JP6192335B2 - microscope - Google Patents

Description

  The present invention relates to a microscope technique for observing a specimen placed on a stage, and more particularly to a microscope suitable for microinsemination and a method for controlling the microscope.

  In recent years, microinsemination in the field of advanced reproductive medicine has been known as an application of a microscope. Microinsemination is a method of fertilizing sperm and ovum under a microscope. In general, sperm injection in the cytoplasm is performed by inserting a micropipette containing sperm into an egg fixed with a holding pipette and injecting sperm into the egg. (Intracytoplasmic Sperm Injection: hereinafter referred to as “ICSI”). In this ICSI, in order to operate a specimen on the stage, it is common to use an inverted microscope having a large working space above the stage.

  In the field of micro insemination, a relief contrast observation method (hereinafter referred to as “RC observation method”) capable of observing an egg three-dimensionally is known in order to improve the insemination rate of the egg (refer to Patent Document 1). .

  In recent years, in the field of micro insemination, in order to improve the insemination rate, a micro insemination method using a microscope while appropriately switching a plurality of observation methods has attracted attention. For example, a method of using an RC observation method, a differential interference observation method (hereinafter referred to as “DIC observation method”) and a polarization observation method (hereinafter referred to as “PO observation method”) while switching depending on the observation purpose is becoming widespread. .

  The DIC observation method can observe an object at a higher magnification than the RC observation method, and is suitable for observing small sperm as compared to an egg, and is therefore used when selecting high-quality sperm.

  In addition, the PO observation method is suitable for observing an ovum spindle having birefringence, so that when the sperm is injected into the ovum, the spindle is prevented from being damaged accidentally, Used when confirming the position. In a microscope that performs such a PO observation method, after adjusting to a crossed Nicol state in which the vibration direction of the light passing through the polarizer and the vibration direction of the light passing through the analyzer are orthogonal to each other, adjusting the retardation of the egg while rotating the compensator The position of the spindle is specified by reversing the contrast of the spindle in the ovum.

JP 51-29149 A

  By the way, in order to set a correct optical condition for each observation method, the compensator needs to change the arrangement in accordance with the movement of another optical unit for each observation method in addition to the PO observation. However, the operation of changing the arrangement of the compensator in accordance with the movement of another optical unit for each observation method has been difficult for a user who has little experience in using a microscope. For this reason, the technique which can arrange | position a compensator in a suitable position according to the movement of another optical unit for every observation method by simple operation was desired.

  The present invention has been made in view of the above, and provides a microscope and a control method capable of arranging a compensator at an appropriate position in accordance with the movement of another optical unit for each observation method with a simple operation. For the purpose.

In order to solve the above-described problems and achieve the object, a microscope according to the present invention includes a light source that generates light for irradiating a specimen, and a light source that is disposed on the optical path of the light and collects light emitted from the light source. A condenser lens that irradiates the specimen, an objective lens disposed at a position on the optical path opposite to the condenser lens across the specimen, and the optical path between the light source and the condenser lens, A first polarizing plate that is rotatably arranged with an optical axis that coincides with at least a part of the optical path as a rotation axis, and that transmits only the polarization component in one direction of the light emitted from the light source, and is switched according to the observation method. A condenser turret that includes any one of the plurality of optical elements on the optical path between the condenser lens and the first polarizing plate; and A compensator arranged on the optical path between the let and the first polarizing plate so as to be rotatable about the optical axis and changing the retardation of the light transmitted through the first polarizing plate, and the objective lens A unidirectional polarization component of light transmitted through the sample according to a relative positional relationship with the first polarizing plate, which is rotatably arranged with the optical axis as a rotation axis on the optical path on the observation side in the subsequent stage A second polarizing plate that transmits only the light, setting information indicating the positions of the objective lens, the first polarizing plate, the condenser turret, and the compensator arranged on the optical path for each observation method, and a polarization observation method. Recording a plurality of retardation values in the sample and retardation information in which the driving ranges of the compensators of the plurality of retardation values are associated with each other. A recording unit, the objective lens, the first polarizing plate, a driving unit for moving or rotating the respective said capacitor turret and the compensator, an input of an instruction signal for instructing the driving range of the compensator when performing polarization observation method an operation input unit that receives the movement by referring to the setting information in which the recording unit to record, the objective lens and the front Symbol capacitor turret their respective to the drive unit, to the position of the optical path in accordance with the observation method A control unit that changes the observation method by performing the observation method, when the observation unit performs an observation method other than the polarization observation method, the compensator is placed at a position that does not affect the required vibration direction of the light. When the polarization observation method is performed while rotating by the driving unit, the retardation information recorded by the recording unit is referred to and the driving unit is The compensator is rotated in a driving range including a position where the retardation value becomes 0 as a reference, and in the driving range according to the instruction signal received by the operation input unit.

In the microscope according to the present invention, in the above invention, the operation input unit can accept an input of an instruction signal instructing an observation method, and the control unit instructs the observation method from the operation input unit. When an instruction signal is input, the objective lens, the first polarizing plate, the condenser turret, and the compensator are moved or rotated to positions corresponding to the instruction signal instructing the observation method in the driving unit. Features.

  The microscope according to the present invention is characterized in that, in the above-mentioned invention, the microscope is used for an intracytoplasmic sperm injection method in which sperm is injected into an egg.

  According to the present invention, the control unit switches and arranges each optical unit to a position where the optimal optical condition is achieved. As a result, the operation time of the microscope can be shortened and the observation method can be switched with a simple operation.

FIG. 1 is a conceptual diagram showing a configuration of a microscope according to the first embodiment of the present invention. FIG. 2 is a block diagram illustrating a schematic configuration of the microscope according to the first embodiment of the present invention. FIG. 3 is a diagram illustrating a configuration of the condenser turret of the microscope according to the first embodiment of the present invention. FIG. 4 is a plan view of a petri dish including a specimen. 5 is a cross-sectional view taken along line AA in FIG. FIG. 6 is a diagram illustrating a configuration of the operation input unit of the microscope according to the first embodiment of the present invention. FIG. 7 is a diagram showing setting information recorded by the setting information recording unit of the microscope according to the first embodiment of the present invention. FIG. 8 is a diagram schematically illustrating the arrangement of each optical unit when the microscope according to the first embodiment of the present invention performs bright field observation. FIG. 9 is a diagram schematically illustrating the arrangement of each optical unit when the microscope according to the first embodiment of the present invention performs RC observation. FIG. 10 is a diagram schematically illustrating the arrangement of the optical units when the microscope according to the first embodiment of the present invention performs PO observation. FIG. 11 is a diagram schematically illustrating the arrangement of each optical unit when the microscope according to the first embodiment of the present invention performs DIC observation. FIG. 12 is a flowchart showing an outline of processing executed by the microscope according to the first embodiment of the present invention. FIG. 13 is a block diagram illustrating a schematic configuration of the microscope according to the second embodiment of the present invention. FIG. 14 is a diagram illustrating a configuration of the operation input unit of the microscope according to the second embodiment of the present invention. FIG. 15 is a diagram illustrating retardation information recorded by the retardation recording unit of the recording unit of the microscope according to the second embodiment of the present invention. FIG. 16 is a flowchart illustrating an outline of processing executed by the microscope according to the second embodiment of the present invention. FIG. 17 is a flowchart showing an outline of processing executed by the microscope according to the third embodiment of the present invention. FIG. 18 is a diagram schematically showing a position from the reference position of the RC observation aperture plate of the condenser turret of the microscope according to the third embodiment of the present invention. FIG. 19 is a diagram schematically illustrating the reference position of the compensator of the microscope according to the third embodiment of the present invention. FIG. 20 is a diagram schematically illustrating the position of the first crossed Nicol relative to the RC observation aperture plate from the reference position of the microscope compensator according to the third embodiment of the present invention. FIG. 21 is a diagram schematically illustrating the position of the second crossed Nicol relative to the RC observation aperture plate from the reference position of the microscope compensator according to the third embodiment of the present invention.

  DESCRIPTION OF EMBODIMENTS Hereinafter, embodiments for carrying out the present invention (hereinafter referred to as “embodiments”) will be described with reference to the drawings. The present invention is not limited to the embodiments described below. Further, in the description of the drawings, the same portions will be described with the same reference numerals.

(Embodiment 1)
FIG. 1 is a conceptual diagram showing a configuration of a microscope according to the first embodiment of the present invention. FIG. 2 is a block diagram illustrating a schematic configuration of the microscope according to the first embodiment of the present invention. 1 and 2, the plane on which the microscope 1 is placed will be described as an XY plane, and a direction perpendicular to the XY plane will be described as a Z direction.

  The microscope 1 shown in FIGS. 1 and 2 has a microscope main body 2 that observes the petri dish 100 in which the specimen Sp is accommodated, an operation input unit 3 that receives inputs of various operations of the microscope 1, and the microscope main body 2 captures images. A display unit 4 for displaying an image corresponding to the image data, a recording unit 5 for recording various programs and parameters for driving the microscope 1, and a control unit 6 for controlling the microscope main body unit 2 and the display unit 4. Prepare. The microscope main body 2, the operation input unit 3, the display unit 4, the recording unit 5, and the control unit 6 are connected by wire or wireless so that data can be transmitted and received.

  First, the microscope main body 2 will be described in detail. The microscope main body 2 includes a light source 10, a polarizer 11, a compensator 12, a condenser turret 13, a condenser lens 14, a stage 15, a stage position detector 16, a revolver 17, an objective lens 18, and a revolver position. A detection unit 19, a DIC prism 20, an analyzer 21, an imaging lens 22, an optical path splitting prism 23, an imaging unit 24, a mirror 25, an eyepiece lens 26, and a drive control unit 27 are provided.

  The light source 10 generates light that irradiates the specimen Sp under the control of the control unit 6. The light source 10 includes a halogen lamp, a xenon lamp, an LED (Light Emitting Diode), or the like.

  The polarizer 11 is disposed on the optical path between the light source 10 and the compensator 12 and transmits only the polarization component in one direction of the illumination light emitted by the light source 10. The polarizer 11 is disposed on the optical path between the light source 10 and the condenser lens 14. The polarizer 11 has an optical axis XA that coincides with at least a part of the optical path, and is disposed on the optical path so as to be rotatable about the optical axis XA as a rotation axis. The polarizer 11 is configured using a polarizing plate that is one of optical elements such as a filter. Further, the polarizer 11 is rotated around the optical axis XA by a motor 11 a configured by a stepping motor, a DC motor, or the like under the drive control of the drive control unit 27. In the first embodiment, the polarizer 11 functions as the first polarizing plate.

  The compensator 12 is an optical element for measuring the phase difference due to the anisotropy of the specimen Sp, and has a structure for changing the retardation of the specimen Sp. The compensator 12 changes the retardation of the light transmitted through the polarizer 11. The compensator 12 is disposed on the optical path between the polarizer 11 and the condenser turret 13. The compensator 12 has an optical axis XA that coincides with at least a part of the optical path, and is arranged on the optical path so as to be rotatable about the optical axis XA as a rotation axis.

  The compensator 12 is configured using a liquid crystal or a wave plate. Specifically, the compensator 12 is configured using a bereak compensator, a senal mon type compensator, a brace scaler compensator, a quartz wedge compensator, and a liquid crystal modulation element. The compensator 12 is preferably a liquid crystal modulation element, a senalmon type compensator, or a brace scaler compensator because it is desirable that the retardation of the visual field be substantially uniform when performing PO observation for observing the ovum spindle. When a liquid crystal modulation element is used as the compensator 12, the retardation can be changed by electrically controlling the liquid crystal molecules.

  When a Senalmon type compensator is used as the compensator 12, the retardation of the compensator 12 can be changed by the rotation of the polarizer 11 with respect to the wave plate in the compensator 12. Further, when a brace scaler compensator is used as the compensator 12, the retardation of the compensator 12 can be changed by the rotation of the prism in the compensator 12. Furthermore, the compensator 12 is rotated around the optical axis XA by a motor 12a configured by a stepping motor, a DC motor, or the like under the drive control of the drive control unit 27.

  The condenser turret 13 has a plurality of optical elements that are used by switching according to the observation method and the magnification of the objective lens 18, and is rotatably arranged on the optical axis XA between the polarizer 11 and the condenser lens 14. The condenser turret 13 is rotated in accordance with the observation method, so that one of the optical elements is detachably disposed on the optical path. Further, the capacitor turret 13 is rotated by a motor 13 a configured by a stepping motor, a DC motor, or the like under the drive control of the drive control unit 27.

  FIG. 3 is a diagram illustrating a configuration of the capacitor turret 13. As shown in FIG. 3, the capacitor turret 13 includes an opening 130, an RC observation aperture plate 131, an RC observation aperture plate 132, and a DIC prism 133.

  The opening 130 is arranged on the optical path by the rotation of the condenser turret 13. The opening 130 together with the capacitor turret 13 constitutes an opening plate (hole). The opening 130 is formed with a sufficient size that does not block the illumination light from the light source 10, and realizes illumination with a high numerical aperture. The opening 130 is used, for example, when the microscope 1 performs the bright field observation method (BF observation method) or the PO observation method. Specifically, when the microscope 1 performs the bright field observation method, the opening 130 is used by a user to search for a place in the petri dish 100 or a manipulator using a 4x or 10x objective lens 18 in preparation for microinsemination. It is used when positioning the tip of a micropipette to be operated.

  The RC observation aperture plate 131 is disposed on the optical path by the rotation of the condenser turret 13. The RC observation aperture plate 131 is an aperture plate used for RC observation, and is disposed on a part of the aperture 131a formed at a position shifted (eccentric) from the optical axis XA when arranged on the optical path. (Third polarizing plate). The opening 131a is formed at a position shifted from the center of the RC observation opening plate 131, thereby realizing oblique illumination. The polarizing plate 131b transmits only the polarization component in one direction of the light transmitted through the polarizer 11. The RC observation aperture plate 131 configured in this way is used, for example, when the microscope 1 performs a 20-fold RC observation method.

  The RC observation aperture plate 132 is arranged on the optical path by the rotation of the condenser turret 13. The RC observation aperture plate 132 is an aperture plate used for RC observation, and when placed on the optical path, a polarizing plate 132b (fourth polarizing plate) is formed on a part of the aperture 132a formed at a position eccentric from the optical path. Have The opening 132a is formed at a position shifted from the center of the RC observation opening plate 132, thereby realizing oblique illumination. The polarizing plate 132b transmits only the polarization component in one direction of the light transmitted through the polarizer 11. The thus configured RC observation aperture plate 132 is used, for example, when the microscope 1 performs a 40-fold RC observation method.

  The DIC prism 133 is arranged on the optical path by the rotation of the condenser turret 13. The DIC prism 133 is paired with a DIC prism 20 disposed on the image side on the objective lens 18 side, which will be described later, and constitutes a differential interference optical system. The DIC prism 133 is configured using a Nomarski prism or the like. The DIC prism 133 is used, for example, when the microscope 1 performs a 60-fold DIC observation method.

  In the thus configured condenser turret 13, the optical element disposed on the optical path is switched by rotating the condenser turret 13 by the motor 13a according to the observation method. Specifically, when performing the RC observation method, the condenser turret 13 has the RC observation aperture plate 131 or the RC observation aperture plate 132 disposed on the optical path. When performing the DIC observation method, the DIC prism 133 is disposed on the optical path. When the bright field observation method or the PO observation method is performed, the opening 130 is disposed on the optical path.

  The condenser lens 14 collects the illumination light emitted from the light source 10 and uniformly irradiates the region including the sample Sp in the petri dish 100. The condenser lens 14 may include a field stop that can adjust the amount of illumination light emitted from the light source 10 and a field stop operation unit that changes the diameter of the field stop.

  The stage 15 is configured to be movable in the XYZ directions. The stage 15 is moved in the XY plane or in the Z direction by a motor 15 a configured by a stepping motor, a DC motor, or the like under drive control by the drive control unit 27. On the stage 15, the petri dish 100 on which the specimen Sp is placed is placed. Under the control of the control unit 6, the stage 15 detects a predetermined origin position on the XY plane by the stage position detection unit 16, and calculates the driving amount of the motor 15 a using the origin position as a base point. Move to the desired observation location (observation region) above. Further, the stage 15 is detected by the stage position detection unit 16 under the control of the control unit 6, and the drive amount of the drive control unit 27 is limited based on this position, whereby the specimen Sp To the position where the condenser lens 14 and the objective lens 18 are in focus (focus position). The stage 15 may be provided with a heating unit that holds the petri dish 100 at a constant temperature.

  Here, the petri dish 100 in which the specimen Sp is arranged will be described in detail. FIG. 4 is a plan view of the petri dish 100 including the specimen Sp. 5 is a cross-sectional view taken along line AA in FIG.

  As shown in FIGS. 4 and 5, the petri dish 100 used for microinsemination is an ICSI drop R1 (culture solution) for fertilizing sperm Sp11 to an egg Sp10 and a sperm selection drop R2 (culture solution) for selecting sperm Sp11. And is covered with mineral oil Wa that prevents each drop from touching the air and infecting bacteria. Note that the number of drops in the petri dish 100 can be changed as appropriate.

  The stage position detection unit 16 detects the stage position of the stage 15 in the XY plane and the Z direction of the stage 15 and outputs the detection result to the control unit 6. The stage position detection unit 16 is configured using an encoder, an optical photo interrupter, or the like. The stage position detection unit 16 detects the stage position of the stage 15 based on the number of pulses of the motor 15a that is driven according to the drive signal output from the drive control unit 27 to the motor 15a, and controls the detection result. You may output to the part 6.

  The revolver 17 is equipped with a plurality of objective lenses 18. The revolver 17 is provided to be rotatable with respect to the optical path, and the objective lens 18 is disposed below the sample Sp. The revolver 17 is configured using a swing revolver or the like. The revolver 17 is rotated by a motor 17 a configured by a stepping motor, a DC motor, or the like under the drive control of the drive control unit 27. A focusing mechanism that moves the revolver 17 in the optical path direction may be provided separately.

  The objective lens 18 is disposed at a position on the optical path facing the condenser lens 14 with the sample Sp interposed therebetween. The objective lens 18 includes an objective lens 181, an objective lens 182, and an objective lens 183.

  The objective lens 181 is an objective lens having a magnification suitable for observing an egg, such as 20 times or 40 times, and is used for the RC observation method. The objective lens 181 has a modulator 1811 having three regions with different transmittances at the pupil position of the objective lens 181. The modulator 1811 includes a region 1811a having a transmittance of 100%, a region 1811b having a transmittance of approximately 25%, and a region 1811c having a transmittance of 0%. The modulator 1811 has an optically conjugate relationship with the RC observation aperture plate 131 and the RC observation aperture plate 132 disposed at the pupil position of the condenser lens 14. The objective lens 181 is also applied to a PO observation method in which an entire image of an ovum is a main observation target and approximately the same magnification is required.

  The objective lens 182 is an objective lens having a magnification suitable for sperm observation, for example, a high magnification such as 60 times or 100 times, and is used in the DIC observation method.

  The objective lens 183 is an objective lens having a magnification suitable for observing the tip of a micropipette, for example, a low magnification of 4 times, and is used for a bright field observation method.

  The revolver position detector 19 detects the position of the revolver 17 in the Z direction and outputs the detection result to the controller 6. The revolver position detector 19 is configured using a magnetic sensor, an optical photo interrupter, or the like. The revolver position detection unit 19 detects the position of the revolver 17 in the Z direction based on the number of pulses of the drive control unit 27 driven according to the drive signal input from the control unit 6, and controls the detection result. You may output to the part 6.

  The DIC prism 20 is paired with the DIC prism 133 and constitutes a differential interference optical system. The DIC prism 20 is configured using a Nomarski prism or the like. The DIC prism 20 is detachably disposed on the optical path between the objective lens 18 and the analyzer 21. Further, the DIC prism 20 is arranged on the optical path by a motor 20 a configured by a stepping motor, a DC motor, or the like under the drive control of the drive control unit 27.

  The analyzer 21 is disposed on the optical path on the observation side after the objective lens 18. The analyzer 21 is disposed on the optical path so as to be rotatable about the optical axis XA as a rotation axis. The analyzer 21 transmits only the polarization component in one direction of the light transmitted through the sample Sp according to the relative positional relationship with the polarizer 11. Further, the polarizer 11 and the analyzer 21 are arranged so as to be in a crossed Nicol state in which the polarization directions are orthogonal to each other when the microscope 1 performs PO observation. Further, the analyzer 21 is rotated about the optical axis as a rotation axis by a motor 21 a configured by a stepping motor, a DC motor, or the like under the drive control of the drive control unit 27. The analyzer 21 may be arranged to be detachable on the optical path. In this case, the analyzer 21 may be arranged so as to be in a 45-degree direction with respect to the vibration direction of the polarizing plate 131b of the RC observation aperture plate 131 of the capacitor turret 13 in order to perform the RC observation method without hindering observation. preferable. In the first embodiment, the analyzer 21 functions as the second polarizing plate.

  The imaging lens 22 condenses the light emitted from the objective lens 18 and forms an observation image. The imaging lens 22 is configured using one or a plurality of lenses.

  The optical path dividing prism 23 divides the light of the observation image formed by the imaging lens 22 into the imaging unit 24 and the mirror 25. The optical path splitting prism 23 is configured by using a prism having a coating for splitting light on the joint surface.

  The imaging unit 24 captures an observation image of the specimen Sp incident through the imaging lens 22 and the optical path dividing prism 23 to generate image data, and outputs the image data to the control unit 6. The imaging unit 24 is configured using an imaging element such as a charge coupled device (CCD) or a complementary metal oxide semiconductor (CMOS).

  The mirror 25 reflects the observation image emitted from the imaging lens 22 toward the eyepiece lens 26. A plurality of relay lenses may be provided on the optical path between the mirror 25 and the eyepiece lens 26.

  The eyepiece 26 enlarges the observation image incident through the imaging lens 22, the optical path splitting prism 23, and the mirror 25. The eyepiece 26 is configured using one or a plurality of lenses.

  The drive control unit 27 is configured using a drive driver, a CPU (Central Processing Unit), and the like, and moves or rotates each optical unit of the microscope main body 2 under the control of the control unit 6. Specifically, the drive control unit 27 drives the polarizer 11, the motor 11 a, the motor 12 a, the motor 13 a, the motor 15 a, the motor 18 a, the motor 20 a, and the motor 21 a under the control of the control unit 6. The compensator 12, the condenser turret 13, the stage 15, the revolver 17, the DIC prism 20, and the analyzer 21 are rotated or moved to predetermined positions.

  The operation input unit 3 receives input of various operations of the microscope 1. The operation input unit 3 is configured using a keyboard, a mouse, a joystick, a touch panel, various buttons, and the like, and outputs instruction signals according to operations of various switches to the control unit 6.

  FIG. 6 is a diagram illustrating a configuration of the operation input unit 3. As shown in FIG. 6, the operation input unit 3 includes buttons B <b> 1 to B <b> 5 that receive input of instruction signals for instructing each observation method, buttons B <b> 6 and B <b> 7 that receive input of instruction signals for adjusting contrast, and the objective lens 18. Buttons B8 to B13 for receiving an input of an instruction signal for instructing the magnification.

  The display unit 4 displays an image corresponding to the image data input from the imaging unit 24 via the control unit 6. The display part 4 is comprised using the display panel which consists of a liquid crystal or organic EL (Electro Luminescence).

  The recording unit 5 records various programs to be executed by the microscope 1 and various data used during the execution of the programs. The recording unit 5 is configured using a semiconductor memory such as a flash memory and a RAM (Random Access Memory). In addition, the recording unit 5 includes a setting information recording unit 51 that records setting information indicating the position of each of the plurality of optical units arranged on the optical path for each observation method.

  FIG. 7 is a diagram showing the setting information recorded by the setting information recording unit 51. As shown in FIG. 7, in the setting information T1, position information of each optical unit corresponding to each observation method is recorded. As shown in FIG. 7, when performing the bright field observation method, the polarizer 11 is arranged in a paranicol state with respect to the analyzer 21, the compensator 12 is on the optical path, and the hole (opening 130) of the capacitor turret 13 is formed. It is described that the objective lens 18 is arranged 4 times, the DIC prism 20 is arranged outside the optical path, the analyzer 21 is arranged on the optical path, and the stage 15 is arranged in the ICSI drop R1.

  The control unit 6 is configured using a CPU or the like, and comprehensively controls the operation of each unit configuring the microscope 1. The control unit 6 outputs an instruction signal corresponding to the operation signal input from the operation input unit 3 to the drive control unit 27, thereby driving the motor under the drive control by the drive control unit 27, thereby driving the microscope 1. The optical units constituting the are moved. Specifically, the control unit 6 refers to the setting information T1 recorded by the setting information recording unit 51, and causes the drive control unit 27 to move the plurality of optical units to positions on the optical path according to the observation method. To output an instruction signal for changing the observation method.

  The microscope 1 configured as described above switches the position and angle of each of the polarizer 11, the compensator 12, the condenser turret 13, the revolver 17, the DIC prism 20, and the analyzer 21 on the optical path under the control of the control unit 6. Bright field observation method, RC observation method, PO observation method and DIC observation method can be performed. Specifically, when an instruction signal instructing the bright field observation method is input from the operation input unit 3, the control unit 6 refers to the setting information T <b> 1 recorded by the setting information recording unit 51, and the drive control unit 27. By driving the motor under the driving control of, the polarizer 11, the compensator 12, the condenser turret 13, the revolver 17, the DIC prism 20, and the analyzer 21 are moved to positions on the optical path according to the bright field observation method. Thereby, the control part 6 changes the observation method of the microscope 1 to the bright field observation method (refer FIG. 8).

  Further, when an instruction signal instructing the RC observation method is input from the operation input unit 3, the control unit 6 refers to the setting information T <b> 1 recorded by the setting information recording unit 51 and drives the drive control unit 27. The revolver 17 is rotated to place the objective lens 181 on the optical path, and the condenser turret 13 is rotated to place the RC observation aperture plate 131 on the optical path. Further, the control unit 6 arranges the DIC prism 20 outside the optical path (see FIG. 9). Thereby, the control part 6 can switch the observation method of the microscope 1 to RC observation method. In the case of the RC observation method, the DIC prism 20 may be disposed on the optical path.

  In addition, when an instruction signal for instructing the PO observation method is input from the operation input unit 3, the control unit 6 refers to the setting information T <b> 1 recorded by the setting information recording unit 51, and performs drive control by the drive control unit 27. Then, by driving the motor, the revolver 17 is rotated to place the objective lens 181 on the optical path, and the condenser turret 13 is rotated to place a hole in the optical path. Further, the control unit 6 causes the polarizer 11 and the analyzer 21 to be in a crossed Nicols state by rotating the polarizer 11 about the optical axis as a rotation axis (see FIG. 10). Thereby, the control part 6 can switch the observation method of the microscope 1 to PO observation method. Further, in the case of the PO observation method, the compensator 12 is rotated at an arbitrary angle around the optical axis passing through the specimen Sp.

  In addition, when an instruction signal instructing the DIC observation method is input from the operation input unit 3, the control unit 6 refers to the setting information T <b> 1 recorded by the setting information recording unit 51, and performs the drive control by the drive control unit 27. By driving the motor, the revolver 17 is rotated to dispose the objective lens 182 on the optical path, and the condenser turret 13 is rotated to dispose the DIC prism 133 on the optical path and transmit through the polarizer 11. The compensator 12 is rotated in a paranicol state in which the vibration direction of the polarization component of the transmitted light and the vibration direction of the polarization component transmitted through the compensator 12 are parallel to each other. Further, the control unit 6 places the DIC prism 20 on the optical path and rotates the polarizer 11 to bring the polarizer 11 and the analyzer 21 into a crossed Nicols state (see FIG. 11). Thereby, the control unit 6 can switch the observation method of the microscope 1 to the DIC observation method.

  Next, processing executed by the microscope 1 will be described. FIG. 12 is a flowchart showing an outline of processing executed by the microscope 1.

  As shown in FIG. 12, the control unit 6 determines whether or not an instruction signal for instructing an observation method is input from the operation input unit 3 (step S101). Specifically, the control unit 6 determines whether or not an instruction signal instructing the observation method is input from any of the buttons B1 to B5 of the operation input unit 3. When the control unit 6 determines that an instruction signal for instructing an observation method is input from the operation input unit 3 (step S101: Yes), the microscope 1 moves to step S102. On the other hand, when the control unit 6 determines that the instruction signal for instructing the observation method is not input from the operation input unit 3 (step S101: No), the control unit 6 selects the observation method from the operation input unit 3. Wait until an instruction signal to be instructed is input.

  In step S <b> 102, the control unit 6 acquires setting information T <b> 1 corresponding to the observation method of the instruction signal input from the operation input unit 3 from the setting information recording unit 51.

  Subsequently, the control unit 6 drives each optical unit based on the setting information T1 acquired from the setting information recording unit 51 (step S103). Specifically, when an instruction signal instructing the DIC observation method is input from the operation input unit 3, the control unit 6 drives the motor under the drive control by the drive control unit 27, thereby The analyzer 21 is arranged in a crossed Nicol state, and the compensator 12 is arranged in a crossed Nicol state with respect to the analyzer 21.

  In addition, when an instruction signal for instructing the RC observation method is input from the operation input unit 3, the control unit 6 drives the motor under the drive control by the drive control unit 27, so that the compensator 12 and the capacitor turret are The vibration direction of the thirteenth RC observation aperture plate 131 and the polarizing plate 131b is arranged in a paranicol state, and the analyzer 21 is moved out of the optical path.

  In addition, when an instruction signal instructing bright field observation (BF) is input from the operation input unit 3, the control unit 6 drives the motor under the drive control by the drive control unit 27, so that the polarizer 11 The analyzer 21 is placed in a paranicol state, and the compensator 12 is placed in a paranicol state with respect to the analyzer 21.

  In addition, when an instruction signal instructing the PO observation method is input from the operation input unit 3, the control unit 6 drives the motor under the drive control by the drive control unit 27, thereby causing the polarizer 11, the analyzer 21, Is placed in the crossed Nicol state, and the compensator 12 is moved to the position of the previous end state.

  In step S104, the control unit 6 determines whether or not all the optical units have been driven. When the control unit 6 determines that the driving of all the optical units is finished (step S104: Yes), the microscope 1 finishes this process. On the other hand, when the control unit 6 determines that the driving of all the optical units has not been completed (step S104: No), the control unit 6 continues this determination.

  According to the first embodiment of the present invention described above, the control unit 6 switches each optical unit to a position where the optimum optical condition is obtained in accordance with the instruction signal instructing the observation method input from the operation input unit 3. Arrange. As a result, the operation time of the microscope 1 can be shortened and the observation method can be switched with a simple operation.

  In the first embodiment of the present invention, the drive control unit 27 may perform drive control of each optical unit in accordance with an instruction signal from the operation input unit 3.

  In Embodiment 1 of the present invention, when the microscope 1 is set to the RC observation method, the compensator 12 may be rotated around the optical axis XA at the same time while maintaining the state of the polarizer 11 and the paranicol.

  In Embodiment 1 of the present invention, an optical unit that is not required for each observation method may be moved out of the optical path.

(Embodiment 2)
Next, a second embodiment of the present invention will be described. The microscope according to the second embodiment has a configuration different from that of the microscope 1 according to the first embodiment described above and a process to be executed. For this reason, below, after demonstrating the structure of the microscope concerning this Embodiment 2, the process which this Embodiment 2 performs is demonstrated. In addition, the same code | symbol is attached | subjected and demonstrated to the structure same as the microscope 1 concerning Embodiment 1 mentioned above.

  FIG. 13 is a block diagram illustrating a schematic configuration of the microscope according to the second embodiment of the present invention. In FIG. 13, a plane on which the microscope 200 is placed is defined as an XY plane, and a direction perpendicular to the XY plane is defined as a Z direction.

  A microscope 200 illustrated in FIG. 13 includes a microscope main body unit 2, a display unit 4, a control unit 6, an operation input unit 300, and a recording unit 400.

  The operation input unit 300 receives input of various operations of the microscope 200. The operation input unit 300 is configured by using a keyboard, a mouse, a joystick, a touch panel, a rotary switch, various buttons, and the like, and outputs instruction signals according to operations of the various switches to the control unit 6.

  FIG. 14 is a diagram illustrating a configuration of the operation input unit 300. As illustrated in FIG. 14, the operation input unit 300 includes buttons B1 to B5 that receive input of instruction signals instructing each observation method, buttons B6 and B7 that receive input of instruction signals to adjust contrast, and the magnification of the objective lens 18. Buttons B8 to B13 for receiving an input of an instruction signal for instructing, and a retardation switch B100 for receiving an input of an instruction signal for regulating the driving range of the compensator 12 when the microscope 200 performs the PO observation method. The retardation switch B100 is configured using a rotary switch, and outputs an instruction signal for instructing a driving range of the compensator 12 to the control unit 6 according to a selected position.

  The recording unit 400 records various programs to be executed by the microscope 200 and various data used during the execution of the programs. The recording unit 400 is configured using a semiconductor memory such as a flash memory and a RAM. The recording unit 5 includes a setting information recording unit 51 and a retardation information recording unit 401 that records the retardation value of the sample Sp and the driving range of the compensator 12 in association with each other when the microscope 200 performs PO observation. . The retardation value of the specimen Sp in the second embodiment is the retardation value of the spindle of the egg Sp10.

  FIG. 15 is a diagram illustrating the retardation information recorded by the retardation information recording unit 401. As shown in FIG. 15, in the retardation information T2, the driving range of the compensator 12 set according to the retardation value of each sample Sp according to the instruction signal input from the retardation switch B100 of the operation input unit 300 is described. Has been. For example, in the retardation information T2, when the retardation value of the spindle of the ovum Sp10 is 5 nm, the driving range of the compensator 12 is described as ± 5 ° with respect to the position where the retardation of the compensator 12 becomes 0. Here, the position where the retardation of the compensator 12 becomes 0 is a state in which the polarizer 11 and the analyzer 21 are arranged in a crossed Nicol state, and the compensator 12 is in a crossed Nicol state with respect to the polarizer 11 or the analyzer 21. It is a position that becomes a dark spot. Note that the driving range of the compensator 12 is appropriately set according to the retardation value of the sample Sp.

  Processing executed by the microscope 200 having the above configuration will be described. FIG. 16 is a flowchart illustrating an outline of processing executed by the microscope 200.

  As shown in FIG. 16, the control unit 6 sets the direction of the button in which one of the button B6 or the button B7 is pressed in the DIR variable (step S201).

  Subsequently, when the observation method is the PO observation method (step S202: Yes), the control unit 6 acquires the setting information of the PO observation method from the setting information recording unit 51 (step S203).

  Then, the control part 6 acquires the drive information of the compensator 12 at the time of performing retardation from the retardation information recording part 401 (step S204). Specifically, the control unit 6 acquires the driving range of the compensator 12 corresponding to the position selected by the retardation switch B100 from the retardation information recording unit 401.

  Subsequently, the control unit 6 sets the DIR side of the drive range of the compensator 12 to the drive instruction variable MOV_POS in the drive control unit 27 (step S205), and drives the motor under the drive control by the drive control unit 27. Thus, the compensator 12 is driven to rotate about the optical axis XA (step S206).

  Thereafter, the control unit 6 determines whether or not the driving of the compensator 12 has been completed (step S207). When the control unit 6 determines that the driving of the compensator 12 has ended (step S207: Yes), the microscope 200 ends this process. On the other hand, when the control unit 6 determines that the driving of the compensator 12 is not completed (step S207: No), the control unit 6 continues this determination.

  In step S202, when the observation method is not PO observation (step S202: No), the control unit 6 refers to the setting information T1 recorded by the setting information recording unit 51 and drives each optical unit for each observation method ( Step S208). After step S208, the microscope 200 ends this process.

  According to the second embodiment of the present invention described above, it is possible to observe in the range of the optimum retardation for the specimen Sp.

(Embodiment 3)
Next, a third embodiment of the present invention will be described. The microscope according to the third embodiment has the same configuration as the microscope 200 according to the second embodiment described above, and the processing to be executed is different. Specifically, in the third embodiment, the adjustment range of the crossed Nicols of the compensator is controlled in accordance with the polarization characteristics of the compensator. For this reason, below, the process which the microscope concerning this Embodiment 3 performs is demonstrated. In addition, the same code | symbol is attached | subjected and demonstrated to the structure same as the microscope 1 concerning Embodiment 2 mentioned above.

  FIG. 17 is a flowchart illustrating an outline of processing executed by the microscope 200 according to the third embodiment of the present invention. In the following, a process for limiting the rotation range to ± 100 degrees by adding a predetermined adjustment range ± 10 degrees to the interval 180 degrees (± 90 degrees) of the crossed Nicols position of the compensator 12 will be described.

  As shown in FIG. 17, when one of the buttons B6 and B7 for changing the contrast of the operation input unit 3 is pressed and the pressed button is the button B6 (step S301: Yes), the control unit 6 Sets +1 to the variable MOV of the drive control unit 27 (step S302). On the other hand, when one of the buttons B6 and B7 for changing the contrast of the operation input unit 3 is pressed and the pressed button is not the button B6 (step 301: No), the control unit 6 The variable MOV of the drive control unit 27 is set to −1 (step S303).

  After step S302 or step S303, the control unit 6 sets the result of adding the variable MOV to the current position CUR_POS to the target position TGT_POS of the compensator 12 (step S304).

  Subsequently, when the target position TGT_POS of the compensator 12 is +101 or −101 (step S305: Yes), the control unit 6 determines that the compensator 12 is out of the crossed Nicols adjustment range. The process ends. On the other hand, when the target position TGT_POS of the compensator 12 is +101 or not −101 (step S305: No), the control unit 6 determines that the compensator 12 is in the crossed Nicols adjustment range, and the microscope 200 The process proceeds to S306.

  In step S306, the control unit 6 instructs the drive control unit 27 to drive the compensator 12, and drives the motor 12a under the drive control by the drive control unit 27, thereby driving the compensator 12 around the optical axis XA. Are driven to rotate by the relative movement amount of the variable MOV (step S306).

  Thereafter, the control unit 6 determines whether or not the driving of the compensator 12 has been completed (step S307). When the control unit 6 determines that the driving of the compensator 12 has been completed (step S307: Yes), the microscope 200 proceeds to step S308. On the other hand, when the control unit 6 determines that the driving of the compensator 12 is not completed (step S307: No), the control unit 6 stands by until a response indicating that the driving of the compensator 12 is completed.

  In step S308, the control unit 6 updates the current position CUR_POS with the target position TGT_POS. After step S308, the microscope 200 ends this process.

  In the microscope 200 configured as described above, the position at which the image becomes darkest while rotating the compensator 12 by operating the operation input unit 3 while the user observes the brightness of the image through the eyepiece 26. Find it as a crossed Nicol position.

  However, the polarization characteristic of the compensator 12 has positions of crossed Nicols at intervals of 180 degrees, and there are two positions within the adjustment range of 360 degrees. For example, as shown in FIG. 18, the user rotates the RC observation aperture plate 131 around the optical axis XA from a position where a predetermined reference position is 0 degree, and the polarizing plate 131b of the RC element is placed at a position of 260 degrees. When fixed to the condenser turret 13, when the compensator 12 is rotated from a position where the predetermined reference position shown in FIG. 19 is 0 degrees as shown in FIG. 19, the rotation is 80 degrees from the reference position as shown in FIG. The RC element polarizing plate 131b and the compensator 12 are in the first crossed Nicol position. In this case, it is difficult for the user to determine crossed Nicols based on changes in image brightness. Therefore, by further operating the operation input unit 3, the user rotates the RC element polarizing plate 131b and the compensator 12 to the second crossed Nicol position by rotation of 260 degrees from the reference position as shown in FIG. Adjust until As a result, the compensator 12 may be operated unnecessarily.

  On the other hand, according to the third embodiment, the control unit 6 limits the rotation range of the compensator 12 to the minimum angle from the reference position when adjusting the crossed Nicols. Thereby, it is possible to prevent the compensator 12 from rotating excessively.

  According to the third embodiment of the present invention described above, it is possible to prevent the compensator 12 from rotating excessively during the adjustment of the crossed Nicols.

  Further, according to the third embodiment of the present invention, by restricting the position of the crossed Nicol to one position within the adjustment range of the rotation angle of the compensator 12, a user unfamiliar with the microscope 200 can easily operate the compensator 12 with a simple operation. It can be adjusted to the position of crossed Nicols.

  Furthermore, according to Embodiment 3 of the present invention, the user adjusts the tone of each optical unit or optical element by operating the operation input unit 3 while observing the image of the specimen Sp through the eyepiece 26. Even so, it is possible to prevent the current positions of the optical units and optical elements from being excessively rotated.

  In the present invention, a microscope provided with a microscope main body, an operation input unit, a display unit, a recording unit, and a control unit has been described as an example. For example, an objective lens for enlarging a sample, and imaging a sample via the objective lens The present invention can also be applied to an imaging apparatus having an imaging function and a display function for displaying an image, such as a video microscope.

  In the present invention, an inverted microscope apparatus used for ICSI for injecting sperm into an egg is described as an example of the microscope apparatus. However, for example, an upright microscope apparatus can also be applied. Furthermore, the present invention can be applied to various systems such as a line apparatus incorporating a microscope apparatus.

  In the present invention, the polarizer is rotated in order to bring the polarizer and the analyzer into a crossed Nicol state. However, the present invention can be applied even when the analyzer is rotated, for example. Of course, the present invention can be applied even when the polarizer and the analyzer are rotated.

DESCRIPTION OF SYMBOLS 1,200 Microscope 2 Microscope main body part 3,300 Operation input part 4 Display part 5,400 Recording part 6 Control part 10 Light source 11 Polarizer 11a, 12a, 13a, 15a, 17a, 20a, 21a Motor 12 Compensator 13 Capacitor turret 14 Capacitor Lens 15 Stage 16 Stage position detection unit 17 Revolver 18 Objective lens 19 Revolver position detection unit 20 DIC prism 21 Analyzer 22 Imaging lens 23 Optical path division prism 24 Imaging unit 25 Mirror 26 Eyepiece 27 Drive control unit 51 Setting information recording unit 100 Petri dish 130 aperture 131, 132 RC observation aperture plate 131b, 132b polarizing plate 133 DIC prism 181, 182, 183 objective lens 401 retardation information recording section Sp specimen Sp10 egg Sp11 Sperm R1 Drop for ICSI R2 Drop for sperm sorting T1 Setting information T2 Retardation information XA Optical axis

Claims (3)

A light source that generates light to illuminate the specimen;
A condenser lens that is disposed on the optical path of the light and collects the light emitted from the light source and irradiates the specimen;
An objective lens disposed at a position on the optical path facing the condenser lens across the sample;
On the optical path between the light source and the condenser lens, the optical axis coinciding with at least a part of the optical path is rotatably arranged, and only the polarization component in one direction of the light emitted from the light source is transmitted. A first polarizing plate,
A condenser turret having a plurality of optical elements that are switched according to an observation method, and disposing any one of the plurality of optical elements on the optical path between the condenser lens and the first polarizing plate; ,
A compensator that is disposed on the optical path between the condenser turret and the first polarizing plate so as to be rotatable about the optical axis and changes the retardation of light transmitted through the first polarizing plate;
1 of the light transmitted through the sample according to the relative positional relationship with the first polarizing plate, arranged rotatably on the optical path on the observation side after the objective lens with the optical axis as a rotation axis. A second polarizing plate that transmits only the polarization component in the direction;
Setting information indicating the positions of the objective lens, the first polarizing plate, the condenser turret, and the compensator arranged on the optical path for each observation method, and a plurality of retardation values in the specimen when performing the polarization observation method And a retardation information that associates the driving range of the compensator with each of the plurality of retardation values, and a recording unit for recording
A drive unit that moves or rotates each of the objective lens, the first polarizing plate, the condenser turret, and the compensator;
An operation input unit for receiving an input of an instruction signal for instructing a driving range of the compensator when performing polarization observation;
By referring to the setting information in which the recording unit to record, the objective lens and the front Symbol capacitor turret their respective to the driving unit, the observation method by moving the position of the optical path in accordance with the observation method A control unit to be changed;
With
The controller is
When performing an observation method other than the polarization observation method, the compensator is rotated by the drive unit to a position that does not affect the required vibration direction of the light,
When the polarization observation method is performed, referring to the retardation information recorded by the recording unit, the driving unit includes a driving range including the position where the retardation value becomes 0 as a reference, and the operation input unit receives A microscope characterized in that the compensator is rotated in the driving range according to the instruction signal. The operation input unit can accept an input of an instruction signal for instructing an observation method,
When an instruction signal for instructing the observation method is input from the operation input unit, the control unit supplies the objective lens, the first polarizing plate, the condenser turret, and the compensator to the driving unit, respectively. 2. The microscope according to claim 1, wherein the microscope is moved or rotated to a position corresponding to an instruction signal for instructing.   3. The microscope according to claim 1, wherein the microscope is used for an intracytoplasmic sperm injection method in which sperm is injected into an egg.

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