JP2012181341A - Microscope device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To perform automatic focusing with high accuracy without being influenced by the quantity of light that irradiates a sample for taking an image.SOLUTION: In a microscope device, a first optical system 107 that is irradiated with light from a first light source 105 and a second light source 106, and irradiates light having a spectrum in which a first spectrum and a second spectrum overlap; a third optical system 110 which is irradiated with light from a second optical system 109, splits into light having a first wavelength region and light having a second wavelength region and emits the split light; an imaging section 112 on which the light having the first wavelength region emitted from the third optical system 110 is incident, and which takes an image of a sample slide 101 due to the light having the first wavelength region; and an AF section 114 on which the light having the second wavelength region emitted from the third optical system 110 is incident, and which focuses the imaging section 112 by adjusting a stage 102 or the second optical system 109 according to at least a part of contrast of the image of the sample slide 101 due to the light having the second wavelength region.

Description

  The present invention relates to a microscope apparatus.

  A pathological specimen or biological tissue (specimen sample) is placed on the slide glass, and the image is acquired by capturing the transmitted light of the slide glass, that is, the optical image of the specimen sample with an imaging device such as an electronic camera. There is a digital microscope apparatus that observes an image on a monitor. In recent years, white LEDs (Light Emitting Diodes) have begun to be widely used as illumination light sources for microscopes because of their low power consumption, low heat generation, long life, and easy maintenance. (For example, see Patent Document 1 and Non-Patent Document 1).

  FIG. 10 is a schematic diagram showing a configuration of a biological microscope using a white LED as an illumination light source, which is conventionally known. Usually, a microscope used in the biological field uses a transmissive slide specimen having a low reflectance, so that a so-called passive AF method that detects an optical image contrast and performs an autofocus (hereinafter referred to as AF) operation is used. Used.

  The illustrated microscope apparatus 1000 includes a stage 1002 on which a sample 1001 is placed, and a stage driving unit 1003 that drives the stage 1002 in the horizontal direction and the optical axis direction. In addition, the microscope apparatus 1000 includes a white LED 1004 that illuminates the sample 1001, a condenser lens 1005 that condenses the light of the white LED 1004, an objective lens 1006 that includes a plurality of lenses so as to face the sample 1001, and an objective lens. An imaging lens 1007 disposed along the optical axis 1006 and a camera 1008 for capturing an image of the sample 1001 are provided.

  The microscope apparatus 1000 also includes an AF unit 1009 that generates a focusing signal necessary for focusing the image of the sample 1001 on the camera 1008, and a stage driving unit 1003 based on the focusing signal generated by the AF unit 1009. And a stage control unit 1010 for controlling. The microscope apparatus 1000 includes a beam splitter 1011 that distributes light from the sample 1001 and guides the light to the AF unit 1009 on the optical axis of the objective lens 1006. Further, the microscope apparatus 1000 includes a condenser lens 1012 that condenses the light from the beam splitter 1011 and guides it to the AF unit 1009.

  Next, a method for imaging the sample 1001 using the microscope apparatus 1000 will be described. First, the stage driving unit 1003 drives the stage 1002 in the horizontal direction, and moves a predetermined imaging region of the sample 1001 placed on the stage 1002 to the visual field range of the camera 1008. Next, the AF unit 1009 detects the contrast of the light image from the sample 1001 and generates a focus signal based on the detected contrast. The AF unit 1009 inputs the generated focus signal to the stage control unit 1010. The stage control unit 1010 analyzes the input focusing signal, controls the stage driving unit 1003 so that an image of the sample 1001 is formed on the imaging element surface of the camera 1008, and moves the stage 1002 to the optical axis. . When an image of the sample 1001 is formed on the image sensor surface of the camera 1008, the camera 1008 acquires an image of a predetermined shooting area of the sample 1001.

  As described above, a microscope apparatus 1000 that splits light from the sample 1001 and guides the light to the camera 1008 and the AF unit 1009 is known (see, for example, Patent Document 2).

JP 2005-148296 A US Pat. No. 7,576,307

“Nikon Home Page for Biological Microscopes for Research and Inspection”, [online], [Search on February 22, 2011], Internet <URL: http://www.nikon-instruments.jp/jpn/page/products/ 50i55i.aspx>

  In order to perform AF with high accuracy, it is necessary to supply sufficient light to the AF sensor provided in the AF unit to generate a sufficiently large AF signal against noise generated by the AF sensor. Further, the amount of light of the white LED is smaller than that of a halogen lamp that is a typical light source of a conventional microscope apparatus. Therefore, when a white LED is used as the light source of the microscope apparatus, if the light quantity distribution to the photographing camera is increased in order to capture a high-quality image, the light to the AF unit becomes insufficient, and high-precision AF is performed. There is a problem that can not be.

  In recent years, in the field of pathology such as cell and tissue diagnosis, a specimen slide is divided and photographed at a high resolution using an imaging device such as an area camera or a line camera, and the photographed divided images are pasted together to assemble the entire specimen slide. A virtual microscope apparatus that can be converted into a digital image, displayed on a PC monitor, and operated as if observing a sample with an actual microscope is becoming widespread. In such a virtual microscope apparatus, there is a strong demand for an apparatus capable of high-speed image reading in order to improve the efficiency of pathological diagnosis. Therefore, since the exposure time of the imaging device cannot be increased, there is a problem that the light to the AF unit becomes insufficient and high-precision AF cannot be performed.

  The present invention has been made to solve the above problems, and provides a microscope apparatus that can perform automatic focusing with high accuracy without being affected by the amount of light that irradiates a sample for imaging. For the purpose.

  The present invention includes a first light source that emits light having a first spectrum having an intensity distribution in a first wavelength region, and a first light source having an intensity distribution in a second wavelength region different from the first wavelength region. A second light source that emits light having a spectrum of 2, a light that is irradiated with light from the first light source and the second light source, and a spectrum obtained by superimposing the first spectrum and the second spectrum A first optical system for irradiating light, a stage for holding the sample so that the light from the first optical system is incident on the sample, and the sample by the light from the first optical system. A second optical system for enlarging an image, and a third light that is incident on the light from the second optical system and emits the light in the first wavelength region and the light in the second wavelength region separately. The light in the first wavelength region from the optical system and the third optical system is incident. An imaging unit that captures an image of the sample with light in the first wavelength region and light in the second wavelength region from the third optical system are incident, and the light in the second wavelength region is incident And an autofocus unit that adjusts the stage or the second optical system according to the contrast of at least a part of the image of the sample to adjust the focus of the imaging unit. .

  In the microscope apparatus of the present invention, the first optical system has a first dichroic mirror, and the third optical system has a second dichroic mirror.

  Further, in the microscope apparatus according to the present invention, the autofocus unit generates an autofocus signal corresponding to a contrast of at least a part of the sample image by the light in the second wavelength region, and the autofocus unit generates And a light amount adjusting unit that controls the amount of light emitted from the second light source based on an autofocus signal.

  In the microscope apparatus of the present invention, the first light source is a white LED, the first wavelength region is a visible light region, the second light source is an infrared irradiation device, and the second wavelength region is It is an infrared region.

  In the microscope apparatus of the present invention, the stage is disposed between the first optical system and the second optical system, and the second optical system enlarges a transmission image of the sample. It is characterized by.

  According to the present invention, the first light source emits light having a first spectrum having an intensity distribution in the first wavelength region. The second light source emits light having a second spectrum having an intensity distribution in a second wavelength region different from the first wavelength region. The first optical system is irradiated with light from the first light source and the second light source, and emits light having a spectrum obtained by superimposing the first spectrum and the second spectrum. The stage holds the sample so that light from the first optical system is incident on the sample. The second optical system enlarges the image of the sample by the light from the first optical system. The third optical system receives light from the second optical system and emits light in the first wavelength region and light in the second wavelength region separately. The imaging unit receives light of the first wavelength region from the third optical system, and captures an image of the sample by the light of the first wavelength region. The autofocus unit receives light of the second wavelength region from the third optical system, and the stage or the second according to the contrast of at least a part of the image of the sample by the light of the second wavelength region. The image pickup unit is focused by adjusting the optical system.

  Thereby, the microscope apparatus can perform the AF process using only the light having the second spectrum having the intensity distribution in the second wavelength region different from the first wavelength region irradiated by the second light source. Therefore, automatic focusing can be performed with high accuracy without being affected by the amount of light emitted from the first light source and having the first spectrum having the intensity distribution in the first wavelength region.

It is the schematic which showed the structure of the microscope apparatus in the 1st Embodiment of this invention. It is the graph which showed the relationship between the wavelength characteristic of the 1st light source in the 1st Embodiment of this invention, and a 2nd light source, and the wavelength characteristic of a 1st optical system and a 3rd optical system. It is the schematic which showed the structure of AF part in the 1st Embodiment of this invention. It is a top view of the line sensor in the 1st embodiment of the present invention. 6 is a graph showing changes in the front pin contrast signal and the rear pin contrast signal when the position of the specimen slide is changed in the optical axis direction of the second optical system in the first embodiment of the present invention. In the 1st Embodiment of this invention, it is the graph which showed the change of the difference contrast signal value when the position of a sample slide is changed to an optical axis direction. It is the schematic which showed the structure of the microscope apparatus in the 2nd Embodiment of this invention. It is the schematic which showed arrangement | positioning with the 1st light source with which the light source unit in the 2nd Embodiment of this invention is equipped, and a 2nd light source. It is the schematic which showed the structure of the microscope apparatus in the 3rd Embodiment of this invention. It is the schematic which showed the structure of the biological microscope using white LED as an illumination light source known conventionally.

(First embodiment)
Hereinafter, a first embodiment of the present invention will be described with reference to the drawings. FIG. 1 is a schematic diagram illustrating a configuration of a microscope apparatus 1 according to the present embodiment. In the illustrated example, the microscope apparatus 1 includes a stage 102, a stage driving unit 103, a stage control unit 104, a first light source 105, a second light source 106, a first optical system 107, and a condenser lens. 108, second optical system 109, third optical system 110, first imaging lens 111, imaging unit 112, second imaging lens 113, and AF (Autofocus, autofocus) unit 114.

  The stage 102 is a stage for placing a specimen slide 101 (sample) on which a pathological specimen or a biological tissue is placed on a slide glass. The stage 102 is disposed between the first optical system 107 and the second optical system 109. The stage driving unit 103 drives the stage 102 in the horizontal and vertical directions. The stage control unit 104 controls the stage driving unit 103 based on the AF signal (autofocus signal) output from the AF unit 114. The first light source 105 is, for example, a white LED or the like, and visible light (light having a first spectrum having an intensity distribution in the first wavelength region) used by the imaging unit 112 to capture an image of the specimen slide 101. It is a light source that generates The second light source 106 is, for example, an infrared irradiation device (infrared LED) or the like, and invisible light (the first wavelength region is used by the AF unit 114 to generate an AF signal based on the image of the specimen slide 101). A light source that generates light having a second spectrum having an intensity distribution in a different second wavelength region.

  The first optical system 107 is, for example, a dichroic mirror, and is a combination of visible light from the first light source 105 and invisible light from the second light source 106 (mixed light, first spectrum and second light). The condenser lens 108 is irradiated with light having a spectrum superposed on the spectrum. In the illustrated example, the first optical system 107 transmits visible light from the first light source 105 and reflects invisible light from the second light source 106 in the direction of the condenser lens 108, thereby The condenser lens 108 is irradiated with light that is a combination of visible light from the light source 105 and invisible light from the second light source 106.

  The condenser lens 108 collects the mixed light irradiated by the first optical system 107 and irradiates the sample slide 101. The second optical system 109 is an objective lens composed of a plurality of lenses, and is disposed so as to face the specimen slide 101. The second optical system 109 condenses the light flux from the specimen slide 101 and irradiates the third optical system 110 with the condensed mixed light. As described above, the second optical system 109 enlarges the transmission image of the specimen slide 101.

  The third optical system 110 is, for example, a dichroic mirror, and irradiates the first imaging lens 111 with visible light included in the mixed light from the second optical system 109. The invisible light included in the mixed light is irradiated to the second imaging lens 113. In the illustrated example, the third optical system 110 transmits visible light included in the mixed light from the second optical system 109 and converts invisible light included in the mixed light from the second optical system 109 to the second. By reflecting in the direction of the imaging lens 113, visible light included in the mixed light from the second optical system 109 is irradiated to the first imaging lens 111, and from the second optical system 109. The invisible light included in the mixed light is irradiated to the second imaging lens 113. An optical path of visible light that is transmitted through the third optical system 110 and incident on the first imaging lens 111 is an optical path A. The optical path of invisible light that is reflected by the third optical system 110 and incident on the second imaging lens 113 is referred to as an optical path B.

  The first imaging lens 111 is disposed along the optical axis of the second optical system 109. In addition, the first imaging lens 111 is formed on the imaging surface of the imaging element provided in the imaging unit 112 with visible light transmitted through the third optical system 110 out of the mixed light collected by the second optical system 109. Make an image. Thereby, visible light included in the mixed light from the specimen slide 101 is guided to the imaging unit 112. The imaging device included in the imaging unit 112 receives visible light from the specimen slide 101, and photoelectrically converts the received visible light into an electrical signal corresponding to the intensity of the received visible light. The imaging unit 112 generates image data of the specimen slide 101 based on the electrical signal photoelectrically converted by the imaging device.

  The second imaging lens 113 forms invisible light reflected by the third optical system 110 out of the mixed light collected by the second optical system 109 on the light receiving surface of the line sensor provided in the AF unit 114. Let Thereby, the invisible light included in the mixed light from the specimen slide 101 is guided to the AF unit 114. The AF unit 114 includes a line sensor that photoelectrically converts invisible light, receives the invisible light from the specimen slide 101, and photoelectrically converts the invisible light into an electrical signal corresponding to the intensity of the received invisible light. Further, the AF unit 114 generates an AF signal based on the electrical signal photoelectrically converted by the line sensor.

  In FIG. 1, the direction parallel to the optical path A is the Z axis, the direction parallel to the optical path B is the X axis, and the direction perpendicular to both the X axis and the Z axis is the Y axis. The microscope apparatus 1 includes a CPU (Central Processing Unit), a ROM (Read Only Memory), a RAM (Random Access Memory), an external storage device, and the like. Including a computer system (not shown). The process performed by the above-described stage control unit 104, imaging unit 112, and AF unit 114 is controlled by a computer system. For example, the stage drive unit 103, the stage control unit 104, and the AF unit 114 correspond to the autofocus unit of the present invention.

  Next, the wavelength characteristics of the first light source 105 and the second light source 106 and the wavelength characteristics of the first optical system 107 and the third optical system 110 will be described. FIG. 2 is a graph showing the relationship between the wavelength characteristics of the first light source 105 and the second light source 106 and the wavelength characteristics of the first optical system 107 and the third optical system 110 in the present embodiment. In the graph shown in the figure, the first light source 105 is a white LED, the second light source 106 is an infrared LED, and the first optical system 107 and the third optical system 110 are dichroic mirrors. The characteristics are shown.

  The horizontal axis of the graph shown shows the wavelength. In addition, the vertical axis of the illustrated graph indicates the intensity of light and whether light is transmitted or reflected. A curve 201 indicates the intensity of each wavelength component of visible light generated by the first light source 105. A curve 202 indicates the intensity of each wavelength component of the invisible light generated by the second light source 106. A broken line 203 indicates whether the first optical system 107 and the third optical system 110 transmit or reflect light.

  As shown in the figure, the light generated by the first light source 105 has a strong intensity of visible light component (wavelength of visible light). In addition, the light generated by the second light source 106 has a high intensity of invisible light component (wavelength of invisible light). Further, the first optical system 107 and the third optical system 110 transmit light having a wavelength of visible light component and reflect light having a wavelength of invisible light component. That is, the first optical system 107 and the third optical system 110 transmit visible light and reflect invisible light.

  Next, the configuration of the AF unit 114 will be described. FIG. 3 is a schematic diagram illustrating the configuration of the AF unit 114 in the present embodiment. In the illustrated example, the AF unit 114 includes a half mirror 1141, a mirror 1142, a line sensor 1143, and a contrast detection unit 1144. The half mirror 1141 is disposed between the second imaging lens 113 and the line sensor 1143. The half mirror 1141 transmits a part of the invisible light from the second imaging lens 113 and reflects a part thereof in the direction of the mirror 1142. Thereby, the half mirror 1141 divides the invisible light from the second imaging lens 114 into two in the direction of the line sensor 1143 and the direction of the mirror 1142. The mirror 1142 reflects the invisible light reflected by the half mirror 1141 toward the line sensor 1143.

  An optical path of invisible light that is transmitted through the half mirror 1141 and incident on the line sensor 1143 is referred to as an optical path C. An optical path of invisible light that is reflected by the half mirror 1141 and reflected by the mirror 1142 and incident on the line sensor 1143 is defined as an optical path D. The mirror 1142 is arranged so that the reflected optical path of invisible light is substantially parallel to the optical path C. Further, since the optical path C is an optical path that is directly guided from the half mirror 1141 to the line sensor 1143, and the optical path D is an optical path that is guided from the half mirror 1141 to the line sensor 1143 via the mirror 1142, the optical path length of the optical path D is It is longer than the optical path length of the optical path C.

  The line sensor 1143 is disposed on the optical path C and the optical path D, and receives invisible light from the specimen slide 101 guided to the optical path C and invisible light from the specimen slide 101 guided to the optical path D. Photoelectrically converted into an electrical signal corresponding to the intensity of the invisible light. A plane that is a light detection surface of the line sensor 1143 is a light receiving surface. The contrast detection unit 1144 is based on the electrical signal output from the line sensor 1143, and the contrast of the invisible light from the specimen slide 101 that is guided to the optical path C and incident on the line sensor 1143 and the optical sensor D is guided to the line sensor 1143. A difference from the contrast of the invisible light from the specimen slide 101 incident on 1143 is detected. The contrast detection unit 1144 generates a contrast signal indicating the detected contrast difference, and inputs the generated contrast signal to the stage control unit 104. In FIG. 3, the direction from the line sensor 1143 to the half mirror 1141 is the z axis, the direction from the half mirror 1141 to the mirror 1142 is the x axis, and the direction perpendicular to both the x axis and the z axis is the y axis. .

  Next, the configuration of the line sensor 1143 will be described. FIG. 4 is a top view of the line sensor 1143 in the present embodiment. As illustrated, the line sensor 1143 includes a plurality of light receiving elements 401 arranged in a straight line in a row. FIG. 4 shows an area 410 in which a projection image by invisible light guided to the optical path C displayed on the line sensor 1143 is displayed and a projection image by invisible light guided to the optical path D in the present embodiment. An area 411 to be displayed is shown. In the present embodiment, a projection image by invisible light (light transmitted through the half mirror 1141) guided to the optical path C is projected onto the area 410. Further, a projection image by light guided to the optical path D (light reflected by the half mirror 1141 and reflected by the mirror 1142) is projected onto the region 411. As described above, the optical path length of the optical path D is longer than the optical path length of the optical path C. Therefore, the imaging point of the invisible light guided to the optical path C is behind the line sensor 1143 (FIG. 3, rear pin position 301 on the plane m), and the imaging point of the invisible light guided to the optical path D is a line. It is the front of the sensor 1143 (FIG. 3, front pin position 302 on the plane n).

  Next, signals output from the line sensor 1143 will be described. The line sensor 1143 receives the invisible light that has been introduced through the optical path C and the invisible light that has been introduced through the optical path D, and photoelectrically converts the invisible light into an electrical signal corresponding to the intensity of the received light. Here, the electric signal obtained by photoelectrically converting the image (invisible light) guided to the optical path C by the line sensor 1143 is defined as a rear pin signal, and the line sensor 1143 photoelectrically converts the image guided to the optical path D. The electrical signal is defined as the front pin signal.

  Next, a contrast signal generated by the contrast detection unit 1144 will be described. The front pin signal and the rear pin signal output from the line sensor 1143 are input to the contrast detection unit 1144. The contrast detection unit 1144 detects a contrast difference between the front pin signal and the rear pin signal, and generates a difference contrast signal. Specifically, the contrast detection unit 1144 first calculates the absolute value of the output difference between the pixels of the light receiving element 401 in each of the front pin signal and the rear pin signal, and sums the absolute value of the front pin contrast signal and the rear pin signal. Generate a pin contrast signal. Next, the contrast detection unit 1144 calculates a difference between the front pin contrast signal and the rear pin contrast signal, and generates a difference contrast signal.

  FIG. 5 is a graph showing changes in the front pin contrast signal and the rear pin contrast signal when the position of the specimen slide 101 is changed in the optical axis direction of the second optical system 109 (FIG. 1, Z-axis direction). is there. The horizontal axis of the illustrated graph indicates the distance (defocus amount) between the specimen slide 101 and the second optical system 109, and the vertical axis indicates the contrast signal value. A curve 501 indicates the relationship between the defocus amount and the front pin contrast signal value. A curve 502 indicates the relationship between the defocus amount and the rear pin contrast signal value. When the specimen slide 101 placed on the stage 102 moving in the Z-axis direction is moved from a position (−Z) sufficiently far from the second optical system 109 to a position (+ Z) sufficiently close, first, The front pin contrast signal value becomes maximum at the projected image focus position (front pin position) along the optical path D and then decreases. Subsequently, the rear pin contrast signal value becomes maximum at the projected image focusing position (rear pin position) along the optical path C and then decreases.

  FIG. 6 is a graph showing changes in the differential contrast signal value when the position of the specimen slide 101 is changed in the optical axis direction (Z-axis direction). The horizontal axis of the illustrated graph indicates the distance (defocus amount) between the specimen slide 101 and the second optical system 109, and the vertical axis indicates the difference contrast signal value. A curve 601 indicates the relationship between the defocus amount and the difference contrast signal value. When the specimen slide 101 placed on the stage 102 moving in the Z-axis direction is moved from a position (−Z) sufficiently far from the second optical system 109 to a position (+ Z) sufficiently close to the second optical system 109, a difference contrast is obtained. The signal value has a maximum at the projected image focus position by the optical path D, and then decreases sharply to zero, and draws an S-shaped curve that is minimum at the projected image focus position by the optical path C.

  In the present embodiment, the imaging surface of the imaging device included in the imaging unit 112 is disposed at a position where the image of the sample slide 101 is formed when the differential contrast signal value is zero. Therefore, the stage control unit 104 moves the position of the sample slide 101 (position of the stage 102) in the Z-axis direction so that the difference contrast signal value input from the AF unit 114 (contrast detection unit 1144) becomes zero. Thus, the focus of the image sensor provided in the imaging unit 112 can be controlled so as to match the specimen slide 101 (optical path difference AF). Note that the process of controlling the focus of the image sensor included in the imaging unit 112 so as to match the specimen slide 101 is referred to as a focusing process.

  Specifically, when the input differential contrast signal value is positive, the stage control unit 104 determines that the focus of the image sensor included in the imaging unit 112 is in the front focus state, and causes the stage drive unit 103 to detect the differential contrast. The stage 102 is moved in the −Z-axis direction by a movement amount according to the magnitude of the signal. Further, when the input differential contrast signal value is negative, the stage control unit 104 determines that the focus of the image sensor included in the imaging unit 112 is in the rear focus state, and causes the stage drive unit 103 to detect the magnitude of the differential contrast signal. The stage 102 is moved in the + Z-axis direction by a movement amount according to the above. The stage control unit 104 repeats this operation, and when the input difference contrast signal value becomes zero, the stage control unit 104 determines that the image pickup device included in the image pickup unit 112 is in a focused state, and ends the focusing process.

  Next, the operation of the microscope apparatus 1 in the present embodiment will be described. First, the stage control unit 104 causes the stage driving unit 103 to drive the stage 102 in the horizontal direction, and moves a predetermined region to be photographed on the specimen slide 101 on the optical axis of the second optical system 109. Next, the first light source 105 generates visible light that irradiates the specimen slide 101, and the second light source 106 generates invisible light that irradiates the specimen slide 101. The first optical system 107 irradiates the condenser lens 108 with light (mixed light) that is a combination of visible light generated by the first light source 105 and invisible light generated by the second light source 106.

  The condenser lens 108 collects the mixed light irradiated by the first optical system 107 and irradiates the sample slide 101. The mixed light that has passed through the specimen slide 101 is collected by the second optical system 109, the visible light is divided in the direction of the first imaging lens 111 by the third optical system 110, and the invisible light is the second invisible light. Are divided in the direction of the imaging lens 113.

  Visible light divided in the direction of the first imaging lens 111 is irradiated to the imaging surface of the imaging element included in the imaging unit 112 via the first imaging lens 111. On the other hand, the invisible light divided in the direction of the second imaging lens 113 is incident on the AF unit 114 via the second imaging lens 113. The invisible light that has entered the AF unit 114 enters the half mirror 1141. The invisible light incident on the half mirror 1141 is divided into the direction of the line sensor 1143 and the direction of the mirror 1142.

  The invisible light divided in the direction of the line sensor 1143 is guided to the optical path C and projected onto the line sensor 1143. The projected image of the light guided to the optical path C is projected onto the area 410 shown in FIG. On the other hand, the light divided in the direction of the mirror 1142 is guided to the optical path D through the mirror 1142, that is, projected onto the line sensor 1143. The projected image of the light guided to the optical path D is projected onto the area 411 shown in FIG.

  Next, the line sensor 1143 receives light that has been guided to the optical path C and entered the region 410 and light that has been guided by the optical path D and entered the region 411, and the front pin according to the intensity of the received light. A signal and a rear pin signal are output. The front pin signal and the rear pin signal output from the line sensor 1143 are input to the contrast detection unit 1144. The contrast detection unit 1144 detects the contrast difference between the front pin signal and the rear pin signal, generates an AF signal (differential contrast signal), and inputs the AF signal to the stage control unit 104. The stage control unit 104 moves the position of the sample slide 101 (position of the stage 102) in the Z-axis direction so that the AF signal value input from the contrast detection unit 1144 becomes zero, and the image sensor included in the imaging unit 112 Control is performed so that the focus is adjusted to the specimen slide 101. Then, when the AF signal value input from the contrast detection unit 1144 becomes zero, the stage control unit 104 determines that the focus of the image sensor included in the imaging unit 112 is in focus, and performs the focusing process. finish. After the focusing process is completed, the imaging unit 112 captures the specimen slide 101 and generates image data of the specimen slide 101 according to a command from a computer system (not shown).

  As described above, according to the present embodiment, the first light source 105 generates visible light, and the second light source 106 generates invisible light. Then, the first optical system 107 irradiates the sample slide 101 with mixed light in which visible light and invisible light are combined. In addition, the third optical system 110 divides the mixed light transmitted through the specimen slide 101 into visible light and invisible light. The AF unit 114 performs AF processing using invisible light, and the imaging unit 112 images the specimen slide 101 using visible light. Thereby, since the microscope apparatus 1 can perform AF processing using only invisible light generated by the second light source 106, it is not affected by the amount of visible light generated by the first light source 105. Automatic focusing can be performed with high accuracy.

  Moreover, since the imaging device 1 can supply all visible light generated by the first light source 105 to the imaging unit 112, it can acquire a high-quality image. In addition, since the imaging apparatus 1 can supply all invisible light generated by the second light source 106 to the AF unit 114, automatic focusing can be performed with high accuracy.

(Second Embodiment)
Next, a second embodiment of the present invention will be described with reference to the drawings. The difference between the configuration of the microscope apparatus 2 in the present embodiment and the configuration of the microscope apparatus 1 in the first embodiment is that the configuration of the light source that irradiates the specimen slide 101 and light that combines visible light and invisible light ( It is the structure of the 1st optical part which irradiates (mixed light).

  FIG. 7 is a schematic diagram showing the configuration of the microscope apparatus 2 in the present embodiment. In the illustrated example, the microscope apparatus 2 includes a stage 102, a stage driving unit 103, a stage control unit 104, a light source unit 701, a first optical system 702, a condenser lens 108, and a second optical system 109. A third optical system 110, a first imaging lens 111, an imaging unit 112, a second imaging lens 113, and an AF unit 114.

  Stage 102, stage drive unit 103, stage control unit 104, condenser lens 108, second optical system 109, third optical system 110, first imaging lens 111, and imaging unit 112 The configurations of the second imaging lens 113 and the AF unit 114 are the same as the configurations of the respective units in the first embodiment.

  The light source unit 701 includes a plurality of first light sources that generate visible light and a plurality of second light sources that generate invisible light. FIG. 8 is a schematic view showing the arrangement of the first light source and the second light source provided in the light source unit 701 in the present embodiment. In the illustrated example, the light source unit 701 has a plurality of first light sources 7011 and a plurality of second light sources 7012 arranged alternately. The first light source 7011 generates visible light. The second light source 7012 generates invisible light. Thereby, the light source unit 701 can generate visible light and invisible light.

  The first optical system 702 is, for example, a diffusing plate, an optical fiber, or a fly-eye lens. Visible light generated by a plurality of first light sources 7011 included in the light source unit 701, and a plurality of second light sources 7012 Then, the invisible light generated is uniformly diffused and mixed, and the condenser lens 108 is irradiated with the diffused and mixed light (mixed light).

  The mixed light applied to the condenser lens 108 passes through the specimen slide 101 and is collected by the second optical system 109 as in the first embodiment. The invisible light is divided in the direction of the first imaging lens 111, and the invisible light is divided in the direction of the second imaging lens 113. Then, the visible light divided in the direction of the first imaging lens 111 is irradiated to the imaging surface of the imaging element included in the imaging unit 112 via the first imaging lens 111. On the other hand, the invisible light divided in the direction of the second imaging lens 113 is incident on the AF unit 114 via the second imaging lens 113. The AF unit 114 performs AF processing using invisible light, and the imaging unit 112 images the specimen slide 101 using visible light.

  Therefore, the imaging apparatus 2 can supply all visible light generated by the plurality of first light sources 7011 included in the light source unit 701 to the imaging unit 112, and thus can acquire a high-quality image. Further, since the imaging apparatus 2 can supply all the invisible light generated by the plurality of second light sources 7012 included in the light source unit 701 to the AF unit 114, it can perform automatic focusing with high accuracy. In addition, since the imaging apparatus 2 can perform AF processing using only invisible light generated by the plurality of second light sources 7012 included in the light source unit 701, the plurality of first light sources 7011 included in the light source unit 701 are included. Automatic focusing can be performed with high accuracy without being affected by the amount of visible light generated.

(Third embodiment)
Next, a third embodiment of the present invention will be described with reference to the drawings. The difference between the configuration of the microscope apparatus according to the present embodiment and the configuration of the microscope apparatus 1 according to the first embodiment is that the microscope apparatus according to the present embodiment has a first function according to the intensity of the AF signal output from the AF unit 114. It is the point provided with the light quantity adjustment part which adjusts the light quantity of the invisible light which the 2 light source 106 generate | occur | produces.

  FIG. 9 is a schematic diagram showing the configuration of the microscope apparatus 3 in the present embodiment. In the illustrated example, the microscope apparatus 3 includes a stage 102, a stage driving unit 103, a stage control unit 104, a first light source 105, a second light source 106, a first optical system 107, and a condenser lens. 108, a second optical system 109, a third optical system 110, a first imaging lens 111, an imaging unit 112, a second imaging lens 113, an AF unit 114, and a light amount adjustment unit. 901.

  Stage 102, stage drive unit 103, stage control unit 104, first light source 105, second light source 106, first optical system 107, condenser lens 108, and second optical system 109 The configuration of the third optical system 110, the first imaging lens 111, the imaging unit 112, the second imaging lens 113, and the AF unit 114 is the same as the configuration of each unit in the first embodiment. It is the same composition.

  The light amount adjustment unit 901 acquires the AF signal output from the AF unit 114 and adjusts the amount of invisible light generated by the second light source 106 based on the intensity of the acquired AF signal. Specifically, when the signal intensity of the AF signal output from the AF unit 114 is so small that the stage control unit 104 cannot accurately drive the stage drive unit 103, the light amount adjustment unit 901 includes the second light intensity adjustment unit 901. The second light source 106 is controlled so that the amount of invisible light generated by the light source 106 increases. In addition, when the signal intensity of the AF signal output from the AF unit 114 is so large that the stage control unit 104 cannot drive the stage drive unit 103 with high accuracy, the light amount adjustment unit 901 includes the second light source 106. The second light source 106 is controlled so that the amount of invisible light that occurs is reduced.

  For example, when the specimen slide 101 is difficult to transmit light, the amount of invisible light incident on the AF unit 114 is reduced and the output intensity of the AF signal output from the AF unit 114 is reduced. When the output intensity of the AF signal output by 114 is small, the light amount adjustment unit 901 controls the second light source 106 so that the amount of invisible light generated by the second light source 106 is increased. A stable AF operation can be performed. When the specimen slide 101 easily transmits light, the amount of invisible light incident on the AF unit 114 increases and the output intensity of the AF signal output from the AF unit 114 increases. In the present embodiment, the AF unit When the output intensity of the AF signal output from 114 is high, the light amount adjustment unit 901 controls the second light source 106 so that the amount of invisible light generated by the second light source 106 is small. A stable AF operation can be performed.

  As described above, according to the present embodiment, the imaging device 3 can supply all visible light generated by the first light source 105 to the imaging unit 112, and thus can acquire a high-quality image. Further, since the imaging device 3 can supply all invisible light generated by the second light source 106 to the AF unit 114, it can perform automatic focusing with high accuracy. In addition, since the imaging apparatus 3 can perform the AF process using only the invisible light generated by the second light source 106, the imaging apparatus 3 is not affected by the amount of visible light generated by the first light source 105. Automatic focusing can be performed with accuracy. Furthermore, since the imaging device 3 can adjust the second light source 106 so as to generate invisible light having an optimum light amount for the AF operation according to the state of the specimen slide 101, a highly accurate and stable AF operation can be performed. It can be performed.

  Although the first to third embodiments of the present invention have been described in detail with reference to the drawings, the specific configuration is not limited to this embodiment and does not depart from the gist of the present invention. Range design etc. are also included.

  For example, the microscope apparatus 2 according to the second embodiment includes the light amount adjustment unit 901 as in the microscope apparatus 3 according to the third embodiment, and the light source unit 701 includes the light source unit 701 according to the intensity of the AF signal output from the AF unit 114. The light emission amount of the plurality of second light sources 7012 may be adjusted.

  Also, for example, in the first to third embodiments, the position of the specimen slide 101 (stage 102) is set so that the AF signal value input from the contrast detection unit 1144 becomes zero when performing autofocus processing. However, the present invention is not limited to this. For example, when performing autofocus processing, the second optical system 109 (objective lens) may be moved in the Z-axis direction so that the AF signal value input from the contrast detection unit 1144 becomes zero. .

  1, 2, 3 ... Microscope device, 102 ... Stage, 103 ... Stage drive unit, 104 ... Stage control unit, 105, 7011 ... First light source, 106, 7012 ... Second light source 107, 702 ... first optical system 108 ... condenser lens 109 ... second optical system 110 ... third optical system 111 ... first Imaging lens, 112 ... imaging unit, 113 ... second imaging lens, 114 ... AF unit, 401 ... light receiving element, 701 ... light source unit, 901 ... light amount adjustment 1141,... Half mirror, 1142 ... Mirror, 1143 ... Line sensor, 1144 ... Contrast detector

Claims (5)

A first light source that irradiates light having a first spectrum having an intensity distribution in a first wavelength region;
A second light source for irradiating light having a second spectrum having an intensity distribution in a second wavelength region different from the first wavelength region;
A first optical system that irradiates light from the first light source and the second light source and emits light having a spectrum obtained by superimposing the first spectrum and the second spectrum;
A stage for holding the sample so that light from the first optical system is incident on the sample;
A second optical system for enlarging an image of the sample by light from the first optical system;
A third optical system that receives light from the second optical system and emits the light in the first wavelength region and the light in the second wavelength region separately;
An imaging unit that receives the light of the first wavelength region from the third optical system and captures an image of the sample by the light of the first wavelength region;
Light of the second wavelength region from the third optical system is incident, and the stage or the second optical according to the contrast of at least a part of the sample image by the light of the second wavelength region An autofocus unit for adjusting the focus of the imaging unit by adjusting the system;
A microscope apparatus characterized by comprising: The first optical system has a first dichroic mirror,
The microscope apparatus according to claim 1, wherein the third optical system includes a second dichroic mirror. The autofocus unit generates an autofocus signal corresponding to the contrast of at least a part of the image of the sample by the light in the second wavelength region,
The microscope apparatus according to claim 1, further comprising: a light amount adjustment unit that controls a light amount of light emitted from the second light source based on an autofocus signal generated by the autofocus unit. The first light source is a white LED, and the first wavelength region is a visible light region;
The microscope device according to any one of claims 1 to 3, wherein the second light source is an infrared irradiation device, and the second wavelength region is an infrared region. The stage is disposed between the first optical system and the second optical system,
The microscope apparatus according to any one of claims 1 to 4, wherein the second optical system enlarges a transmission image of the sample.

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