WO2018190125A1 - Unité confocal, système confocal et microscope confocal - Google Patents

Unité confocal, système confocal et microscope confocal Download PDF

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Publication number
WO2018190125A1
WO2018190125A1 PCT/JP2018/012712 JP2018012712W WO2018190125A1 WO 2018190125 A1 WO2018190125 A1 WO 2018190125A1 JP 2018012712 W JP2018012712 W JP 2018012712W WO 2018190125 A1 WO2018190125 A1 WO 2018190125A1
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Prior art keywords
line
detector
light source
linear
shaped
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PCT/JP2018/012712
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English (en)
Japanese (ja)
Inventor
山宮　広之
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横河電機株式会社
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Priority claimed from JP2017249617A external-priority patent/JP6677238B2/ja
Application filed by 横河電機株式会社 filed Critical 横河電機株式会社
Priority to EP18784330.5A priority Critical patent/EP3611550B1/fr
Priority to CN201880021124.4A priority patent/CN110462483B/zh
Priority to US16/604,292 priority patent/US11506878B2/en
Publication of WO2018190125A1 publication Critical patent/WO2018190125A1/fr

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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes

Definitions

  • the present invention relates to a confocal scanner, a microscope system, and a confocal microscope.
  • Patent Document 2 discloses another example of a line scanning confocal microscope.
  • the line-shaped illumination light is projected onto the observation object, and the return light from the observation object is detected by the line-shaped detector.
  • the entire optical system is translated and moved in one direction with respect to the object.
  • Patent Documents 3 and 4 disclose an example in which a slit array is provided between an objective lens, a light branching unit, and an imaging lens, and scanning is performed by reciprocating the slit array.
  • Japanese Patent No. 4905356 Japanese Patent No. 2660613 Japanese Patent Laid-Open No. 2001-13445 JP 2003-247817 A
  • the scan mirror used in Patent Document 1 is capable of high-speed scanning, but generally has a drawback that the system is expensive and the system is expensive.
  • the pupil position of the objective lens is within the objective lens or near the mounting position, and is suitable for placement of the scan mirror.
  • the scan mirror needs to transfer the pupil position of the objective lens by a relay lens, which has a drawback that the entire optical system becomes large.
  • Patent Document 2 it is necessary to translate the entire optical system as a scanning method, so that it is difficult to attach it to a manual microscope or the like already owned. There was a drawback that required.
  • a slit is arranged on the image plane magnified by the microscope, and the light passing through the slit is projected onto the focal plane of the microscope and returned.
  • the light that has passed through the slit in the light is imaged by an image sensor using a relay lens system.
  • a relay lens system for transferring an image on the imaging plane of the microscope is required, and there is a drawback that the optical system becomes large.
  • One aspect of the present invention has been made in view of the above circumstances, and is to provide a confocal scanner, a microscope system, and a confocal microscope that can obtain a confocal microscope image with a simple configuration.
  • one embodiment of the present invention is a confocal scanner mounted on a microscope, which includes a line-shaped light source that emits line-shaped light and a line that detects incident light for each line.
  • a linear detector having a linear detector, and a moving mechanism that translates the linear light source and the linear detector with respect to the microscope, the linear light source and the linear detector, are confocal scanners that are arranged in mutually corresponding positional relationships in the imaging plane at a position conjugate to the focal plane of the microscope.
  • the linear light source and the linear detector are disposed between an imaging lens provided in the microscope and an image plane of the imaging lens.
  • the light is branched by the branched optical system, and each is directly arranged on the imaging surface of the imaging lens.
  • the moving mechanism maintains the positional relationship between the line-shaped light source and the line-shaped detector, and the imaging plane of the microscope. Translate the top.
  • the line light source and the line detector include a longitudinal direction of the line light source and an optical axis direction of light emitted from the line light source.
  • a plane determined from a longitudinal direction of the linear detector and an optical axis direction of light incident on the linear detector are arranged in a positional relationship such that they are the same plane or planes parallel to each other,
  • the moving mechanism translates the linear light source and the linear detector in a direction perpendicular to the same plane or the plane parallel to each other.
  • the line light source and the line detector include a longitudinal direction of the line light source and an optical axis direction of light emitted from the line light source.
  • a positional relationship in which a plane that is determined and a plane that is determined from the longitudinal direction of the linear detector and the optical axis direction of light incident on the linear detector are the same plane, and the optical axis of the linear light source It arrange
  • the moving mechanism integrally translates the line light source and the line detector.
  • the confocal scanner includes a plurality of the same number of the line light sources and the line detector, and the line light sources and the line detectors that are paired with each other. Are arranged so as to correspond to each other.
  • each of the pair of line light sources and the line detector is emitted from a longitudinal direction of each line light source and the line light source.
  • the plane determined from the optical axis direction of the light to be transmitted and the plane determined from the longitudinal direction of each of the line detectors and the optical axis direction of the light incident on the line detector are the same plane or parallel to each other.
  • the moving mechanism translates each of the pair of linear light sources and the linear detectors in a direction perpendicular to the same plane or the plane parallel to each other. Move.
  • One embodiment of the present invention is the above confocal scanner, wherein the confocal scanner includes a plurality of the line-shaped light sources and the line-shaped detectors, and the line-shaped light sources and the line-shaped detectors are relatively moved.
  • the line-shaped light source and the line-shaped detector are selectively arranged so as to be compatible.
  • the confocal scanner includes a plurality of the line light sources and the one line detector, and relatively moves the line light source and the line detector.
  • One line-shaped light source and one line-shaped detector are selectively arranged from among the plurality of line-shaped light sources.
  • the linear light source in the confocal scanner, includes a plurality of light sources and slits, and the plurality of light sources and the slits include the plurality of light sources with respect to the slits.
  • the wavelength of one light source is selectively arranged from the plurality of light sources so as to be the wavelength of light emitted as the line light source.
  • a stage on which an observation target is installed an objective lens, an imaging lens that forms an image of the observation target incident through the objective lens, and the imaging lens
  • a line-shaped light source that emits line-shaped light that illuminates the observation object via the objective lens, and a line that emits light emitted by the observation object and incident via the objective lens and the imaging lens
  • a linear detector having a linear detection unit for detecting each line, and a moving mechanism that translates the linear light source and the linear detector with respect to the imaging lens.
  • the line-shaped detectors are confocal microscopes arranged in corresponding positions in the imaging plane at a position conjugate to the focal plane of the own microscope.
  • a confocal microscope image can be obtained with a simple configuration.
  • FIG. 2 The figure which shows an example of a structure of the microscope system which concerns on 1st Embodiment. 2 is a detailed view of a confocal scanner according to the first embodiment.
  • FIG. The figure which shows the detailed example of the drive mechanism which concerns on 1st Embodiment.
  • the side view which shows the detailed example of the linear light source which concerns on 1st Embodiment.
  • FIG. 6 is a flowchart illustrating an example of line scanning processing according to the first embodiment.
  • FIG. 10 is a diagram illustrating a configuration example of a confocal scanner according to a fifth embodiment.
  • the figure which shows an example of the drive mechanism which concerns on 6th Embodiment The figure which shows an example of the drive mechanism which concerns on 6th Embodiment.
  • the figure which shows an example of the drive mechanism which concerns on 7th Embodiment The figure which shows an example of the drive mechanism which concerns on 7th Embodiment.
  • the figure which shows an example of the two drive mechanisms which concern on 8th Embodiment The figure which shows an example of the drive mechanism which concerns on 9th Embodiment.
  • FIG. 1 is a diagram illustrating an example of a configuration of a microscope system 1 according to the first embodiment.
  • the illustrated microscope system 1 includes a microscope 10 that is a microscope main body, a confocal scanner 20 attached to the microscope 10, a processing device 30 that controls the confocal scanner 20, and a monitor 40 attached to the processing device 30. ing.
  • the microscope 10 includes a transmission illumination 11, a sample stage 12, an objective lens 13, a bending mirror 14, and an imaging lens 15.
  • the transmitted illumination 11 irradiates the sample stage 12 with light for observation.
  • the transmitted illumination 11 is irradiated from a line light source 22 included in the confocal scanner 20. Since line-shaped light is used, it is not necessary.
  • the microscope 10 also shows a transmission illumination 11 as an example of a general-purpose microscope.
  • a sample to be observed with the microscope 10 is placed on the sample stage 12.
  • the objective lens 13 enlarges the image of the observation target installed on the sample stage 12.
  • the bending mirror 14 guides the parallel light of the observation target image magnified by the objective lens 13 to the imaging lens 15.
  • the parallel light guided from the objective lens 13 through the bending mirror 14 is imaged by the imaging lens 15.
  • the image of the observation object imaged by the imaging lens 15 is also referred to as an observation image.
  • the eyepiece lens is not mounted after the imaging lens 15.
  • the confocal scanner 20 includes a linear light source 22 that emits linear light, a dichroic mirror 21 that guides the light to the imaging lens 15 of the microscope 10, and light from the microscope 10 via the dichroic mirror 21 for each line. And a line-shaped detector 23 for detecting (imaging).
  • the dichroic mirror 21 is disposed between the imaging lens 15 and the image plane of the imaging lens 15.
  • the confocal scanner 20 scans the observation target in a direction perpendicular to the line direction (longitudinal direction of the line-shaped light), thereby sequentially detecting (imaging) the light of the observation image for each line. Then, a captured image of the entire observation image is constructed.
  • symbol K shows in this figure has shown the optical path (optical axis), and is the same also in another figure.
  • FIG. 2 is a detailed view of the confocal scanner 20 as seen from the direction of the arrow 100 in FIG.
  • the confocal scanner 20 includes a dichroic mirror 21, an excitation filter 24, and a fluorescence filter 25, which are arranged on the optical path.
  • the excitation filter 24 transmits only light having a specific wavelength.
  • the dichroic mirror 21 is arranged as a branching optical system, and a dichroic mirror 21 is selected that reflects the wavelength of light (excitation light) transmitted through the excitation filter 24 and transmits fluorescence having a longer wavelength than the excitation light. Has been.
  • the dichroic mirror 21 reflects the excitation light in the direction of the imaging lens 15 of the microscope 10.
  • the fluorescent filter 25 transmits only the fluorescent light out of the light transmitted through the dichroic mirror 21.
  • the fluorescence transmitted through the fluorescence filter 25 reaches the line detector 23.
  • the line-shaped light source 22 and the line-shaped detector 23 are arranged at positions corresponding to each other in the image forming plane of the image forming lens 15.
  • the line-shaped light source 22 and the line-shaped detector 23 are branched by the dichroic mirror 21 and are directly disposed on the imaging surface of the imaging lens 15.
  • the confocal scanner 20 includes a drive mechanism 26 (moving mechanism) that can translate the line-shaped light source 22 and the line-shaped detector 23 integrally in the imaging plane. That is, the line-shaped light source 22 and the line-shaped detector 23 can translate on the image plane of the microscope 10 (image forming lens 15) while maintaining a corresponding positional relationship.
  • FIGS. 2, 3A, and 3B are diagrams showing a detailed example of the drive mechanism 26.
  • FIG. FIG. 3A is a diagram illustrating an example of the drive mechanism 26 viewed from the direction of the arrow 101 in FIG.
  • FIG. 3B is a diagram showing a part of the drive mechanism 26 as seen from the direction of the arrow 102 in FIG. 3A.
  • the drive mechanism 26 includes a linear guide 261, a stage portion 262, a connecting member 263, a fixed block 264, a stepping motor 265, and a ball screw 266.
  • the line light source 22 and the line detector 23 are a plane determined by the line direction (longitudinal direction) of the line light source 22 and the optical axis direction of the light emitted from the line light source 22, and the line of the line detector 23.
  • the connecting member 263 has a positional relationship in which the plane determined from the direction (longitudinal direction) and the optical axis direction of the light incident on the line detector 23 is the same plane, and the optical axes are orthogonal to each other. It is fixed to.
  • the line-shaped light source 22 and the line-shaped detector 23 are a plane determined from the line direction (longitudinal direction) of the line-shaped light source 22 and the optical axis direction of the light emitted from the line-shaped light source 22, and the line-shaped detector 23.
  • the plane determined by the line direction (longitudinal direction) and the optical axis direction of the light incident on the line detector 23 may be fixed to the connecting member 263 so that the planes are parallel to each other.
  • the connecting member 263 is fixed to the stage portion 262 of the linear guide 261, and can be translated in the direction of the arrow 103 (scanning direction) in FIG. 3A.
  • a fixed block 264 is connected to the linear guide 261, and a stepping motor 265 is fixed to the fixed block 264. Further, a ball screw 266 is rotatably supported on the fixed block 264 by a bearing (not shown). The ball screw 266 is connected to the stepping motor 265 by a coupling (not shown). A ball nut (not shown) is disposed inside the stage portion 262 and is screwed with the ball screw 266.
  • the direction orthogonal to the line direction (longitudinal direction) of each of the line light source 22 and the line detector 23 is orthogonal to the plane defined by the line light source 22 and the line detector 23 described above. It corresponds to the direction to do.
  • the line light source 22, the line detector 23, the stepping motor 265, and the like are connected to the processing device 30 so as to be controllable by the processing device 30.
  • FIG. 4A and 4B are diagrams showing detailed examples of the line-shaped light source 22.
  • FIG. 4A is a side view of the line light source 22
  • FIG. 4B is a front view of the line light source 22 as viewed from the direction of the arrow 104 in FIG. 4A.
  • the line-shaped light source 22 includes LED (Light Emitting Diode) elements 221 arranged in a line, a light guide plate 222 disposed in front of the LED elements 221, and a slit 223 disposed in front of the light guide plate 222. .
  • the light emitted from the LED element 221 is guided to the slit 223 by the light guide plate 222 and emitted as line-shaped light from the slit-shaped emission region 22M opened in the slit 223.
  • FIG. 5A and 5B are diagrams showing a detailed example of the line detector 23.
  • FIG. 5A is a side view of the line detector 23
  • FIG. 5B is a front view of the line detector 23 viewed from the direction of the arrow 105 in FIG. 5A.
  • CMOS Complementary MOS
  • one row of CMOS elements is arranged in the vertical direction in the detection region 23M of the line detector 23, and detects (images) light in the line region.
  • the microscope system 1 only the necessary wavelength of the light emitted from the line light source 22 of the confocal scanner 20 is transmitted by the excitation filter 24, and this transmitted light is reflected by the dichroic mirror 21 and travels to the microscope 10.
  • This light is converted into parallel light by the imaging lens 15 inside the microscope 10, converted into a right angle (in the direction of the objective lens 13) by the bending mirror 14, and is not shown on the sample stage 12 by the objective lens 13.
  • a focused image is projected onto the sample to be observed.
  • the illumination light from the confocal scanner 20 has a line shape
  • the light projected on the sample to be observed also has a line shape.
  • the fluorescence emitted from the sample returns to the opposite direction in the same optical path, enters the objective lens 13, is turned by the bending mirror 14, and is imaged by the imaging lens 15.
  • the fluorescence from this sample passes through the dichroic mirror 21, and only the necessary wavelength passes through the fluorescent filter 25 and is imaged and projected onto the detection region 23 ⁇ / b> M of the line detector 23.
  • the line-shaped light source 22 and the line-shaped detector 23 are arranged in a corresponding positional relationship in the imaging plane that is in a conjugate positional relationship with respect to the focal plane of the microscope. For this reason, fluorescence other than the vicinity of the line-shaped illumination light projected on the sample is hardly detected by the line-shaped detector 23.
  • the processing device 30 is a computer device used by a user, and applies a PC (Personal Computer), a tablet PC, a mobile phone such as a smartphone or a feature phone, a personal digital assistant (PDA: Personal Digital Assistant), or the like. it can.
  • PC Personal Computer
  • PDA Personal Digital Assistant
  • the processing device 30 controls the line light source 22, the line detector 23, and the stepping motor 265 of the confocal scanner 20 so that the line light source 22 and the line detector 23 are integrated in the scanning direction.
  • the image to be observed is detected (imaged) for each line while being translated.
  • the processing device 30 constructs a captured image of the observation image based on the image detected (captured) for each line.
  • FIG. 6 is a flowchart illustrating an example of line scanning processing according to the first embodiment.
  • Step S100 First, the processing device 30 sets the positions of the line light source 22 and the line detector 23 of the confocal scanner 20 to the initial state, and proceeds to the process of step S102.
  • the initial state is a state in which, for example, the detection target is the start position (one end of the scanning region) when line scanning is performed on the sample to be observed.
  • the light emitted from the line light source 22 of the confocal scanner 20 passes through the excitation filter 24, is reflected by the dichroic mirror 21, and travels toward the microscope 10.
  • Light incident on the microscope 10 from the dichroic mirror 21 is focused and projected onto a sample to be observed on the sample stage 12 via the imaging lens 15, the bending mirror 14, and the objective lens 13 inside the microscope 10.
  • the fluorescence emitted from the sample returns to the opposite direction in the same optical path, and travels toward the confocal scanner 20 via the objective lens 13, the bending mirror 14, and the imaging lens 15.
  • the light transmitted through the imaging lens 15 is imaged and projected onto the detection region 23M of the line detector 23 via the dichroic mirror 21 and the fluorescence filter 25.
  • Step S102 The processing device 30 transmits the transmission image (line-shaped observation image, also referred to as “line image”) that has passed through the dichroic mirror 21 and the fluorescence filter 25 to the line-shaped detector 23 of the confocal scanner 20. To image. Then, the process proceeds to step S104.
  • line image line-shaped observation image, also referred to as “line image”
  • Step S104 The processing device 30 drives the stepping motor 265 to translate the line light source 22 and the line detector 23 by one line in a direction (scanning direction) orthogonal to the respective line direction (longitudinal direction). Move.
  • One line is a movement amount set in advance based on the line width (width in the short direction) of the emission region 22M of the line light source 22 and the detection region 23M of the line detector 23. Thereby, the line-shaped light projected on the sample to be observed moves by one line in the scanning direction, and the line image detected by the line-shaped detector 23 also moves by one line in the scanning direction. Then, the process proceeds to step S106.
  • Step S106 The processing device 30 determines whether or not the scanning line is the final line. When determining that the line is not the final line (NO), the processing device 30 returns to the process of step S102, causes the line detector 23 of the confocal scanner 20 to capture a line image, and further performs a line light source by the process of step S104. 22 and the line-shaped detector 23 are further moved by one line in the scanning direction. On the other hand, if the processing device 30 determines in step S106 that it is the last line (YES), it proceeds to the processing of step S108.
  • Step S108 The processing device 30 causes the line detector 23 of the confocal scanner 20 to capture a line image (line image of the final line) that has passed through the dichroic mirror 21 and the fluorescent filter 25. Then, the process proceeds to step S110.
  • Step S110 The trajectory of the line-shaped light that has been scanned up to the final line is a plane orthogonal to the optical axis.
  • the processing device 30 constructs an image of an observation image corresponding to the scanned surface from the position information for each line of the stepping motor 265 that is drive-controlled and the line image for each line captured by the line detector 23. To do. This image becomes a confocal microscope image containing almost no fluorescence from the vicinity of the focal plane. Further, the processing device 30 displays the constructed image of the observation image on the monitor 40 and saves it in a storage device (not shown).
  • the storage device may be built in the processing device 30 or may be connected to the outside.
  • the sample stage 12 may be appropriately driven to select a desired location in the sample to be observed, or to focus the sample or position the observation surface in the depth direction.
  • the transmission illumination 11 may be illuminated as necessary, and the line detector 23 may be moved in translation to capture a line image.
  • the processing device 30 may acquire the line image of each line from the confocal scanner 20 every time the line is scanned, or collectively acquire from the confocal scanner 20 after the scanning to the last line is completed. May be.
  • processing device 30 may stop the line light source 22 and the line detector 23 for each line during the line scanning process, and then capture each line image, or the line light source 22 and the line detection.
  • the line image of each line may be taken while moving the device 23 without stopping for each line.
  • the one-color LED elements 221 may be arranged in a line or a plurality of color LED elements 221 may be arranged regularly.
  • the dichroic mirror 21, the excitation filter 24, and the fluorescence filter 25 may be selected corresponding to a plurality of colors. In this case, it is preferable to sequentially capture line images by turning on one color of the LED element 221.
  • the line light source 22 includes an LED element 221, a light guide plate 222 disposed in front of the LED element 221, and a slit 223 disposed in front of the light guide plate 222.
  • an LED element 221 a light guide plate 222 disposed in front of the LED element 221, and a slit 223 disposed in front of the light guide plate 222.
  • another optical system may be used instead of the light guide plate 222, and at least an LED element and a slit may be included without using an optical system such as a light guide plate.
  • CMOS elements are arranged in the detection region 23M as the line detector 23.
  • the present invention is not limited to this.
  • a plurality of rows of CMOS elements may be arranged in the detection region 23M, or a line-shaped CCD (Charge Coupled Device) may be arranged.
  • a CMOS element or CCD capable of capturing a two-dimensional image is arranged in the line detector 23 so that the detection result (imaging result) of the CMOS element or CCD in the column corresponding to the scanning position of the line-shaped illumination light is read. It may be.
  • the excitation light is reflected by the dichroic mirror 21 and the fluorescence is transmitted.
  • the present invention is not limited to this.
  • the arrangement of the line light source 22 and the excitation filter 24 and the line detector 23 and the fluorescence filter 25 may be switched using a dichroic mirror that transmits the excitation light and reflects the fluorescence.
  • the confocal scanner 20 attached to the microscope 10 has a line-like light source 22 that emits line-shaped light and a line-like shape that detects incident light for each line.
  • a line-shaped detector 23 having a CMOS element (detection unit) disposed, and a drive mechanism 26 (moving mechanism) that translates the line-shaped light source 22 and the line-shaped detector 23 relative to the microscope 10 are provided.
  • the line-shaped light source 22 and the line-shaped detector 23 are arranged in a positional relationship corresponding to each other in the imaging plane at a position conjugate to the focal plane of the microscope 10.
  • the confocal scanner 20 translates the line-shaped light source 22 and the line-shaped detector 23 in the imaging plane of the microscope 10 while maintaining the corresponding positional relationship.
  • a confocal microscope image can be taken with a general-purpose microscope such as the microscope 10. Therefore, according to the present embodiment, a confocal microscope image can be obtained with a simple configuration, and the introduction cost can be reduced. Further, it is possible to make the optical system smaller than using a scan mirror as described in Japanese Patent No. 4905356.
  • the line light source 22 and the line detector 23 include a plane determined from the line direction (longitudinal direction) of the line light source 22 and the optical axis direction of light emitted from the line light source 22, and the line detector 23.
  • the plane determined by the line direction (longitudinal direction) and the optical axis direction of the light incident on the line detector 23 are arranged in a positional relationship such that they are the same plane or planes parallel to each other.
  • the drive mechanism 26 (moving mechanism) can translate the line light source 22 and the line detector 23 in a direction (scanning direction) orthogonal to the same plane or the plane parallel to each other.
  • the confocal scanner 20 can link the line-shaped light source 22 and the line-shaped detector 23 with one drive mechanism 26, and can have a simple configuration.
  • the line direction of the line light source 22 and the line detector 23 can be parallel to the scanning direction.
  • the line light source is used for scanning. Since the moving direction of 22 and the moving direction of the line detector 23 are different, the same drive mechanism cannot be used.
  • the drive mechanism 26 (moving mechanism) can translate the line light source 22 and the line detector 23 integrally.
  • the confocal scanner 20 can link the line light source 22 and the line detector 23 with the same drive mechanism, it is not necessary to take synchronization required when a plurality of drive mechanisms are used. It is convenient. Further, since the positional relationship between the line-shaped light source 22 and the line-shaped detector 23 is fixed by the integration, the accuracy can be ensured more easily than when driven separately.
  • the line light source 22 and the line detector 23 are integrally translated.
  • the line light source 22 and the line detector 23 are separately translated, May be moved in synchronization.
  • the line light source 22 and the line detector 23 are a plane determined from the line direction (longitudinal direction) of the line light source 22 and the optical axis direction of the light emitted from the line light source 22, and the line detection.
  • the positional relationship in which the plane determined from the line direction (longitudinal direction) of the detector 23 and the optical axis direction of the light incident on the line detector 23 is the same plane, and the center optical axis of the line light source 22 and the line detection
  • the central optical axis of the device 23 is arranged so as to intersect with the dichroic mirror 21 (branching optical system).
  • the linear light source 22 and the linear detector 23 By arranging the linear light source 22 and the linear detector 23 in this way, the change in the optical axis direction is minimized, so that the optical system can be downsized and configured with a small space.
  • the present embodiment is a modification of the linear light source 22 (see FIGS. 4A and 4B) described in the first embodiment.
  • the example of the line light source 22 in which the LED elements 221 are arranged in a line is shown, but the line light source may be configured by using a surface-emitting light source.
  • 7A and 7B are diagrams illustrating a detailed example of the line light source 22A according to the second embodiment.
  • 7A is a side view of the linear light source 22A
  • FIG. 7B is a front view of the linear light source 22A as viewed from the direction of the arrow 106 in FIG. 7A.
  • the illustrated linear light source 22A includes a surface emitting LED 221A as a light source and a slit 223A arranged to cover the light emitting surface of the surface emitting LED 221A.
  • Light emitted from the surface emitting LED 221A is emitted in a line shape from a slit (exit region 22M) opened in the slit 223A.
  • a slit exit region 22M
  • the linear light source 22A when the linear light source 22A is translated in the direction of the arrow 107 (scanning direction), the entire linear light source 22A is not moved, but only the slit 223A is moved in the direction of the arrow 107 (scanning direction). ) May be translated.
  • a confocal microscope image can be obtained with a simple configuration as in the first embodiment, and the optical system can be reduced in size and cost of introduction. Can be suppressed.
  • FIGS. 5A and 5B are diagrams illustrating a detailed example of the line detector 23B according to the third embodiment.
  • 8A is a side view of the line detector 23B
  • FIG. 8B is a front view of the line detector 23B viewed from the direction of the arrow 108 in FIG. 8A.
  • the light detected by the line detector 23B passes through the slit 231B and forms an image on the line detector 23B via the relay lens 232B.
  • the line detector 23B includes a line-shaped CMOS element, a line-shaped CCD, or a CMOS element or CCD that can capture a two-dimensional image, and a CMOS element that can capture a two-dimensional image.
  • the line image formed through the slit 231B and the relay lens 232B is detected (captured).
  • the line detector 23B is configured as described above, a confocal microscope image can be obtained with a simple configuration as in the first embodiment, and the downsizing and introduction cost of the optical system can be suppressed.
  • the width of the slit 231B can be easily changed.
  • FIG. 9A is a diagram illustrating an example of the drive mechanism 26C when viewed from the same direction as in FIG. 3A.
  • FIG. 9B is a diagram showing a part of the drive mechanism 26C viewed from the direction of the arrow 109 in FIG. 9A.
  • the drive mechanism 26C is different from the first embodiment in that two line light sources 22a and 22b and two line detectors 23a and 23b are provided on the connecting member 263C.
  • the line-shaped light source 22a and the line-shaped detector 23a are paired and arranged so as to correspond to each other in the imaging plane of the microscope 10 respectively. Further, the line-shaped light source 22b and the line-shaped detector 23b are paired and arranged so as to correspond to each other in the imaging plane of the microscope 10 respectively.
  • the connecting member 263C is fixed to the stage portion 262 of the linear guide 261, and can be translated in the direction of the arrow 110 (scanning direction) of 9A. That is, the two line light sources 22a and 22b and the two line detectors 23a and 23b are arranged so as to be aligned in the translation direction. Specifically, among the two line light sources 22a and 22b and the two line detectors 23a and 23b, each of the paired line light source and the line detector is a line of the respective line light source.
  • the two linear light sources 22a and 22b are mounted with LED elements having different wavelengths, respectively, and excitation filters (not shown) corresponding to the respective wavelengths are provided on the entire surface.
  • the corresponding line detectors 23a and 23b are provided with fluorescent filters (not shown) corresponding to the respective fluorescence wavelengths.
  • each of the line-shaped light source and the line-shaped detector paired out of the two line-shaped light sources 22a and 22b and the two line-shaped detectors 23a and 23b is the line direction (longitudinal direction) of each line-shaped light source.
  • the optical axis direction of the light emitted from the line light source, and the plane determined from the line direction (longitudinal direction) of each line detector and the optical axis direction of the light incident on the line detector. May be arranged in a positional relationship that forms planes parallel to each other.
  • FIG. 10 is a diagram illustrating a configuration example of a confocal scanner 20D according to the fifth embodiment.
  • FIG. 10 shows a modified example of the confocal scanner 20 shown in FIG. 2, and corresponding portions are denoted by the same reference numerals and description thereof is omitted.
  • the two linear light sources 22a and 22b are equipped with LED elements having different wavelengths, and excitation filters 24a and 24b corresponding to the respective wavelengths are arranged in front of the LED elements. Further, one light is transmitted through the excitation dichroic mirror 21b, and the other light is reflected by the excitation dichroic mirror 21b. As a result, the two lights overlap each other.
  • fluorescent filters 25a and 25b are arranged in front of the two line-shaped detectors 23a and 23b according to the wavelength assigned to each detector. Further, one light is transmitted through the fluorescent dichroic mirror 21c, and the other light is reflected by the fluorescent dichroic mirror 21c. As a result, the two lights overlap each other.
  • the dichroic mirror 21 a reflects the wavelengths of the excitation light that is transmitted through the excitation dichroic mirror 21 b and the excitation light that is reflected, and transmits the fluorescence from the imaging lens 15 of the microscope 10.
  • the line-shaped light source 22a and the line-shaped detector 23a are arranged so as to correspond to each other in the imaging plane of the microscope 10 respectively. Further, the line-shaped light source 22b and the line-shaped detector 23b are arranged so as to have a corresponding positional relationship in the imaging plane of the microscope 10, respectively.
  • each of the two linear light sources and the two detectors has a plane determined from the line direction (longitudinal direction) of each linear light source and the optical axis direction of the light emitted from the linear light source, It is fixed to the connecting member 263D so that the plane defined by the line direction (longitudinal direction) of the line detector and the optical axis direction of the light incident on the line detector is in the same plane. .
  • the line light source 22a and the line detector 23a, and the line light source 22b and the line detector 23b are not limited to this, and may be arranged so as to be shifted in the vertical direction (scanning direction) with respect to the same plane. Good. In this case, since the light of a mutual wavelength does not mix, measurement with a higher sensitivity is possible.
  • the line-shaped light source 22a and the line-shaped detector 23a are in a corresponding positional relationship in the imaging plane of the microscope 10, and the line-shaped light source 22b and the line-shaped detector 23b are respectively in the imaging plane of the microscope 10. Any arrangement is possible as long as they correspond to each other.
  • each of the paired line-shaped light source and line-shaped detector is the line direction (longitudinal direction) of each line-shaped light source.
  • a plane determined from the optical axis direction of the light emitted from the linear light source, and a plane determined from the line direction (longitudinal direction) of each linear detector and the optical axis direction of the light incident on the linear detector may be arranged in a positional relationship in which the planes are parallel to each other.
  • the line detector 23 is expensive, it may be difficult to provide a plurality of detectors. Further, when the line-shaped light source 22 is configured with a plurality of colors, it may be preferable to switch the plurality of light sources. A configuration example in which one line detector 23 and a plurality of line light sources 22 correspond will be described with reference to FIGS. 11A and 11B.
  • FIG. 11A and FIG. 11B are diagrams illustrating an example of a drive mechanism 26E according to the sixth embodiment.
  • FIG. 11A is a diagram illustrating an example of the drive mechanism 26E when viewed from the same direction as FIG. 9A.
  • FIG. 11B is a diagram showing a part of the drive mechanism 26E viewed from the direction of the arrow 109 in FIG. 11A.
  • one line detector 23a is arranged on the connecting member 263E of the drive mechanism 26E.
  • the connecting member 263E is provided with a stage portion 301 via a linear guide 300, and four line light sources 22a, 22b, 22c, and 22d are arranged on the stage portion 301.
  • the stage unit 301 can be moved in the direction of an arrow 302 (direction orthogonal to the longitudinal direction of the line-shaped detector 23a and the line-shaped light sources 22a, 22b, 22c, and 22d) by an actuator (not shown).
  • the four linear light sources 22a, 22b, 22c, and 22d move with respect to one linear detector 23a. That is, the four line-shaped light sources 22a, 22b, 22c, and 22d and the line-shaped detector 23a move relatively, and selectively one of the four line-shaped light sources 22a, 22b, 22c, and 22d
  • the line-shaped detectors 23a are in a positional relationship corresponding to each other in the imaging plane of the microscope 10.
  • the line light source 22c is selected, and the line light source 22c and the line detector 23a are in a corresponding positional relationship.
  • the processing device 30 drives the stepping motor 265
  • the entire image of the optical system attached to the connecting member 263E is translated in the scanning direction, and the image to be observed is detected line by line ( Imaging).
  • the stage unit 301 is moved as needed to move the desired line light source to the corresponding position.
  • one line detector 23 for example, 23a
  • a plurality of line light sources 22 for example, 22a, 22b, 22c, and 22d
  • a state detector 23 and one line light source 22 may be provided.
  • the plurality of line-shaped detectors 23 are selectively moved by moving the plurality of line-shaped detectors 23.
  • one line-shaped detector 23 and one line-shaped light source 22 can have a corresponding positional relationship in the imaging plane of the microscope 10.
  • a plurality of the line light sources 22 and the line detectors 23 are provided, and the line light sources are selectively moved by relatively moving the line light sources 22 and the line detectors 23.
  • 22 and the line-shaped detector 23 are arranged so as to correspond to each other.
  • the plurality of line light sources 22 is moved by relatively moving the one line detector 23 and the plurality of line light sources 22.
  • One line-shaped light source 22 and one line-shaped detector 23 are arranged so as to correspond to each other selectively. Accordingly, it is possible to capture line images of a plurality of wavelengths by selectively using the line-shaped light sources 22 having a plurality of wavelengths while suppressing the cost as compared with providing a plurality of line-shaped detectors 23.
  • This embodiment is a configuration example in which one line detector 23 corresponds to a plurality of line light sources 22 as in the sixth embodiment, but instead of moving the plurality of line light sources 22, The difference is that one line detector 23 is moved.
  • FIG. 12A and 12B are diagrams illustrating an example of a drive mechanism 26F according to the seventh embodiment.
  • FIG. 12A is a diagram illustrating an example of the drive mechanism 26F when viewed from the same direction as in FIG. 11A.
  • FIG. 12B is a diagram showing a part of the drive mechanism 26F viewed from the direction of the arrow 109 in FIG. 12A.
  • four line light sources 22a, 22b, 22c, and 22d are arranged on the connecting member 263F of the drive mechanism 26F.
  • a stage portion 304 is provided on the connecting member 263F via a linear guide 303, and one line detector 23a is arranged on the stage portion 304.
  • the stage unit 304 is movable in the direction of an arrow 305 (a direction orthogonal to the longitudinal direction of the line detector 23a and the line light sources 22a, 22b, 22c, and 22d) by an actuator (not shown). By moving the stage unit 304 in the direction of the arrow 305, one line-shaped detector 23a moves with respect to the four line-shaped light sources 22a, 22b, 22c, and 22d.
  • the four line-shaped light sources 22a, 22b, 22c, and 22d and the line-shaped detector 23a move relatively, and selectively one of the four line-shaped light sources 22a, 22b, 22c, and 22d
  • the line-shaped detectors 23a are in a positional relationship corresponding to each other in the imaging plane of the microscope 10.
  • the line light source 22c is selected, and the line light source 22c and the line detector 23a are in a corresponding positional relationship.
  • the processing device 30 drives the stepping motor 265
  • the entire optical system provided in the connecting member 263E is integrally translated in the scanning direction, and the image to be observed is detected line by line ( Imaging).
  • the stage unit 301 is moved as needed to move the desired line light source to the corresponding position.
  • one line detector 23 for example, 23a
  • a plurality of line light sources 22 for example, 22a, 22b, 22c, and 22d
  • a state detector 23 and one line light source 22 may be provided.
  • moving one line detector 23 for example, 23a
  • one of the plurality of line detectors 23 is selectively moved.
  • the line detector 23 and one line light source 22 can be in a positional relationship corresponding to each other in the imaging plane of the microscope 10.
  • any one of the line-shaped light source 22 and the line-shaped detector 23 is provided, and the line-shaped light source 22 and the line-shaped detector 23 are relative to each other.
  • the line-shaped light source 22 and the line-shaped detector 23 are arranged so that they can be selectively supported by the movement.
  • the line-shaped light sources 22 having a plurality of wavelengths can be reduced while suppressing the cost compared to the case where the plurality of line-shaped detectors 23 are provided.
  • By selectively using it it is possible to capture line images of a plurality of wavelengths.
  • FIG. 13 is a diagram illustrating an example of two drive mechanisms according to the eighth embodiment.
  • the drive mechanism 26G includes a linear guide 261, a stage unit 310, a fixed block 264, a stepping motor 265, and a ball screw 266.
  • the connecting member 263E shown in FIGS. 11A and 11B is not provided, and four line light sources 22a, 22b, 22c, and 22d are arranged on the stage portion 310.
  • the drive mechanism 26H basically includes a linear guide 306, a stage portion 311, a fixed block 307, a stepping motor 308, and a ball screw 308, similarly to the drive mechanism 26G.
  • the line-shaped detector 23 a is arranged on the stage portion 311 via the mounting base 312 so as to face the same direction as the example shown in FIGS. 11A and 11B.
  • the line light source 22c is selected, and the line light source 22c and the line detector 23a are in a corresponding positional relationship.
  • the processing device 30 drives the stepping motor 265 or the stepping motor 308, so that the plurality of line light sources 22 and the line detectors are driven.
  • the desired line-shaped light source 22 and the line-shaped detector 23a can be associated with each other by relatively moving the line 23a.
  • the processing device 30 drives the two stepping motors 265 and 308 synchronously to maintain the positional relationship between the line light source 22 and the line detector 23a. As it is, it is translated in the scanning direction.
  • one line detector 23 for example, 23a
  • a plurality of line light sources 22 for example, 22a, 22b, 22c, and 22d
  • a state detector 23 and one line light source 22 may be provided.
  • the linear light source 22 including the optical system and the slit is not moved as described in the sixth embodiment, but the light source is selectively moved by moving the light source with respect to the optical system and the slit.
  • FIG. 14A and 14B are diagrams illustrating an example of a drive mechanism 26I according to the ninth embodiment.
  • FIG. 14A is a diagram illustrating an example of the drive mechanism 26I when viewed from the same direction as FIG. 11A.
  • FIG. 14B is a diagram showing a part of the drive mechanism 26I as seen from the direction of the arrow 109 in FIG. 14A.
  • one line detector 23a is arranged on the connecting member 263I of the drive mechanism 26I.
  • the connecting member 263I is provided with a stage portion 316 via a linear guide 315, and four light sources 22a ′, 22b ′, 22c ′, and 22d ′ are arranged on the stage portion 316.
  • the stage unit 316 is movable in the direction of the arrow 317 by an actuator (not shown).
  • the illumination optical system 314 guides light emitted from one selected light source among the four light sources 22 a ′, 22 b ′, 22 c ′, and 22 d ′ to the slit 313.
  • the light source 22c ′ is selected, and light emitted from the light source 22c ′ is emitted as line-shaped light from the slit-shaped opening of the slit 313 via the illumination optical system 314.
  • the light is incident on the plane determined from the slit direction (longitudinal direction) of the slit 313 and the optical axis direction of the light emitted from the slit 313, the line direction (longitudinal direction) of the line detector 23a, and the line detector 23a.
  • the slits 313 are fixed to the connecting member 263I so that the plane relationship determined from the optical axis direction of the light is the same plane, and the optical axes are orthogonal to each other.
  • the slit 313 may be fixed to the connecting member 263I such that the plane determined from the optical axis direction of the light is a plane relationship parallel to each other. That is, one selected light source among the four light sources 22 a ′, 22 b ′, 22 c ′, and 22 d ′ and the line-shaped detector 23 a each have a corresponding positional relationship in the imaging plane of the microscope 10. Become.
  • the light source 22c ′ is selected, and the light source 22c ′ and the line-shaped detector 23a have a corresponding positional relationship, but excitation by any one of the light sources 22a ′, 22b ′, and 22d ′ is necessary.
  • a desired light source is moved to a corresponding position by moving the stage unit 316 at any time.
  • the processing device 30 drives the stepping motor 265, so that the entire optical system provided in the connecting member 263I is translated and moved in the scanning direction integrally, and the observation target An image can be detected (captured) line by line.
  • the light source when using a plurality of light sources, the light source can be selectively used by moving only the light source element without moving the optical system or the slit. It can be constructed at a lower cost than when a plurality of line-shaped light sources are moved.
  • FIG. 15 is a diagram illustrating an example of a drive mechanism 26J according to the tenth embodiment. This figure corresponds to FIG. 14B and shows a part of the connecting member 263J of the drive mechanism 26J.
  • the connecting member 263J shown is different from the example shown in FIG. 14B in that an excitation filter unit 319 in which four excitation filters are connected and a stage unit 316 are connected via a filter connecting member 318.
  • Each of the four excitation filters is an excitation filter corresponding to each of the four light sources 22a ′, 22b ′, 22c ′, and 22d ′, and is an excitation filter that passes only a desired excitation wavelength from each light source. .
  • Each of the four excitation filters and each of the four light sources 22 a ′, 22 b ′, 22 c ′, and 22 d ′ are arranged in a positional relationship that corresponds (opposes) with the slit 313 and the illumination optical system 314 interposed therebetween.
  • the stage unit 316 moves, it moves relative to the slit 313 integrally while maintaining the positional relationship between each light source and each excitation filter.
  • an excitation filter that passes only a desired excitation wavelength is selected corresponding to the light source selected by moving the stage unit 316.
  • the confocal scanner 20 (20 ⁇ / b> D) in each embodiment described above may be configured as a confocal microscope integrated with the microscope 10.
  • the confocal microscope includes, for example, a sample stage 12 on which an observation target is set, an objective lens 13, an imaging lens 15 that forms an image of the observation target incident through the objective lens 13, and an imaging lens.
  • the line-shaped light source 22 that emits line-shaped light that illuminates the observation target via the objective lens 13 and the line-shaped light source 22 are arranged in a corresponding positional relationship in the imaging plane of the microscope 10.
  • a line-shaped detector 23 having a line-shaped CMOS element (detection unit) for detecting, on a line-by-line basis, the light emitted by the observation object through the objective lens 13 and the imaging lens 15; At least a drive mechanism 26 that translates the linear light source 22 and the line detector 23 relative to the microscope.
  • the processing apparatus 30 in embodiment mentioned above may make it implement
  • a program for realizing the above-described function is recorded on a computer-readable recording medium, and the program recorded on the recording medium is read into a computer system and executed to realize the above-described function.
  • the “computer system” is a computer system built in the processing device 30 and includes an OS and hardware such as peripheral devices.
  • the “computer system” includes a homepage providing environment (or display environment) if a WWW system is used.
  • the “computer-readable recording medium” refers to a storage device such as a flexible medium, a magneto-optical disk, a portable medium such as a ROM or a CD-ROM, and a hard disk incorporated in a computer system.
  • the “computer-readable recording medium” is a medium that dynamically holds a program for a short time, such as a communication line when transmitting a program via a network such as the Internet or a communication line such as a telephone line, In this case, a volatile memory inside a computer system that serves as a server or a client may be included that holds a program for a certain period of time.
  • the program may be a program for realizing a part of the functions described above, and may be a program capable of realizing the functions described above in combination with a program already recorded in a computer system.
  • a part or all of the processing device 30 in the above-described embodiment may be realized as an integrated circuit such as an LSI (Large Scale Integration).
  • LSI Large Scale Integration
  • Each functional block of the processing device 30 may be individually made into a processor, or a part or all of them may be integrated into a processor.
  • the method of circuit integration is not limited to LSI, and may be realized by a dedicated circuit or a general-purpose processor.
  • an integrated circuit based on the technology may be used.
  • unit is used to indicate a component, unit, hardware, or a portion of software programmed to perform a desired function.
  • Typical examples of hardware are devices and circuits, but are not limited to these.

Abstract

Selon la présente invention, un scanner confocal fixé à un microscope comprend : une source de lumière linéaire qui émet une lumière linéaire ; un détecteur linéaire ayant une unité de détection linéaire qui détecte une lumière incidente à chaque ligne ; et un dispositif de déplacement qui traduit la source de lumière linéaire et le détecteur linéaire par rapport au microscope. La source de lumière linéaire et le détecteur linéaire sont agencés au niveau de relations de position correspondantes dans le plan d'imagerie à des positions conjuguées par rapport au plan focal du microscope.
PCT/JP2018/012712 2017-04-13 2018-03-28 Unité confocal, système confocal et microscope confocal WO2018190125A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP18784330.5A EP3611550B1 (fr) 2017-04-13 2018-03-28 Unité confocal, système confocal et microscope confocal
CN201880021124.4A CN110462483B (zh) 2017-04-13 2018-03-28 共聚焦扫描仪、显微镜系统以及共聚焦显微镜
US16/604,292 US11506878B2 (en) 2017-04-13 2018-03-28 Confocal scanner, microscope system, and confocal microscope

Applications Claiming Priority (4)

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JP2017079986 2017-04-13
JP2017-079986 2017-04-13
JP2017249617A JP6677238B2 (ja) 2017-04-13 2017-12-26 共焦点スキャナ、及び共焦点顕微鏡
JP2017-249617 2017-12-26

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JPS495356B1 (fr) 1970-05-08 1974-02-06
JPH03172815A (ja) * 1989-12-01 1991-07-26 Fuji Photo Film Co Ltd 共焦点走査型顕微鏡
JPH04409A (ja) * 1989-09-22 1992-01-06 Fuji Photo Film Co Ltd 共焦点走査型顕微鏡
JP2660613B2 (ja) 1991-08-23 1997-10-08 富士写真フイルム株式会社 共焦点走査型顕微鏡
JP2001013445A (ja) 1999-06-28 2001-01-19 Yokogawa Electric Corp 共焦点光スキャナ
JP2003247817A (ja) 2002-02-27 2003-09-05 Takaoka Electric Mfg Co Ltd リニアスキャン型共焦点表面形状計測装置
JP2005292839A (ja) * 2004-04-05 2005-10-20 Fujifilm Electronic Imaging Ltd スリット共焦点顕微鏡およびその作動方法
WO2007055082A1 (fr) * 2005-11-11 2007-05-18 Nikon Corporation Microscope confocal

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JPS495356B1 (fr) 1970-05-08 1974-02-06
JPH04409A (ja) * 1989-09-22 1992-01-06 Fuji Photo Film Co Ltd 共焦点走査型顕微鏡
JPH03172815A (ja) * 1989-12-01 1991-07-26 Fuji Photo Film Co Ltd 共焦点走査型顕微鏡
JP2660613B2 (ja) 1991-08-23 1997-10-08 富士写真フイルム株式会社 共焦点走査型顕微鏡
JP2001013445A (ja) 1999-06-28 2001-01-19 Yokogawa Electric Corp 共焦点光スキャナ
JP2003247817A (ja) 2002-02-27 2003-09-05 Takaoka Electric Mfg Co Ltd リニアスキャン型共焦点表面形状計測装置
JP2005292839A (ja) * 2004-04-05 2005-10-20 Fujifilm Electronic Imaging Ltd スリット共焦点顕微鏡およびその作動方法
WO2007055082A1 (fr) * 2005-11-11 2007-05-18 Nikon Corporation Microscope confocal

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