAWING: Figure 2

Description

  The present invention relates to an optical observation apparatus for irradiating a minute observation object such as a cell tissue with light and observing the light.

  2. Description of the Related Art Conventionally, an optical observation apparatus that irradiates a minute biological sample such as a cell tissue with light and magnifies and observes the biological sample like an optical microscope has been widely used. In such an optical observation device, for example, as shown in Patent Document 1, a fluorescent substance is generated by causing an enzyme to act on a physiologically active substance released from a biological sample, and the biological sample is irradiated with excitation light. There is an optical observation apparatus that can create a fluorescence image by capturing generated fluorescence into an imaging unit. According to this optical observation apparatus, it is possible to observe the concentration distribution of the physiologically active substance in the biological sample. However, in this optical observation apparatus, since background light that is light other than the target fluorescence from the biological sample is also taken into the imaging unit, the observation image is often blurred and fine information cannot be obtained in many cases. Further, not only in this optical observation apparatus, but also in an optical observation apparatus that performs irradiation of light on the specimen and observation of the specimen on the same side, such as an episcopic microscope, the target light from the specimen (hereinafter referred to as target light) ), The observation image is blurred and a fine image is often not obtained.

  In order to cope with this problem, the inventors of the present invention invented an optical observation apparatus capable of reducing the influence of background light included in the observation image and acquiring a good observation image, as shown in Patent Document 2. did. In this optical observation apparatus, two observation images are obtained by irradiating the sample with excitation light having different incident angles, and the ratio of the target light component and the background light component in each observation image is different. Utilizing this, background light components are removed by image processing. According to this optical observation device, a clear observation image can be obtained.

JP 2008-139280 A JP2015-114630A

  However, the optical observation device disclosed in Patent Document 2 has a structure in which excitation light is incident at two incident angles from the side surface side of a flat transparent substrate on which a sample is placed, and the sample is irradiated from the transparent substrate surface. If the apparatus is configured such that the excitation light irradiator is appropriately disposed in the vicinity of the side surface of the flat transparent substrate, there is a problem that the structure of the apparatus becomes complicated. In addition, since it is necessary to place the specimen on a flat transparent substrate, the living tissue cannot be observed as it is, and it is inefficient when optically observing a plurality of specimens continuously. There's a problem.

  The present invention has been made to solve this problem, and an object of the present invention is to irradiate light at a plurality of incident angles on an observation object and process a plurality of obtained captured images to create a clear image. In the observation apparatus, the apparatus has a simple structure, and it is not necessary to place the observation object on a specific instrument, and it is possible to observe the living tissue as it is or to continuously observe different observation objects. The object is to provide an optical observation device.

  In order to achieve the above object, a feature of the present invention includes an imaging unit that images an observation object, and a light irradiation unit that is disposed in a housing having the imaging unit and that irradiates light to the observation object. In the optical observation apparatus, the light irradiation means has a surface facing the object to be observed on one end side of the housing, and a lateral light guide member that guides light in the surface, and a lateral light guide member In addition to being provided in series, a longitudinal light guide member that guides light incident from a part of the casing to the lateral light guide member, and a longitudinal light guide member that reflects incident light. A plurality of light beams having different reflection angles so that the light beams are guided to the lateral light guide member and incident on the surface of the lateral light guide member facing the observation object at different incident angles according to the reflection position of the light. The reason is that a reflection part is provided.

  According to this, when the light incident from a part of the housing is guided from the vertical light guide member to the horizontal light guide member, the light is reflected by the reflecting portion provided in the vertical light guide member. It is reflected and guided. Here, since a plurality of reflecting portions having different reflection angles are provided, light is incident on the surface of the lateral light guide member facing the observation object at different incident angles according to the reflection position. It becomes. That is, an optical path for irradiating the observation object facing the surface of the horizontal light guide member with light having a plurality of incident angles with a simple configuration in which a plurality of reflection portions having different reflection angles are provided on the vertical light guide member Formation can be realized. In addition, since the light irradiation means can be provided integrally with the imaging unit in the housing, the entire apparatus can be reduced in size and handleability can be improved as compared with the case where the light irradiation means is provided separately. it can. Furthermore, since it is not necessary to place the observation object on a specific instrument, the living tissue can be observed as it is, and a plurality of observation objects can be observed efficiently and continuously.

  Another feature of the present invention is that the lateral light guide member has a plate-like portion having a surface facing the object to be observed and a back surface opposite to the surface, and is provided in the longitudinal light guide member. A part of the plurality of reflecting portions is configured to guide light toward the back surface side of the plate-shaped portion, and the light guided to the back surface side of the plate-shaped portion is reflected on the back surface side of the plate-shaped portion. After reflecting, it is configured to be incident on the surface facing the object to be observed.

  According to this, since the surface facing the observation target object of the lateral light guide member can be formed by one plate-like portion, the surface facing the observation target object and the observation target object of the horizontal light guide member when visually observed The interval between Furthermore, since the lateral light guide member can be formed with a simple shape, the processing of the lateral light guide member can be facilitated and the component cost can be reduced.

  Another feature of the present invention is that the light incident on the surface facing the object to be observed after reflecting on the back side of the plate-like portion of the lateral light guide member has an incident angle of 75 to 85 °. It is in that.

  As a result of experiments, the inventors have confirmed that in the light incident on the surface of the lateral light guide member facing the object to be observed, the incident angle of one light is close to 90 ° and the incident angle of the other light is 80. It was found that the difference between the ratio of the target light and the background light is the largest when the temperature is around 0 °, and a clear image can be obtained by image processing. Therefore, a clear image can be obtained if the incident angle of at least a part of the light, that is, the light reflected on the back side of the plate-like portion is 75 to 85 °.

  According to another feature of the present invention, the longitudinal light guide member includes a first longitudinal light guide member and a second longitudinal light guide member, and each of the plurality of longitudinal light guide members is arranged laterally. Light is guided to the directional light guide member, and a first light emitter that makes the light incident on the first longitudinal light guide member and a second that makes the light incident on the second longitudinal light guide member. A light emitter, and the reflecting portion is provided on each of the first vertical light guide member and the second vertical light guide member, and the reflective portion of the first vertical light guide member and the second light guide member. The reflection angle of the vertical light guide member is different from that of the reflection portion.

  According to this, since the first vertical light guide member and the second vertical light guide member can independently enter light, the influence of disturbance on the light propagating through the horizontal light guide member Can be suppressed. Furthermore, since the optical path can be adjusted in each of the first light emitter and the first vertical light guide member, and the second light emitter and the second vertical light guide member, maintenance is facilitated. Can do.

  Another feature of the present invention is that the front end of the imaging unit is an objective lens, a peripheral wall portion in which a vertical light guide member is disposed, and a horizontal light guide member disposed on the bottom surface of the peripheral wall portion. The bottomed enclosure is formed by the above-described configuration, and the objective lens is included in the bottomed enclosure.

  According to this, even when the observation object is immersed in the liquid, the surface facing the observation object of the lateral light guide member is not observed in the liquid without the objective lens being immersed in the liquid. It can be in the vicinity of an object. Further, if the objective lens is sealed with a bottomed enclosure, the objective lens can be inserted into the living tissue and the living tissue can be observed as it is.

  Another feature of the present invention in the formation of the bottomed envelope is that the longitudinal light guide member includes a plate-shaped first longitudinal light guide member and a second longitudinal light guide member, The light irradiating means includes two plate-like members connected to the vertical light guide member and the horizontal light guide member, and the vertical light guide member and the horizontal light guide member are integrally bottomed with the two plate members. This is because an outer enclosure is formed.

  According to this, the same effect as described above can be obtained, and the processing is facilitated. In particular, if the vertical light guide member, the horizontal light guide member, and the plate-like member are all plate-shaped, the processing is most facilitated and the manufacturing cost can be suppressed.

  Another feature of the present invention is that the optical observation device includes a plate-shaped light guide member that is detachably formed on a surface of the lateral light guide member that faces the observation object. In the state of being mounted on the lateral light guide member, the light is guided integrally with the lateral light guide member.

  The lateral light guide member may be contaminated or contaminated by contact with the observation object or chemical substance, or may be damaged or lost due to an operational error or unexpected situation during the observation operation. If a plate-shaped light guide member is mounted on the facing surface of the observation object that is likely to be defective, even if the portion is contaminated or defective, only the plate-shaped light guide member needs to be replaced. According to this, the maintenance of the optical observation device can be facilitated, and the useful life of the entire light guide member can be improved. In addition, for example, as shown in the prior art document, in order to obtain a fluorescent image, it is necessary to react an observation object with an enzyme to generate a fluorescent material. If it is carried, it is not necessary to add an enzyme to the place where the observation object is present and observe with an optical observation device, and the efficiency of observation can be improved. That is, if a plate-like light guide member is attached to the lateral light guide member, and the surface of the plate-like light guide member is brought close to the object to be observed, the fluorescent material can be generated. Observation can be carried out as it is with the observation device.

  Another feature of the present invention is that the optical observation apparatus is provided with a drive mechanism that changes the distance from the tip of the imaging unit to the observation object, and a stage that sets an instrument on which the observation object is placed. The mechanism is that the distance between the tip of the imaging unit and the stage is changed.

  According to this, when the observation object is set on the stage of the optical observation apparatus, the tip of the imaging unit can be observed with respect to the observation object by the driving mechanism. In addition, the drive mechanism that changes the distance between the tip of the imaging unit and the stage can have a simple structure like a normal microscope. Here, for example, when an objective lens is provided in the imaging unit, in order to obtain an observation image by enlarging a minute observation object, it is necessary to bring the tip of the imaging unit close to the vicinity of the observation object. However, if the distance from the tip of the imaging unit to the observation object can be controlled by the drive mechanism on the order of micrometers, the imaging unit is arranged at an optimal position for observation of the observation object, and the surface of the observation object And a clear observation image of the fluorescent substance emitted from the surface of the observation object can be obtained.

  Another feature of the present invention is an optical observation apparatus including a stage and a drive mechanism, which is attached to the housing and faces the observation object of the lateral light guide member from the surface of the instrument set on the stage. A plurality of distance detection means for detecting the distance to the imaging unit, the distance between the plurality of distance detection means arranged around the optical axis of the imaging unit, and the surface of the lateral light guide member facing the observation object and the stage And a change command is output to the plurality of interval change means arranged at a position in the normal direction of the stage surface from the distance detection position of the plurality of distance detection means and the plurality of interval change means. When a change command is input from the interval change command means in a state where the plurality of distances detected by the plurality of distance detection means are equal to or less than the set distance by the interval change command means and the drive mechanism, A plurality of distances detected by the separation detecting means, and a plurality of interval changing means for controlling the plurality of interval changing means so that the plurality of interval changing means change to the commanded value or the commanded positive / negative side. That is.

  According to this, even if the surface of the instrument is slightly deviated from parallel to the stage surface, the instrument surface and the surface facing the observation object of the lateral light guide member are always parallel, The distance from the surface of the lateral light guide member facing the observation object to the observation object can be changed to be optimal. In this case, the optimal distance is the distance at which the captured image of the imaging unit becomes the clearest, and this optimal distance changes the distance from the tip of the imaging unit to the observation object by the output from the interval change command means. Each time it is done, it can be determined by checking the captured image of the imaging unit.

  Another feature of the present invention is that in an optical observation apparatus including a stage and a drive mechanism, the observation target is a living tissue or a cell, and is within the imaging range of the imaging unit based on spatial resolution information. Calculate the distance from the surface facing the observation object of the lateral light guide member to the observation object, and add the setting value calculated based on the information of the observation object to the distance to make the optimum distance the optimum A distance calculating means, and a distance detecting means for detecting a distance from a surface of an instrument attached to the housing and set on the stage to a surface facing the observation target of the lateral light guide member within the imaging range of the imaging unit; The distance detected by the distance detecting means by the distance changing means for changing the distance between the surface facing the observation target of the lateral light guide member within the imaging range of the imaging unit and the stage, and the drive mechanism When the optimum distance calculated by the optimum distance calculation means is input in a state where the distance is equal to or less than the set distance, the interval control for controlling the interval changing means so that the distance detected by the distance detection means becomes the optimum distance. And means.

  According to this, the distance from the surface facing the observation target of the lateral light guide member within the imaging range of the imaging unit to the observation target can be shortened by using the value calculated by the optimum distance calculation means. Therefore, it is possible to obtain an optimum distance, so that a captured image of the observation object can be obtained in a short time, and work efficiency can be improved.

1 is an overall schematic diagram of an optical observation apparatus according to an embodiment of the present invention. It is a fragmentary sectional view which shows the structure of the objective lens and stage vicinity of the optical observation apparatus of FIG. It is the figure which looked around the objective lens of the optical observation apparatus of FIG. 1 from the stage side. It is a figure which shows the structure of the distance sensor and distance detection apparatus of the optical observation apparatus of FIG. It is a figure which shows the control system of the optical observation apparatus of FIG. It is a flowchart of the program which the stage height control apparatus in the control system of FIG. 5 performs. It is the figure which showed a mode that the plate-shaped light guide member was mounted | worn with the optical observation apparatus in another embodiment of this invention. It is a fragmentary sectional view which shows the structure of the objective lens and stage vicinity of the optical observation apparatus which concerns on another embodiment of this invention. It is the figure which looked around the objective lens of the optical observation apparatus which concerns on another embodiment of this invention from the stage side. 2 is an overall schematic view of an optical observation apparatus according to another embodiment of the present invention. It is a fragmentary sectional view which shows the structure of the front-end | tip of an imaging unit at the time of applying this invention to the optical observation apparatus which inserts in a biological tissue and images.

  A configuration of an optical observation apparatus according to an embodiment of the present invention will be described with reference to FIGS. FIG. 1 is an overall schematic diagram of an optical observation apparatus. This optical observation apparatus performs imaging by irradiating a specimen S, which is an observation object placed on the inner bottom surface of a glass dish P set on a stage 30, with an excitation light, and is disclosed in Patent Document 2. A clear image is created and displayed by the image processing of the computer device 80, which is an image processing for removing the background light component by utilizing the different mixing ratio of the target light component and the background light component in different excitation lights. Device. As shown in FIG. 1, the overall appearance is almost the same as that of an optical microscope having a photographing function, but a light guide member 20 is attached to the apparatus main body 1 so as to surround the objective lens 10, and the light guide member 20 has an inside. A feature point is that it includes a system for accurately controlling the distance between the point where the excitation light is incident and the bottom surface of the specimen S and the light guide member 20 (the lower surface of the lateral light guide member 25 described later). is there. In addition, as shown in Patent Document 1 and Patent Document 2 of the prior art document, an enzyme and a coenzyme for reacting with a substance included in the specimen S to generate a fluorescent substance are added inside the glass dish P. . Therefore, the specimen S adhering to the inner surface of the glass plate P generates a fluorescent material, and fluorescence is generated when the specimen S is irradiated with excitation light. The specimen S may be a product cultured in a glass dish P, or may be collected from another container or specimen and transferred to the glass dish P.

  The optical observation apparatus has the same function as an optical microscope having a normal photographing function. FIG. 2 is a partial cross-sectional view showing the structure of the apparatus main body 1 near the objective lens 10 and the stage 30. The structure from the objective lens 10 shown in FIG. 2 to the camera 2 shown in FIG. The structure of the optical microscope with is applied as it is. Hereinafter, this portion is referred to as an imaging unit. Similarly, the structure from the objective lens 10 to the eyepiece 3 is the same as that of a normal optical microscope. That is, this optical observation apparatus can perform observation by visual observation of an observation region (photographing region) near the focal point of the objective lens 10 and photographing by the camera 2 in the same manner as an optical microscope having a normal photographing function. The optical observation apparatus has an elevating mechanism similar to a normal optical microscope, and the base 5 moves in the vertical direction by rotating the elevating handle 4.

  As shown in FIG. 2, the structure for attaching the objective lens 10 to the apparatus main body 1 is also the same as that of a normal optical microscope, and the male screw of the objective lens 10 is a cylindrical shape formed on the bottom wall 6 of the apparatus main body 1. It is attached by screwing into a female screw produced inside the attachment portion 7. Above the cylindrical mounting portion 7, an optical system portion 12 having a cylindrical tip is attached, and a plurality of optical components are arranged up to the camera 2 and eyepiece 3 as in an optical microscope having a normal photographing function.

  A light guide member 20 is attached around the objective lens 10. The light guide member 20 includes a first vertical light guide member 21, a second vertical light guide member 22, a first reflecting portion 23, a second reflecting portion 24, a horizontal light guide member 25, and a third vertical light guide member. 26, a fourth vertical light guide member 27 and a screw portion 28, and surrounds the objective lens 10. For this reason, even if the light guide member 20 is put in the liquid, the liquid does not enter the inside of the light guide member 20, and the objective lens 10 is not contaminated with the liquid. As with the objective lens 10, the light guide member 20 is detachable from the apparatus main body 1, and is a disk in which a female screw is formed in a central hole on a male screw formed on the outer peripheral tip of the cylindrical mounting portion 7. It is attached by screwing the screw-shaped portion 28.

  The material of the light guide member 20 is glass such as BK7 except for the screw portion 28. However, the location where the light is reflected on the top surfaces of the first reflecting portion 23, the second reflecting portion 24, and the lateral light guide member 25 is deposited with a metal having a reflectance close to 100%, such as aluminum or silver, on the surface. Is supposed to be reflected. The light guide member 20 is coated with a light shielding material or a light shielding sheet on the surface of the light guide member 20 so that light does not leak outside except for the upper end of the light guide member 20 and below the tip of the objective lens 10. ing. This is because the light applied to the specimen S is limited to the light traveling along the optical axis indicated by the one-dot chain line in FIG. In addition, with such a configuration, the observation light from the specimen S enters the objective lens 10. In addition, about the upper surface of the horizontal direction light guide member 25, it is good also as a glass surface, without requiring processes, such as the above metal vapor deposition.

  The first vertical light guide member 21 and the second vertical light guide member 22 are formed in a plate shape, and the first LED light irradiator 13 and the second LED light irradiator 14 of the apparatus body 1 are cylindrical. The inner diameter of the exit port is smaller than the thickness of the first longitudinal light guide member 21 and the second longitudinal light guide member 22. Then, the light guide member 20 is attached to the apparatus main body 1 by screwing the female screw of the screw portion 28 to the male screw formed at the outer peripheral tip of the cylindrical attachment portion 7, and the rotation of the screw portion 28 is stopped. When the female screw is screwed in, the upper ends of the first longitudinal light guide member 21 and the second longitudinal light guide member 22 are poles of the exit port of the first LED light irradiator 13 and the exit port of the second LED light illuminator 14. Located in the vicinity. The central axes of the first LED light irradiator 13 and the second LED light irradiator 14 are at the center positions of the first vertical light guide member 21 and the second vertical light guide member 22 in the plate thickness direction. Thereby, when the light guide member 20 is attached to the apparatus main body 1 and the LED light is emitted from the first LED light irradiator 13 and the second LED light irradiator 14, most of the LED light is the first longitudinal light guide member 21 and The light enters the second vertical light guide member 22 and travels through the light guide member 20.

  In the first LED light irradiator 13 and the second LED light irradiator 14, the first LED light source 15 and the second LED light source 16 and the collimating lenses 17 and 18 are attached to a cylindrical frame, and the LED light is emitted from each of them. When emitted, approximately parallel LED light enters the light guide member 20 from the end surfaces of the first vertical light guide member 21 and the second vertical light guide member 22. Hereinafter, the LED light emitted from the first LED light irradiator 13 is referred to as first LED light, and the LED light emitted from the second LED light irradiator 14 is referred to as second LED light.

  The first LED light and the second LED light incident from the end surfaces of the first vertical light guide member 21 and the second vertical light guide member 22 are reflected by the first reflection unit 23 and the second reflection unit 24, respectively, and are guided in the horizontal direction. Proceed to the optical member 25. The lateral light guide member 25 is also formed in a plate shape, and the upper and lower planes are perpendicular to the optical axis of the objective lens 10 (the optical axis of the imaging unit). Further, the optical axis of the objective lens 10 (the optical axis of the imaging unit) is at the lateral center of the lateral light guide member 25. When the light guide member 20 is attached to the apparatus main body 1, the center of the imaging region of the imaging unit is substantially coincident with the lower surface of the lateral light guide member 25, and the camera 2 is near the lower surface of the lateral light guide member 25. Can be obtained. Hereinafter, the center of the imaging region of the imaging unit is referred to as the imaging center.

  The first reflecting portion 23 is at an angle close to 45 ° with respect to the central axis of the first vertical light guide member 21, and the first LED light is reflected at an angle close to 90 ° with respect to the central axis, and the horizontal light guide member It is incident on the lower surface of the lateral center position of 25 at an incident angle close to 90 °. The second reflector 24 is at an angle of about 40 ° with respect to the central axis of the second longitudinal light guide member 22, and the second LED light is reflected at an angle of about 80 ° with respect to the central axis, and incident at about 80 °. The light is incident on the upper surface of the lateral light guide member 25 at an angle and is reflected, and is incident on the lower surface of the lateral center position of the lateral light guide member 25 at an incident angle of about 80 °. That is, the first LED light is incident on the photographing center of the lateral light guide member 25 at an incident angle close to 90 °, and the second LED light is incident on the photographing center of the lateral light guide member 25 at an incident angle of about 80 °.

  The inventors have taken an image with the incident angle of the first LED light being close to 90 ° and changing the incident angle of the second LED light in various ways, and when the incident angle of the second LED light is around 80 °, the most is the case. The inventors have found that there is a difference in the ratio of the target light and the background light between the photographed image by the first LED light and the photographed image by the second LED light, and a clear image can be obtained by image processing. Therefore, the angle of the second reflecting portion 24 and the thickness of the lateral light guide member 25 may be set so that the incident angle of the second LED light is in the range of 75 to 85 °. In FIG. 2, in order to emphasize that the first LED light and the second LED light are incident on the lower surface of the lateral light guide member 25 at different incident angles, the incident angles are significantly smaller than 90 ° and 80 °. It is.

  The third vertical light guide member 26 and the fourth vertical light guide member 27 are also formed in a plate shape, and the plane thereof is parallel to the paper surface of FIG. The screwed portion 28 is a disc-shaped member fixed to the first to fourth longitudinal light guide members 21, 22, 26, 27, and the cylindrical mounting portion 7 of the apparatus main body 1 is in the central hole. A female thread is formed that mates with a male thread formed at the outer tip. As described above, the light guide member 20 is attached to the apparatus main body 1 by screwing the female screw of the screw portion 28 in accordance with the cylindrical attachment portion 7.

  2 are distance sensor mounting portions 19-1 and 19-2, which are imaginary lines because they are in front of the cross-sectional position in FIG. Further, the distance sensor mounting portion 19-3 is not visible by the objective lens 10 and the third longitudinal light guide member 26 in FIG. FIG. 3 is a view of the state when the light guide member 20 is attached to the apparatus main body 1 as viewed from the stage 30 side. The objective lens 10, the cylindrical mounting portion 7, the screwed portion 28, and the like are shown by dotted lines because they are not visible by the lateral light guide member 25 and the first and second longitudinal light guide members 21 and 22. . As shown in FIG. 3, the three distance sensor mounting portions 19-1, 19-2 and 19-3 are arranged at intervals of 120 ° with the optical axis of the objective lens 10 (the optical axis of the imaging unit) as the center. It is formed at a position slightly outward from the light guide member 20 in the radial direction from the optical axis of the objective lens 10. As shown in FIG. 2, the distal ends of the distance sensor mounting portions 19-1, 19-2 and 19-3 are slightly above the lower surface of the lateral light guide member 25. This interval is, for example, in the range of 2 to 5 mm, and even if the lateral light guide member 25 is about to come into contact with the inner bottom surface of the glass dish P, the distance sensor mounting portions 19-1, 19-2, 19-3 The tip is not in contact and is a sufficiently safe interval. Note that the actual interval is a known value, and is stored as a default value so as to be used by the data processing device 62 described later. Further, even if the lower surface of the lateral light guide member 25 is about 10 mm above the inner bottom surface of the glass dish P, the distance sensor 40-1 attached to the distance sensor attachment portions 19-1, 19-2, 19-3. , 40-2, 40-3 and the distance detection device 50 are distances that allow distance measurement from the tips of the distance sensors 40-1, 40-2, 40-3 to the inner bottom surface of the glass plate P.

  FIG. 4 is a diagram illustrating the structure of the distance sensors 40-1, 40-2, and 40-3 attached to the distance sensor attaching portions 19-1, 19-2, and 19-3 and the structure of the distance detecting device 50. is there. The distance detection device 50 is a device that uses the measurement principle of optical coherence tomography (OCT). Optical coherence tomography is often used as a device for detecting a cross-sectional image of a human body (especially the fundus). It can also be used as a detection device. Hereinafter, although the structure and structure of the distance sensors 40-1, 40-2, and 40-3 and the distance detection device 50 will be described, only known portions will be described briefly.

  The distance detecting device 50 includes a laser light source 51, a collimating lens 52, a condenser lens 53, an optical coupler 55, a light receiving sensor 59, and optical fibers 54, 56, 57, 58 connected to the optical coupler 55. The laser light source 51 is composed of a super luminescent diode (SLD) or LED, and emits a laser beam with low coherence. The collimating lens 52 converts this laser light into parallel light, and the condensing lens 53 condenses the parallel light from the collimating lens 52 and makes it incident on the optical fiber 54. The focal length of the condensing lens 53 is set so that the laser light incident in the optical fiber 54 is totally reflected in the optical fiber 54, and the laser light incident on the optical fiber 54 is guided to the optical coupler 55.

  The optical coupler 55 splits the laser light incident via the optical fiber 54 into two, one is incident on the optical fiber 57 leading to the optical changeover switch 60 described later, and the other is the optical fiber 56 connected to the optical path length varying device 61 described later. To enter. The optical coupler 55 receives the reflected light guided from the distance sensors 40-1, 40-2, and 40-3 via the optical fiber 57 and the reflected light guided from the optical path length variable device 61 via the optical fiber 56, respectively. The light is branched into two, and one of them is guided to the light receiving sensor 59 through the optical fiber 58. In this embodiment, the incident light is branched into two using the optical coupler 55. However, the incident light is converted into parallel light having a small cross-sectional diameter, and then branched into two using a beam splitter. You may let them.

  The light receiving sensor 59 outputs a signal having a magnitude representing the intensity of the received laser beam. Since the two reflected lights incident on the light receiving sensor 59 interfere and the laser light has low coherence, the signal output from the light receiving sensor 59 is returned to the optical coupler 55 by the two laser lights branched by the optical coupler 55. The intensity increases only when the optical path lengths until the arrival are equal.

  The distance detecting device 50 further includes an optical changeover switch 60, an optical path length varying device 61, a data processing device 62, a laser driving circuit 63, and a controller 64. When a switching command is input from the controller 64, the optical switch 60 connects the optical fiber 57 and the commanded optical fiber among the optical fibers 65-1, 65-2, and 65-3. The ends of the optical fibers 65-1, 65-2, and 65-3 are connected to the distance sensors 40-1, 40-2, and 40-3, and the laser light is transmitted to the distance sensors 40-1, 40-2, and 40-3. Out of the distance sensor commanded from the controller 64. Then, the laser light emitted from the commanded distance sensor among the distance sensors 40-1, 40-2, 40-3 is reflected by the inner bottom surface of the glass plate P, and travels backward in the same optical path to the optical coupler 55. Return to. Therefore, when the command from the controller 64 is changed in order at every minute time interval and the commanded optical fiber is sequentially changed and input to the optical changeover switch 60, the laser light traveling from the optical coupler 55 to the optical fiber 57 is transmitted to the inner bottom surface of the glass plate P. Are sequentially reflected at different reflection positions and returned to the optical coupler 55. When viewed in a plane perpendicular to the optical axis of the objective lens 10, this reflection position is the position of the distance sensor mounting portions 19-1, 19-2, 19-3 shown in FIG.

  The distance sensors 40-1, 40-2, 40-3 are mounted inside cylindrical distance sensor mounting portions 19-1, 19-2, 19-3. Since the distance sensors 40-1, 40-2, and 40-3 all have the same structure, FIG. 4 shows only the structure of the distance sensor 40-1, and the distance sensors 40-2 and 40-3 are omitted. Yes. The distance sensor 40-1 includes a collimating lens 41, an objective lens 42, and a reference translucent body 43. The collimating lens 41 converts the laser light emitted from the optical fiber 65-1 into parallel light and guides it to the objective lens 42. The objective lens 42 condenses the laser light that is parallel light, It emits toward the inner bottom surface of the glass dish P. By passing through the reference light-transmitting body 43, the laser light is reflected by the incident surface of the reference light-transmitting body 43 and the exit surface of the reference light-transmitting body 43, and further the liquid level of the liquid put in the glass plate P, the glass plate Reflected on the inner bottom surface of P and the outer bottom surface of glass dish P. Therefore, when the optical path length of the light returning to the optical coupler 55 becomes equal when the optical path length is changed from the minimum to the maximum by the optical path length variable device 61 described later, there are five cases where the above-mentioned reflecting portions are different, A change curve when the optical path length is changed from the minimum to the maximum with the horizontal axis indicating the amount of change in the optical path length by the optical path length varying device 61 and the vertical axis indicating the signal intensity output from the light receiving sensor 59 (hereinafter, the change in received light intensity). (Referred to as a curve) produces five peak points. In addition, the front-end | tip of each distance sensor attaching part 19-1, 19-2, 19-3 and the front-end | tip of each distance sensor 40-1, 40-2, 40-3 are formed so that it may become flush | level. .

  The distal ends of the distance sensor mounting portion 19-1 and the distance sensor 40-1 are waterproofed. The sensor mounting portion 19-1 and the distance sensor 40-1 do not enter inside. Therefore, even if there is a lot of liquid in the glass dish P, the lower surface of the lateral light guide member 25 can be brought close to just before contacting the inner bottom surface of the glass dish P. If the tip of the distance sensor 40-1 enters the liquid, the laser beam incident on the distance sensor 40-1 is reflected at four locations, but the peak when the optical path length is the shortest in the received light intensity change curve. Since the reflection location corresponding to the peak when the optical path length is the longest is determined, it is possible to associate each peak with the reflection location in the received light intensity change curve.

  Further, if the numerical aperture of the objective lens 42 is small and the lower surface of the lateral light guide member 25 is in a state of approaching the inner bottom surface of the glass dish P, the distance from the objective lens 42 to the inner bottom surface of the glass dish P changes. Even so, the inner bottom surface of the glass pan P is always within the focal depth of the objective lens 42. The reference light transmitting body 43 may be made of any material as long as the light transmittance is high, for example, glass.

  As the optical path length variable device 61, any device can be used as long as it changes the optical path length and outputs a signal representing the change amount of the optical path length. For example, as disclosed in Japanese Patent Application Laid-Open No. 2013-221811, a device in which a triangular reflector is disposed on a disk-shaped plate and the disk-shaped plate is rotated to reflect laser light with the triangular reflector. It is preferable because the optical path length can be changed at high speed and there is no backlash. When an operation command is input from the controller 64, the optical path length varying device 61 starts to operate, changes the optical path length, and outputs a signal representing the amount of change in the optical path length to the data processing device 62 and the controller 64 described later.

  The data processing device 62 starts to operate when an operation command is input from the controller 64, and a signal indicating a change in the optical path length from the optical path length variable device 61 while the start command and the end command are input from the controller 64. A signal corresponding to the received light intensity is input from the light receiving sensor 59, and the digital data of the change amount of the optical path length and the digital data of the signal intensity are stored as a pair. As will be described later, the controller 64 receives digital data when the laser light is incident on one of the distance sensors 40-1, 40-2, and 40-3 and the optical path length changes from the minimum to the maximum. Since the start command and the end command are output so as to be stored in 62, the data processing device 62 stores digital data of the received light intensity change curve. The controller 64 also outputs a signal indicating which of the distance sensors 40-1, 40-2, and 40-3 the laser beam is incident upon when the start command is given. A signal is also stored.

When an end command is input from the controller 64, the data processing device 62 uses the stored digital data of the received light intensity change curve to determine the minimum and maximum optical path length peak points among all the peak points of the received light intensity curve. The optical path length of the peak point is detected. And the distance from the output surface of the reference | standard translucent body 43 to the inner bottom face of the glass dish P using the refractive index n (the refractive index of water is sufficient) and the refractive index n0 of air which are put into the glass dish P D is calculated. Assuming that the detected optical path lengths are P1, P2, and P3 in ascending order, the formula for calculating the distance D is as follows. Dividing by 2 is that the laser beam is reflected and returns to the original, so the difference in optical path length between the two points is the difference due to the round trip of the laser beam, and the distance between the two points is half of the difference in optical path length. Because it becomes.
(Equation 1)
D = {(P2-P1) + (P3-P2). (N0 / n)} / 2
When the tips of the distance sensors 40-1, 40-2, and 40-3 are in the liquid, the received light intensity change curve has four peaks, and the optical path that is detected except for the peak points of the minimum and maximum optical path lengths. The length becomes two, P1 and P2, in ascending order. At that time, the calculation formula is the following formula 2.
(Equation 2)
D = {(P2-P1). (N0 / n)} / 2

  When the distance D is calculated, the data processing device 62 performs the lateral direction from the exit surface of the reference translucent body 43 for each of the distance sensors 40-1, 40-2, and 40-3 that are obtained and stored with high accuracy in advance. The distance from the lower surface of the light guide member 25 to the inner bottom surface of the glass dish P is calculated by subtracting the distance to the lower surface of the light guide member 25. Then, the digital data of the calculated distance W is output to the controller 64. The data processing device 62 performs the above-described arithmetic processing every time a start command and an end command are input from the controller 64 until a stop command is input from the controller 64, and the controller 64 outputs the digital data of the calculated distance W to the controller 64. Output.

  The laser drive circuit 63 starts operating when an operation command is input from the controller 64, and outputs a voltage and a current for emitting a laser beam with a certain intensity to the laser light source 51. The controller 64 is a microcomputer, and when an operation command is input from a stage height control device 75 described later, the operation command is output to the optical path length varying device 61, the data processing device 62, and the laser driving circuit 63, and the optical path length varying device 61 is output. From the signal representing the change amount of the optical path length input from, the timing when the minimum optical path length and the maximum optical path length are reached is detected, and a start command and an end command are output to the data processor 62. Further, each time a start command is output, the laser light that is incident on one of the optical fibers 65-1, 65-2, 65-3 to the optical changeover switch 60 is changed to the optical fibers 65-1, 65-2. , 65-3, a command for switching to another one is output, and a signal that can identify the optical fiber on which the laser beam is incident is output to the data processing device 62. This switching command is such that the laser light is sequentially incident on the optical fibers 65-1, 65-2, 65-3, that is, the laser light is emitted in order from the distance sensors 40-1, 40-2, 40-3. Is done. The controller 64 repeatedly executes the above-described processing until a stop command is input from the stage height control device 75, and performs lateral guidance at the positions of the distance sensors 40-1, 40-2, 40-3 at minute time intervals. The output of the distance from the lower surface of the optical member 25 to the inner bottom surface of the glass dish P is repeated. When a stop command is input from a stage height control device 75 described later, a stop command is output to the optical path length varying device 61, the data processing device 62, and the laser driving circuit 63, and the above-described processing is stopped.

  FIG. 5 is a diagram illustrating a control system of the optical observation apparatus. The control device 70 provided in the apparatus main body 1 includes a first LED drive circuit 73, a second LED drive circuit 74, a stage height control device 75, and drive signal output circuits 76-1, 76-2, 76-3. I have. Further, an input device 71 that inputs a command to the control device 70 and lamps 77 and 78 that are turned on and off by a signal from the control device 70 are provided on the outer surface of the apparatus body 1.

  When a lighting command is input from the input device 71, the first LED driving circuit 73 outputs a voltage and a current for emitting LED light having a light amount set in the first LED light source 15, and the first LED driving circuit 73 is turned off from the input device 71. When a command is input, output of voltage and current is stopped. The second LED drive circuit 74 also outputs the same output to the second LED light source 16 in response to an input from the input device 71. Accordingly, by operating the input device 71, the sample S can be irradiated with the first LED light or the second LED light through the light guide member 20.

  Similarly, the first LED drive circuit 73 and the second LED drive circuit 74 also perform voltage and current output and output stop in response to a command from the computer device 80. Further, the camera 2 outputs captured image data to the computer device 80 when a command is input from the computer device 80. The computer device 80 is installed with a program for sequentially driving the first LED driving circuit 73 and the second LED driving circuit 74 and sequentially inputting photographed image data from the camera 2, and this program is executed by input from the input device 81. Is done. That is, captured image data when the sample S is irradiated with the first LED light and the second LED light by the input from the input device 81 is taken into the computer device 80, respectively. The image processing shown is executed, and a clear image of the specimen S is displayed on the display device 82.

  The drive signal output circuits 76-1, 76-2, and 76-3 store a relation table or relational expression between the stretch width and the signal intensity, and the stretch width digital data input from the stage height control device 75 described later. On the basis of the above, a drive signal having an intensity with which the piezoelectric actuators 31-1, 31-2, and 31-3 input the extension width is output. The stretch width means the length that the piezoelectric actuators 31-1, 31-2, 31-3 extend from the state where no drive signal is input. In the description of the program flow of FIG. The width is used in that sense. The piezoelectric actuators 31-1, 31-2, 31-3 are attached between the stage 30 and the base 5, and are slightly expanded and contracted by the input voltage. The attachment positions of the piezoelectric actuators 31-1, 31-2, 31-3 on the stage 30 are positions where the central axes of the distance sensors 40-1, 40-2, 40-3 intersect the stage 30, and the piezoelectric actuator 31 When -1, 31-2, 31-3 expands and contracts, the expansion / contraction amount is added to the distances detected by the distance sensors 40-1, 40-2, 40-3 and the distance detection device 50 as they are. The distance between the stage 30 and the base 5 and the inclination of the stage 30 with respect to the base 5 change due to the expansion and contraction of the piezoelectric actuators 31-1, 31-2, and 31-3. The interval between the lower surfaces of the optical members 25 and the inclination of the lateral light guide member 25 with respect to the inner bottom surface of the glass dish P also change.

  The position detection sensor 72 is attached to an elevating mechanism of the base 5 of the apparatus main body 1, and the base 5 is raised by rotating the elevating handle 4 to raise the base 5 to a height higher than a set height. Then, a signal indicating the start of operation is output to a stage height controller 75 described later. Further, the base 5 is lowered, and when the base 5 is below the set height, a signal indicating an operation stop is output to the stage height control device 75.

  The stage height control device 75 is a microcomputer, and when an operation start signal is input from the position detection sensor 72, the installed program of the flow shown in FIG. In accordance with a command input from 80, the distance between the stage 30 and the base 5 is controlled and the inclination of the stage 30 with respect to the base 5 is controlled to be optimum. This controls the distance between the inner bottom surface of the glass dish P and the lower surface of the lateral light guide member 25 according to an input command, and controls the inner bottom surface of the glass dish P and the lower surface of the lateral light guide member 25 to be parallel. Is. Hereinafter, the control performed by the stage height control device 75 when the operator rotates the lifting handle 4 to raise the base 5 will be described in accordance with the flow program shown in FIG.

  When an operation start command is input from the position detection sensor 72, the program starts in step S10, and the operation command is output to the controller 64 of the distance detection device 50 in step S12. Accordingly, as described above, the distance W from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P at the positions of the distance sensors 40-1, 40-2, and 40-3 from the distance detection device 50. However, it keeps inputting in order at minute time intervals. Next, in step 14, time measurement is started. Although the time measurement will be described later, output to the drive signal output circuits 76-1, 76-2, 76-3 is performed at time intervals, and the glass plate P is removed from the lower surface of the lateral light guide member 25. This is to change the distance W to the inner bottom surface at a low speed. Next, in step S16, the stretch width (1) to the stretch width (3) are set to "0", and in step S17, the stretch width (1) to the stretch width (3) are changed to the drive signal output circuits 76-1, 76. Output to -2, 76-3. However, since the expansion width of the piezoelectric actuators 31-1, 31-2, 31-3 is “0” at the start of the program, there is no change.

  In step S18, the distance W input from the distance detection device 50 is fetched. In step S18, all the distances W at the positions of the distance sensors 40-1, 40-2, and 40-3 are captured. Hereinafter, these three distances W are referred to as distance (1) to distance (3). Next, in step S20, it is determined whether the maximum value of the distance (1) to the distance (3) is equal to or less than the set value A. If not, it is determined as No and the process passes through steps S22, S23, and S24. Then, the process returns to step S18. In step S22, a lamp 77 (hereinafter, adjustable display) indicating that the distance from the lower surface of the lateral light guide member 25 attached to the outer surface of the apparatus main body 1 to the inner bottom surface of the glass dish P can be adjusted. This process is not related to this case because the lamp 77 is turned off when the program starts. Step S23 determines whether or not an operation stop signal has been input from the position detection sensor 72. At the stage where the program is started, the lift handle 4 is rotated to raise the base 5. It determines with No and goes to step S24. Step S24 is a process of returning to step 16 when the maximum value of the stretch width (1) to the stretch width (3) is not “0”, but at the stage where the program is started, the stretch width (1) to the stretch width Since all the widths (3) are “0”, it is determined Yes and the process goes to step S24.

  In the processing of step S18 and step S20, the distance from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P is reduced to a distance that can be adjusted by the expansion and contraction of the piezoelectric actuators 31-1, 31-2, 31-3. This is a process for determining whether or not. Since the base 5 continues to rise at the stage where the program is started, if the processes of Step S18 and Step S20 are repeated, the distance (1) to the distance (3) continue to decrease, and eventually the distance (1 ) To the distance (3) is equal to or less than the set value A, it is determined Yes in step S20, and the process proceeds to step S26. At this time, the distance from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P becomes a distance that can be adjusted by the expansion and contraction of the piezoelectric actuators 31-1, 31-2, 31-3.

  In step S26, the adjustable display lamp 77 attached to the outer surface of the apparatus main body 1 is turned on. This indicates that the adjustment can be made by the expansion and contraction of the piezoelectric actuators 31-1, 31-2, 31-3 as described above. Stop rotation. In other words, the operator raises the base 5 by rotating the lifting handle 4 until the adjustable display lamp 77 is lit.

  Next, in step S28, it is determined whether the minimum value of the distance (1) to the distance (3) is equal to or less than the set value B. The set value B is a distance at which the lower surface of the lateral light guide member 25 may come into contact with the inner bottom surface of the glass dish P when the distance (1) to the distance (3) is further reduced. That is, it is a value for determining the upper limit of the elongation amount of the piezoelectric actuators 31-1, 31-2, 31-3. In addition, since the sample S exists between the lower surface of the lateral light guide member 25 and the inner bottom surface of the glass dish P, when the sample S is a delicate substance, it is not crushed. When S is rigid, the set value B may be determined in consideration of a gap in order to avoid damage to the distance sensors 40-1, 40-2, and 40-3. Since the minimum value of the distance (1) to the distance (3) does not become the set value B or less at the time when the adjustable display lamp 77 is lit, it is determined No in step S28, and the process proceeds to step S32. In step S32, a lamp 78 (hereinafter, limit display) indicating that the distance from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P, which is attached to the outer surface of the apparatus main body 1, is the minimum limit. The limit display lamp 78 is turned off at this point, and the process proceeds to step S34 as it is.

  In step S34, differences Δdc (1) to Δdc (3) between the values of distance (1) to distance (3) and the minimum value of distance (1) to distance (3) are calculated. There are three types: Δdc (1) = {distance (1) −minimum value}, Δdc (2) = {distance (2) −minimum value}, Δdc (3) = {distance (3) −minimum value}. The value is calculated, and naturally one value becomes zero. Next, in step S36, Δdc (1) to Δdc (3) are added to the stretch width (1) to the stretch width (3), and in step S38, the stretch width (1) to the stretch width (3) are driven. The signal is output to the signal output circuits 76-1, 76-2, 76-3. Thereby, the difference in the distance (1) to the distance (3) is eliminated by the extension of the piezoelectric actuators 31-1, 31-2, 31-3, and the lower surface of the lateral light guide member 25 is made of the glass dish P. Parallel to the inner bottom surface. If all values of distance (1) to distance (3) match in step S34, the matching value is set as the minimum value. In this case, Δdc (1) to Δdc (3) are 0, and steps S36 and S38 are not substantially performed. Therefore, after the lower surface of the horizontal light guide member 25 becomes parallel to the inner bottom surface of the glass dish P, the processes in steps S34 to S38 are not performed unless the posture of the glass dish P is changed.

  The process of step S40 to step S58 is performed by expanding or contracting the piezoelectric actuators 31-1, 31-2, 31-3 in accordance with an input from the input device 71 or an input from the computer device 80, and the lower surface of the lateral light guide member 25. It is a process which changes the distance from the inner bottom face of the glass pan P. The input device 71 has an input for moving the stage 30 upward (input for extending the piezoelectric actuators 31-1, 31-2, 31-3) and an input for moving the stage 30 downward (piezoelectric actuators 31-1, 31-2 and 31-3 are input), and an instruction for upward movement or downward movement is output to the stage height control device 75 in accordance with this input. Moreover, the computer apparatus 80 calculates | requires and outputs the optimal distance from the lower surface of the horizontal direction light guide member 25 to the inner bottom face of the glass plate P by arithmetic processing so that it may mention later. Hereinafter, processing will be described for each case.

  When an input of an upward movement is input from the input device 71, it is determined No in step S40, Yes is determined in step S42, and the current stretch width (1) to stretch width (3) are determined in step S44. Is added to the preset minute length Δdt, and the extension width (1) to the extension width (3) are output to the drive signal output circuits 76-1, 76-2, and 76-3 in step S58. As a result, the piezoelectric actuators 31-1, 31-2, 31-3 extend by Δdt, and the stage 30 rises by Δdt. In other words, the lower surface of the lateral light guide member 25 approaches the inner bottom surface of the glass dish P by Δdt.

  When the input of the downward movement is input from the input device 71, it is determined No in step S40, No is determined in step S42, Yes is determined in step S46, and the current expansion width is determined in step S48. It is determined whether the minimum value of (1) to elongation width (3) is equal to or greater than Δdt. Then, if it is larger, it is determined as Yes, and in step S50, a preset minute length Δdt is subtracted from the stretch width (1) to the stretch width (3), and in step S58, the stretch width (1) to the stretch width is subtracted. The width (3) is output to the drive signal output circuits 76-1, 76-2, and 76-3. As a result, the piezoelectric actuators 31-1, 31-2, 31-3 contract by Δdt, and the stage 30 moves down by Δdt. In other words, the lower surface of the lateral light guide member 25 is moved away from the inner bottom surface of the glass plate P by Δdt. If the minimum value of the current extension width (1) to extension width (3) is smaller than Δdt in step S48, it is impossible to shrink Δdt, so it is determined No, and the piezoelectric actuators 31-1, 31 are used. The process returns to step S18 without expanding and contracting -2, 31-3. For example, in the movement of the first stage 30 immediately after the adjustable indicator lamp 77 is turned on in step S26, any one of the extension width (1) to the extension width (3) is “0”. Even if the downward movement is input from the device 71, nothing is changed and the process returns to the step S18.

  When the optimal distance from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P is input from the computer device 80, it is determined Yes in step S40, and the optimal distance is determined from the set value B in step S52. If it is larger, it is determined as Yes, the difference ds from the optimum distance at the minimum value of the distance (1) to the distance (3) is calculated in Step S54, and the current stretch width (1) to stretch width ( The difference ds is added to 3), and the extension width (1) to the extension width (3) are output to the drive signal output circuits 76-1, 76-2, 76-3 in step S58. At this time, if all the values of the distance (1) to the distance (3) match in step S54, the matching value is set as the minimum value. Thereby, the distance from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P becomes the optimum distance input from the computer device 80. When the optimum distance is equal to or smaller than the set value B in step S52, the determination is No, the process goes to step S42, and the movement by the input from the input device 71 described above is performed. This is to prevent the stage 30 from being lifted even if a distance at which there is a risk that the lower surface of the lateral light guide member 25 contacts the inner bottom surface of the glass dish P is input from the computer device 80 as the optimum distance. is there.

  When output to the drive signal output circuits 76-1, 76-2, 76-3 having the stretch width (1) to the stretch width (3) is output in step S58, the measurement time is reset in step S60 to 0. In step S62, the process returns to step S18 after waiting for ΔT to elapse. When returning to step S18, the processing from step S18 to step S62 described above is performed again, so that the processing from step S18 to step S62 is repeated unless it is determined Yes in step S23. Thus, the horizontal light guide member 25 is controlled by an input from the input device 71 or an input from the computer device 80 after the lower surface of the horizontal light guide member 25 and the inner bottom surface of the glass dish P are controlled to be parallel. The distance from the lower surface to the inner bottom surface of the glass dish P changes. The reason for waiting for the measurement time to elapse in step S62 is that the stage 30 is moved when the input device 71 is continuously input with the upward movement and the downward movement unless the time has elapsed. This is because it is performed at once.

  If the upward movement is continuously input to the input device 71, the minimum value of the distance (1) to the distance (3) may be less than or equal to the set value B. In this case, in step S28 It determines with Yes, goes to step S30, turns on the limit display lamp 78, and goes to step S46. Thereby, when the limit display lamp 78 is lit, the stage 30 is moved only by the downward movement input from the input device 71. Then, when the stage 30 moves downward and the minimum value of the distances (1) to (3) becomes larger than the set value B, it is determined No in step S28, and the process goes to step S32. Turns off.

  Further, when the distance from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P is being adjusted, if the elevator handle 4 is rotated to lower the base 5, the distance (1) to the distance ( Since the maximum value of 3) is larger than the set value A, it is determined No in step S20, the process proceeds to step S22, the adjustable display lamp 77 is turned off in step S22, No in step S23, and No in step S24. And go to step S16. In step S16, the extension width (1) to the extension width (3) are set to 0 and output in step S17, so that the extension widths of the piezoelectric actuators 31-1, 31-2, 31-3 are " 0 ”. Thereafter, when the lifting handle 4 is rotated to raise the base 5, the same processing as described above is performed from the beginning.

  Further, when the lifting handle 4 is rotated and the base 5 is kept lowered, an operation stop signal is input from the position detection sensor 72, it is determined Yes in step S23, the process proceeds to step S64, and the distance is determined in step S64. The operation stop output is output to the controller 64 of the detection device 50, the time measurement is stopped in step S66, and the program is stopped in step S68. Thereby, it returns to the same state as before the stage height control device 75 is operated.

  The computer device 80 performs calculation processing and outputs commands or calculation results to the camera 2, the stage height control device 75, and the display device 82 in accordance with input from the input device 81. When a captured image display command is input from the input device 81, an operation command is output to the camera 2 to operate, display image data is generated from the captured image data input from the camera 2, and output to the display device 82. A photographed image is displayed on 82. When a processing image display command is input from the input device 81, a preinstalled program is started, the first LED driving circuit 73 and the second LED driving circuit 74 are sequentially driven, and the captured image data is sequentially transmitted from the camera 2. The image data created by executing the image processing shown in Patent Document 2 of the prior art document is output to the display device 82, and the display device 82 displays a clear image of the specimen S.

  Further, when the computer device 80 receives the characteristic data of the sample S and information on the spatial resolution from the input device 81 and inputs an instruction for calculating the optimum distance, the computer device 80 calculates the set value and the sample from the lower surface of the lateral light guide member 25 by an arithmetic process. The distance to S is calculated, and a value obtained by adding the calculated set value to the calculated distance is output to the stage height controller 75 as an optimum distance. For example, when the specimen S is a living tissue or a cell, the optimum distance output to the stage height control device 75 is obtained by the following calculation.

  The characteristic data of the specimen S includes, for example, the standard thickness (height) of the cell when the specimen S is a living tissue or a cell. The set value calculated by the calculation process is the distance from the inner bottom surface of the glass plate P to the upper surface of the sample S, in other words, the distance from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass plate P. Thus, the lower surface of the lateral light guide member 25 is a value that substantially contacts the upper surface of the specimen S. Here, in order to avoid that the specimen S is extremely strongly pressed by the lower surface of the lateral light guide member 25 when the specimen S has a standard thickness or more, the characteristic data of the specimen S is used as characteristic data of the specimen S. An input of a value indicating a thickness variation (for example, a value to be added or a magnification) is also required. The distance from the inner bottom surface of the glass plate P to the upper surface of the sample S is calculated as a set value from the value indicating the standard thickness of the sample S and the variation in thickness by a calculation process of the computer device 80.

  The distance from the lower surface of the lateral light guide member 25 to the sample S is a value determined according to the required spatial resolution. Here, when irradiating excitation light and observing the fluorescent substance emitted from the specimen S, the emitted fluorescent substance diffuses away from the emission point, and the concentration decreases. Accordingly, when the distance between the specimen S and the lower surface of the lateral light guide member 25 is increased, the spatial resolution of fluorescence observation is lowered and the sensitivity is also lowered. Assuming that the emitted fluorescent material simply diffuses radially, the spatial resolution is the extent of the distance from the sample S, and the sensitivity decreases approximately in inverse proportion to the square of the distance. Therefore, it is necessary to appropriately set the distance between the sample S and the lower surface of the lateral light guide member 25 in accordance with the required resolution. Specifically, for example, in order to realize a spatial resolution of 10 μm, the distance from the sample S to the lower surface of the lateral light guide member 25 needs to be about 10 μm or less, which is the same as the spatial resolution. The relationship between the spatial resolution and the distance is stored in advance in the computer device 80 as a default value. When the spatial resolution is designated by the operator's input operation, the distance corresponding to the designated spatial resolution is calculated as the distance from the lower surface of the lateral light guide member 25 to the sample S. Then, the optimum distance is derived by adding the set value calculated by the above calculation process to the calculated distance. As described above, when the optimum distance is output to the stage height controller 75, the distance W from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P is controlled to be the optimum distance, The distance from the lower surface of the lateral light guide member 25 to the specimen S is a distance corresponding to the designated spatial resolution. As described above, since the first LED light is incident on the lower surface of the lateral center position of the lateral light guide member 25 at an incident angle close to 90 °, the lateral light guide member 25 as described above. When the distance from the lower surface to the specimen S is set, the LED light is irradiated to a shallow position on the surface (upper surface) of the specimen S, and a clear observation image of the surface (upper surface) of the specimen S can be obtained.

  When a clear image of the specimen S is obtained by the optical observation device configured as described above, the operator performs the following operation. First, the objective lens 10 is screwed and fixed to the apparatus main body 1, and then the screw portion 28 of the light guide member 20 is screwed into the apparatus main body 1 to fix the light guide member 20 to the apparatus main body 1. Next, the specimen S adhering to the inner bottom surface of the glass dish P is placed on the stage 30. Subsequently, the first LED light or the second LED light is irradiated by the input from the input device 71 of the apparatus main body 1, and the captured image is displayed on the display device 82 by the input from the input device 81 of the computer device 80. Next, the elevating handle 4 is rotated to raise the base 5, and the lower surface of the lateral light guide member 25 of the light guide member 20 is visually close to the inner bottom surface of the glass dish P. At this time, the program of the stage height control device 75 is started and the adjustable display lamp 77 is lit, so that the stage is determined by the input from the input device 71 of the apparatus body 1 while viewing the photographed image displayed on the display device 82. 30 is slightly moved up and down (the distance from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass plate P is slightly changed), and the position is determined to be optimal. At this time, when the characteristic data and the spatial resolution of the sample S are input from the input device 81 of the computer device 80 and an instruction for calculating the optimum distance is input, the position of the stage 30 (lateral direction guide) is calculated by the optimum distance calculated by the computer device 80. The distance from the lower surface of the optical member 25 to the inner bottom surface of the glass dish P) is determined. Therefore, in this case, an observation image of autofluorescence on the surface of the specimen S can be obtained by simply irradiating the first LED light or the second LED light without adjusting the position of the stage 30 while viewing the captured image. Furthermore, when it is desired to observe the release state of a specific physiologically active substance possessed by the specimen S, an enzyme and a coenzyme as shown in Patent Document 2 of the prior art document are added to the inside of the glass dish P, and specified. If a fluorescent substance is generated from a coenzyme according to a specific enzyme reaction with the physiologically active substance, a two-dimensional spatial distribution image of a specific physiologically active substance on the surface of the specimen S can be obtained. When the specimen S is a living tissue or a cell, the irradiation of the first LED light and the second LED light may be performed after the position of the stage 30 is determined in order to reduce the influence of phototoxicity.

  Next, when the operator inputs a processing image display command from the input device 81 of the computer device 80, an image created by executing the image processing shown in Patent Document 2 of the prior art document is displayed on the display device 82. Is done. Thus, the operator can perform processing such as analyzing the image or storing the image in the computer device 80. When the lifting handle 4 is rotated to lower the base 5, the adjustable display lamp 77 is extinguished, and the voltage output to the piezoelectric actuators 31-1, 31-2, and 31-3 becomes “0”. The operation of the distance detecting device 50 is stopped. If the worker stops the irradiation of the first LED light or the second LED light by the input from the input device 71 of the apparatus main body 1, the optical observation device is returned to the original state. Thereafter, the above-described operation can be repeatedly performed on the glass dish P to which another specimen S is attached.

  As can be understood from the above description, in the above-described embodiment, the optical observation apparatus is arranged in the imaging unit including the objective lens 10 that images the sample S, the camera 2, and the optical member therebetween, and the apparatus main body 1 including the imaging unit. And an LED light irradiation mechanism that irradiates the sample S with LED light. The LED light irradiation mechanism has a surface facing the sample S on one end side of the apparatus main body 1, and the LED light is irradiated in the surface. A lateral light guide member 25 that guides light, and a vertical light guide that is connected to the lateral light guide member 25 and guides LED light incident from a part of the apparatus body 1 to the lateral light guide member 25. The light guide members 21 and 22 and the vertical light guide members 21 and 22 are provided to reflect the incident LED light and guide it to the lateral light guide member 25, and according to the reflection position of the LED light. For the specimen S of the lateral light guide member 25 As at an incident angle different plane direction, and a plurality of reflection portions 23, 24 of different reflection angle.

  According to this, when the LED light incident from a part of the apparatus main body 1 is guided to the horizontal light guide member 25 via the vertical light guide members 21 and 22, the vertical light guide member. The LED light is reflected and guided by the reflecting portions 23 and 24 provided at 21 and 22. Here, since the plurality of reflecting portions 23 and 24 having different reflection angles are provided, the LED light has different incident angles on the surface facing the sample S of the lateral light guide member 25 according to the reflection position. Will be incident. That is, with a simple configuration in which a plurality of reflecting portions 23 and 24 having different reflection angles are provided on the longitudinal light guide members 21 and 22, a plurality of incident angles on the sample S facing the surface of the lateral light guide member 25. The optical path formation for irradiating the LED light can be realized. In addition, since the LED light irradiation mechanism can be integrally provided in the apparatus main body 1 together with the imaging unit including the objective lens 10, the camera 2, and the optical member therebetween, compared with the case where the LED light irradiation mechanism is provided separately, The entire apparatus can be reduced in size, and the handleability can be improved. In addition, since it is not necessary to place the specimen S on a specific instrument, if a plurality of specimens S are placed on different instruments, the observation can be performed efficiently by changing the instrument placed on the stage 30 and repeating the observation. The sample S can be observed.

  Moreover, in the said embodiment, the horizontal direction light guide member 25 has a plate-shaped part which has the surface which opposes the sample S, and the back surface on the other side of this surface, and is provided in the vertical direction light guide members 21 and 22. Among the plurality of reflection portions 23 and 24 provided, the reflection portion 24 guides the LED light toward the back side of the plate-like portion of the lateral light guide member 25 and guides it to the back side of the plate-like portion. The reflected LED light is reflected on the back side of the plate-like portion and then incident on the surface facing the specimen S.

  According to this, since the surface facing the specimen S of the lateral light guide member 25 can be formed by one plate-like portion, the specimen S when viewed and the surface facing the specimen S of the lateral light guide member 25, The interval of becomes easier to understand. Furthermore, since the horizontal light guide member 25 can be formed with a simple shape, the processing of the horizontal light guide member 25 can be facilitated, and the component cost can be reduced.

  Moreover, in the said embodiment, after reflecting on the back surface side of the plate-shaped part of the horizontal direction light guide member 25, LED light which injects into the surface which opposes the sample S is made so that an incident angle is 75-85 degrees. ing.

  As a result of experiments, the inventors have confirmed that in the LED light incident on the surface of the lateral light guide member 25 facing the sample S, the incident angle of one LED light is close to 90 °, and the other LED light is incident. It has been found that when the angle is around 80 °, the difference in the ratio between the target light and the background light is the largest, and a clear image can be obtained by image processing. Therefore, a clear image can be obtained if the incident angle of at least a part of the LED light, that is, the LED light reflected on the back side of the plate-like portion of the lateral light guide member 25 is 75 to 85 °. Can do.

  Moreover, in the said embodiment, the vertical direction light guide members 21 and 22 are provided with the 1st vertical direction light guide member 21 and the 2nd vertical direction light guide member 22, and these several vertical direction light guide members 21 and 22 are included. LED light is guided to the lateral light guide member 25 by each of the first LED light irradiator 13 that causes the LED light to enter the first longitudinal light guide member 21, and the second longitudinal light guide member. The second LED light irradiator 14 that causes the LED light to enter 22 is provided, and the reflectors 23 and 24 are provided in the first vertical light guide member 21 and the second vertical light guide member 22, respectively, and the first The first reflecting portion 23 of the longitudinal light guide member 21 and the second reflecting portion 24 of the second longitudinal light guide member 22 have different reflection angles.

  According to this, since the 1st vertical direction light guide member 21 and the 2nd vertical direction light guide member 22 can each make LED light enter independently, with respect to the LED light which propagates the horizontal direction light guide member 25, The influence of disturbance can be suppressed. Furthermore, the optical path can be adjusted in each of the first LED light irradiator 13 and the first vertical light guide member 21, and the second LED light irradiator 14 and the second vertical light guide member 22, thereby facilitating maintenance. can do.

  In the above embodiment, the tip of the imaging unit is the objective lens 10, and the peripheral wall portion in which the vertical light guide members 21 and 22 are disposed and the lateral light guide disposed on the bottom surface of the peripheral wall portion. A bottomed envelope is formed by the member 25, and the objective lens 10 is included in the bottomed envelope.

  According to this, even when the specimen S is immersed in the liquid, the surface of the lateral light guide member 25 that faces the observation object is not immersed in the liquid while the objective lens 10 is immersed in the liquid. It can be in the vicinity of S.

  Moreover, in the said embodiment, the raising / lowering mechanism which changes the distance from the front-end | tip of the objective lens 10 to the sample S, and the stage 30 which sets the instrument which mounted the sample S are provided, and the raising / lowering mechanism is the objective lens 10's. The distance between the tip and the stage 30 is changed.

  According to this, when the sample S is set on the stage 30, observation can be performed with the tip of the objective lens 10 at an appropriate position with respect to the sample S by the lifting mechanism. In addition, the lifting mechanism that changes the distance between the tip of the objective lens 10 and the stage 30 can have a simple structure like a normal microscope. That is, when an observation image is obtained by enlarging the minute specimen S by the imaging unit provided with the objective lens 10 as in the above embodiment, it is necessary to bring the tip of the objective lens 10 close to the vicinity of the specimen S. However, if the distance from the tip of the objective lens 10 to the specimen S can be controlled by the lifting mechanism on the order of micrometers, the objective lens 10 is disposed at an optimum position for observing the specimen S, and the characteristics of the specimen S And a clear observation image of the fluorescent substance emitted from the surface of the specimen S can be obtained.

  Moreover, in the said embodiment, the sensor and apparatus which are attached to the apparatus main body 1, and detect the distance from the inner bottom face of the glass pan P set to the stage 30 to the surface facing the sample S of the horizontal direction light guide member 25. The distance sensors 40-1, 40-2, 40-3 and the distance detection device 50 arranged around the optical axis of the objective lens 10, the surface of the lateral light guide member 25 facing the sample S, and the stage The piezoelectric actuators 31-1, 31-2, and 31-3 that change the distance between the distance sensor 30 and the distance 30 from the distance detection position of the distance sensors 40-1, 40-2, and 40-3. Piezoelectric actuators 31-1, 31-2, 31-3 arranged at positions in the linear direction, an input device 71 for outputting change commands to the piezoelectric actuators 31-1, 31-2, 31-3, When a change command is input from the input device 71 in a state where the distance detected by the distance sensors 40-1, 40-2, 40-3 is equal to or less than the set distance by the mechanism, the distance sensors 40-1, The piezoelectric actuators 40-2, 40-3 and the plurality of distances detected by the distance detecting device 50 become equal and the piezoelectric actuators 31-1, 31-2, 31-3 change to the commanded positive / negative side. And a stage height control device 75 for controlling 31-1, 31-2, and 31-3.

  According to this, even if the surface of the instrument on which the sample S is placed is slightly deviated from parallel to the surface of the stage 30, the surface of the instrument on which the sample S is placed and the sample S of the lateral light guide member 25. It is possible to change the distance from the surface facing the sample S of the lateral light guide member 25 to the sample S so that the distance between the surface and the sample S is always parallel. In this case, the optimum distance is a distance at which the image captured by the camera 2 becomes the clearest, and this optimum distance changes the distance from the tip of the objective lens 10 to the sample S due to the output from the input device 71. Each can be determined by checking the captured image of the camera 2.

  In the above embodiment, the specimen S is a living tissue or a cell, and the distance from the surface facing the specimen S of the lateral light guide member 25 to the observation location of the specimen S is calculated based on the spatial resolution information. Then, the computer apparatus 80 having a value obtained by adding a set value calculated based on the information of the sample S to the distance and the apparatus body 1 attached to the apparatus body 1 and laterally guided from the surface of the instrument set on the stage 30. Distance sensors 40-1, 40-2, 40-3 for detecting the distance to the surface of the optical member 25 facing the sample S, and the distance detection device 50, and the surface of the lateral light guide member 25 facing the sample S The distances detected by the distance sensors 40-1, 40-2, and 40-3 are set by the piezoelectric actuators 31-1, 31-2, and 31-3 that slightly change the distance from the stage 30 and the lifting mechanism. The When the optimum distance calculated by the computer device 80 is input in a state where the distance is less than or equal to the distance, the distances detected by the distance sensors 40-1, 40-2, 40-3 and the distance detection device 50 become the optimum distance. And a stage height control device 75 for controlling the piezoelectric actuators 31-1, 31-2, 31-3.

  According to this, the distance from the surface facing the specimen S of the lateral light guide member 25 to the specimen S can be made the optimum distance in a short time by using the value calculated by the computer device 80. The captured image of the specimen S can be obtained in a short time, and the working efficiency can be improved.

  In carrying out the present invention, the present invention is not limited to the above embodiment, and various modifications can be made without departing from the object of the present invention.

  In the above embodiment, the light guide in the lateral direction is performed only by the lateral light guide member 25 formed in a plate shape. However, as shown in FIG. 7, a plate-shaped light guide member 130 formed so as to be detachable is attached to the lower surface of the horizontal light guide member 25 so as to guide the light in the horizontal direction. The light guide member 130 may be configured so as to be performed. In this case, in order to prevent refraction and scattering from occurring at the joint surface between the light guide members and to allow the plate light guide member 130 to be easily attached to and detached from the lateral light guide member 25, the plate guide is used. The material of the optical member 130 may be the same as that of the lateral light guide member 25 and vaseline having a refractive index similar to that of the lateral light guide member 25 may be applied to the joint surface. Alternatively, a matching oil having a refractive index similar to that of the lateral light guide member 25 may be applied. Alternatively, a refractive index bonding film having the same refractive index as that of the lateral light guide member 25 may be attached to the bonding surface. Further, as shown in FIG. 7, the lower surface of the horizontal light guide member 25 has a width that is a minute amount larger than either the vertical or horizontal width of the plate light guide member 130 and has the same depth as the thickness of the plate light guide member 130. The groove 133 may be formed so as to cut out a part of the lower surface of the lateral light guide member 25 that serves as an imaging location. When the lateral light guide member 25 has this shape, the plate-like light guide member 130 can be mounted so as to be fitted into the imaging location, and can be easily removed. Further, the optical path of the LED light in the lateral light guide member 25 can be made the same as in the above embodiment. Thereby, the opposing surface of the observation object that is likely to be contaminated or damaged can be always maintained in a good state by appropriate replacement of the plate-shaped light guide member 130.

  Furthermore, the plate-shaped light guide member 130 is preliminarily attached to the plate-shaped light guide member 130 with an enzyme 132 or a coenzyme that reacts with a specific substance of the specimen S to generate a fluorescent substance. An observation image may be obtained as in the above embodiment. In this case, in order to attach the enzyme 132 or the coenzyme to the plate-shaped light guide member 130, the plate-shaped light guide member 130 was placed on a clean cloth with the mounting surface side facing down, and covered. What is necessary is just to spray the enzyme solution made into the mist form. Note that the enzyme 132 may be bound to the plate-shaped light guide member 130 by a reaction using a silane coupling agent of an aminosilane compound. When the plate-like light guide member 130 is attached to the horizontal light guide member 25, first, the light guide member 20 is removed from the apparatus body 1, and the horizontal light guide member 25 is faced upward. Apply petroleum jelly or matching oil to the central portion of the groove 133 or attach a refractive index bonding film. Next, the end portion of the plate-shaped light guide member 130 to which the enzyme 132 is attached is picked with tweezers or the like, and is fitted into the groove 133 of the lateral light guide member 25 and slid to an appropriate position. Then, the light guide member 20 may be attached to the apparatus main body 1. Further, after removing the light guide member 130 from the lateral light guide member 25 after obtaining the observation image, the glass plate P attached to the lateral light guide member 25 after removing the light guide member 20 from the apparatus main body 1. The liquid inside is washed away with pure water, the lateral light guide member 25 is faced upward, and the joint is removed by tweezing and opening the end of the joint between the plate light guide member 130 and the lateral light guide member 25 with tweezers or the like. Good. Thereby, the plate-shaped light guide member 130 can be appropriately replaced. If petrolatum or matching oil or refractive index bonding film remains in the groove of the lateral light guide member 25, wipe it off with a cloth soaked with a cleaning liquid such as alcohol, and if necessary, immerse it in a container containing a cleaning liquid such as alcohol. And hold for a predetermined time.

  In addition, when not obtaining the groove | channel 133 in the horizontal direction light guide member 25, but attaching the plate-shaped light guide member 130 and obtaining an observation image, the plate-shaped light guide member 130 is created as thinly as possible, and it is the same as that of the said embodiment. What is necessary is just to apply | coat petrolatum or matching oil to the center part of the horizontal direction light guide member 25, or to make it attach through a refractive index joining film. In this case, instead of the distance from the lateral light guide member 25 to the inner bottom surface of the glass dish P, it is necessary to control the distance from the surface of the plate-shaped light guide member 130 to the inner bottom surface of the glass dish P. . To this end, the distance sensors 40-1, 40-2, 40-3 stored in the data processing device 62, from the exit surface of the reference translucent body 43 to the lower surface of the lateral light guide member 25, are stored. What is necessary is just to change a distance into the distance which added the thickness of the plate-shaped light guide member 130. FIG. Since the plate-shaped light guide member 130 may or may not be mounted, when the presence / absence of the plate-shaped light guide member 130 is input from the input device 81, the computer device 80 installs the data processing device 62. When the information is input and the data processor 62 is mounted, the stored distance may be changed to a distance obtained by adding the thickness of the plate-shaped light guide member 130. When the plate-shaped light guide member 130 is attached to the horizontal light guide member 25 similar to the above embodiment, the optical path of the LED light changes. However, if the thickness of the plate-shaped light guide member 130 is made as thin as possible, the effect is as follows. Almost negligible.

  Further, not only in the case of generating a fluorescent material, but also in the case of generating another material by reacting an observation object with a specific material, the plate-shaped light guide member 130 is loaded with a necessary reactive material and similarly Observations may be made.

  Further, in the above embodiment, the second LED light is reflected by the upper surface of the lateral light guide member 25 and then enters the vicinity of the location where the optical axes of the objective lens 10 on the lower surface of the lateral light guide member 25 intersect. did. However, if it is not necessary to consider the manufacturing cost and ease of handling of the light guide member 20, the light is reflected by the second reflecting portion 24 and then directly incident on the same portion of the lateral light guide member 25 as shown in FIG. Thus, the lower surface of the lateral light guide member 25 may be formed of two planes. Also by this, since the effect that the apparatus main body 1 can be made into a simple structure and the specimen S need not be placed on a specific instrument, the stage 30 can be obtained by placing a plurality of specimens S on different instruments. By repeating the observation while changing the instrument placed on the head, it is possible to obtain an effect that a plurality of specimens S can be observed efficiently.

  Moreover, in the said embodiment, the light guide member 20 is the 1st vertical direction light guide member 21, the 2nd vertical direction light guide member 22, the horizontal direction light guide member 25, the 3rd vertical direction light guide member 26, and the 4th vertical direction. The directional light guide member 27 is formed so as to surround the objective lens 10. However, since LED light does not enter the third vertical light guide member 26 and the fourth vertical light guide member 27, the first vertical light guide member 21, the second vertical light guide member 22, and the horizontal light guide member 27. A plate-like object made of different materials may be bonded to the direction light guide member 25 so as to surround the objective lens 10. Further, the liquid depth of the liquid placed in the instrument such as the glass dish P placed on the stage 30 is limited to be small, and if the objective lens 10 does not contact the liquid, the third longitudinal light guide member 26 and the fourth vertical light guide member 27 are eliminated, and the light guide member 20 is formed by only the first vertical light guide member 21, the second vertical light guide member 22, and the horizontal light guide member 25. Also good.

  Moreover, in the said embodiment, the light guide member 20 is the 1st vertical direction light guide member 21, the 2nd vertical direction light guide member 22, the horizontal direction light guide member 25, the 3rd vertical direction light guide member 26, and the 4th vertical direction. A rectangular parallelepiped shape is formed by the direction light guide member 27. However, if the LED light incident on the first vertical light guide member 21 and the second vertical light guide member 22 is reflected by the first reflection unit 23 and the second reflection unit 24 and guided to the horizontal light guide member 25. For example, the light guide member 20 may have a shape other than a rectangular parallelepiped shape. For example, the light guide member 20 may have a cylindrical shape as shown in FIG. 9 which is a view of the state when the light guide member 20 is attached to the apparatus main body 1 as viewed from the stage 30 side. In this case, the first to fourth vertical light guide members 21, 22, 26, and 27 become one vertical light guide member 29. In this case, the sectional view corresponding to FIG. 2 is the same as FIG. Also by this, the same effect as the above embodiment can be obtained. In addition, since the distance sensors 40-1, 40-2, and 40-3 can be made closer to the light guide member 20 than in the above embodiment, the specimen S can be placed even on an instrument such as a glass dish P having a small diameter. Can be used.

  Moreover, you may make the light guide member 20 into the shape which made a part of column side surface flat. For example, the first vertical light guide member 21 and the second vertical light guide member 22 are formed in a columnar shape as in FIG. 9, and the third vertical light guide member 26 and the fourth vertical light guide member 27 are formed in the above embodiment. Similarly to the above, it may be plate-shaped. Also by this, the same effect as the said embodiment can be acquired, and distance sensor 40-1, 40-2, 40-3 is brought closer to the light guide member 20 from the said embodiment like the shape shown in FIG. Can be structured.

  In the above embodiment, the three piezoelectric actuators 31-1, 31-2, 31-3 and the three distance sensors 40-1, 40-2, 40-3 are provided, and the distance sensors 40-1, 40- are provided. 2, 40-3, the extension width of the piezoelectric actuators 31-1, 31-2, 31-3 is controlled so that the distances detected by the two are constant, The inner bottom surface and the lower surface of the lateral light guide member 25 were made parallel. However, the appliance such as the glass pan P is limited, and when it is placed on the stage 30, if the inner bottom surface is parallel to the lower surface of the lateral light guide member 25 with high accuracy, one piezoelectric actuator and one distance sensor are provided. The height of the stage 30 (the extension width of the piezoelectric actuator) may be adjusted. In this case, if the height of the stage 30 can be adjusted with the same degree of accuracy as the piezoelectric actuator, the piezoelectric actuator may be changed to another lifting mechanism. As another elevating mechanism, an automatic or manual elevating mechanism that slides one of two wedge-shaped rigid bodies can be considered.

  Further, in the above embodiment, by inputting two commands of upward movement and downward movement from the input device 71, the expansion width (stage) of the piezoelectric actuators 31-1, 31-2, 31-3 for each Δdt. (30 height) was changed. However, what can be input from the input device 71 may be other than the moving direction of the stage 30. For example, the movement direction and the movement distance may be input, or the distance from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P may be input.

  Moreover, in the said embodiment, distance sensor 40-1, 40-2, 40-3 and the distance detection apparatus 50 which detect the distance from the lower surface of the horizontal direction light guide member 25 to the inner bottom face of the glass dish P are light. The apparatus used the measurement principle of coherence tomography (OCT). However, the distance from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P can be detected with high accuracy, the distance detection range is 10 mm or more, and the distance sensors 40-1, 40-2, 40-3 are An apparatus using any measurement principle may be adopted as long as it can be miniaturized to such an extent that it can be attached to the distance sensor attaching portions 19-1, 19-2, 19-3.

  In the above embodiment, the optical observation apparatus to which the present invention is applied is an optical microscope having a photographing function, but the present invention can also be applied to other optical observation apparatuses. For example, as shown in FIG. 10, the present invention can also be applied to a system that automatically photographs and displays a sample placed on a glass plate P that is successively sent by a moving stage 91. This system is similar to the above-described embodiment in that the camera 2, the objective lens 10, the light guide member 20, the first and second LED light irradiators 13 and 14, the distance sensors 40-1 to 3, and the control device 70 are provided with the lifting device 101. Added imaging device 100, distance detection device 50 similar to the above embodiment, computer device 80, input device 81 and display device 82 similar to the above embodiment, moving stage 91, drive device 92 and drive control device 93. And a moving device 90 composed of Then, by executing a program installed in the computer device 80 by an input from the input device 81, a command is output from the computer device 80 to the imaging device 100 and the drive control device 93, and the movement stage 91 is moved and stopped. Then, the elevating device 101 is moved up and down, the first and second LED light irradiators 13 and 14 are turned on and off, and photographing by the camera 2 is performed. Then, image processing is performed by the computer device 80 as in the above embodiment, and an image is displayed on the display device 82. In this case, since there is no stage that can adjust the height and inclination in a minute amount unlike the above embodiment, the lifting device 101 includes a minute height adjustment mechanism and a minute inclination adjustment mechanism. A stage height control device 75 in the control device 70 controls the minute height adjustment mechanism and the minute inclination adjustment mechanism instead of the piezoelectric actuator, and from the lower surface of the lateral light guide member 25 to the inner bottom surface of the glass dish P. Is controlled so as to be a preset value. According to such a modification, a plurality of specimens S can be efficiently observed continuously.

  Further, the present invention can also be applied to an optical observation apparatus that performs imaging by inserting it into a living tissue like an endoscope. As shown in FIG. 11, the tip of the image pickup unit of this apparatus has the structure of the tip of the image pickup unit in a normal endoscope, and the light guide member 20 and the first and second LED light irradiators 13 and 14 of the above embodiment are compact. The light guide member 120 and the first and second LED light irradiators 113 and 114 are added. The objective lens 110 at the tip of the imaging unit is sealed with a light guide member 120 so that liquid does not enter between the objective lens 110 and the light guide member 120. In addition, this apparatus detects the distance to the biological tissue Bi as in the above embodiment, controls the distance between the lower surface of the lateral light guide member 125 and the biological tissue Bi, and guides the lateral direction with respect to the surface of the biological tissue Bi. Since the inclination of the lower surface of the optical member 125 cannot be adjusted, the position and posture of the distal end of the imaging unit with respect to the biological tissue Bi are adjusted while viewing the captured image. According to such a modification, it is not necessary to place the specimen S on a specific instrument, so that the living tissue can be observed as it is.

  In the above embodiment, the excitation light incident on the light guide member 20 is LED light. The light may be fluorescent light or lamp light. In addition, when observing a fluorescent substance, it is necessary to irradiate excitation light, but the present invention is not limited to fluorescence observation, and may be applied to optical observation with normal light.

  Moreover, in the said embodiment, although the excitation light made to inject into the light guide member 20 was made into 1st LED light and 2nd LED light, three or more LED light was changed in the incident angle, and the light guide member 125 of the horizontal direction light guide member 125 was changed. You may make it inject into a lower surface. In this case, the third vertical light guide member 26 and the fourth vertical light guide member 27 are provided with the third reflective portion and the fourth reflective portion, and the third vertical light guide member 26 and the fourth vertical light guide member are provided. Excitation light may also be incident from 27.

  Moreover, in the said embodiment, the light guide member 20 is made into left-right symmetry, and the location where the optical axis of the objective lens 10 on the lower surface of the lateral light guide member 25 where the first LED light and the second LED light are incident is Although the vicinity of the center of the light guide member 20 is used, the first LED light and the second LED light may be incident on the lower surface of the lateral light guide member 25 near the portion where the optical axes of the objective lens 10 intersect. It may be shifted from the center.

  In the above embodiment, the lower surface of the lateral light guide member 25 is a single plane. However, only the portion where the first LED light and the second LED light propagate is made one plane, and the other lateral direction guides are used. The lower surface of the optical member 25 may be a plane or a curved surface that forms an angle with this plane.

  In the above embodiment, the lateral light guide member 25 is shaped like a plate whose lower surface and upper surface are parallel. However, after the LED light is reflected from the upper surface of the lateral light guide member 25, the lateral light guide member 25 is guided. The upper surface of the lateral light guide member 25 may be deviated from being parallel to the lower surface as long as the light enters the vicinity of the location where the optical axes of the objective lens 10 intersect on the lower surface of the optical member 25.

  In the embodiment, the LED light emitted from the first LED light irradiator 13 and the second LED light irradiator 14 is incident on the light guide member 20. However, if each of the plurality of LED lights is incident on the light guide member 20 by different optical paths independently and is incident on the lateral light guide member 25 at different incident angles as in the above embodiment or the embodiment shown in FIG. For example, the configuration in which the LED light is incident on the light guide member 20 may be any configuration. For example, a single LED light irradiator is used, the LED light is divided by a beam splitter or the like, and a shutter is provided in the optical path of each LED light so that each LED light enters the light guide member 20 through a different optical path. May be.

  In the above embodiment, the lower surface of the lateral light guide member 25 intersects the optical axis of the objective lens 10 substantially perpendicularly. The lower surface of the lateral light guide member 25 may form an angle with a plane perpendicular to the optical axis of the objective lens 10 in accordance with the inclination of the surface of the instrument on which the lens is placed.

DESCRIPTION OF SYMBOLS 1 ... Apparatus main body, 2 ... Camera, 3 ... Eyepiece lens, 4 ... Lifting handle, 5 ... Base, 6 ... Bottom wall, 7 ... Mounting part, 10 ... Objective lens, 12 ... Optical system part, 13 ... 1st LED light Illuminator, 14 ... 2nd LED light irradiator, 15 ... 1st LED light source, 16 ... 2nd LED light source, 19-1, 19-2, 19-3 ... Distance sensor attaching part, 20 ... Light guide member, 21 ... 1st Longitudinal light guide member, 22... Second longitudinal light guide member, 23... First reflection portion, 24... Second reflection portion, 25 .. Lateral light guide member, 26. 4th longitudinal direction light guide member, 28 ... screwing part, 30 ... stage, 31-1, 31-2, 31-3 ... piezoelectric actuator, 40-1, 40-2, 40-3 ... distance sensor, 50 ... Distance detecting device 51 ... Laser light source 55 ... Optical coupler 59 ... Light receiving sensor 60 ... Optical changeover switch DESCRIPTION OF SYMBOLS 1 ... Optical path length variable apparatus, 62 ... Data processing apparatus, 63 ... Laser drive circuit, 64 ... Controller, control apparatus ... 70, 71 ... Input device, 72 ... Position detection sensor, 73 ... 1st LED drive circuit, 74 ... 2nd LED Drive circuit, 75... Stage height control device, 76-1, 76-2, 76-3 ... drive signal output circuit, 77 ... adjustable display lamp, 78 ... limit display lamp, 80 ... computer device, 81 ... input device , 82 ... Display device, 90 ... Moving device, 91 ... Moving stage, 92 ... Driving device, 93 ... Drive control device, 100 ... Imaging device, 101 ... Lifting device, 110 ... Objective lens, 113 ... First LED light irradiator, 114 ... second LED light irradiator, 120 ... light guide member, 130 ... plate-like light guide member, 132 ... enzyme, 133 ... groove, P ... glass dish, S ... specimen, Bi ... biological tissue

Claims (10)

In an optical observation apparatus comprising an imaging unit that images an observation object, and a light irradiation unit that is disposed in a housing having the imaging unit and that irradiates light to the observation object.
The light irradiation means includes
A lateral light guide member that has a surface facing the observation object on one end side of the housing and guides the light in the surface;
A longitudinal light guide member that is connected to the lateral light guide member and guides light incident from a part of the housing to the lateral light guide member;
The observation object of the horizontal light guide member is provided on the vertical light guide member, reflects the incident light and guides it to the horizontal light guide member, and according to the reflection position of the light. An optical observation apparatus comprising a plurality of reflecting portions having different reflection angles so as to be incident at different incident angles on a surface facing the surface. The optical observation apparatus according to claim 1,
The lateral light guide member has a plate-like portion having a surface facing the object to be observed and a back surface on the opposite side of the surface,
A part of the plurality of reflecting portions provided in the longitudinal light guide member guides light toward the back side of the plate-like portion,
Optical observation characterized in that light guided to the back side of the plate-like part is reflected on the back side of the plate-like part and then enters the surface facing the object to be observed apparatus. The optical observation device according to claim 2,
An optical observation apparatus characterized in that light incident on the surface facing the object to be observed after being reflected on the back side of the plate-like portion of the lateral light guide member has an incident angle of 75 to 85 °. . In the optical observation apparatus according to claim 1 to claim 3,
The vertical light guide member includes a first vertical light guide member and a second vertical light guide member, and each of the plurality of vertical light guide members transmits light to the horizontal light guide member. Is to guide light,
A first light emitter for making light incident on the first longitudinal light guide member, and a second light emitter for making light incident on the second longitudinal light guide member,
The reflective portion is provided in each of the first vertical light guide member and the second vertical light guide member, and the reflective portion of the first vertical light guide member and the second vertical direction. An optical observation apparatus characterized in that a reflection angle is different from that of a reflection portion of a light guide member. The optical observation apparatus according to claim 1, wherein:
The tip of the imaging unit is an objective lens,
A bottomed enclosure is formed by the peripheral wall portion provided with the vertical light guide member and the horizontal light guide member provided on the bottom surface of the peripheral wall portion. An optical observation apparatus characterized in that the objective lens is included in the interior of the optical observation apparatus. The optical observation apparatus according to claim 5,
The longitudinal light guide member includes a plate-shaped first longitudinal light guide member and a second longitudinal light guide member,
The light irradiation means includes two plate-like members connected to the vertical light guide member and the horizontal light guide member, and the vertical light guide member and the horizontal light guide member include the two plate-like members. An optical observation apparatus characterized by integrally forming a bottomed enclosure. The optical observation device according to any one of claims 1 to 6,
A plate-shaped light guide member formed detachably on the surface of the lateral light guide member facing the object to be observed;
The optical observation apparatus, wherein the plate-shaped light guide member guides light integrally with the horizontal light guide member when the plate light guide member is attached to the horizontal light guide member. The optical observation device according to claim 1, wherein:
A drive mechanism for changing the distance from the tip of the imaging unit to the object to be observed;
A stage for setting an instrument on which the observation object is placed;
The optical observation apparatus, wherein the drive mechanism changes a distance between a tip of the imaging unit and the stage. The optical observation apparatus according to claim 8, wherein
A plurality of distance detecting means for detecting a distance from the surface of the instrument set on the stage to the surface of the lateral light guide member facing the object to be observed; A plurality of distance detection means arranged around the optical axis;
A position in the normal direction of the surface of the stage from the distance detection position of the plurality of distance detection means while changing a slight distance between the surface of the lateral light guide member facing the object to be observed and the stage A plurality of interval changing means arranged in
Interval change command means for outputting a change command to the plurality of interval change means;
When a change command is input from the interval change command means in a state where the plurality of distances detected by the plurality of distance detection means are equal to or less than a set distance by the drive mechanism, the plurality of distance detection means Interval control means for controlling the plurality of interval change means so that the detected plurality of distances become equal and the plurality of interval change means change to a commanded value or a commanded positive / negative side. An optical observation device. The optical observation apparatus according to claim 8, wherein
The observation object is a living tissue or a cell,
Based on the information of the spatial resolution, the distance from the surface facing the observation object of the lateral light guide member within the imaging range of the imaging unit to the observation object is calculated, and the observation object is calculated as the distance. An optimum distance calculating means for setting the optimum distance to a value obtained by adding a set value calculated based on the information of the object;
Distance detection means for detecting a distance from a surface of the instrument set on the stage and set on the stage to a surface facing the observation target of the lateral light guide member within the imaging range of the imaging unit; ,
Interval changing means for changing the interval between the surface facing the observation object of the lateral light guide member within the imaging range of the imaging unit and the stage to a minute amount;
When the optimum distance calculated by the optimum distance calculating means is input in a state where the distance detected by the distance detecting means is equal to or less than a set distance by the driving mechanism, the distance detecting means detects the distance. An optical observation apparatus comprising: an interval control unit that controls the interval changing unit so that the distance becomes the optimum distance.

Images