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WO2018016010A1 - Unité spectroscopique et dispositif spectroscopique - Google Patents

Unité spectroscopique et dispositif spectroscopique Download PDF

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Publication number
WO2018016010A1
WO2018016010A1 PCT/JP2016/071185 JP2016071185W WO2018016010A1 WO 2018016010 A1 WO2018016010 A1 WO 2018016010A1 JP 2016071185 W JP2016071185 W JP 2016071185W WO 2018016010 A1 WO2018016010 A1 WO 2018016010A1
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WIPO (PCT)
Prior art keywords
unit
filter
signal
spectroscopic
pixel
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Ceased
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PCT/JP2016/071185
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English (en)
Japanese (ja)
Inventor
卓二 堀江
正法 三井
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Olympus Corp
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Olympus Corp
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Priority to PCT/JP2016/071185 priority Critical patent/WO2018016010A1/fr
Publication of WO2018016010A1 publication Critical patent/WO2018016010A1/fr
Anticipated expiration legal-status Critical
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/12—Generating the spectrum; Monochromators
    • G01J3/26—Generating the spectrum; Monochromators using multiple reflection, e.g. Fabry-Perot interferometer, variable interference filters
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01J—MEASUREMENT OF INTENSITY, VELOCITY, SPECTRAL CONTENT, POLARISATION, PHASE OR PULSE CHARACTERISTICS OF INFRARED, VISIBLE OR ULTRAVIOLET LIGHT; COLORIMETRY; RADIATION PYROMETRY
    • G01J3/00—Spectrometry; Spectrophotometry; Monochromators; Measuring colours
    • G01J3/46—Measurement of colour; Colour measuring devices, e.g. colorimeters
    • G01J3/50—Measurement of colour; Colour measuring devices, e.g. colorimeters using electric radiation detectors
    • G01J3/51—Measurement of colour; Colour measuring devices, e.g. colorimeters using electric radiation detectors using colour filters

Definitions

  • the present invention relates to a spectroscopic unit and a spectroscopic device that split incident light.
  • Patent Document 1 discloses a first filter (linear variable filter) in which a transmission wavelength of transmitted light continuously changes from a first wavelength to a second wavelength longer than the first wavelength along a predetermined direction, a light source, The intensity of the light from the light source is adjusted for each wavelength so that the intensity of the transmitted light that is disposed between the first filter and the first filter is substantially the same in the section from the first wavelength to the second wavelength.
  • a spectroscopic measurement device including a second filter (flattening filter) is described.
  • the present invention has been made in view of the above, and an object of the present invention is to provide a spectroscopic unit and a spectroscopic device capable of accurately measuring spectral sensitivity even for light having greatly different intensities according to wavelengths. To do.
  • the spectroscopic unit includes a flat plate-like first filter whose transmission center wavelength of light continuously changes along a preset direction, Is a flat plate-like second filter in which the transmission center wavelength continuously changes along a preset direction, and at least a part of the continuously changing transmission center wavelength is the transmission center wavelength of the first filter.
  • the first filter is arranged on the projection plane in which the direction of change of the transmission center wavelength is orthogonal to the thickness direction.
  • a second filter that intersects with the direction of change of the transmission center wavelength.
  • the spectroscopic unit according to the present invention is provided on a side closer to a position where light enters than the first and second filters along the thickness direction, and diffuses and uniformizes the incident light.
  • a diffusion optical system is further provided.
  • the spectroscopic unit according to the present invention is characterized in that, in the above-described invention, an intersection angle between the change direction of the transmission center wavelength of the first filter and the change direction of the transmission center wavelength of the second filter is variable.
  • the spectroscopic device includes the spectroscopic unit described above, an image pickup unit that generates an image pickup signal by picking up light transmitted through the spectroscopic unit, a pixel position and transmission characteristics with respect to the image pickup signal, And a calculation unit that performs a calculation based on the above correspondence.
  • the calculation unit includes an image analysis unit that extracts a signal of a predetermined pixel from the imaging signal, and a signal of each pixel extracted by the image analysis unit. And a spectrum calculation unit that calculates a spectrum of the imaging signal based on a transmission wavelength band corresponding to the position of each pixel.
  • the image analysis unit passes through the centers of the first and second filters on the projection plane, and the transmission wavelength bands of the first and second filters match.
  • a pixel signal corresponding to a point on a straight line passing through the position when the position is projected onto the projection plane is extracted.
  • the image analysis unit outputs the saturated signal value when the extracted pixel signal has a saturated signal value, and outputs the saturated signal.
  • a pixel located in the vicinity of the target pixel and corresponding on the projection plane is replaced with an unsaturated signal value output by a pixel not located on the straight line.
  • the image analysis unit performs correction to match the bandwidth of the unsaturated signal with the bandwidth of a signal output by a pixel corresponding to a point located on the straight line.
  • the spectrum calculation unit calculates the spectrum using the unsaturated signal value after correction.
  • the spectroscopic device is the spectroscopic device according to the above-described invention, wherein one of the first and second filters is passed through a center of the first and second filters and around a central axis orthogonal to the straight line.
  • a filter drive unit that rotationally drives the image by an angle; and a control unit that controls the filter drive unit, wherein the image analysis unit has a direction perpendicular to the straight line in pixels that form a transmission pattern by the imaging signal.
  • the maximum value of the signal value of the edge pixel located at the edge of the edge is calculated and compared with a reference value.When the maximum value is larger than the reference value as a result of the comparison by the image analysis unit, the control unit, One of the first and second filters is rotated by the filter driving unit by the predetermined angle.
  • the present invention it is possible to accurately measure the spectral sensitivity even for light having greatly different intensities depending on the wavelength.
  • FIG. 1 is a block diagram showing the configuration of the spectroscopic device according to Embodiment 1 of the present invention.
  • FIG. 2 is a diagram schematically illustrating the characteristics of the linear variable filter included in the spectroscopic device according to Embodiment 1 of the present invention.
  • FIG. 3 is a diagram schematically illustrating the light transmittance at typical positions of the linear variable filter provided in the spectroscopic device according to Embodiment 1 of the present invention.
  • FIG. 4 is a diagram schematically showing the configuration of the filter unit provided in the spectroscopic device according to Embodiment 1 of the present invention.
  • FIG. 5 is a diagram schematically showing a spectrum of light transmitted at the point P shown in FIG. FIG.
  • FIG. 6 is a diagram schematically showing a spectrum of light transmitted at the point Q shown in FIG.
  • FIG. 7 is a diagram schematically showing a spectrum of light transmitted at the point R shown in FIG.
  • FIG. 8 is a block diagram showing a configuration of a microscope system that is a spectroscopic device according to Embodiment 2 of the present invention.
  • FIG. 9 is a diagram showing a configuration of a diffusion optical system provided in the spectroscopic unit according to Embodiment 2 of the present invention.
  • FIG. 10 is a diagram for explaining the operation when fluorescence observation is performed using a microscope system that is a spectroscopic device according to Embodiment 2 of the present invention.
  • FIG. 11 is a diagram for explaining the outline of the spectrum calculated by the spectrum calculation unit when the stained specimen is observed in the bright field in the modification of the second embodiment of the present invention.
  • FIG. 12 is a block diagram showing a configuration of an imaging apparatus that is a spectroscopic apparatus according to Embodiment 3 of the present invention.
  • FIG. 13 is a block diagram showing a configuration of a spectroscopic device according to Embodiment 4 of the present invention.
  • FIG. 14 is a flowchart showing an outline of the intersection angle determination process performed by the spectroscopic device according to Embodiment 4 of the present invention.
  • FIG. 12 is a block diagram showing a configuration of an imaging apparatus that is a spectroscopic apparatus according to Embodiment 3 of the present invention.
  • FIG. 13 is a block diagram showing a configuration of a spectroscopic device according to Embodiment 4 of the present invention.
  • FIG. 14 is a flowchart showing an outline of the intersection angle determination process performed by the spectr
  • FIG. 15 is a diagram schematically illustrating an example of distribution of pixel signal values in a transmission pattern acquired when the spectroscopic device according to Embodiment 4 of the present invention has an intersection angle of two linear variable filters of 90 °. is there.
  • FIG. 16 is a diagram schematically illustrating a distribution example of pixel signal values in a transmission pattern acquired when the spectroscopic device according to Embodiment 4 of the present invention has an acute angle between two linear variable filters. is there.
  • FIG. 1 is a block diagram showing the configuration of the spectroscopic device according to Embodiment 1 of the present invention.
  • the spectroscopic device 1 shown in the figure is a device that detects the spectral characteristics of a light source.
  • the spectroscopic device 1 performs an operation on a spectroscopic unit 2 that is a spectroscopic unit, an image capturing unit 3 that captures an image of light imaged by the spectroscopic unit 2 and generates an image signal, and an imaging signal generated by the image capturing unit 3.
  • a storage unit 7 and a control unit 8 that controls the overall operation of the spectroscopic device 1 are provided.
  • the spectroscopic unit 2 includes a filter unit 21 and an imaging optical system 22.
  • the filter unit 21 includes flat linear variable filters (hereinafter referred to as LVF) 23 and 24 in which the transmission center wavelength of light continuously changes along a preset direction in a manner in which the thickness direction is aligned. It is lined up along the thickness direction.
  • the LVF 23 and the LVF 24 are projection planes orthogonal to the thickness direction, and the direction of change of the center wavelength (hereinafter referred to as the transmission center wavelength) that passes through each LVF intersects.
  • the LVF 23 and the LVF 24 have the same square main surface, and are arranged so that their centers coincide with each other along the thickness direction. In the first embodiment, it is assumed that the LVF 23 and the LVF 24 have the same light transmission characteristics.
  • the LVF 23 and the LVF 24 do not have to have the same characteristics of transmitting light, and it is sufficient that at least a part of each transmission wavelength band is common. Further, the LVF 23 and the VFE 24 may be arranged so as to be in contact with each other in the thickness direction, or may be arranged apart from each other. However, in the case where they are arranged apart from each other, no other component is provided between the LVF 23 and the LVF 24.
  • One of the LVF 23 and the LVF 24 is a first filter, and the other is a second filter.
  • FIG. 2 is a diagram schematically illustrating the characteristics of the LVF 23, and is a diagram illustrating a relationship between a position in the filter and a transmission center wavelength having the highest transmittance at the position.
  • the four vertical lines described in the LVF 23 are virtual lines connecting positions where the transmitted wavelength bands are the same. Therefore, the four vertical lines are orthogonal to the direction in which the transmission center wavelength changes, that is, the x-axis direction in FIG.
  • FIG. 3 is a diagram schematically showing the light transmittance at typical positions of the LVF 23.
  • the spectrum of transmitted light at each position of the LVF 23 has substantially uniform transmittance at the transmission center wavelength and substantially uniform bandwidth.
  • FIG. 4 is a diagram schematically illustrating the configuration of the filter unit 21, and is a diagram schematically illustrating the positional relationship between the LVF 23 and the LVF 24 when viewed on the projection plane orthogonal to the thickness directions of the LVF 23 and the LVF 24.
  • FIG. 4 as with the vertical line in FIG. 2, a virtual straight line connecting positions where the transmitted wavelength bands are the same is shown.
  • the centers of the LVF 23 and the LVF 24 are made coincident with each other, and the LVF 24 is rotated by an angle ⁇ counterclockwise with respect to the central axis of each LVF relative to the LVF 23 and perpendicular to the paper surface.
  • this angle ⁇ is referred to as a crossing angle.
  • the crossing angle ⁇ can be set arbitrarily.
  • intersection angle ⁇ it is also possible to change the intersection angle ⁇ manually or automatically.
  • the crossing angle ⁇ is 90 °
  • the same transmission characteristics overlap on the diagonal lines of the LVFs 23 and 24.
  • the filter unit 21 can realize the most combinations of transmission characteristics in 0 ° ⁇ ⁇ 90 °.
  • the intersection angle ⁇ is 90 °
  • the amount of transmitted light in the narrow band is small and the signal level is low.
  • a point P shown in FIG. 4 is a point that is the center of the LVF 23 and the LVF 24 and both of the transmission center wavelengths are ⁇ P.
  • a point Q shown in FIG. 4 is a point where the transmission center wavelength of the LVF 23 is ⁇ P , while the transmission center wavelength of the LVF 24 is ⁇ Q ( ⁇ P ).
  • the point R shown in FIG. 4 is that the transmission center wavelength of the LVF 23 is ⁇ P , while the transmission center wavelength of the LVF 24 is ⁇ R ( ⁇ Q ).
  • the transmission characteristics at each position of the filter unit 21 are the products in the common wavelength region of the transmission characteristics of the LVF 23 and the transmission characteristics of the LVF 24.
  • FIG. 5 is a diagram schematically showing a spectrum of light transmitted at the point P shown in FIG.
  • the spectrum S P shown in FIG. 5 has a transmission center wavelength of ⁇ P , and the transmission obtained by the product of the spectrum S 1P that gives the transmission characteristic at the point P of the LVF 23 and the spectrum S 2P that gives the transmission characteristic at the point P of the LVF 24. It is a characteristic. In this case, the spectrum S 1P and the spectrum S 2P match.
  • Transmission wavelength band [Delta] [lambda] PP of the spectrum S P is substantially the same spectrum S 1P, the transmission wavelength band of the S 2P.
  • a straight line L1 passing through the point P is a set of points where the transmission center wavelengths of the LVF 23 and the LVF 24 are equal. Spectrum of the transmitted light at each point on the straight line L1, while the transmission wavelength band are different from each other, have the same transmittance distribution and the spectrum S P shown in FIG.
  • FIG. 6 is a diagram schematically showing a spectrum of light transmitted at the point Q shown in FIG.
  • the spectrum S Q shown in FIG. 6 is a transmission characteristic obtained by the product of the spectrum S 1Q that gives the transmission characteristic at the point Q of the LVF 23 and the spectrum S 2Q that gives the transmission characteristic at the point Q of the LVF 24.
  • the transmission center wavelength of the spectrum S Q is ( ⁇ P + ⁇ Q ) / 2, and the transmission wavelength band ⁇ PQ is narrower than the transmission wavelength band ⁇ PP of the spectrum S P shown in FIG.
  • the transmittance at the transmission center wavelength ( ⁇ P + ⁇ Q ) / 2 is smaller than the transmittance at the transmission center wavelength ⁇ P of the spectrum S P.
  • a straight line L2 passing through the point Q is a set of points where the difference between the transmission center wavelengths of the LVF 23 and the LVF 24 is ⁇ P ⁇ Q.
  • the spectrum of the transmitted light at each point on the straight line L2 has the same transmittance distribution as the spectrum S Q shown in FIG.
  • FIG. 7 is a diagram schematically showing a spectrum of light passing at the point R shown in FIG.
  • the spectrum S R shown in FIG. 7 is a transmission characteristic obtained by the product of the spectrum S 1R that gives the transmission characteristic at the point R of the LVF 23 and the spectrum S 2R that gives the transmission characteristic at the point R of the LVF 24.
  • the transmission center wavelength of the spectrum S R is ( ⁇ P + ⁇ R ) / 2, and the transmission wavelength band ⁇ PR is narrower than the transmission wavelength band ⁇ PQ shown in FIG. Further, the transmittance at the transmission center wavelength ( ⁇ P + ⁇ R ) / 2 is smaller than the transmittance at the transmission center wavelength ⁇ Q of the spectrum S Q.
  • a straight line L3 passing through the point R is a set of points where the difference between the transmission center wavelengths of the LVF 23 and the LVF 24 is ⁇ P ⁇ R.
  • the transmittance and the transmission wavelength band at the transmission center wavelength become smaller as the absolute value of the difference between the transmission center wavelength of the LVF 23 and the transmission center wavelength of the LVF 24 at the same point increases. For this reason, the farther the distance from the straight line L1 shown in FIG. 4, the narrower the transmission wavelength band and the smaller the transmitted light amount. Note that the transmittance of the filter unit 21 as a whole is zero at a position where the transmission wavelength band of the LVF 23 and the transmission wavelength band of the LVF 24 do not overlap. Transmission characteristics including information related to the spectrum of transmitted light at each position of the filter unit 21 are stored in advance in the filter information storage unit 71 of the storage unit 7.
  • the filter unit 21 having the configuration described above, it is possible to obtain transmitted light having a different spectrum depending on the incident position of light, and to realize various transmission characteristics on the same plane. Further, even when the transmission wavelength band at each point of one LVF is wide, transmitted light having a narrow transmission wavelength band can be obtained by superimposing the two LVFs. By providing such a filter unit 21, even when the light source 9 emits light including bright lines and having greatly different intensities for each wavelength, the signal intensity of each wavelength is made uniform by selectively using transmitted light. Can be.
  • the imaging unit 3 images the light that has passed through the filter unit 21 and has been imaged by the imaging optical system 22.
  • the imaging unit 3 includes an imaging element 31 that generates and outputs an imaging signal by photoelectrically converting the imaged light.
  • the image pickup device 31 is configured using an image sensor such as a CCD (Charge Coupled Device) or a CMOS (Complementary Metal Oxide Semiconductor).
  • the image sensor 31 captures an image indicating the transmission pattern of the filter unit 21 and generates an image signal corresponding to the image.
  • the calculation unit 4 includes an image analysis unit 41 that analyzes the imaging signal output from the imaging unit 3 and a spectrum calculation unit 42 that calculates the spectrum of the imaging signal based on the analysis result of the image analysis unit 41.
  • the image analysis unit 41 refers to the filter information stored in the filter information storage unit 71 of the storage unit 7 and the sensitivity information of the image sensor 31 stored in the sensitivity information storage unit 72 of the storage unit 7, and the imaging generated by the imaging unit 3 A signal of a predetermined pixel is extracted from the signal.
  • the image analysis unit 41 passes the center of the LVF 23 and the LVF 24 (point P in FIG. 4) on the projection plane orthogonal to the thickness direction of the LVF 23 and the LVF 24, and sets the position where the transmission wavelength bands of the LVF 23 and the LVF 24 coincide with each other.
  • a signal of a pixel corresponding to a point on a straight line passing through the position when projected onto is extracted.
  • this straight line is referred to as a center line.
  • the straight line L1 is the center line.
  • the image analysis unit 41 When the signal extracted by the image analysis unit 41 has a saturated signal value, the image analysis unit 41 is a pixel located in the vicinity of a pixel (saturated pixel) that outputs a saturated signal value. The signal value of the saturated pixel is replaced with the unsaturated signal value output by the pixel whose corresponding point on the projection plane is not located on the center line. At this time, the image analysis unit 41 searches in order from the pixel with the shortest distance from the saturated pixel, and replaces the signal value of the saturated pixel with the unsaturated signal value found first. The unsaturated signal has a narrower band than the pixel signal extracted first by the image analysis unit 41. Therefore, the image analysis unit 41 performs correction to match the bandwidth of the replaced signal with the bandwidth of the pixel signal to be extracted first. Even after the signal value is replaced, the position of the pixel that output the signal is the position before the replacement.
  • the filter information storage unit 71 individually stores information regarding the transmission characteristics of the LVFs 23 and 24, and the image analysis unit 41 calculates the characteristics for each pixel in accordance with the intersection angle between the LVF 23 and the LVF 24. Also good.
  • the spectrum calculation unit 42 calculates the spectrum of the imaging signal based on the transmission characteristics such as the transmission wavelength band according to the signal of each pixel extracted by the image analysis unit 41 and the position of each pixel.
  • the spectrum calculation unit 42 refers to various information such as the transmission center wavelength of the spectrum at each position of the filter unit 21 stored in the storage unit 7, the filter characteristics of the filter unit 21, and the sensitivity characteristics of the imaging unit 3. The relative intensity for each is calculated, and the spectrum indicating the spectral sensitivity is calculated and output.
  • the arithmetic unit 4 is configured using a general-purpose processor such as a CPU (Central Processing Unit) or a dedicated integrated circuit that executes a specific function such as an ASIC (Application Specific Integrated Circuit) or FPGA (Field Programmable Gate Array). Various arithmetic processes are executed by reading various programs stored in the storage unit 7.
  • a general-purpose processor such as a CPU (Central Processing Unit) or a dedicated integrated circuit that executes a specific function such as an ASIC (Application Specific Integrated Circuit) or FPGA (Field Programmable Gate Array).
  • ASIC Application Specific Integrated Circuit
  • FPGA Field Programmable Gate Array
  • the input unit 5 includes an input device such as a keyboard, various buttons, and various switches, and a pointing device such as a mouse and a touch panel provided on the display screen of the display unit 6. The input of the corresponding signal is received and input to the control unit 8.
  • an input device such as a keyboard, various buttons, and various switches
  • a pointing device such as a mouse and a touch panel provided on the display screen of the display unit 6. The input of the corresponding signal is received and input to the control unit 8.
  • the display unit 6 is configured by using a display panel made of liquid crystal, organic EL (Electro Luminescence), or the like, and displays various types of information including a calculation result by the spectrum calculation unit 42.
  • the storage unit 7 stores a filter information storage unit 71 that stores information about the LVFs 23 and 24, a sensitivity information storage unit 72 that stores sensitivity information of the image sensor 31, and spectral information of the light source 9 calculated by the spectroscopic device 1. And a spectrum information storage unit 73.
  • the filter information storage unit 71 stores transmission characteristics including a spectrum of transmitted light at each position in the filter unit 21.
  • the filter information storage unit 71 stores the transmission characteristics at each position according to the value of the intersection angle.
  • the storage unit 7 also stores an imaging signal acquired by the imaging unit 3, a plurality of programs and various setting information respectively executed by the calculation unit 4 and the control unit 8.
  • the program may be written and stored in a computer-readable recording medium. Writing of the program to the storage unit 7 or the recording medium may be performed when the computer or the recording medium is shipped as a product, or may be performed by downloading via a communication network.
  • the storage unit 7 includes a volatile memory such as a RAM (Random Access Memory) and a nonvolatile memory such as a ROM (Read Only Memory). Note that the storage unit 7 may be configured using a computer-readable recording medium such as a memory card that can be externally mounted.
  • a volatile memory such as a RAM (Random Access Memory)
  • a nonvolatile memory such as a ROM (Read Only Memory).
  • the storage unit 7 may be configured using a computer-readable recording medium such as a memory card that can be externally mounted.
  • the control unit 8 is configured using a general-purpose processor such as a CPU or a dedicated integrated circuit that executes a specific function such as an ASIC or FPGA. Note that the control unit 8 and the calculation unit 4 can be configured by the same general-purpose processor or integrated circuit.
  • the white plate 10 having a uniform diffuse reflection surface is irradiated with light emitted from the light source 9, and the white plate 10
  • the light having a spatially uniform spectrum is incident on the spectroscopic device 1.
  • the imaging unit 3 captures the transmitted light and generates an imaging signal.
  • An image pickup signal in an appropriate region is extracted from the image pickup signal, and the spectrum (spectral sensitivity characteristic) of the light source 9 is calculated.
  • the calculation result of the light source 9 spectrum is displayed on the display unit 6.
  • the direction of change of the transmission center wavelength in the two flat plate-like LVFs intersects at the projection plane orthogonal to the thickness direction. Therefore, it is possible to transmit light with different transmission characteristics, and it is possible to accurately measure the spectral sensitivity even for light having greatly different intensities according to wavelengths.
  • the spectral sensitivity characteristic can be accurately obtained even when the incident light has a component such as a bright line.
  • the flattening filter has no effect unless it reflects the characteristics of the light source to be flattened. It is not suitable for applications that detect spectral sensitivity characteristics.
  • the first embodiment is suitable for detecting the spectral sensitivity characteristics of various light sources since no flattening filter is used.
  • FIG. 8 is a block diagram showing a configuration of a microscope system that is a spectroscopic device according to Embodiment 2 of the present invention.
  • a microscope system 100 shown in FIG. 1 includes a microscope apparatus 101, a control apparatus 102, an input apparatus 103, and a display apparatus 104.
  • the input device 103 and the display device 104 have the same functions as the input unit 5 and the display unit 6 described in the first embodiment, respectively.
  • components having the same functions as those described in the first embodiment will be described with the same reference numerals as those in the first embodiment.
  • the microscope apparatus 101 includes a stage 111, an objective optical system 112, an imaging optical system 113, a microscope imaging unit 114, an epi-illumination light source 115, an epi-illumination optical system 116, a fluorescent cube 117, a transmission illumination light source 118, a transmission illumination optical system 119, And an apparatus for observing a specimen SMP placed on a stage 111, having a mirror 120.
  • the objective optical system 112 is arranged to face the surface on which the specimen SMP of the stage 111 is placed.
  • the objective optical system 112 includes a plurality of interchangeable objective lenses, and the magnification can be changed.
  • the light that has passed through the objective optical system 112 is imaged by the imaging optical system 113 and enters the microscope imaging unit 114.
  • the imaging optical system 113 is configured by using a plurality of lenses, and the zoom magnification can be changed.
  • the microscope imaging unit 114 includes an imaging element that photoelectrically converts received light to generate an image signal.
  • the microscope imaging unit 114 outputs the generated image signal to the control device 102. Note that a configuration may be adopted in which light is further branched on the observation optical path between the objective optical system 112 and the imaging optical system 113, and an eyepiece is provided on the branched optical path so that the user can directly observe.
  • the epi-illumination optical system 116 includes various optical members (filter unit, shutter, field stop, aperture stop, etc.) that collect the epi-illumination light emitted from the epi-illumination light source 115 and guide it to the observation optical path.
  • the transmission illumination optical system 119 includes various optical members (collector lens, filter unit, field stop, shutter, aperture stop, etc.) that collect the transmission illumination light emitted from the transmission illumination light source 118 and guide it to the mirror 120. The light that has passed through the transmission illumination optical system 119 is reflected by the mirror 120 and then propagates along the observation optical path.
  • the fluorescent cube 117 includes an excitation filter 117a that selectively transmits light (excitation light) in a specific wavelength band out of the light emitted from the incident illumination light source 115 and passed through the incident illumination optical system 116, and the excitation filter 117a.
  • a dichroic mirror 117b that reflects the selected excitation light and transmits the fluorescence generated by the sample SMP, and an absorption filter 117c that selectively transmits only the fluorescence of light incident from the direction of the sample SMP are formed in a cube shape. Combined.
  • the fluorescent cube 117 is inserted into the observation optical path when performing fluorescence observation. On the other hand, when performing bright field observation, the fluorescent cube 117 is retracted from the observation optical path.
  • the microscope apparatus 101 further includes a spectroscopic unit 2A, an imaging unit 3A, and a mirror 121 that reflects light from the sample SMP and guides it to the spectroscopic unit 2A.
  • the mirror 121 is detachably provided on the observation optical path between the fluorescent cube 117 and the imaging optical system 113.
  • the mechanism for inserting and removing the mirror 121 with respect to the observation optical path may be manual or electric.
  • FIG. 8 shows a state where the mirror 121 is inserted.
  • the mirror 121 reflects the light that has passed through the objective optical system 112 while being inserted in the observation optical path, and causes the light to enter the spectroscopic unit 2A.
  • the spectroscopic unit 2A which is a spectroscopic unit according to the second embodiment, includes an imaging optical system 25, a diffusion optical system 26, a filter unit 21, and an imaging optical system 22 in order from the light incident side.
  • FIG. 9 is a diagram showing a configuration of the diffusion optical system 26.
  • the diffusion optical system 26 is an optical system that diffuses incident light to make it uniform.
  • the diffusion optical system 26 includes a field stop 261 disposed at a position where an image formed by the imaging optical system 25 is formed, a diffusion plate 262 that diffuses a light beam that has passed through the field stop 261, and a diffused light beam. And a light beam homogenizing optical element 263 for homogenizing.
  • the diffusion plate 262 is configured using, for example, a material obtained by processing one side of flat glass into a sand surface, or a material in which a light diffusing substance is dispersed in the glass.
  • the light beam homogenizing optical element 263 is made of a light-transmitting member formed using glass, plastic, or the like, and emits light while reflecting at least part of the light beam incident from the incident surface 263a one or more times by the side surface 263b. The light is guided to 263c and emitted.
  • the beam homogenizing optical element 263 has a shape whose width increases from the incident side to the emission side, and the area of the emission surface 263c is larger than the area of the incident surface 263a. Further, the side surface 263b of the light beam homogenizing optical element 263 has a curved surface whose inclination angle gradually decreases from the incident surface 263a to the exit surface 263c.
  • the side surface 263b does not need to be a curved surface, and may be configured by a flat surface.
  • the cross section perpendicular to the optical axis of the light beam homogenizing optical element 263 may be rectangular or circular, and the incident surface 263a may be circular, while the output surface 263c may be rectangular.
  • the diffusing optical system 26 having the above configuration can ensure the measurement accuracy of the spectrum by making the illuminance distribution of the light beam irradiated to the image sensor 31 uniform.
  • a more detailed configuration of the diffusion optical system 26 is disclosed in, for example, Japanese Patent Application Laid-Open No. 2013-29322.
  • the imaging unit 3 ⁇ / b> A includes the imaging element 31 similarly to the imaging unit 3, generates an imaging signal corresponding to an image indicating the transmission pattern of the filter unit 21, and outputs the imaging signal to the control device 102.
  • the mirror 121, the spectroscopic unit 2A, and the imaging unit 3A have been described as single components, but these may be configured as one unit. Further, the spectroscopic unit 2A and the imaging unit 3A may be configured as one unit.
  • the control device 102 includes a calculation unit 4A, a storage unit 7A, and a control unit 8A.
  • the calculation unit 4A includes an image analysis unit 41 and a spectrum calculation unit 42.
  • the spectrum calculation unit 42 calculates the number of fluorescent spectra, that is, the number of fluorescent components, from the peak position of the narrowband signal included in the imaging signal, and the position of the pixel that outputs the narrowband signal and the transmission wavelength band.
  • the fluorescence spectrum is calculated on the basis of the correspondence.
  • the calculation unit 4A performs image processing including optical black subtraction processing, white balance adjustment processing, color matrix calculation processing, ⁇ correction processing, color reproduction processing, and edge enhancement processing on the image signal generated by the microscope imaging unit 114. Do.
  • FIG. 10 is a diagram for explaining the operation when fluorescence observation is performed using the microscope system 100.
  • the microscope apparatus 101 turns on the epi-illumination light source 115 and inserts the fluorescent cube 117 on the observation optical path, while turning off the power of the transmission illumination light source 118.
  • the absorption filter 117c of the fluorescent cube 117 is a filter that transmits a fluorescent component in the green (G) wavelength band
  • the sample SMP emits fluorescence of two spectra S1 and S2 shown in FIG.
  • Such a phenomenon can occur, for example, when at least two fluorescent substances exist in the specimen SMP and the wavelengths of the two fluorescent substances are close to each other. In this case, an image including the components of the microscope imaging unit 114 and the two spectra S1 and S2 is captured. As a result, the microscope system 100 cannot detect only the fluorescence (either one of the spectra S1 and S2) that is originally desired to be detected from the image captured by the microscope imaging unit 114.
  • the microscope imaging unit 114 before the microscope imaging unit 114 performs imaging, the mirror 121 is inserted into the observation optical path, and the light emitted from the sample SMP using the spectroscopic unit 2A and the imaging unit 3A. Measure the spectrum of. As a result of the measurement, when the presence of two fluorescent components close to each other in wavelength band is detected as in the spectra S1 and S2 shown in FIG. 10, the user separates the two fluorescent components and extracts only the desired fluorescent component. The absorption filter 117c can be replaced. As a result, the microscope imaging unit 114 can capture a fluorescent image showing only a desired fluorescent component.
  • the calculated spectral data may be used when adjusting the white balance (WB) of the image captured by the microscope imaging unit 114.
  • WB white balance
  • the second embodiment it is possible to acquire an observation image including only a desired fluorescent component using an appropriate absorption filter by measuring in advance the fluorescence emitted from the specimen.
  • the epi-illumination light source 115 is turned off, the fluorescent cube 117 is retracted from the observation optical path, and the transmitted illumination light source 118 is turned on.
  • examples of staining of a specimen for bright field observation include non-fluorescent staining such as hematoxylin and eosin staining (HE staining) using two pigments of hematoxylin and eosin, and Papanicolaou staining (Pap staining). .
  • FIG. 11 is a diagram for explaining an outline of a spectrum calculated by the spectrum calculation unit 42 from an imaging signal transmitted by the spectroscopic unit 2A and generated by the imaging unit 3A when the stained specimen is observed in the bright field in the microscope apparatus 101. It is. Of the three curves C, C1, and C2 shown in FIG. 11, the curve C represents a spectrum that serves as a reference when performing the color homogenization process. On the other hand, the curves C1 and C2 show spectra calculated when different facilities or engineers create specimens with the same staining. As shown in FIG. 11, a dyed specimen may have different dye characteristics for each dyeing, for each facility, and for each engineer.
  • the imaging unit 3A captures the light separated using the spectroscopic unit 2A and calculates the spectrum for each stained specimen, it is possible to accurately estimate the difference (characteristic change) from the reference spectrum. it can. As a result, it is possible to improve the accuracy of the color homogenization process when the microscope imaging unit 114 images a stained specimen for bright field observation.
  • the spectrum of the entire stained specimen is calculated using a narrow-band transmission spectrum, so that the transmission wavelength band is wide and the transmission wavelength band between bands is the same as that measured by a conventional multiband sensor (Triton).
  • the spectrum can be calculated with high accuracy compared to the method with many overlaps.
  • FIG. 12 is a block diagram showing a configuration of an imaging apparatus that is a spectroscopic apparatus according to Embodiment 3 of the present invention.
  • An imaging apparatus 200 shown in the figure includes a main body 201 and a spectroscopic unit 2B that can be attached to and detached from the main body 201. Also in the third embodiment, components having the same functions as those described in the first embodiment will be described with the same reference numerals as those in the first embodiment.
  • the spectroscopic unit 2B which is a spectroscopic unit according to the third embodiment includes an objective optical system 27, an imaging optical system 25, a diffusion optical system 26, a filter unit 21 and a filter unit 21 in order from the front side (light incident side) facing the subject.
  • An imaging optical system 22 is included.
  • the spectroscopic unit 2 ⁇ / b> B diffuses the light collected from the dotted area and enters the filter unit 21.
  • the spectroscopic unit 2B corresponds to a configuration in which an objective optical system is added to the spectroscopic unit 2A described in the second embodiment.
  • the spectroscopic unit 2B is attached to the main body unit 201 instead of the lens unit for normal photographing.
  • the main body unit 201 includes an imaging unit 3B, a calculation unit 4B, an input unit 5B, a display unit 6B, a storage unit 7B, and a control unit 8B.
  • the imaging unit 3B includes an imaging element 31.
  • the imaging element 31 generates an imaging signal corresponding to an image indicating the transmission pattern of the filter unit 21 of the spectroscopic unit 2B.
  • the calculation unit 4B includes an image analysis unit 41 and a spectrum calculation unit 42.
  • the calculation unit 4B also has a function of performing various image processing on the image pickup signal generated by the image pickup unit 3B.
  • the calculation unit 4B like the calculation unit 4A described in the second embodiment, performs optical black subtraction processing, white balance adjustment processing, color matrix calculation processing, ⁇ correction processing, color reproduction on an image signal. Image processing including processing and edge enhancement processing is performed.
  • the storage unit 7B includes a filter information storage unit 71, a sensitivity information storage unit 72, and a spectrum information storage unit 73.
  • the storage unit 7B stores data of an image captured by the imaging unit 3B and subjected to image processing by the calculation unit 4B.
  • the calculated spectral data is used when adjusting the white balance of the image captured by the imaging unit 3B, so that even when a light source such as a bright line is captured, white The balance can be adjusted with high accuracy.
  • the spectroscopic device according to the third embodiment can be realized by installing the spectroscopic unit 2B in the main body of a normal imaging device and downloading software for realizing the function of the arithmetic unit 4B. Highly accurate spectral sensitivity can be measured easily.
  • the imaging device 200 When the imaging device 200 is applied as an inspection device such as a coating plate color or a light source spectrum, the reference transmission pattern obtained by the calculation unit 4B based on the characteristics of the filter unit 21 and the transmission in the image captured by the imaging unit 3B. Inspection and determination may be performed by matching the pattern.
  • FIG. 13 is a block diagram illustrating a configuration of the spectroscopic device according to the fourth embodiment.
  • the spectroscopic device 1A shown in the figure includes a filter driving unit 11 that rotates the LVF 24 around a central axis parallel to the thickness direction in addition to the same configuration as the spectroscopic device 1 described in the first embodiment.
  • the filter driving unit 11 is configured by using, for example, an actuator that rotates the LVF 24. By rotating the LVF 24 under the control of the control unit 8, the filter driving unit 11 on the projection plane described in the first embodiment with reference to FIG. The intersection angle with the LVF 23 is changed.
  • the image analysis unit 41 of the calculation unit 4 extracts and extracts all the signal values of the edge pixels located at the edge in the direction perpendicular to the center line on the projection plane among the pixels that form the transmission pattern with the imaging signal.
  • the maximum value of the signal value of the edge pixel is calculated, and this maximum value is compared with a predetermined reference value.
  • the reference value is preferably set so that the number of pixels equal to or greater than the reference value is maximized with respect to the determined exposure condition.
  • Such a reference value can be set to about 10% of the saturation signal value of the pixel, for example.
  • the reference value can be set to a value that exceeds the noise level.
  • the filter drive unit 11 controls the LVF 24 by a predetermined angle under the control of the control unit 8 when the maximum value of the signal value of the edge pixel is equal to or less than the reference value.
  • the predetermined angle is, for example, about 0.1 ° to 30 °, and more preferably about 1 ° to 5 °.
  • FIG. 14 is a flowchart showing an outline of the intersection angle determination process performed by the spectroscopic apparatus 1A.
  • the filter drive unit 11 rotationally drives the LVF 24 so that the intersection angle between the LVF 23 and the LVF 24 is 90 ° (step S1).
  • the image analysis unit 41 acquires an imaging signal from the imaging unit 3 (step S2), and among the pixels forming the transmission pattern, the signal value of the edge pixel positioned at the edge in the direction perpendicular to the center line is obtained. All are extracted (step S3).
  • the image analysis unit 41 calculates the maximum signal value of the extracted edge pixels (step S4).
  • the image analysis unit 41 determines whether or not the calculated maximum value is equal to or less than a predetermined reference value (step S5).
  • the control unit 8 outputs a control signal to the filter driving unit 11, and causes the filter driving unit 11 to rotationally drive the LVF 24 by a predetermined angle.
  • Step S6 the filter drive unit 11 rotates the LVF 24 clockwise by a predetermined angle on the projection plane shown in FIG.
  • step S7: Yes When the crossing angle ⁇ after the rotation in step S6 is 0 ° or less (step S7: Yes), the control unit 8 outputs a control signal to the filter driving unit 11 to drive the LVF 24 in rotation and is preset.
  • the intersection angle ⁇ 0 is set (step S8).
  • the intersection angle ⁇ 0 is 1 °, for example, but this is only an example.
  • step S7: No when the crossing angle ⁇ after the rotation in step S6 is larger than 0 ° (step S7: No), the spectroscopic device 1A returns to step S2.
  • step S5 when the maximum value is larger than the reference value (step S5: No), the spectroscopic device 1A ends the process. Thereby, the intersection angle of LVF23 and LVF24 is determined.
  • FIG. 15 is a diagram schematically showing an example of pixel signal value distribution in the transmission pattern acquired from the light of the specific light source 9 by the spectroscopic device 1A when the crossing angle between the LVF 23 and the LVF 24 is 90 °.
  • FIG. 15 shows an example of the distribution of signal values when viewed from the incident side of the spectroscopic device 1A.
  • the transmissive pixel region B where the transmissive pattern exists has a symmetrical shape with respect to the center line M.
  • the signal value of the pixel passing through the center line M is the highest, and the signal value decreases as the distance from the center line M increases. ing.
  • the difference in signal value is schematically represented by a pattern.
  • the signal values of the edge pixels located at the edge in the direction perpendicular to the center line M of the partial pixel region Bm are all the same (shown in black).
  • the signal value of the edge pixel is assumed to the reference value.
  • the maximum signal value of the edge pixel calculated by the image analysis unit 41 in step S4 is equal to the reference value. Therefore, the image analysis unit 41 determines that the maximum value is equal to or less than the reference value (step S5: Yes), and the filter driving unit 11 rotationally drives the LVF 24 by a predetermined angle (step S6).
  • the spectroscopic device 1A returns to step S2 to acquire an imaging signal.
  • FIG. 16 schematically illustrates an example of pixel signal value distribution in the transmission pattern obtained from the light of the same light source 9 as that in FIG. 15 when the crossing angle between the LVF 23 and the LVF 24 is an acute angle ( ⁇ 90 °).
  • FIG. 16 is a diagram schematically illustrating an example of signal value distribution when a transmission pattern of light from the same light source 9 as in FIG. 15 is acquired. Also in FIG. 16, similarly to FIG. 15, an example of the distribution of signal values when viewed from the incident side of the spectroscopic device 1A is shown.
  • the transmissive pixel region B ′ shown in FIG. 16 has a symmetric shape with respect to the center line M ′ as in the case shown in FIG. 15.
  • the signal value of the pixel passing through the center line M ′ is the highest, and the signal value increases as the distance from the center line M ′ increases. It is low.
  • the transmissive pixel region B ' is wider than the transmissive pixel region B shown in FIG. 15, and the number of extracted pixels is large.
  • the signal value of the edge pixel is larger than the reference value (displayed by a mesh line). Therefore, in the case shown in FIG. 16, the maximum value of the signal value of the edge pixel calculated by the image analysis unit 41 in step S4 is larger than the reference value. Therefore, the image analysis unit 41 determines that the maximum value is larger than the reference value (step S5: No). In this case, the spectroscopic device 1A ends the intersection angle determination process between the LVF 23 and the LVF 24.
  • the initial value of the crossing angle ⁇ may be set to 0 °, and the crossing angle may be gradually increased, or the crossing angle may be increased or decreased according to a predetermined rule starting from a preset initial value. Good.
  • the fourth embodiment by optimizing the crossing angle, the number of candidates for replacing a pixel having a saturated signal value with another pixel increases. Therefore, the accuracy when reducing noise is further increased. Can be improved.
  • a mechanism for manually rotating the LVF 24 by the operator may be provided instead of providing the filter driving unit 11, a mechanism for manually rotating the LVF 24 by the operator.
  • the filter driving unit 11 may rotate the LVF 23, or both the LVF 23 and the LVF 24 may be rotated.
  • the filter driving unit described in the fourth embodiment may be added so that the intersection angle of the two LVFs on the projection plane can be changed.
  • an optimal intersection angle may be determined using the signal value for each pixel of the imaging signal.
  • the shape and thickness of the surfaces of the two LVFs do not have to be the same.
  • the shape of the LVF that is rotationally driven may be relatively small or the thickness may be thin.
  • the number of LVFs constituting the filter unit is not necessarily two, and the filter unit may be configured using a plurality of LVFs.
  • the thickness direction of the plurality of LVFs may be aligned, and the plurality of LVFs may be arranged side by side along the thickness direction.
  • two adjacent LVFs may be arranged in contact with each other, or may be arranged apart from each other. In the latter case, no other component is provided between two adjacent LVFs.
  • the present invention can include various embodiments not described herein, and various design changes can be made within the scope of the technical idea specified by the claims. It is.

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  • Spectroscopy & Molecular Physics (AREA)
  • General Physics & Mathematics (AREA)
  • Spectrometry And Color Measurement (AREA)

Abstract

L'unité spectroscopique selon la présente invention est pourvue de : un premier filtre plan dans lequel la longueur d'onde centrale de transmission de lumière varie en continu le long d'une direction prédéfinie ; et un deuxième filtre plan dans lequel la longueur d'onde centrale de transmission de lumière varie de façon continue le long d'une direction prédéfinie, au moins une partie de la longueur d'onde du centre de transmission variant en continu étant en commun avec la longueur d'onde centrale de transmission du premier filtre, le deuxième filtre étant agencé le long de la direction de l'épaisseur de celui-ci dans un état dans lequel la direction de l'épaisseur de celui-ci est alignée avec celle du premier filtre, et la direction de variation de la longueur d'onde centrale de transmission du deuxième filtre coupant la direction de variation de la longueur d'onde centrale de transmission du premier filtre au niveau d'une surface de projection orthogonale à la direction de l'épaisseur.
PCT/JP2016/071185 2016-07-19 2016-07-19 Unité spectroscopique et dispositif spectroscopique Ceased WO2018016010A1 (fr)

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WO2023032256A1 (fr) * 2021-09-03 2023-03-09 ソニーセミコンダクタソリューションズ株式会社 Dispositif de mesure de sensibilité spectrale et procédé de mesure de sensibilité spectrale

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US20020140935A1 (en) * 1998-11-26 2002-10-03 Carter Christopher Frederick Versatile filter based spectrophotometer
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JP2011253078A (ja) * 2010-06-03 2011-12-15 Nikon Corp 光学部品及び分光測光装置
JP2013029322A (ja) * 2011-07-26 2013-02-07 Olympus Corp 波長分布測定装置
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US5784507A (en) * 1991-04-05 1998-07-21 Holm-Kennedy; James W. Integrated optical wavelength discrimination devices and methods for fabricating same
US5703357A (en) * 1993-09-27 1997-12-30 Shih; Ishiang Methods for wavelength discrimination of monochromatic light beams
US20020140935A1 (en) * 1998-11-26 2002-10-03 Carter Christopher Frederick Versatile filter based spectrophotometer
US20040032584A1 (en) * 2002-08-15 2004-02-19 Tokuyuki Honda Optical channel monitoring device
JP2007527516A (ja) * 2003-07-02 2007-09-27 ヴェリティー インストルメンツ，インコーポレイテッド 電荷結合装置に基づく分光計のダイナミックレンジ拡張装置および方法
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