JP5904737B2 - Microscope system - Google Patents

Description

  The present invention relates to a microscope system that performs differential interference observation that converts phase difference (retardation) into color contrast using light interference, for example.

  2. Description of the Related Art Conventionally, in the fields of medicine and biology, a microscope system having a microscope device that illuminates and observes a specimen is used for observation of cells and the like. In the industrial field, microscope apparatuses are used for various purposes such as quality control of metal structures, research and development of new materials, inspection of electronic devices and magnetic heads, and the like. As a microscope system for observing a specimen, there is known a microscope system having a configuration in which a specimen image is captured using an imaging element such as a CCD camera and displayed on a monitor in addition to a visual observation.

  Also, in the manufacturing process of silicon wafers used in electronic devices, differential interference observation method that converts phase difference (retardation) into color contrast using light interference, for example, scratches on transparent film or transparent Inspections for the presence of manufacturing defects, such as variations in film thickness, are performed.

  In the differential interference observation method, a differential interference observation optical system is configured using an ordinary optical system (illumination lens, objective lens, imaging lens) provided in an optical microscope and optical elements such as a polarizer, a Nomarski prism, and an analyzer. Form. The polarizer is inserted into the optical path of the illumination light, and converts the illumination light into linearly polarized light in a specific direction (single vibration plane).

  The Nomarski prism splits the linearly polarized light that has passed through the polarizer into two linearly polarized lights whose vibration directions are orthogonal to each other (having vibration surfaces orthogonal to each other), and emitted the two linearly polarized lights. This is a differential interference prism that integrates and interferes with two light beams from a specimen on a single vibration surface. Two lights from the specimen interfere with each other by a differential interference prism, and generate an interference image. Of the light interfered by the differential interference prism, light of a certain wavelength is intensified, and light of a certain wavelength is canceled out. The color that appears due to the composition change of light is called interference color.

  The analyzer passes only the integrated light through the Nomarski prism and blocks the other light. The analyzer is arranged in a direction parallel to the vibration direction of the interference image, that is, a direction orthogonal to the polarizer.

  As a microscope system based on differential interference observation, there is an optical microscope that includes an actuator for electrically driving a polarizer, analyzer, or Nomarski prism, and an observer can drive these optical elements by operating the operation unit. It is disclosed (see, for example, Patent Document 1). In this optical microscope, when the observer selects the differential interference observation method, the control unit issues a drive instruction to the actuator to insert these optical elements into the optical path. Thereafter, when the observer gives an instruction to adjust the retardation by operating the operation unit, the control unit issues a drive instruction to the actuator to move the Nomarski prism, and the retardation is adjusted.

  In addition, it is equipped with an actuator for electrically driving the Nomarski prism, and by an operation by an observer, it is automatically moved to a representative interference color with reference to a table storing the correspondence between position information and interference color, An optical microscope that can finely adjust the interference color and easily reproduce the retardation position for each sample is disclosed (for example, see Patent Document 2). With this optical microscope, it becomes possible to adjust the retardation more easily as compared with Patent Document 1.

Japanese Patent Laid-Open No. 9-325281 JP 2008-286871 A

  By the way, in the inspection using the differential interference observation method, depending on the inspection object, there is a shape that can be detected only with a specific interference color, and therefore setting the retardation position (setting the origin position) is important. However, since the optical microscope disclosed in Patent Document 2 described above moves the Nomarski prism with reference to a table storing the correspondence relationship between the position information and the interference color, the correspondence relationship between the position information and the interference color is Nomarski. There may be cases where the difference between the devices which occurs when the prism is assembled to the device or the difference in solids from sample to sample is not dealt with. As a result, the position of the Nomarski prism moved by the actuator is not arranged at a desired position, and fine adjustment has to be performed.

  The present invention has been made in view of the above, and an object of the present invention is to provide a microscope system capable of setting an optimum retardation position with high accuracy in retardation adjustment of a microscope apparatus that performs differential interference observation.

In order to solve the above-described problems and achieve the object, a microscope system according to the present invention includes a stage on which a specimen is placed, an objective lens that collects observation light from at least the specimen on the stage, and An illumination light source that emits illumination light that irradiates the specimen, and the illumination light emitted from the illumination light source is split to irradiate the specimen, and light from the specimen that is irradiated with the split light Differential interference observation optical system for causing interference, an imaging unit for capturing an interference image generated by the differential interference observation optical system, an image data storage unit for storing image data of an interference image captured by the imaging unit , and interference of the interference color appearing by obtains a storage unit having a representative interference color storage unit that stores a representative interference color is a typical interference colors, the representative interference color image of the representative interference color, said surrogate table interference A color bar creation unit that creates a color bar by arranging images, a display unit that displays the color bar, and selection instruction information of a representative interference color image to be selected for the color bar on the display unit are input. An input unit; and a retardation adjustment unit that performs retardation adjustment according to selection instruction information from the input unit , wherein the color bar creating unit is configured such that the image data stored in the image data storage unit is the representative It is determined whether or not the representative interference color stored in the interference color storage unit, and when the image data is determined to be the representative interference color, the representative corresponding to each determined image data A color bar is created by arranging interference color images .

Further, the microscope system according to the present invention is the above-described invention, wherein the differential interference observation optical system is inserted into an optical path of the illumination light, and the polarizer converts the illumination light into linearly polarized light having a single vibration surface, The linearly polarized light is movable in a direction perpendicular to the optical axis of the objective lens, and is decomposed and emitted into two linearly polarized lights having vibration planes orthogonal to each other, and the emitted two linearly polarized lights A Nomarski prism that integrates and interferes with the two linearly polarized lights from the specimen irradiated with a single vibration surface, and an analyzer that passes only the light that has passed through the Nomarski prism. And a drive control unit that controls movement of the Nomarski prism in a direction perpendicular to the optical axis of the objective lens, and the color bar creating unit includes the representative interference Characterized by sequences in association with image in the moving direction of the Nomarski prism.

In the microscope system according to the present invention as set forth in the invention described above, the storage unit includes a position information storage unit that stores position information of the Nomarski prism driven by the drive control unit. The image processing unit further includes an image processing unit that determines whether the interference color of the interference image is the darkest image data based on the image data of the interference image sequentially acquired along with the drive control unit. When the image data is determined to be the darkest image data, the position of the Nomarski prism from which the image data was acquired is set as the origin.
In the microscope system according to the present invention, in the above invention, the storage unit includes a position information storage unit that stores position information of the Nomarski prism driven by the drive control unit, and the retardation adjustment unit includes: The origin in the movement of the Nomarski prism is set based on the position information obtained by acquiring the image of the color bar selected by the input unit.

  The microscope system according to the present invention is characterized in that, in the above invention, the input unit is a touch panel that receives an input according to a contact position of an object from the outside.

  The microscope system according to the present invention further includes a display control unit that controls a display mode of the display unit in the above invention, the display control unit including the representative interference color image instructed by the input unit, An interference color image of an interference color near the representative interference color corresponding to the representative interference color image instructed by the input unit is displayed on the display unit.

  According to the present invention, the retardation adjustment is performed based on the selection information of the representative interference color image in the color bar created by arranging the images corresponding to the set representative interference color. In the retardation adjustment of the microscope apparatus, the optimum retardation position can be set with high accuracy.

FIG. 1 is a conceptual diagram showing an example of the configuration of a microscope system according to an embodiment of the present invention. FIG. 2 is a block diagram showing a functional configuration of the microscope system according to the embodiment of the present invention. FIG. 3 is a diagram illustrating an example of a display input unit of the microscope system according to the embodiment of the present invention. FIG. 4 is a flowchart showing an outline of processing performed by the microscope system according to the embodiment of the present invention. FIG. 5 is a schematic diagram illustrating one aspect of display processing performed by the microscope system according to the embodiment of the present invention. FIG. 6 is a flowchart showing an outline of processing performed by the microscope system according to the embodiment of the present invention. FIG. 7 is a flowchart showing an outline of processing performed by the microscope system according to the embodiment of the present invention. FIG. 8 is a flowchart showing an outline of processing performed by the microscope system according to the embodiment of the present invention. FIG. 9 is a schematic diagram illustrating one aspect of display processing performed by the microscope system according to the embodiment of the present invention. FIG. 10 is a schematic diagram illustrating one aspect of display processing performed by the microscope system according to the modification of the embodiment of the present invention.

  DESCRIPTION OF EMBODIMENTS Hereinafter, embodiments for carrying out the present invention will be described in detail with reference to the drawings. In addition, this invention is not limited by the following embodiment. The drawings referred to in the following description only schematically show the shape, size, and positional relationship so that the contents of the present invention can be understood. That is, the present invention is not limited only to the shape, size, and positional relationship illustrated in each drawing.

  FIG. 1 is a conceptual diagram showing an example of the configuration of the microscope system according to the present embodiment. FIG. 2 is a block diagram showing a functional configuration of the microscope system according to the present embodiment. 1 and 2, the plane on which the microscope system 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.

  As shown in FIGS. 1 and 2, the microscope system 1 images the specimen S through the microscope apparatus 2 that observes the specimen S, the microscope control unit 3 that drives and controls the microscope apparatus 2, and the microscope apparatus 2. The image pickup apparatus 4 that generates image data, an image pickup control unit 5 that controls driving of the image pickup apparatus 4, and an image corresponding to the image data picked up by the image pickup apparatus 4 via the control terminal 7 are displayed, and the microscope system 1. A display input unit 6 that receives inputs of various operations, a microscope control unit 3, an imaging control unit 5, and a control terminal 7 that controls the display input unit 6. The microscope device 2, the microscope control unit 3, the imaging device 4, the imaging control unit 5, the display input unit 6, and the control terminal 7 are connected by wire or wireless so that data can be transmitted and received.

  The microscope apparatus 2 includes a stage 21 on which a specimen S is placed, a substantially C-shape in side view, a stage 21 that supports the stage 21 and holds an objective lens 23 via a revolver 22, a specimen And an epi-illumination light source 25 that irradiates the sample S with light.

  The stage 21 is configured to be movable in the XYZ directions. The stage 21 is movable in the XY plane by a motor 211. The stage 21 detects a predetermined origin position on the XY plane by an XY position origin sensor (not shown) under the control of the microscope control unit 3, and the drive amount of the motor 211 is controlled based on this origin position. The observation location on the specimen S is moved. Further, the stage 21 is movable in the Z direction by a motor 212. Under the control of the microscope control unit 3, the stage 21 detects a predetermined origin position in the Z direction of the stage 21 by a Z position origin sensor (not shown), and the driving amount of the motor 212 is controlled based on this origin position. Thus, the specimen S is focused and moved to an arbitrary Z position within a predetermined height range. The focusing movement may be performed not only by moving the stage in the Z direction but also by moving the objective lens 23 or the observation optical system including the objective lens 23 up and down.

  The revolver 22 is provided so as to be slidable or rotatable with respect to the microscope body 24, and the objective lens 23 is disposed above the sample S. The revolver 22 is configured using an electric revolver or the like. The revolver 22 holds a plurality of objective lenses 23 having different magnifications (observation magnifications) by the mounter 221. The revolver 22 is inserted into the optical path of the observation light and selectively switches the objective lens 23 used for observing the specimen S. Therefore, the revolver driving unit 222 that slides or rotates the mounter 221 and the connection state of the revolver 22 are changed. And a revolver detector 223 for detecting.

  The revolver driving unit 222 slides or rotates the mounter 221 under the control of the microscope control unit 3. The revolver detection unit 223 includes a revolver connection sensor (not shown) that detects that the revolver 22 is connected to the microscope main body 24, and the type of the objective lens 23 in which the objective lens 23 is inserted in the optical path of the observation light. And a movement completion sensor (not shown) for detecting that the objective lens 23 is inserted on the optical path of the observation light. The revolver detection unit 223 outputs detection results detected by various sensors to the microscope control unit 3.

  Instead of the revolver 22, a nose piece may be used. The nosepiece can dispose the desired objective lens 23 above the sample S via a slider that is slidable in a direction perpendicular to the optical axis of the objective lens. The nosepiece holds a plurality of objective lenses 23 having different magnifications (observation magnifications) depending on the slider. Under the control of the microscope control unit 3, the nosepiece can selectively switch the objective lens 23 inserted in the optical path of the observation light and used for observing the specimen S by sliding the slider in the X direction. it can.

  The objective lens 23 includes, for example, an objective lens 231 with relatively low magnification of 1 ×, 2 ×, and 4 × (hereinafter referred to as “low magnification objective lens 231”), and a 10 ×, 20 ×, and 40 × low magnification objective lens. At least one objective lens 232 (hereinafter referred to as “high-magnification objective lens 232”) having a high magnification relative to the magnification of 231 is attached to the mounter 221. Note that the magnifications of the low-magnification objective lens 231 and the high-magnification objective lens 232 are examples, and it is sufficient that the high-magnification objective lens 232 is higher than the low-magnification objective lens 231.

  The microscope main body 24 collects the illumination lens 241 that collects the illumination light L1 emitted from the epi-illumination light source 25 through the fiber 251 (hereinafter referred to as “epi-illumination light L1”), and the optical path of the epi-illumination light L1. A half mirror 242 that deflects along the optical axis of the objective lens 23, and an imaging lens 243 that focuses the reflected light of the sample S incident through the objective lens 23 and the half mirror 242, and forms an observation image; Is provided inside.

  The microscope main body 24 includes a polarizer 244 provided between the irradiation lens 241 and the half mirror 242, and a Nomarski prism 245 provided between the half mirror 242 and the objective lens 23 (hereinafter referred to as “prism 245”). And an analyzer 246 provided between the half mirror 242 and the imaging lens 243. The polarizer 244, the prism 245, and the analyzer 246 form a differential interference observation optical system. The polarizer 244 is inserted in the optical path of the epi-illumination light L1, and makes the epi-illumination light L1 linearly polarized light in a specific direction (single vibration plane). The polarizer 244 moves by driving the motor 244a under the control of the control terminal 7.

  The prism 245 transmits the polarized light that is transmitted through the polarizer 244 and deflected by the half mirror 242 into two linearly polarized lights whose vibration directions are orthogonal to each other (having vibration surfaces orthogonal to each other), and then emitted. This is a differential interference prism that integrates and interferes with the two light beams from the specimen S irradiated with the two linearly polarized light on a single vibration surface. The two lights from the sample S interfere with each other by the differential interference prism to generate an interference image. The prism 245 moves by driving the motor 245 a under the control of the control terminal 7. The microscope body 24 is provided with a prism detection unit 245b that detects the prism 245 when the prism 245 is disposed at the detection position. In the case of epi-illumination, one Nomarski prism serves as both illumination light and an observation method, but in the case of transmitted illumination, a pair of Nomarski prisms are required.

  The analyzer 246 passes only the integrated light passing through the prism 245 and blocks other light. The analyzer 246 is arranged in a direction parallel to the vibration direction of the interference image, that is, in a direction orthogonal to the polarizer 244. The analyzer 246 moves under the control of the control terminal 7 by driving the motor 246a.

  A zoom lens mechanism for enlarging the specimen S may be provided between the half mirror 242 and the prism 245. The zoom lens mechanism is configured using, for example, a zoom lens group and a zoom lens motor that drives the zoom lens group. The zoom lens motor can change the magnification of the zoom lens mechanism by moving the zoom lens group along the optical axis direction under the control of the microscope control unit 3.

  The epi-illumination light source 25 includes a halogen lamp, a xenon lamp, an LED (Light Emitting Diode), or the like. The epi-illumination light source 25 emits epi-illumination light L1 for forming an observation image of the specimen S through the fiber 251.

  The epi-illumination light L1 is applied to the specimen S through the illumination lens 241, the half mirror 242, the differential interference observation optical system, and the objective lens 23. Reflected light L <b> 2 (hereinafter referred to as “observation light L <b> 2”) reflected by the sample S enters the imaging device 4 through the objective lens 23, the half mirror 242, and the imaging lens 243.

  The microscope control unit 3 is configured using a CPU (Central Processing Unit) or the like, and comprehensively controls the operation of each unit configuring the microscope apparatus 2 under the control of the control terminal 7. Specifically, the microscope control unit 3 drives the revolver driving unit 222 to slide or rotate the mounter 221 to switch the objective lens 23 arranged on the optical path of the observation light L2, the motor 211 or the motor 212 is driven to drive the stage 21, the motor 244a, the motor 245a, and the motor 246a are driven to drive the differential interference observation optical system including the polarizer 244, the prism 245, and the analyzer 246, and the specimen S Adjustment processing for adjusting each part of the microscope apparatus 2 accompanying observation is performed. The microscope control unit 3 outputs the state of each part constituting the microscope apparatus 2, for example, position information (XY position, Z position) of the stage 21, type information of the objective lens 23 attached to the revolver 22, and the like to the control terminal 7. .

  The imaging device 4 is configured using an imaging device 41 such as a CCD (Charge Coupled Device) or a CMOS (Complementary Metal Oxide Semiconductor). Under the control of the imaging control unit 5, the imaging device 4 captures an observation image of the specimen S incident through the imaging lens 243 and continuously generates image data of the specimen S. The imaging device 4 outputs the image data of the specimen S generated via the camera cable to the control terminal 7. In the present embodiment, the imaging device 4 functions as an imaging unit.

  The imaging control unit 5 is configured using a CPU or the like, and controls the operation of the imaging device 4. Specifically, the imaging control unit 5 controls the photographing operation of the imaging device 4 by performing ON / OFF switching processing of automatic gain control of the imaging device 4, gain setting processing, frame rate setting processing, and the like. The imaging control unit 5 includes an AE processing unit 51 and an AF processing unit 52.

  The AE processing unit 51 automatically sets the exposure condition of the imaging device 4 based on the image data generated by the imaging device 4. Specifically, the AE processing unit 51 calculates the luminance from the image data acquired via the control terminal 7, and determines the exposure condition of the imaging device 4, for example, the exposure time based on the calculated luminance. 4 automatic exposure.

  The AF processing unit 52 automatically adjusts the focus of the imaging device 4 based on the image data generated by the imaging device 4. Specifically, the AF processing unit 52 automatically adjusts the focus of the imaging device 4 by evaluating the contrast included in the image data and detecting the in-focus position (focal position). . Note that the AF processing unit 52 may detect a focused focus position (Z position) by evaluating the contrast of the image at each Z position of the stage 21 based on the image data.

  The display input unit 6 includes a display communication unit 61 that performs communication with the control terminal 7, a display unit 62 that displays an image, and a touch panel 63 that outputs a position signal corresponding to an external object contact.

  The display communication unit 61 is a communication interface for performing communication with the control terminal 7. The display communication unit 61 outputs the image data output from the control terminal 7 to the display unit 62.

  The display unit 62 is configured using a display panel made of liquid crystal, organic EL (Electro Luminescence), or the like. The display unit 62 displays an image corresponding to the image data input via the display communication unit 61. The display unit 62 displays various operation information of the microscope system 1 and the like.

  The touch panel 63 is provided on the display screen of the display unit 62 and accepts an input according to the contact position of the object from the outside. Specifically, the touch panel 63 detects a position where the user touches (contacts) according to an operation icon displayed on the display unit 62, and sends a position signal corresponding to the detected touch position via the display communication unit 61. Output to the control terminal 7. For example, as shown in FIG. 3, the touch panel 63 is displayed on the display unit 62 by a live image corresponding to various operation information of the microscope system 1 or image data continuously generated by the imaging device 4, or by the imaging device 4. By displaying the captured image in the image display area A1, it functions as a graphical user interface (GUI). In general, the touch panel includes a resistance film method, a capacitance method, an optical method, and the like. In the first embodiment, any type of touch panel is applicable.

  The control terminal 7 includes a control communication unit 71 that communicates with the microscope control unit 3, the imaging control unit 5, and the display input unit 6, a storage unit 73 that stores various types of information of the microscope system 1, and each unit of the microscope system 1. An input unit 72 that receives an input of a drive instruction signal that instructs driving, and a control unit 74 that controls each unit of the microscope system 1 are provided.

  The control communication unit 71 is a communication interface for communicating with each of the microscope control unit 3, the imaging control unit 5, and the display input unit 6. Further, the control communication unit 71 outputs the image data output from the imaging device 4 to the control unit 74 via the camera cable.

  The input unit 72 is configured using a keyboard, a mouse, a joystick, various switches, and the like, and outputs operation signals corresponding to operation inputs of the various switches to the control unit 74.

  The storage unit 73 is realized by using a semiconductor memory such as a flash memory and a RAM (Random Access Memory) fixedly provided inside the control terminal 7. The storage unit 73 stores various programs to be executed by the microscope system 1 and various data used during execution of the programs. The storage unit 73 temporarily stores information being processed by the control unit 74. The storage unit 73 includes an image data storage unit 731 that stores image data captured by the imaging device 4, a position information storage unit 732 that stores position information indicating an imaging position of the obtained image data, and differential interference observation optics. A representative interference color storage unit 733 for storing an interference color set as a representative interference color among the interference colors obtained by the system; In addition, the storage unit 73 temporarily stores a position signal indicating the contact position input from the touch panel 63 of the display input unit 6. In addition, the memory | storage part 73 may be comprised using the memory card etc. which are mounted | worn from the outside.

  The control unit 74 is configured using a CPU or the like, and transfers instructions and data corresponding to each unit constituting the microscope system 1 according to a drive instruction signal, a position signal, a switching signal, and the like from the input unit 72 and the touch panel 63. To perform overall control of the operation of the microscope system 1.

  The control unit 74 includes an image processing unit 741, a drive control unit 742, a display control unit 743, a color bar creation unit 744, and a retardation adjustment unit 745.

  The image processing unit 741 performs predetermined image processing on the image data input via the control communication unit 71 to generate a display image to be displayed on the display unit 62. Specifically, the image processing unit 741 includes optical black subtraction processing, white balance adjustment processing, synchronization processing, color matrix calculation processing, γ correction processing, color reproduction processing, edge enhancement processing, and the like for image data. Perform image processing. The image processing unit 741 compresses the image data by a predetermined method, for example, JPEG (Joint Photographic Experts Group) method, and outputs the compressed image data to the image data storage unit 731.

  The drive control unit 742 performs drive control for driving the electric unit (stage 21, differential interference observation optical system) constituting the microscope apparatus 2 in accordance with, for example, an input from the touch panel 63 or the input unit 72.

  The display control unit 743 controls the display mode of the display unit 62. Specifically, the display control unit 743 displays a live image generated by the imaging device 4 and each of a plurality of image data stored in the image data storage unit 731 via the control communication unit 71 and the display communication unit 61. Display on the display unit 62. The display control unit 743 displays operation information regarding each operation of the microscope system 1, for example, operation information of the stage 21, on the display unit 62 via the control communication unit 71 and the display communication unit 61.

  The color bar creation unit 744 is a color obtained by arranging and synthesizing the images of the representative interference colors (representative interference color images) stored in the representative interference color storage unit 733 from the image data in the order corresponding to the movement amount of the prism 245. Create a bar.

  The retardation adjustment unit 745 performs retardation adjustment according to selection instruction information (input of a selection instruction signal) for the representative interference color image of the color bar created by the color bar creation unit 744.

  The microscope system 1 configured as described above transfers the image data of the specimen S imaged by the imaging device 4 to the display unit 62 via the control communication unit 71 and the display communication unit 61 under the control of the control unit 74. By displaying the image, the user can observe the image of the specimen S. Further, in the microscope system 1, the control unit 74 outputs an instruction signal or a drive signal to each unit of the microscope system 1 based on a signal input from the input unit 72 or the touch panel 63, whereby the microscope device 2 and the imaging device 4. Drive. Further, the microscope system 1 uses the color bar created by the color bar creation unit 744 to drive the differential interference observation optical system based on a signal input from the input unit 72 or the touch panel 63 to adjust the retardation. I do.

  Next, color bar generation processing for adjusting retardation will be described. FIG. 4 is a flowchart showing an outline of color bar generation processing performed by the microscope system according to the present embodiment.

  In the color bar generation process, first, a zero-order dark spot setting process is performed under the control of the control unit 74 (step S101). In this zero-order dark spot setting process, the control unit 74 sets a zero-order dark spot as an origin for the movement of the prism 245 in order to correct the difference for each apparatus. Each interference color (image data) and its coordinates (position information) are associated with each other and stored in the image data storage unit 731 and the position information storage unit 732 of the storage unit 73 by the zero-order dark spot setting process.

  Here, the zero-order dark spot used in the present embodiment refers to an image having the darkest background color (interference color) of the interference image, for example. Note that an image (interference image) obtained by differential interference is a composite image of two bright field images having different phases, and the image obtained as the phase difference of each bright field image approaches ½ of the wavelength. The (composite image) is dark. In the 0th-order dark spot setting process, for example, an image that looks darkest visually or an image with the smallest luminance value as the entire image is set as the 0th-order dark spot.

  The control unit 74 performs color bar creation processing after the 0th-order dark spot setting processing (step S102). In the color bar creation process, the color bar creation unit 744 creates a color bar used for retardation adjustment. In this color bar creation process, it is identified whether the obtained image is the representative interference color stored in the representative interference color storage unit 733, and if it is the representative interference color, the representative interference color image is obtained. A color bar is created by arranging the acquired representative interference color images along the moving direction of the prism 245.

  FIG. 5 is a schematic diagram showing an aspect of color bar display performed by the microscope system according to the present embodiment. As shown in FIG. 5, the color bar 8 is a combination of representative interference color images G <b> 1 to G <b> 8 respectively corresponding to the representative interference colors arranged in a line corresponding to the movement direction of the prism 245. The representative interference color images G1 to G8 are images of the same specimen and the same field of view. In the present embodiment, for example, it is assumed that the feature point P exists in this field of view.

  Here, in the representative interference color image selected as the zero-order dark spot, the representative interference color is compared with the feature points P of the other representative interference color images G1 to G4 and G6 to G8 due to the shape of the feature point P, for example. The brightness of the feature point P of the color image G5 may be different. As described above, the image selected as the zero-order dark spot is determined based on the darkest image or the point with the lowest luminance value in the image. However, the brightness feature is different from that of the other images. It is also possible to select and select the part.

  After the color bar creation process, the control unit 74 displays the obtained color bar on the display screen (step S103). For example, the control unit 74 instructs the display control unit 743 to display the color bar 8 shown in FIG. 5 on the display screen shown in FIG.

  Next, the zero-order dark spot setting process in step S101 will be described with reference to FIG. FIG. 6 is a flowchart showing an outline of the zero-order dark spot setting process performed by the microscope system according to the present embodiment. First, the drive control unit 742 moves the prism 245 to the sensor detection position of the prism detection unit 245b (step S201). When the drive control unit 742 moves the prism 245 to the sensor detection position, the drive control unit 742 sets this position to 0 (step S202).

  After setting the position, the drive control unit 742 moves the prism 245 in a direction perpendicular to the optical axis of the objective lens 23 (a direction parallel to the sample S placement surface of the stage 21) (step S203). At this time, counting of moving pulses is started.

  At this time, the image processing unit 741 determines whether or not it is a zero-order dark spot based on images sequentially obtained with the movement of the prism 245 (step S204). At this time, the drive control unit 742 repeats the movement process of the prism 245 until receiving a signal indicating that the image that is the 0th-order dark spot is determined from the image processing unit 741 (step S204: No).

  In addition, when the drive control unit 742 receives a signal indicating that the image that is the 0th-order dark spot is determined from the image processing unit 741 (step S204: Yes), the drive control unit 742 ends the count of the movement pulses and acquires the number of movement pulses. (Step S205). The control unit 74 stores the number of movement pulses in the storage unit 73 (step S206), and sets the position of the prism 245 (a position corresponding to the number of movement pulses) at the time when the 0th-order dark spot is obtained as an origin ( Step S207).

  Further, the control unit 74 sets the movable range of the prism 245 based on the obtained number of movement pulses (step S208). The control unit 74 stores the set origin and movable range in the position information storage unit 732.

  Next, the color bar creation processing in step S102 will be described with reference to FIG. FIG. 7 is a flowchart showing an outline of color bar creation processing performed by the microscope system according to the present embodiment. First, the drive control unit 742 moves the prism 245 to one end of the movable range set by the 0th-order dark spot setting process (step S301).

  The drive controller 742 moves the prism 245 to one end of the movable range, and then moves the prism 245 to the other end side (step S302). At this time, the moving direction of the prism 245 is a direction orthogonal to the optical axis of the objective lens 23 (a direction parallel to the sample S placement surface of the stage 21) as described above.

  At this time, the drive control unit 742 moves the prism 245 for each set pulse. The control unit 74 causes the imaging control unit 5 to perform imaging processing of the sample S while the prism 245 is stopped, and acquires an image of the sample S (step S303).

  Here, the color bar creating unit 744 determines whether or not the obtained image is a representative interference color (step S304). At this time, when the color bar creating unit 744 determines that the obtained image is not a representative interference color (step S304: No), the control unit 74 proceeds to step S302 to move the prism 245 and the sample S. An imaging process is performed.

  When the obtained image is the representative interference color (step S304: Yes), the control unit 74 stores the image data of the representative interference color image of the representative interference color and the position of the representative interference color in the storage unit 73. (Step S305). Thereafter, the drive control unit 742 determines whether or not the position of the prism 245 is the other end of the movable range (step S306). If not, the process proceeds to step S302 to move the prism 245 (step S306). S306: No).

  On the other hand, when the drive control unit 742 determines that the position of the prism 245 is the other end of the movable range (step S306: Yes), the color bar creation unit 744 refers to the image data storage unit 731 to represent each representative. A representative interference color image of the interference color is acquired, and the acquired representative interference color images of the representative interference colors are arranged and combined in correspondence with the movement direction of the prism 245 to create a color bar (step S307).

  Next, processing for performing retardation adjustment based on the created color bar will be described with reference to FIGS. FIG. 8 is a flowchart showing an outline of the retardation adjustment process performed by the microscope system according to the present embodiment. FIG. 9 is a schematic diagram showing an aspect of color bar display performed by the microscope system according to the present embodiment.

  First, as an instruction input screen for adjusting the retardation, the color bar 8 created by the color bar creation process described above can be moved along the arrangement direction of the representative interference color image in the color bar 8, and an input instruction is given as a GUI. A possible slider 81 is displayed, and a touch panel 63 is provided on the display screen. In this instruction input screen, the observer operates the GUI (slider 81) on the instruction input screen via the touch panel 63 (step S401).

  The observer operates the GUI (slider 81) to select a representative interference color image (in this case, the representative interference color) to be selected as a zero-order dark spot among the images that are the representative interference colors arranged as the color bar 8. The slider 81 is moved to the image G5), and a selection instruction signal indicating that the representative interference color image G5 is the 0th order dark spot is input. When the control unit 74 receives the selection instruction signal indicating that it is the 0th order dark spot, the control unit 74 sets the selected representative interference color image G5 as the 0th order dark point image, and the retardation adjustment unit 745 has the position information storage unit 732. , The retardation adjustment is performed by setting the position corresponding to the image G5 as the origin (step S402). At this time, the movable range of the prism 245 may be reset.

  Thereafter, the drive control unit 742 moves the prism 245 with reference to the origin position after the retardation adjustment, and the image processing unit 741 and the display control unit 743 correspond to the image data captured with the origin position as a reference. Display is performed (step S403).

  Through the above-described processes, based on the color bar created based on the origin position automatically set by the apparatus, the observer confirms this color bar and intuitively selects the 0th-order dark spot and retardation. By performing the adjustment, the retardation adjustment and the origin can be set more accurately.

  According to the above-described embodiment, since the retardation adjustment is performed based on the selection information of the representative interference color image in the color bar created by arranging the images corresponding to the set representative interference color, In the retardation adjustment of the microscope apparatus that performs differential interference observation, the optimum retardation position can be set with high accuracy.

  FIG. 10 is a schematic diagram illustrating an aspect of display processing performed by the microscope system according to the modification of the present embodiment. In the above-described embodiment, it has been described that the observer operates the GUI (slider 81) to select the representative interference color image to be the zero-order dark spot. However, the display control unit 743 selects the selected representative. For the interference color image, an interference color image of an interference color near the representative interference color corresponding to the selected image, for example, adjacent images G51 and G52 adjacent along the moving direction of the prism 245 are displayed on the display unit 62. May be.

  Here, for the adjacent images G51 and G52, the image processing unit 741 may extract an image captured in advance with reference to the position information and the image data storage unit 731 or the image G5 is selected. The control unit 74 that has received the information to the effect instructs the drive control unit 742 to place the prism 245 at a position adjacent to the image G5, and images the adjacent images G51 and G52 to the imaging control unit 5. The adjacent images G51 and G52 may be acquired.

  It is also possible to select the displayed images G51 and G52 as the 0th order dark spot, and the degree of freedom of the image to be selected as the 0th order dark spot can be improved. In the processing relating to this image display and selection of adjacent images, the observer operates the slider 81 to select a representative interference color image on the color bar 8, and touches (contacts) the displayed adjacent images G51 and G52. By doing this, it is possible to input an image selection instruction signal for the 0th-order dark spot. Further, the adjacent image may be displayed above the color bar 8 as shown in FIG. 10, or the selected image G5 and the adjacent images G51 and G52 may be displayed in a pop-up manner.

  In the color bar creating process according to the present embodiment described above, the color bar creating unit 744 has been described as determining whether or not this image is a representative interference color for each obtained image. When the control unit 742 moves the prism 245 from one end to the other end of the movable range, the image processing unit 741 or the color bar creation unit 744 is a CCD camera or the like during this time, and the interference color is the representative interference color. The position information storage unit 732 stores the position information in the case of the representative interference color, moves the prism 245 to the other end side, and then represents the position based on the stored position information. An interference color image may be acquired.

  The retardation adjustment according to the present embodiment described above may be performed for each apparatus or may be performed for each specimen to be observed. Since the color bar changes depending on the transmittance or pupil position of the objective lens and the reflectance of the sample, it is preferable to perform retardation adjustment when the objective lens and the sample are changed. Here, the storage unit 73 stores a table in which the 0th-order dark spot is associated with each objective lens, and the control unit 74 refers to this table to adjust the retardation in the retardation adjustment unit 745 according to the objective lens to be used. May be performed.

  In addition, the color bar according to the present embodiment described above is described as a rectangular image that is a representative interference color image arranged along the movement direction of the prism, but based on the position information of the representative interference color image. , Expressing the relative distance between each representative interference color image and the prism position by changing the length of the arrangement direction in the image according to the distance of the prism position from the adjacent representative interference color image It is good.

  Further, the representative interference color according to the present embodiment described above may be selected from the interference colors stored in the storage unit 73, color information (for example, colors existing in the image), or the like. May be arbitrarily selected by the observer.

  In the above-described embodiment, the representative interference color image in the color bar selected as the origin is described as the 0th-order dark spot that is the darkest background color of the interference image. Is not limited to the 0th-order dark spot, and may be an interference color image arbitrarily selected by the observer. It is also possible to change the origin position by retardation adjustment according to the type of specimen.

  In the above-described embodiment, the input instruction is given through the touch panel 63. However, the input instruction may be given by the input unit 72, and the display screen may be operated by a keyboard, a mouse, a joystick, or the like. An input instruction may be performed by operating the upper pointer.

  In the above-described embodiment, the display input unit and the control terminal are separately configured. However, for example, a portable terminal in which the display input unit and the control terminal are integrally formed may be used.

  In the above-described embodiment, the upright microscope apparatus has been described as an example of the microscope apparatus. However, the present invention can be applied to an inverted microscope apparatus, for example. Furthermore, the present invention can be applied to various systems such as a line apparatus incorporating a microscope apparatus.

  In the above-described embodiment, a microscope system including a microscope apparatus, an imaging apparatus, a display input unit, and a control terminal has been described as an example. For example, an objective lens that forms a differential interference observation optical system and expands a specimen The present invention can also be applied to an imaging apparatus having an imaging function for imaging a specimen through an objective lens and a display function for displaying an image, such as a video microscope.

  As described above, the microscope system according to the present invention is useful for setting an optimum retardation position with high accuracy in the retardation adjustment of a microscope apparatus that performs differential interference observation.

DESCRIPTION OF SYMBOLS 1 Microscope system 2 Microscope apparatus 3 Microscope control part 4 Imaging apparatus 5 Imaging control part 6 Display input part 7 Control terminal 21 Stage 22 Revolver 23 Objective lens 24 Microscope main body 25 Light source for epi-illumination 41 Image pick-up element 51 AE process part 52 AF process Unit 61 Display communication unit 62 Display unit 63 Touch panel 71 Control communication unit 72 Input unit 73 Storage unit 74 Control unit 211, 212, 244a, 245a, 246a Motor 222 Revolver drive unit 223 Revolver detection unit 241 Illumination lens 242 Half mirror 243 Imaging Lens 244 Polarizer 245 Prism 245b Prism detection unit 246 Analyzer 731 Image data storage unit 732 Position information storage unit 733 Representative interference color storage unit 741 Image processing unit 742 Drive control unit 743 Display control unit 744 Color Over creation section 745 retardation adjusting unit

Claims (6)

A stage on which the specimen is placed;
An objective lens that collects at least the observation light from the specimen on the stage;
An illumination light source for emitting illumination light to irradiate the specimen;
A differential interference observation optical system that splits the illumination light emitted from the illumination light source and irradiates the specimen, and interferes light from the specimen irradiated with the dispersed light;
An imaging unit that captures an interference image generated by the differential interference observation optical system;
A storage unit having an image data storage unit that stores image data of an interference image captured by the imaging unit , and a representative interference color storage unit that stores a representative interference color that is a representative interference color among interference colors that appear due to interference. When,
A color bar creating unit that obtains a representative interference color image of the representative interference color and creates a color bar by arranging the representative interference color image;
A display unit for displaying the color bar;
An input unit for inputting selection instruction information of a representative interference color image to be selected for the color bar on the display unit;
In accordance with the selection instruction information from the input unit, a retardation adjustment unit that performs retardation adjustment,
Equipped with a,
The color bar creating unit determines whether the image data stored in the image data storage unit is the representative interference color stored in the representative interference color storage unit, and the image data is the representative data A microscope system characterized in that, when it is determined that the color is an interference color, a color bar is created by arranging the representative interference color images corresponding to the determined image data . The differential interference observation optical system is
A polarizer that is inserted into the optical path of the illumination light and converts the illumination light into linearly polarized light having a single vibration surface;
The linearly polarized light that is movable in a direction orthogonal to the optical axis of the objective lens and is emitted after being decomposed into two linearly polarized light having vibration planes orthogonal to each other, and the two straight lines that are emitted A Nomarski prism that integrates and interferes with two light beams according to the two linearly polarized light from the specimen irradiated with polarized light on a single vibration surface;
An analyzer that passes only light that has passed through the Nomarski prism;
Consists of
A drive controller for controlling movement of the Nomarski prism in a direction perpendicular to the optical axis of the objective lens;
The microscope system according to claim 1 , wherein the color bar creating unit arranges the representative interference color image in association with a moving direction of the Nomarski prism. The storage unit includes a position information storage unit that stores position information of the Nomarski prism driven by the drive control unit;
Further comprising an image processing unit that determines whether the interference color of the interference image is the darkest image data based on the image data of the interference image sequentially acquired along with the movement of the Nomarski prism;
When the image processing unit determines that the interference color of the interference image is the darkest image data, the drive control unit sets the position of the Nomarski prism that acquired the image data as an origin. The microscope system according to claim 2 . The storage unit includes a position information storage unit that stores position information of the Nomarski prism driven by the drive control unit;
The retardation adjusting unit, on the basis of the positional information acquiring an image of the color bar, which is selected by the input unit, the Roh claim, characterized in <br/> setting the origin in the movement of the circle ski prism 2 The microscope system described in 1. Wherein the input unit, the microscope system according to any one of claims 1-4, characterized in that a touch panel for accepting an input in accordance with the contact position of the object from the outside. A display control unit for controlling a display mode of the display unit;
The display control unit
A representative interference color image instructed by the input unit;
An interference color image of an interference color near the representative interference color corresponding to the representative interference color image instructed by the input unit;
Is displayed on the display unit. The microscope system according to any one of claims 1 to 5 .

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