WO2020009220A1 - Microscopic observation apparatus, microscopic observation method, and method for manufacturing microscopic observation apparatus - Google Patents

Microscopic observation apparatus, microscopic observation method, and method for manufacturing microscopic observation apparatus Download PDF

Info

Publication number
WO2020009220A1
WO2020009220A1 PCT/JP2019/026801 JP2019026801W WO2020009220A1 WO 2020009220 A1 WO2020009220 A1 WO 2020009220A1 JP 2019026801 W JP2019026801 W JP 2019026801W WO 2020009220 A1 WO2020009220 A1 WO 2020009220A1
Authority
WO
WIPO (PCT)
Prior art keywords
observation target
unit
light
light receiving
microscopic observation
Prior art date
Application number
PCT/JP2019/026801
Other languages
French (fr)
Japanese (ja)
Inventor
宗一郎 上野
Original Assignee
株式会社Ｉｄｄｋ
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 株式会社Ｉｄｄｋ filed Critical 株式会社Ｉｄｄｋ
Publication of WO2020009220A1 publication Critical patent/WO2020009220A1/en

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • H—ELECTRICITY
    • H04—ELECTRIC COMMUNICATION TECHNIQUE
    • H04N—PICTORIAL COMMUNICATION, e.g. TELEVISION
    • H04N25/00—Circuitry of solid-state image sensors [SSIS]; Control thereof
    • H04N25/70—SSIS architectures; Circuits associated therewith

Definitions

  • the present invention relates to a microscopic observation device, a microscopic observation method, and a method for manufacturing a microscopic observation device.
  • Patent Document 1 an observation method capable of easily observing the entire observation target without adjusting the optical system such as imaging and enlarging and reducing and scanning the observation target.
  • Patent Document 1 does not assume irradiating an observation target with excitation light and observing fluorescence from the observation target.
  • the present invention has been made in view of such problems, and an object of the present invention is to provide a microscopic observation apparatus and a microscopic observation method capable of performing observation using fluorescence from an observation target irradiated with excitation light. Another object of the present invention is to provide a method for manufacturing such a microscopic observation device.
  • a microscopic observation apparatus that irradiates an observation target with excitation light and observes fluorescence generated from the observation target, and a light source that irradiates the observation target with excitation light, a semiconductor substrate, A plurality of light receiving portions formed on the semiconductor substrate, a protection layer covering the semiconductor substrate and the plurality of light receiving portions, a first polarizing means formed on the protection layer and containing a metal, An oxide flattening layer covering metal particles, a resin flattening layer formed on the oxide flattening layer, and a resin flattening layer formed on the resin flattening layer, and each of the plurality of light receiving units is formed from the observation target.
  • the excitation having passed through the second polarizing means Configured such that light is cut by the first polarizing means, microscopic observation device is provided.
  • a microscopic observation apparatus for irradiating an observation target with excitation light and observing fluorescence generated from the observation target, comprising: a light source for irradiating the observation target with excitation light; Unit, a non-imaging lens system for guiding fluorescence from the observation target to each of the plurality of light receiving units, a first polarizing unit disposed between the observation target and the light receiving unit, the light source and the light source A second polarization unit disposed between the first polarization unit and the observation target, wherein the polarization characteristics of the first polarization unit and the second polarization unit are such that the excitation light that has passed through the second polarization unit is the first polarization unit.
  • a microscopic observation device configured to be cut by means is provided.
  • a mounting portion may be provided above the non-imaging lens system and a bottom surface may be the second polarizing means.
  • a method of manufacturing a microscopic observation apparatus that irradiates an observation target with excitation light to observe fluorescence generated from the observation target, and irradiates the observation target with excitation light.
  • a method for manufacturing a microscopic observation device, which is attached so as to be cut by two-polarizing means, is provided.
  • a microscopic observation method for irradiating an observation target with excitation light and observing fluorescence generated from the observation target, the method being arranged above a non-imaging lens system in the microscopic observation apparatus. Placing the observation target on the placed mounting portion, irradiating the observation target with excitation light from the light source via the first polarization unit, and converting the excitation light passing through the first polarization unit into the second polarization unit. Guiding the fluorescence from the observation target to the light receiving unit by the non-imaging lens system while cutting by utilizing the polarization characteristics of the means.
  • FIG. 1 is a sectional view showing a schematic configuration of a microscopic observation apparatus 100 according to one embodiment.
  • FIG. 3B is a system configuration diagram of a microscope observation system including the microscope observation device 100 of FIG. 3A.
  • FIG. 3 is a partially enlarged view of the light receiving unit 60.
  • FIG. 4 is a partially enlarged cross-sectional view of a light receiving unit 60 which is a modified example of FIG. 3.
  • FIG. 4 is a partially enlarged cross-sectional view of a light receiving unit 60 which is another modified example of FIG. 3.
  • FIG. 1 is an electric block diagram schematically showing a microscopic observation system using a microscopic observation device 100.
  • FIG. 1 is a diagram illustrating the concept of microscopic observation according to the present invention.
  • the microscopic observation device 100 includes a light source 1, a light receiving unit 2, and polarizing units 3 and 4.
  • the light source 1 irradiates the observation target T with the excitation light L1.
  • the observation target T is excited by the excitation light L1 and emits fluorescence L2.
  • the light receiving unit 2 receives the fluorescence L2 from the observation target T and converts it into an electric signal.
  • the observation target T can be observed by forming an image based on the electric signal.
  • the fluorescence L2 from the observation target T is often weaker than the excitation light L1 from the light source 1. Therefore, if the excitation light L1 from the light source 1 reaches the light receiving unit 2, the influence of the excitation light L1 is too strong, and accurate observation using the fluorescence L2 becomes difficult.
  • two polarizing means 3 and 4 are provided between the light source 1 and the light receiving section 2.
  • the polarization means 3 and 4 can switch the polarization action electrically or mechanically, for example. Then, by utilizing the polarization characteristics of the polarization units 3 and 4, the excitation light L1 passing through the polarization unit 3 on the light source 1 side is cut by the polarization unit 4 on the light receiving unit 2 side.
  • the polarizing means 3 on the light source 1 side transmits only linearly polarized light in a predetermined direction (vertical direction in FIG.
  • the polarizing means 3 and 4 can be configured and arranged to pass only linearly polarized light (in the horizontal direction in the figure).
  • the polarizing means 3 on the light source 1 side transmits only circularly polarized light in a predetermined direction (clockwise in FIG. 2), and the polarizing means 4 on the light receiving unit 2 side is opposite.
  • the polarization means 3 and 4 may be configured and arranged to pass only circularly polarized light in the direction (counterclockwise in the figure).
  • FIG. 3A is a cross-sectional view illustrating a schematic configuration of the microscopic observation device 100 according to one embodiment.
  • the microscopic observation device 100 includes a light source unit 50 and a light receiving unit 60.
  • the light source unit 50 integrates the light source 1 shown in FIG.
  • the light receiving unit 60 includes a semiconductor substrate 11, a plurality of photodiode layers 12, a protective layer 13, metal particles 14, an oxide planarization layer 15, a resin planarization layer 16, a plurality of microlenses 17, And a mounting portion 18.
  • the semiconductor substrate 11 is a silicon substrate or the like.
  • the photodiode layer 12 is an impurity diffusion layer formed in the semiconductor substrate 11, and forms a pn junction photodiode by injecting a p-type impurity into the n-type semiconductor substrate 11, for example.
  • a plurality of photodiode layers 12 are formed apart from each other in a matrix and constitute the light receiving section 2 in FIG. The light receiving section 2 only needs to perform photoelectric conversion on incident light, and does not necessarily need to be the photodiode layer 12.
  • the protection layer 13 is, for example, a silicon oxide film or a silicon nitride film, and covers the exposed portions of the photodiode layer 12 and the semiconductor substrate 11.
  • the metal particles 14 are smaller than the photodiode layer 12 and are formed on the protective layer 13. Although the metal particles 14 may be formed directly on the semiconductor substrate 11 or the photodiode layer 12, it is preferable to provide the protective layer 13.
  • the plurality of metal particles 14 function as the polarization unit 4 in FIG. As described with reference to FIG. 1, the polarization unit 4 is configured to cut the excitation light that has passed through the polarization unit 3.
  • the oxide planarization layer 15 is, for example, a silicon oxide film, and covers the exposed portions of the metal particles 14 and the protection layer 13.
  • the resin planarization layer 16 is formed on the oxide planarization layer 15.
  • a plurality of microlenses 17 made of resin are formed in a matrix on the resin flattening layer 16. Although the microlenses 17 may be formed directly on the oxide flattening layer 15, the microlenses 17 can be adhered to the resin flattening layer 16 by providing the resin flattening layer 16.
  • Each of the microlenses 17 is formed immediately above each of the photodiode layers 12.
  • One photodiode layer 12 and one corresponding microlens 17 form a pixel.
  • the micro lens 17 is a non-imaging lens system, and does not need to form an image on the photodiode layer 12, and guides light incident from a specific viewing angle ⁇ (described later) to the photodiode layer 12. Note that any layer other than the microlens 17 that can control the viewing angle may be applied.
  • the mounting portion 18 is a portion on which the observation target T is mounted, for example, may be formed on the microlens 17 and may be made of resin and a plate shape, or may be a liquid or solid state embedded in the microlens 17. It may be something.
  • the mounting portion 18, the resin flattening layer 16, the oxide flattening layer 15, and the protective layer 13 can transmit at least the fluorescence from the observation target T. Further, in the microscopic observation apparatus 100, it is not necessary to provide a lens system for imaging or enlargement / reduction between the light source 1 and the photodiode layer 12.
  • the microscopic observation device 100 is manufactured by attaching the light source unit 50 and the light receiving unit 60. More specifically, the light source unit 50 and the light receiving unit are arranged such that the excitation light passing through the polarizing unit 3 is cut by the polarizing unit 4 by the polarization characteristics of the polarizing unit 3 in the light source unit 50 and the polarizing unit 4 in the light receiving unit 60. And 60.
  • FIG. 3B is a system configuration diagram of a microscope observation system including the microscope observation device 100 of FIG. 3A.
  • the microscope observation system includes a light source control unit 71 that controls the light source 1, a polarization control unit 72 that electrically or mechanically switches the polarization operation of the polarization unit 3 (and / or the polarization unit 4), A light receiving unit controller 73 for controlling the unit 60 and an overall controller 70 are provided.
  • the general control device 70 controls the light source control unit 71, the polarization control unit 72, and the light receiving unit control unit 73, and is configured by, for example, a personal computer or a microcomputer.
  • microscopic observation is performed as follows. First, the observation target T is placed on the placement unit 18. Next, excitation light is emitted from the light source 1 to the observation target T via the polarizing means 3. Only the excitation light having a specific polarization state among the excitation lights from the light source 1 passes through the polarization means 3 and reaches the observation target T.
  • the observation target T emits fluorescence when excited by the excitation light. This fluorescence is guided to the corresponding photodiode layer 12 by each micro lens 17.
  • the excitation light transmitted from the light source 1 through the polarizing means 3 is cut by the polarizing means 4 composed of the metal particles 14 and hardly reaches the photodiode layer 12.
  • the light entering the photodiode layer 12 is dominated by the fluorescence from the observation target T, and the observation target T can be observed by converting the light into an electric signal.
  • the excitation is performed. Observation using fluorescence can be performed while suppressing the influence of light.
  • FIG. 4 is a partially enlarged cross-sectional view of the light receiving unit 60.
  • the microscopic observation device 100 can image the observation target T arranged within a predetermined distance L from the vertex of the microlens 17.
  • the predetermined distance L is a distance determined by the viewing angle ⁇ of the microlens 17 and the distance P between the photodiode layers 12.
  • the predetermined distance L will be described.
  • the conditions under which the observation target T can be photographed are as follows, where S is the distance of the imaging surface of the observation target T to be guided by one photodiode layer 12, and P is the interval between the photodiode layers 12. Is represented by 2 ⁇ S ⁇ P (Equation 1)
  • the microscopic observation apparatus 100 photographs the observation target T in a so-called out-of-focus state.
  • the microscopic observation apparatus 100 can image the observation target T whose distance L from the vertex of the microlens 17 is within P / (2 ⁇ tan ⁇ ). In other words, if the observation target T is mounted on the mounting portion 18 so that the distance L from the vertex of the microlens 17 is within P / (2 ⁇ tan ⁇ ), the image is out of focus without performing the imaging operation. None.
  • the microscopic observation apparatus 100 sets the distance L from the vertex of the microlens 17 to the distance L. Can photograph the observation target T arranged within 5 ⁇ m.
  • the microscopic observation device 100 sets the distance L from the vertex of the microlens 17 to 10 ⁇ m. It is possible to photograph the observation target T arranged within the range.
  • the microscopic observation device 100 sets the distance L from the vertex of the microlens 17 to 50 ⁇ m. It is possible to photograph the observation target T arranged within the range.
  • the microscopic observation apparatus 100 captures an image of the observation target T whose distance L from the vertex of the microlens 17 is within P / (2 ⁇ tan ⁇ ). Can be done. Therefore, the thickness of the mounting portion 18 when observing on the surface of the mounting portion 18 needs to be within P / (2 ⁇ tan ⁇ ). Further, when the observation target T is inside the mounting portion 18, the observation target T whose distance L from the vertex of the microlens 17 is within P / (2 ⁇ tan ⁇ ) can be photographed.
  • the metal particles 14 forming the polarizing means 4 are formed between the photodiode layer 12 forming the light receiving unit 2 and the microlens 17.
  • the position of the polarizing means 4 is not particularly limited.
  • a polarization action pattern specifically, a resin or metal pattern having a reflection / absorption action may be applied. If it is a metal pattern, a known semiconductor process can be applied. Further, a material having a polarizing property may be used without using a pattern.
  • FIG. 5 is a partially enlarged cross-sectional view of a light receiving unit 60 which is a modification of FIG.
  • the polarizing means 4 is formed on the resin flattening layer 16 so as to cover the microlenses 17. Then, the mounting portion 18 is arranged on the polarizing means 4.
  • the polarizing means 4 is disposed between the micro lens 17 and the observation target T.
  • FIG. 6 is a partially enlarged cross-sectional view of a light receiving unit 60 which is another modified example of FIG.
  • a petri dish 18 ' is disposed above the microlens 17 as a mounting portion.
  • the bottom surface of the petri dish 18 ' serves as the polarizing means 4.
  • the polarizing means 4 is arranged between the microlens 17 and the observation target T.
  • FIG. 7 is an electric block diagram schematically showing a microscopic observation system using the microscopic observation device 100.
  • the microscopic observation system includes a microscopic observation device 100, a logic circuit unit 200 serving as a signal processing circuit, and a display device 300.
  • the logic circuit unit 200 performs color correction (white balance, color matrix) and noise correction (noise reduction, flaw correction) on a voltage signal (raw @ data) obtained by photoelectric conversion by the photodiode layer 12 of the microscopic observation apparatus 100. And performs predetermined signal processing such as image quality correction (edge enhancement, gamma correction), and outputs the signal-processed voltage signal as an image signal.
  • the logic circuit unit 200 since the microscopic observation apparatus 100 does not include a lens system for imaging or scaling, the logic circuit unit 200 includes a correction circuit for correcting such lens aberration and shading. You don't have to.
  • Such a logic circuit unit 200 may be built in the light receiving unit 60 by being formed around a region where the photodiode layer 12 is formed, for example, on the semiconductor substrate 11, or may be provided on a separate substrate from the light receiving unit 60.
  • the light receiving unit 60 may be provided as a separate component.
  • the display device 300 forms and displays an image of the observation target T based on the image signal output from the logic circuit unit 200.
  • the display device 300 can display the whole observation target T arranged on the mounting portion 18 of the microscopic observation device 100 at one time in real time.
  • the two polarization means 3 and 4 are arranged between the light source 1 and the light receiving unit 2, and the excitation light from the light source 1 passing through the polarization means 3 is cut by the polarization means 4. So that it hardly reaches the light receiving section 2. Therefore, the observation using the fluorescence from the observation target T can be performed while suppressing the influence of the excitation light.
  • a normal microscopic observation device for example, an optical microscope
  • a lens system for image formation and enlargement / reduction Therefore, when the polarizing means is provided in such a microscopic observation apparatus, the fluorescence from the observation target T becomes weaker when passing through the polarizing means, and further weakens when passing through the lens system. Therefore, when observing using fluorescence, there is no need to provide a polarizing means.
  • the microscopic observation device 100 does not include a lens system for imaging or enlargement / reduction, so that the polarization unit 4 can be provided.
  • the entire observation target T can be easily observed without the need to adjust the optical system such as focus and magnification, or scan the observation target T.

Landscapes

  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Optics & Photonics (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Multimedia (AREA)
  • General Health & Medical Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Signal Processing (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Microscoopes, Condenser (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Transforming Light Signals Into Electric Signals (AREA)
  • Studio Devices (AREA)

Abstract

The present invention provides a microscopic observation apparatus and a microscopic observation method which enable observation using fluorescence from an object to be observed irradiated with excitation light, and a method for manufacturing such a microscopic observation apparatus. Provided is a microscopic observation apparatus for observing fluorescence generated from an object to be observed by irradiating the object to be observed with excitation light. The microscopic observation apparatus is provided with: a light source which irradiates the object to be observed with the excitation light; a plurality of light receiving parts; a non-imaging lens system which guides the fluorescence from the object to be observed to each of the plurality of light receiving parts; and a first polarizing means and a second polarizing means which are disposed between the light source and the light receiving parts, wherein the first polarizing means and the second polarizing means have polarization properties configured such that the excitation light that has passed through the first polarizing means is cut by the second polarizing means.

Description

顕微観察装置、顕微観察方法および顕微観察装置の製造方法Microscopic observation device, microscopic observation method, and method of manufacturing microscopic observation device
　本発明は、顕微観察装置、顕微観察方法および顕微観察装置の製造方法に関する。 The present invention relates to a microscopic observation device, a microscopic observation method, and a method for manufacturing a microscopic observation device.
　従来の光学顕微鏡とは異なり、結像や拡大縮小といった光学系の調整および観察対象の走査を要することなく、観察対象の全体を簡易に観察できる観察方法が提案されている（特許文献１）。 Unlike the conventional optical microscope, there has been proposed an observation method capable of easily observing the entire observation target without adjusting the optical system such as imaging and enlarging and reducing and scanning the observation target (Patent Document 1).
特開２０１８－４２２８３号公報JP 2018-42283 A
　特許文献１に記載の観察方法は、観察対象に励起光を照射し、観察対象からの蛍光を観察することまでは想定していない。 観 察 The observation method described in Patent Document 1 does not assume irradiating an observation target with excitation light and observing fluorescence from the observation target.
　本発明はこのような問題点に鑑みてなされたものであり、本発明の課題は、励起光が照射された観察対象からの蛍光を利用した観察を行うことができる顕微観察装置および顕微観察方法、また、そのような顕微観察装置の製造方法を提供することである。 The present invention has been made in view of such problems, and an object of the present invention is to provide a microscopic observation apparatus and a microscopic observation method capable of performing observation using fluorescence from an observation target irradiated with excitation light. Another object of the present invention is to provide a method for manufacturing such a microscopic observation device.
　本発明の一態様によれば、観察対象に励起光を照射して前記観察対象から生じる蛍光を観察する顕微観察装置であって、前記観察対象に励起光を照射する光源と、半導体基板と、前記半導体基板に形成された複数の受光部と、前記半導体基板および前記複数の受光部を覆う保護層と、前記保護層上に形成され、金属を含む第１偏光手段と、前記保護層および前記金属粒子を覆う酸化物平坦化層と、前記酸化物平坦化層上に形成された樹脂平坦化層と、前記樹脂平坦化層上に形成され、前記複数の受光部のそれぞれに前記観察対象からの蛍光を導く複数の樹脂製マイクロレンズと、前記光源と前記観察対象との間に配置される第２偏光手段と、を備え、前記第１偏光手段および前記第２偏光手段の偏光特性は、前記第２偏光手段を通過した励起光が前記第１偏光手段でカットされるように構成される、顕微観察装置が提供される。 According to one aspect of the present invention, a microscopic observation apparatus that irradiates an observation target with excitation light and observes fluorescence generated from the observation target, and a light source that irradiates the observation target with excitation light, a semiconductor substrate, A plurality of light receiving portions formed on the semiconductor substrate, a protection layer covering the semiconductor substrate and the plurality of light receiving portions, a first polarizing means formed on the protection layer and containing a metal, An oxide flattening layer covering metal particles, a resin flattening layer formed on the oxide flattening layer, and a resin flattening layer formed on the resin flattening layer, and each of the plurality of light receiving units is formed from the observation target. A plurality of resin microlenses for guiding the fluorescence, and a second polarizing means disposed between the light source and the observation target, wherein the polarization characteristics of the first polarizing means and the second polarizing means, The excitation having passed through the second polarizing means Configured such that light is cut by the first polarizing means, microscopic observation device is provided.
　本発明の別の態様によれば、観察対象に励起光を照射して前記観察対象から生じる蛍光を観察する顕微観察装置であって、前記観察対象に励起光を照射する光源と、複数の受光部と、前記複数の受光部のそれぞれに前記観察対象からの蛍光を導く非結像レンズ系と、前記観察対象と前記受光部との間に配置される第１偏光手段と、前記光源と前記観察対象との間に配置される第２偏光手段と、を備え、前記第１偏光手段および前記第２偏光手段の偏光特性は、前記第２偏光手段を通過した前記励起光が前記第１偏光手段でカットされるように構成される、顕微観察装置が提供される。 According to another aspect of the present invention, there is provided a microscopic observation apparatus for irradiating an observation target with excitation light and observing fluorescence generated from the observation target, comprising: a light source for irradiating the observation target with excitation light; Unit, a non-imaging lens system for guiding fluorescence from the observation target to each of the plurality of light receiving units, a first polarizing unit disposed between the observation target and the light receiving unit, the light source and the light source A second polarization unit disposed between the first polarization unit and the observation target, wherein the polarization characteristics of the first polarization unit and the second polarization unit are such that the excitation light that has passed through the second polarization unit is the first polarization unit. A microscopic observation device configured to be cut by means is provided.
　前記非結像レンズ系の上方に配置され、底面が前記第２偏光手段である載置部を備えてもよい。 を A mounting portion may be provided above the non-imaging lens system and a bottom surface may be the second polarizing means.
　前記光源と前記受光部との間に、結像用および拡大縮小用のレンズ系が配置されないのが望ましい。 レ ン ズ It is desirable that no lens system for image formation and enlargement / reduction is arranged between the light source and the light receiving unit.
　また、本発明の別の態様によれば、観察対象に励起光を照射して前記観察対象から生じる蛍光を観察する顕微観察装置を製造する方法であって、前記観察対象に励起光を照射する光源、および、前記光源に取り付けられた第１偏光手段を有する光源ユニットと、複数の受光部、前記複数の受光部のそれぞれに前記観察対象からの蛍光を導く非結像レンズ系、および、前記受光部の上方に配置される第２偏光手段を有する受光ユニットと、を、前記第１偏光手段および前記第２偏光手段の偏光特性によって、前記第１偏光手段を通過した前記励起光が前記第２偏光手段でカットされるよう、取り付ける、顕微観察装置の製造方法が提供される。 According to another aspect of the present invention, there is provided a method of manufacturing a microscopic observation apparatus that irradiates an observation target with excitation light to observe fluorescence generated from the observation target, and irradiates the observation target with excitation light. A light source, and a light source unit having a first polarizing unit attached to the light source, a plurality of light receiving units, a non-imaging lens system that guides fluorescence from the observation target to each of the plurality of light receiving units, and A light-receiving unit having a second polarizing means disposed above a light-receiving section, wherein the excitation light having passed through the first polarizing means is changed to the second light by the polarization characteristics of the first polarizing means and the second polarizing means. A method for manufacturing a microscopic observation device, which is attached so as to be cut by two-polarizing means, is provided.
　また、本発明の別の態様によれば、観察対象に励起光を照射して前記観察対象から生じる蛍光を観察する顕微観察方法であって、顕微観察装置における非結像レンズ系の上方に配置された載置部に観察対象を載置することと、光源から第１偏光手段を介して励起光を前記観察対象に照射することと、前記第１偏光手段を通過した励起光を第２偏光手段の偏光特性を利用してカットしつつ、前記観察対象からの蛍光を前記非結像レンズ系によって前記受光部に導くことと、を備える顕微観察方法が提供される。 According to another aspect of the present invention, there is provided a microscopic observation method for irradiating an observation target with excitation light and observing fluorescence generated from the observation target, the method being arranged above a non-imaging lens system in the microscopic observation apparatus. Placing the observation target on the placed mounting portion, irradiating the observation target with excitation light from the light source via the first polarization unit, and converting the excitation light passing through the first polarization unit into the second polarization unit. Guiding the fluorescence from the observation target to the light receiving unit by the non-imaging lens system while cutting by utilizing the polarization characteristics of the means.
　励起光が照射された観察対象からの蛍光を利用した観察を行うことができる。 観 察 Observation using fluorescence from the observation target irradiated with the excitation light can be performed.
本発明に係る顕微観察の概念を説明する図。The figure explaining the concept of the microscopic observation concerning the present invention. 偏光手段３，４の関係を模式的に示す図。The figure which shows the relationship of the polarization means 3 and 4 typically. 偏光手段３，４の関係を模式的に示す図。The figure which shows the relationship of the polarization means 3 and 4 typically. 一実施形態に係る顕微観察装置１００の概略構成を示す断面図。FIG. 1 is a sectional view showing a schematic configuration of a microscopic observation apparatus 100 according to one embodiment. 図３Ａの顕微鏡観察装置１００を含む顕微鏡観察システムのシステム構成図。FIG. 3B is a system configuration diagram of a microscope observation system including the microscope observation device 100 of FIG. 3A. 受光ユニット６０の一部拡大図。FIG. 3 is a partially enlarged view of the light receiving unit 60. 図３の変形例である受光ユニット６０の一部拡大断面図。FIG. 4 is a partially enlarged cross-sectional view of a light receiving unit 60 which is a modified example of FIG. 3. 図３の別の変形例である受光ユニット６０の一部拡大断面図。FIG. 4 is a partially enlarged cross-sectional view of a light receiving unit 60 which is another modified example of FIG. 3. 顕微観察装置１００を用いた顕微観察システムを模式的に示す電気ブロック図。FIG. 1 is an electric block diagram schematically showing a microscopic observation system using a microscopic observation device 100.
　図１は、本発明に係る顕微観察の概念を説明する図である。顕微観察装置１００は、光源１と、受光部２と、偏光手段３，４とを備えている。光源１は観察対象Ｔに対して励起光Ｌ１を照射する。観察対象Ｔは励起光Ｌ１によって励起され蛍光Ｌ２を発する。受光部２は観察対象Ｔからの蛍光Ｌ２を受け、電気信号に変換する。電気信号に基づいて画像を形成することで観察対象Ｔを観察できる。 FIG. 1 is a diagram illustrating the concept of microscopic observation according to the present invention. The microscopic observation device 100 includes a light source 1, a light receiving unit 2, and polarizing units 3 and 4. The light source 1 irradiates the observation target T with the excitation light L1. The observation target T is excited by the excitation light L1 and emits fluorescence L2. The light receiving unit 2 receives the fluorescence L2 from the observation target T and converts it into an electric signal. The observation target T can be observed by forming an image based on the electric signal.
　ここで、光源１からの励起光Ｌ１に比べて観察対象Ｔからの蛍光Ｌ２は弱いことが多い。そのため、仮に光源１からの励起光Ｌ１が受光部２に達すると、励起光Ｌ１の影響が強すぎて、蛍光Ｌ２を利用した正確な観察が困難となる。 Here, the fluorescence L2 from the observation target T is often weaker than the excitation light L1 from the light source 1. Therefore, if the excitation light L1 from the light source 1 reaches the light receiving unit 2, the influence of the excitation light L1 is too strong, and accurate observation using the fluorescence L2 becomes difficult.
　そこで、本発明では、光源１と受光部２との間に２つの偏光手段３，４を設ける。偏光手段３，４は、例えば電気的あるいは機械的に偏光作用を切り替えることができる。そして、偏光手段３，４の偏光特性を利用して、光源１側の偏光手段３を通過した励起光Ｌ１が受光部２側の偏光手段４によってカットされるようにする。 Therefore, in the present invention, two polarizing means 3 and 4 are provided between the light source 1 and the light receiving section 2. The polarization means 3 and 4 can switch the polarization action electrically or mechanically, for example. Then, by utilizing the polarization characteristics of the polarization units 3 and 4, the excitation light L1 passing through the polarization unit 3 on the light source 1 side is cut by the polarization unit 4 on the light receiving unit 2 side.
　一例として、図２Ａに模式図を示すように、光源１側の偏光手段３が所定方向（同図では縦方向）の直線偏光のみを通過させ、受光部２側の偏光手段４が異なる方向（同図では横方向）の直線偏光のみを通過させるように偏光手段３，４を構成・配置することができる。 As an example, as shown in the schematic diagram of FIG. 2A, the polarizing means 3 on the light source 1 side transmits only linearly polarized light in a predetermined direction (vertical direction in FIG. The polarizing means 3 and 4 can be configured and arranged to pass only linearly polarized light (in the horizontal direction in the figure).
　別の例として、図２Ｂに模式図を示すように、光源１側の偏光手段３が所定方向（同図では時計回り）の円偏光のみを通過させ、受光部２側の偏光手段４が反対方向（同図では反時計回り）の円偏光のみを通過させるように偏光手段３，４を構成・配置してもよい。 As another example, as shown in a schematic diagram in FIG. 2B, the polarizing means 3 on the light source 1 side transmits only circularly polarized light in a predetermined direction (clockwise in FIG. 2), and the polarizing means 4 on the light receiving unit 2 side is opposite. The polarization means 3 and 4 may be configured and arranged to pass only circularly polarized light in the direction (counterclockwise in the figure).
　以下、顕微観察装置１００の具体的な構成例を説明する。
　図３Ａは、一実施形態に係る顕微観察装置１００の概略構成を示す断面図である。顕微観察装置１００は、光源ユニット５０と、受光ユニット６０とを備えている。 Hereinafter, a specific configuration example of the microscopic observation apparatus 100 will be described.
FIG. 3A is a cross-sectional view illustrating a schematic configuration of the microscopic observation device 100 according to one embodiment. The microscopic observation device 100 includes a light source unit 50 and a light receiving unit 60.
　光源ユニット５０は、図１に示した光源１と、偏光手段３とを一体化したものである。 The light source unit 50 integrates the light source 1 shown in FIG.
　受光ユニット６０は、半導体基板１１と、複数のフォトダイオード層１２と、保護層１３と、金属粒子１４と、酸化物平坦化層１５と、樹脂平坦化層１６と、複数のマイクロレンズ１７と、載置部１８とを有する。 The light receiving unit 60 includes a semiconductor substrate 11, a plurality of photodiode layers 12, a protective layer 13, metal particles 14, an oxide planarization layer 15, a resin planarization layer 16, a plurality of microlenses 17, And a mounting portion 18.
　半導体基板１１はシリコン基板などである。フォトダイオード層１２は半導体基板１１に形成された不純物拡散層であり、例えばｎ型の半導体基板１１に対してｐ型の不純物を注入することによってｐｎ接合によるフォトダイオードを形成する。複数のフォトダイオード層１２が互いに離間してマトリクス状に形成され、図１における受光部２を構成する。なお、受光部２は入射される光に対して光電変換を行うものであればよく、必ずしもフォトダイオード層１２である必要はない。 The semiconductor substrate 11 is a silicon substrate or the like. The photodiode layer 12 is an impurity diffusion layer formed in the semiconductor substrate 11, and forms a pn junction photodiode by injecting a p-type impurity into the n-type semiconductor substrate 11, for example. A plurality of photodiode layers 12 are formed apart from each other in a matrix and constitute the light receiving section 2 in FIG. The light receiving section 2 only needs to perform photoelectric conversion on incident light, and does not necessarily need to be the photodiode layer 12.
　保護層１３は、例えばシリコン酸化膜あるいはシリコン窒化膜であり、フォトダイオード層１２および半導体基板１１の露出した部分を覆っている。 The protection layer 13 is, for example, a silicon oxide film or a silicon nitride film, and covers the exposed portions of the photodiode layer 12 and the semiconductor substrate 11.
　金属粒子１４はフォトダイオード層１２より小さい粒子であり、保護層１３上に形成される。半導体基板１１上あるいはフォトダイオード層１２上に直接金属粒子１４を形成してもよいが、保護層１３を設けるのが望ましい。複数の金属粒子１４が図１における偏光手段４として機能する。図１を用いて説明したように、偏光手段４は偏光手段３を通過した励起光をカットするように構成される。 The metal particles 14 are smaller than the photodiode layer 12 and are formed on the protective layer 13. Although the metal particles 14 may be formed directly on the semiconductor substrate 11 or the photodiode layer 12, it is preferable to provide the protective layer 13. The plurality of metal particles 14 function as the polarization unit 4 in FIG. As described with reference to FIG. 1, the polarization unit 4 is configured to cut the excitation light that has passed through the polarization unit 3.
　酸化物平坦化層１５は、例えばシリコン酸化膜であり、金属粒子１４および保護層１３の露出した部分を覆っている。樹脂平坦化層１６は酸化物平坦化層１５上に形成される。樹脂平坦化層１６上に樹脂製で複数のマイクロレンズ１７がマトリクス状に形成される。酸化物平坦化層１５上に直接マイクロレンズ１７を形成してもよいが、樹脂平坦化層１６を設けることで、マイクロレンズ１７を樹脂平坦化層１６に密着させることができる。 The oxide planarization layer 15 is, for example, a silicon oxide film, and covers the exposed portions of the metal particles 14 and the protection layer 13. The resin planarization layer 16 is formed on the oxide planarization layer 15. A plurality of microlenses 17 made of resin are formed in a matrix on the resin flattening layer 16. Although the microlenses 17 may be formed directly on the oxide flattening layer 15, the microlenses 17 can be adhered to the resin flattening layer 16 by providing the resin flattening layer 16.
　マイクロレンズ１７のそれぞれはフォトダイオード層１２のそれぞれの直上に形成される。１つのフォトダイオード層１２と、対応する１つのマイクロレンズ１７とで画素が構成される。マイクロレンズ１７は非結像レンズ系であり、フォトダイオード層１２上に結像する必要はなく、特定の視野角θ（後述）から入射する光をフォトダイオード層１２に導くものである。なお、マイクロレンズ１７以外の視野角制御が可能な任意の層を適用してもよい。 Each of the microlenses 17 is formed immediately above each of the photodiode layers 12. One photodiode layer 12 and one corresponding microlens 17 form a pixel. The micro lens 17 is a non-imaging lens system, and does not need to form an image on the photodiode layer 12, and guides light incident from a specific viewing angle θ (described later) to the photodiode layer 12. Note that any layer other than the microlens 17 that can control the viewing angle may be applied.
　載置部１８は観察対象Ｔが載置される部分であり、例えばマイクロレンズ１７上に形成され樹脂製で板状のものであってもよいし、マイクロレンズ１７に埋め込まれる液状あるいは固体状のものであってもよい。 The mounting portion 18 is a portion on which the observation target T is mounted, for example, may be formed on the microlens 17 and may be made of resin and a plate shape, or may be a liquid or solid state embedded in the microlens 17. It may be something.
　なお、載置部１８、樹脂平坦化層１６、酸化物平坦化層１５および保護層１３は、少なくとも観察対象Ｔからの蛍光を透過させることができる。また、顕微観察装置１００には、光源１とフォトダイオード層１２との間に結像用あるいは拡大縮小用のレンズ系を設ける必要はない。 The mounting portion 18, the resin flattening layer 16, the oxide flattening layer 15, and the protective layer 13 can transmit at least the fluorescence from the observation target T. Further, in the microscopic observation apparatus 100, it is not necessary to provide a lens system for imaging or enlargement / reduction between the light source 1 and the photodiode layer 12.
　光源ユニット５０と受光ユニット６０とを取り付けることで顕微観察装置１００が製造される。より具体的には、光源ユニット５０における偏光手段３および受光ユニット６０における偏光手段４の偏光特性によって、偏光手段３を通過した励起光が偏光手段４でカットされるよう、光源ユニット５０と受光ユニット６０とを取り付ける。 顕 The microscopic observation device 100 is manufactured by attaching the light source unit 50 and the light receiving unit 60. More specifically, the light source unit 50 and the light receiving unit are arranged such that the excitation light passing through the polarizing unit 3 is cut by the polarizing unit 4 by the polarization characteristics of the polarizing unit 3 in the light source unit 50 and the polarizing unit 4 in the light receiving unit 60. And 60.
　図３Ｂは、図３Ａの顕微鏡観察装置１００を含む顕微鏡観察システムのシステム構成図である。図示のように、顕微鏡観察システムは、光源１を制御する光源制御部７１と、偏光手段３（および／または偏光手段４）の偏光作用を電気的あるいは機械的に切り替える偏光制御部７２と、受光ユニット６０を制御する受光ユニット制御部７３と、全体制御装置７０とを備えている。全体制御装置７０は、光源制御部７１、偏光制御部７２および受光ユニット制御部７３を制御するものであり、例えばパーソナルコンピュータやマイクロコンピュータによって構成される。 FIG. 3B is a system configuration diagram of a microscope observation system including the microscope observation device 100 of FIG. 3A. As shown in the drawing, the microscope observation system includes a light source control unit 71 that controls the light source 1, a polarization control unit 72 that electrically or mechanically switches the polarization operation of the polarization unit 3 (and / or the polarization unit 4), A light receiving unit controller 73 for controlling the unit 60 and an overall controller 70 are provided. The general control device 70 controls the light source control unit 71, the polarization control unit 72, and the light receiving unit control unit 73, and is configured by, for example, a personal computer or a microcomputer.
　このような顕微観察装置１００を用い、次のようにして顕微観察が行われる。まず、観察対象Ｔを載置部１８上に載置する。次いで、光源１から偏光手段３を介して励起光を観察対象Ｔに向かって照射する。光源１からの励起光のうち特定の偏光状態のもののみが偏光手段３を通過し、観察対象Ｔに到達する。 顕 Using such a microscopic observation apparatus 100, microscopic observation is performed as follows. First, the observation target T is placed on the placement unit 18. Next, excitation light is emitted from the light source 1 to the observation target T via the polarizing means 3. Only the excitation light having a specific polarization state among the excitation lights from the light source 1 passes through the polarization means 3 and reaches the observation target T.
　観察対象Ｔは励起光によって励起されて蛍光を発する。この蛍光は各マイクロレンズ１７によって、対応するフォトダイオード層１２に導かれる。 The observation target T emits fluorescence when excited by the excitation light. This fluorescence is guided to the corresponding photodiode layer 12 by each micro lens 17.
　一方、光源１から偏光手段３を通過した励起光は金属粒子１４から構成される偏光手段４によってカットされ、フォトダイオード層１２にはほとんど到達しない。 On the other hand, the excitation light transmitted from the light source 1 through the polarizing means 3 is cut by the polarizing means 4 composed of the metal particles 14 and hardly reaches the photodiode layer 12.
　これにより、フォトダイオード層１２に入る光は観察対象Ｔからの蛍光が支配的となり、当該光を電気信号に変換することで、観察対象Ｔを観察できる。このように、光源１とフォトダイオード層１２との間に、偏光手段３と、金属粒子１４から構成される偏光手段４と配置された状態で励起光を観察対象Ｔに照射することで、励起光の影響を抑えて蛍光を利用した観察が可能となる。 (4) Thereby, the light entering the photodiode layer 12 is dominated by the fluorescence from the observation target T, and the observation target T can be observed by converting the light into an electric signal. By irradiating the observation target T with the excitation light in a state where the polarizing means 3 and the polarizing means 4 including the metal particles 14 are arranged between the light source 1 and the photodiode layer 12 in this manner, the excitation is performed. Observation using fluorescence can be performed while suppressing the influence of light.
　図４は、受光ユニット６０の一部拡大断面図である。顕微観察装置１００は、マイクロレンズ１７の頂点から所定の距離Ｌ以内に配置される観察対象Ｔを撮影できるものである。ここで、所定の距離Ｌとは、マイクロレンズ１７の視野角θおよびフォトダイオード層１２の間隔Ｐにより決定される距離である。以下に、所定の距離Ｌについて説明する。 FIG. 4 is a partially enlarged cross-sectional view of the light receiving unit 60. The microscopic observation device 100 can image the observation target T arranged within a predetermined distance L from the vertex of the microlens 17. Here, the predetermined distance L is a distance determined by the viewing angle θ of the microlens 17 and the distance P between the photodiode layers 12. Hereinafter, the predetermined distance L will be described.
　観察対象Ｔを撮影できる条件は、図４に示すように、１つのフォトダイオード層１２によって導かれるべき観察対象Ｔの撮影面の距離をＳ、フォトダイオード層１２の間隔をＰとして、以下のように表現される。
　２×Ｓ≦Ｐ・・・（式１） As shown in FIG. 4, the conditions under which the observation target T can be photographed are as follows, where S is the distance of the imaging surface of the observation target T to be guided by one photodiode layer 12, and P is the interval between the photodiode layers 12. Is represented by
2 × S ≦ P (Equation 1)
　撮影面の距離Ｓが上記式１を満足しない場合、観察対象Ｔからの光は、本来受光されるフォトダイオード層１２において受光される他、当該フォトダイオード層１２に隣接する他のフォトダイオード層１２においても受光されてしまうこととなり、顕微観察装置１００は観察対象Ｔを、いわゆるピンボケした状態で撮影してしまう。 When the distance S of the photographing surface does not satisfy the above equation 1, the light from the observation target T is received by the photodiode layer 12 which is originally received, and the other photodiode layers 12 adjacent to the photodiode layer 12 In this case, the microscopic observation apparatus 100 photographs the observation target T in a so-called out-of-focus state.
　ここで、観察対象Ｔの撮影面の距離Ｓは、マイクロレンズ１７の視野角をθ、マイクロレンズ１７の頂点から観察対象Ｔまでの距離をＬとして、以下のように表現される。
　ｔａｎθ＝Ｓ／Ｌ・・・（式２） Here, the distance S between the imaging surface of the observation target T and the viewing angle of the microlens 17 is represented by θ, and the distance from the vertex of the microlens 17 to the observation target T is represented by L as follows.
tan θ = S / L (formula 2)
　以上の式１および式２より、マイクロレンズ１７の頂点から観察対象Ｔまでの距離Ｌは、以下のように表現される。
　Ｌ≦Ｐ／（２×ｔａｎθ）・・・（式３） From the above equations 1 and 2, the distance L from the vertex of the microlens 17 to the observation target T is expressed as follows.
L ≦ P / (2 × tan θ) (Equation 3)
　式３より、本実施形態に係る顕微観察装置１００は、マイクロレンズ１７の頂点からの距離Ｌが、Ｐ／（２×ｔａｎθ）以内に配置される観察対象Ｔを撮影できる。言い換えると、マイクロレンズ１７の頂点からの距離ＬがＰ／（２×ｔａｎθ）以内となるよう観察対象Ｔを載置部１８上に載置すれば、結像操作を行わなくても、ピンボケすることはない。 よ り From Expression 3, the microscopic observation apparatus 100 according to the present embodiment can image the observation target T whose distance L from the vertex of the microlens 17 is within P / (2 × tan θ). In other words, if the observation target T is mounted on the mounting portion 18 so that the distance L from the vertex of the microlens 17 is within P / (2 × tan θ), the image is out of focus without performing the imaging operation. Never.
　例えば、フォトダイオード層１２の間隔Ｐ＝１．７６μｍ（Ｓ＝０．８８μｍ）、θ＝１０ｄｅｇである場合、Ｌ≦５μｍとなるため、顕微観察装置１００は、マイクロレンズ１７の頂点からの距離Ｌが５μｍ以内に配置された観察対象Ｔを撮影することができる。 For example, when the distance P between the photodiode layers 12 is 1.76 μm (S = 0.88 μm) and θ = 10 deg, L ≦ 5 μm, so that the microscopic observation apparatus 100 sets the distance L from the vertex of the microlens 17 to the distance L. Can photograph the observation target T arranged within 5 μm.
　フォトダイオード層１２の間隔Ｐ＝３．５２μｍ（Ｓ＝１．７６μｍ）、θ＝１０ｄｅｇである場合、Ｌ≦１０μｍとなるため、顕微観察装置１００は、マイクロレンズ１７の頂点からの距離Ｌが１０μｍ以内に配置された観察対象Ｔを撮影することができる。 When the interval P between the photodiode layers 12 is 3.52 μm (S = 1.76 μm) and θ = 10 deg, L ≦ 10 μm. Therefore, the microscopic observation device 100 sets the distance L from the vertex of the microlens 17 to 10 μm. It is possible to photograph the observation target T arranged within the range.
　フォトダイオード層１２の間隔Ｐ＝１７．６４μｍ（Ｓ＝８．８２μｍ）、θ＝１０ｄｅｇである場合、Ｌ≦５０μｍとなるため、顕微観察装置１００は、マイクロレンズ１７の頂点からの距離Ｌが５０μｍ以内に配置された観察対象Ｔを撮影することができる。 When the distance P between the photodiode layers 12 is 17.64 μm (S = 8.82 μm) and θ = 10 deg, L ≦ 50 μm. Therefore, the microscopic observation device 100 sets the distance L from the vertex of the microlens 17 to 50 μm. It is possible to photograph the observation target T arranged within the range.
　なお、以上に説明したように、本実施形態に係る顕微観察装置１００は、マイクロレンズ１７の頂点からの距離Ｌが、Ｐ／（２×ｔａｎθ）以内に配置される観察対象Ｔを撮影することができるものである。したがって、載置部１８の表面上で観察する場合の載置部１８の厚さは、Ｐ／（２×ｔａｎθ）以内である必要がある。また、載置部１８の内部に観察対象Ｔがある場合は、マイクロレンズ１７の頂点からの距離Ｌが、Ｐ／（２×ｔａｎθ）以内に配置される観察対象Ｔを撮影できる。 As described above, the microscopic observation apparatus 100 according to the present embodiment captures an image of the observation target T whose distance L from the vertex of the microlens 17 is within P / (2 × tan θ). Can be done. Therefore, the thickness of the mounting portion 18 when observing on the surface of the mounting portion 18 needs to be within P / (2 × tan θ). Further, when the observation target T is inside the mounting portion 18, the observation target T whose distance L from the vertex of the microlens 17 is within P / (2 × tan θ) can be photographed.
　なお、以上説明した図３の受光ユニット６０では、受光部２を構成するフォトダイオード層１２と、マイクロレンズ１７との間に、偏光手段４を構成する金属粒子１４を形成する例を示したが、偏光手段４の位置に特に制限はない。例えば、金属粒子１４に代えて、偏光作用パターン、具体的には反射・吸収作用のある樹脂あるいは金属パターンを適用してもよい。金属パターンであれば、公知の半導体プロセスを適用できる。また、パターンを用いなくとも偏光性を有する材料を用いてもよい。 In the light receiving unit 60 of FIG. 3 described above, an example is shown in which the metal particles 14 forming the polarizing means 4 are formed between the photodiode layer 12 forming the light receiving unit 2 and the microlens 17. The position of the polarizing means 4 is not particularly limited. For example, instead of the metal particles 14, a polarization action pattern, specifically, a resin or metal pattern having a reflection / absorption action may be applied. If it is a metal pattern, a known semiconductor process can be applied. Further, a material having a polarizing property may be used without using a pattern.
　図５は、図３の変形例である受光ユニット６０の一部拡大断面図である。この受光ユニット６０は、マイクロレンズ１７を覆うように樹脂平坦化層１６上に偏光手段４が形成される。そして、偏光手段４上に載置部１８が配置される。この受光ユニット６０では、マイクロレンズ１７と観察対象Ｔとの間に偏光手段４が配置されることとなる。 FIG. 5 is a partially enlarged cross-sectional view of a light receiving unit 60 which is a modification of FIG. In this light receiving unit 60, the polarizing means 4 is formed on the resin flattening layer 16 so as to cover the microlenses 17. Then, the mounting portion 18 is arranged on the polarizing means 4. In the light receiving unit 60, the polarizing means 4 is disposed between the micro lens 17 and the observation target T.
　図６は、図３の別の変形例である受光ユニット６０の一部拡大断面図である。この受光ユニット６０は、シャーレ１８’が載置部としてマイクロレンズ１７の上方に配置される。そして、そのシャーレ１８’の底面が偏光手段４となっている。この場合も、マイクロレンズ１７と観察対象Ｔとの間に偏光手段４が配置されることとなる。 FIG. 6 is a partially enlarged cross-sectional view of a light receiving unit 60 which is another modified example of FIG. In the light receiving unit 60, a petri dish 18 'is disposed above the microlens 17 as a mounting portion. The bottom surface of the petri dish 18 'serves as the polarizing means 4. Also in this case, the polarizing means 4 is arranged between the microlens 17 and the observation target T.
　図７は、顕微観察装置１００を用いた顕微観察システムを模式的に示す電気ブロック図である。顕微観察システムは、顕微観察装置１００、信号処理回路であるロジック回路部２００、および、表示装置３００によって構成される。 FIG. 7 is an electric block diagram schematically showing a microscopic observation system using the microscopic observation device 100. The microscopic observation system includes a microscopic observation device 100, a logic circuit unit 200 serving as a signal processing circuit, and a display device 300.
　ロジック回路部２００は、顕微観察装置１００のフォトダイオード層１２による光電変換により得られた電圧信号（ｒａｗ　ｄａｔａ）に対して色補正（ホワイトバランス、カラーマトリクス）、ノイズ補正（ノイズリダクション、傷補正）、画質補正（エッジ強調、ガンマ補正）など所定の信号処理を施し、信号処理された電圧信号を画像信号として出力する。 The logic circuit unit 200 performs color correction (white balance, color matrix) and noise correction (noise reduction, flaw correction) on a voltage signal (raw @ data) obtained by photoelectric conversion by the photodiode layer 12 of the microscopic observation apparatus 100. And performs predetermined signal processing such as image quality correction (edge enhancement, gamma correction), and outputs the signal-processed voltage signal as an image signal.
　本実施形態においては、顕微観察装置１００に結像用あるいは拡大縮小用のレンズ系が含まれないため、ロジック回路部２００には、このようなレンズ収差の補正やシェーディング補正するための補正回路はなくてよい。 In the present embodiment, since the microscopic observation apparatus 100 does not include a lens system for imaging or scaling, the logic circuit unit 200 includes a correction circuit for correcting such lens aberration and shading. You don't have to.
　このようなロジック回路部２００は、例えば半導体基板１１において、フォトダイオード層１２が形成された領域の周囲に形成することによって受光ユニット６０に内蔵させてもよいし、受光ユニット６０とは別基板に設けられた、受光ユニット６０とは別部品あってもよい。 Such a logic circuit unit 200 may be built in the light receiving unit 60 by being formed around a region where the photodiode layer 12 is formed, for example, on the semiconductor substrate 11, or may be provided on a separate substrate from the light receiving unit 60. The light receiving unit 60 may be provided as a separate component.
　また、表示装置３００はロジック回路部２００から出力される画像信号に基づいて観察対象Ｔの画像を形成し、表示する。表示装置３００は顕微観察装置１００の載置部１８に配置された観察対象Ｔの全体を一度にリアルタイム表示できる。 (4) The display device 300 forms and displays an image of the observation target T based on the image signal output from the logic circuit unit 200. The display device 300 can display the whole observation target T arranged on the mounting portion 18 of the microscopic observation device 100 at one time in real time.
　以上説明したように、本実施形態では、光源１と受光部２との間に２つの偏光手段３，４を配置し、偏光手段３を通過した光源１からの励起光が偏光手段４によってカットされ、受光部２にほとんど達しないようにする。そのため、励起光の影響を抑えて観察対象Ｔからの蛍光を利用した観察を行うことができる。 As described above, in the present embodiment, the two polarization means 3 and 4 are arranged between the light source 1 and the light receiving unit 2, and the excitation light from the light source 1 passing through the polarization means 3 is cut by the polarization means 4. So that it hardly reaches the light receiving section 2. Therefore, the observation using the fluorescence from the observation target T can be performed while suppressing the influence of the excitation light.
　なお、通常の顕微観察装置（例えば、光学顕微鏡）には、結像用や拡大縮小用のレンズ系が設けられる。そのため、このような顕微観察装置に偏光手段を設けると、観察対象Ｔからの蛍光は、偏光手段を通過する際に弱くなり、上記レンズ系を通過する際にさらに弱くなる。そのため、蛍光を利用した観察を行う際には、偏光手段を設けるようなことは行われない。 通常 Note that a normal microscopic observation device (for example, an optical microscope) is provided with a lens system for image formation and enlargement / reduction. Therefore, when the polarizing means is provided in such a microscopic observation apparatus, the fluorescence from the observation target T becomes weaker when passing through the polarizing means, and further weakens when passing through the lens system. Therefore, when observing using fluorescence, there is no need to provide a polarizing means.
　しかしながら、本実施形態の構成によれば、顕微観察装置１００は結像用や拡大縮小用のレンズ系を備えないため、偏光手段４を設けることが可能となる。しかも、ピントや倍率調整といった光学系の調整や、観察対象Ｔの走査を要することなく、観察対象Ｔの全体を簡易に観察できる。 However, according to the configuration of the present embodiment, the microscopic observation device 100 does not include a lens system for imaging or enlargement / reduction, so that the polarization unit 4 can be provided. In addition, the entire observation target T can be easily observed without the need to adjust the optical system such as focus and magnification, or scan the observation target T.
　上述した実施形態は、本発明が属する技術分野における通常の知識を有する者が本発明を実施できることを目的として記載されたものである。上記実施形態の種々の変形例は、当業者であれば当然になしうることであり、本発明の技術的思想は他の実施形態にも適用しうることである。したがって、本発明は、記載された実施形態に限定されることはなく、特許請求の範囲によって定義される技術的思想に従った最も広い範囲とすべきである。 The above embodiments are described for the purpose of allowing anyone having ordinary knowledge in the technical field to which the present invention pertains to implement the present invention. Various modifications of the above embodiment can be naturally made by those skilled in the art, and the technical idea of the present invention can be applied to other embodiments. Accordingly, the invention is not limited to the embodiments described, but is to be accorded the widest scope consistent with the spirit as defined by the appended claims.
１　光源
２　受光部
３，４　偏光手段
１１　半導体基板
１２　フォトダイオード層
１３　保護層
１４　金属粒子
１５　酸化膜平坦化層
１６　樹脂平坦化層
１７　マイクロレンズ
１８　載置部
１８’　シャーレ
５０　光源ユニット
６０　受光ユニット
７０　全体制御装置
７１　光源制御部
７２　偏光制御部
７３　受光ユニット制御部
１００　顕微観察装置
２００　ロジック回路
３００　表示装置 REFERENCE SIGNS LIST 1 light source 2 light receiving sections 3 and 4 polarizing means 11 semiconductor substrate 12 photodiode layer 13 protective layer 14 metal particles 15 oxide film flattening layer 16 resin flattening layer 17 microlens 18 mounting section 18 'petri dish light source unit 60 light receiving unit 70 overall control device 71 light source control unit 72 polarization control unit 73 light receiving unit control unit 100 microscopic observation device 200 logic circuit 300 display device

Claims (6)

  1. 　観察対象に励起光を照射して前記観察対象から生じる蛍光を観察する顕微観察装置であって、
    　前記観察対象に励起光を照射する光源と、
    　半導体基板と、
    　前記半導体基板に形成された複数の受光部と、
    　前記半導体基板および前記複数の受光部を覆う保護層と、
    　前記保護層上に形成され、金属を含む第１偏光手段と、
    　前記保護層および前記金属粒子を覆う酸化物平坦化層と、
    　前記酸化物平坦化層上に形成された樹脂平坦化層と、
    　前記樹脂平坦化層上に形成され、前記複数の受光部のそれぞれに前記観察対象からの蛍光を導く複数の樹脂製マイクロレンズと、
    　前記光源と前記観察対象との間に配置される第２偏光手段と、を備え、
    　前記第１偏光手段および前記第２偏光手段の偏光特性は、前記第２偏光手段を通過した励起光が前記第１偏光手段でカットされるように構成される、顕微観察装置。     A microscopic observation apparatus that irradiates the observation target with excitation light and observes fluorescence generated from the observation target,
    A light source for irradiating the observation target with excitation light,
    A semiconductor substrate;
    A plurality of light receiving units formed on the semiconductor substrate,
    A protective layer covering the semiconductor substrate and the plurality of light receiving units;
    A first polarizing unit formed on the protective layer and including a metal;
    An oxide planarization layer covering the protective layer and the metal particles,
    A resin planarization layer formed on the oxide planarization layer,
    A plurality of resin microlenses formed on the resin flattening layer and guiding fluorescence from the observation target to each of the plurality of light receiving units,
    A second polarizing means disposed between the light source and the observation target,
    The microscopic observation apparatus, wherein the polarization characteristics of the first polarization unit and the second polarization unit are configured such that the excitation light passing through the second polarization unit is cut by the first polarization unit.
  2. 　観察対象に励起光を照射して前記観察対象から生じる蛍光を観察する顕微観察装置であって、
    　前記観察対象に励起光を照射する光源と、
    　複数の受光部と、
    　前記複数の受光部のそれぞれに前記観察対象からの蛍光を導く非結像レンズ系と、
    　前記観察対象と前記受光部との間に配置される第１偏光手段と、
    　前記光源と前記観察対象との間に配置される第２偏光手段と、を備え、
    　前記第１偏光手段および前記第２偏光手段の偏光特性は、前記第２偏光手段を通過した前記励起光が前記第１偏光手段でカットされるように構成される、顕微観察装置。     A microscopic observation apparatus that irradiates the observation target with excitation light and observes fluorescence generated from the observation target,
    A light source for irradiating the observation target with excitation light,
    A plurality of light receiving sections,
    A non-imaging lens system that guides fluorescence from the observation target to each of the plurality of light receiving units,
    A first polarizing unit disposed between the observation target and the light receiving unit;
    A second polarizing means disposed between the light source and the observation target,
    The microscopic observation apparatus, wherein the polarization characteristics of the first polarization unit and the second polarization unit are configured such that the excitation light that has passed through the second polarization unit is cut by the first polarization unit.
  3. 　前記非結像レンズ系の上方に配置され、底面が前記第２偏光手段である載置部を備える、請求項２に記載の顕微観察装置。 The microscopic observation device according to claim 2, further comprising a mounting portion disposed above the non-imaging lens system and having a bottom surface serving as the second polarizing means.
  4. 　前記光源と前記受光部との間に、結像用および拡大縮小用のレンズ系が配置されない、請求項１乃至３のいずれかに記載の顕微観察装置。 The microscope observation apparatus according to any one of claims 1 to 3, wherein a lens system for imaging and enlargement / reduction is not arranged between the light source and the light receiving unit.
  5. 　観察対象に励起光を照射して前記観察対象から生じる蛍光を観察する顕微観察装置を製造する方法であって、
    　前記観察対象に励起光を照射する光源、および、前記光源に取り付けられた第１偏光手段を有する光源ユニットと、
    　複数の受光部、前記複数の受光部のそれぞれに前記観察対象からの蛍光を導く非結像レンズ系、および、前記受光部の上方に配置される第２偏光手段を有する受光ユニットと、を、
    　前記第１偏光手段および前記第２偏光手段の偏光特性によって、前記第１偏光手段を通過した前記励起光が前記第２偏光手段でカットされるよう、取り付ける、顕微観察装置の製造方法。     A method for manufacturing a microscopic observation apparatus for irradiating an observation target with excitation light and observing fluorescence generated from the observation target,
    A light source for irradiating the observation target with excitation light, and a light source unit having a first polarizing unit attached to the light source,
    A plurality of light receiving units, a non-imaging lens system that guides fluorescence from the observation target to each of the plurality of light receiving units, and a light receiving unit having a second polarizing unit disposed above the light receiving unit,
    A method for manufacturing a microscopic observation device, comprising: mounting the excitation light having passed through the first polarizing means so as to be cut by the second polarizing means by the polarization characteristics of the first polarizing means and the second polarizing means.
  6. 　観察対象に励起光を照射して前記観察対象から生じる蛍光を観察する顕微観察方法であって、
    　顕微観察装置における非結像レンズ系の上方に配置された載置部に観察対象を載置することと、
    　光源から第１偏光手段を介して励起光を前記観察対象に照射することと、
    　前記第１偏光手段を通過した励起光を第２偏光手段の偏光特性を利用してカットしつつ、前記観察対象からの蛍光を前記非結像レンズ系によって前記受光部に導くことと、を備える顕微観察方法。     A microscopic observation method of irradiating the observation target with excitation light and observing fluorescence generated from the observation target,
    Placing the observation target on a mounting portion arranged above the non-imaging lens system in the microscopic observation device;
    Irradiating the observation target with excitation light from the light source via the first polarizing means;
    Guiding the fluorescence from the observation target to the light receiving unit by the non-imaging lens system while cutting the excitation light that has passed through the first polarizing unit using the polarization characteristics of the second polarizing unit. Microscopic observation method.
PCT/JP2019/026801 2018-07-06 2019-07-05 Microscopic observation apparatus, microscopic observation method, and method for manufacturing microscopic observation apparatus WO2020009220A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2018129018A JP2020008682A (en) 2018-07-06 2018-07-06 Microscopic observation device, microscopic observation method and microscopic observation device manufacturing method
JP2018-129018 2018-07-06

Publications (1)

Publication Number Publication Date
WO2020009220A1 true WO2020009220A1 (en) 2020-01-09

Family

ID=69060465

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2019/026801 WO2020009220A1 (en) 2018-07-06 2019-07-05 Microscopic observation apparatus, microscopic observation method, and method for manufacturing microscopic observation apparatus

Country Status (2)

Country Link
JP (1) JP2020008682A (en)
WO (1) WO2020009220A1 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11378444B2 (en) 2020-08-05 2022-07-05 Visera Technologies Company Limited Biosensor and bio detection system

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08145888A (en) * 1994-11-16 1996-06-07 Olympus Optical Co Ltd Fluorescence calorimeter
JP2002055226A (en) * 2000-08-07 2002-02-20 Nippon Sheet Glass Co Ltd Polarizing element and method for manufacturing the same
JP2005099662A (en) * 2002-12-27 2005-04-14 Olympus Corp Confocal microscope
US20110300490A1 (en) * 2009-01-24 2011-12-08 Bastien Rachet High-resolution microscopy and photolithography devices using focusing micromirrors
JP2012529025A (en) * 2009-06-02 2012-11-15 コミッサリアータ レネルジー アトミック エ オゼネルジー アルテルナティーブ Microlens imaging system and sample detection system attached device
JP2017134191A (en) * 2016-01-27 2017-08-03 一般社団法人 日本薬理評価機構 Microscope observation-purpose container and microscope observation system
JP2018042283A (en) * 2017-12-01 2018-03-15 株式会社東芝 Observation method using microscopic imaging apparatus
JP2018098343A (en) * 2016-12-13 2018-06-21 ソニーセミコンダクタソリューションズ株式会社 Imaging device, metal thin film filter, and electronic equipment

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH03190167A (en) * 1989-12-19 1991-08-20 Sharp Corp Solid-state image sensing device
US7591980B2 (en) * 2004-03-01 2009-09-22 Mesosystems Technology, Inc. Biological alarm
JP2009075281A (en) * 2007-09-20 2009-04-09 Nikon Corp Microscope objective lens
JP2009156659A (en) * 2007-12-26 2009-07-16 Olympus Corp Measuring apparatus and method
WO2014192734A1 (en) * 2013-05-30 2014-12-04 新日鐵住金株式会社 Film thickness measurement method, film thickness measurement device, and recording medium

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08145888A (en) * 1994-11-16 1996-06-07 Olympus Optical Co Ltd Fluorescence calorimeter
JP2002055226A (en) * 2000-08-07 2002-02-20 Nippon Sheet Glass Co Ltd Polarizing element and method for manufacturing the same
JP2005099662A (en) * 2002-12-27 2005-04-14 Olympus Corp Confocal microscope
US20110300490A1 (en) * 2009-01-24 2011-12-08 Bastien Rachet High-resolution microscopy and photolithography devices using focusing micromirrors
JP2012529025A (en) * 2009-06-02 2012-11-15 コミッサリアータ レネルジー アトミック エ オゼネルジー アルテルナティーブ Microlens imaging system and sample detection system attached device
JP2017134191A (en) * 2016-01-27 2017-08-03 一般社団法人 日本薬理評価機構 Microscope observation-purpose container and microscope observation system
JP2018098343A (en) * 2016-12-13 2018-06-21 ソニーセミコンダクタソリューションズ株式会社 Imaging device, metal thin film filter, and electronic equipment
JP2018042283A (en) * 2017-12-01 2018-03-15 株式会社東芝 Observation method using microscopic imaging apparatus

Also Published As

Publication number Publication date
JP2020008682A (en) 2020-01-16

Similar Documents

Publication Publication Date Title
KR102206218B1 (en) Backside illumination image sensor, manufacturing method thereof and image-capturing device
US11282882B2 (en) Focus detecting device and electronic device
US9100639B2 (en) Light field imaging device and image processing device
TWI531851B (en) Camera device and filter
JP5274299B2 (en) Imaging device
CN101395926A (en) Fused multi-array color image sensor
JP2013512470A (en) Optical imaging device
TW200841462A (en) Solid-state image capturing apparatus, method for manufacturing the same, and electronic information device
US9678324B2 (en) Microscope system
US20160363673A1 (en) Method of fabricating integrated digital x-ray image sensor, and integrated digital x-ray image sensor using the same
JP2015015296A (en) Solid-state imaging device and electronic equipment
US9425229B2 (en) Solid-state imaging element, imaging device, and signal processing method including a dispersing element array and microlens array
JP2014003116A (en) Image pickup device
JP6462837B2 (en) Observation method using microscope
EP3264745B1 (en) Scanning imaging system with a novel imaging sensor with gaps for electronic circuitry
WO2020009220A1 (en) Microscopic observation apparatus, microscopic observation method, and method for manufacturing microscopic observation apparatus
JP6320870B2 (en) Observation method using microscope
KR20140090548A (en) Image pickup device, method of manufacturing same, and electronic apparatus
JP5434121B2 (en) Back-illuminated image sensor and imaging apparatus
JP2009188461A (en) Imaging apparatus
JP2022133325A (en) Microscope observation device, detector, and microscope observation method
JP6692404B2 (en) Observation system
KR101205128B1 (en) Optical examination apparatus adjusting resolution per pixel using image sensor binning
JP7383876B2 (en) Imaging element and imaging device
WO2016103315A1 (en) Solid-state image pickup device, and image pickup device

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19831480

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19831480

Country of ref document: EP

Kind code of ref document: A1