WO2019131376A1 - Procédé et système d'observation d'échantillon organique - Google Patents

Procédé et système d'observation d'échantillon organique Download PDF

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Publication number
WO2019131376A1
WO2019131376A1 PCT/JP2018/046769 JP2018046769W WO2019131376A1 WO 2019131376 A1 WO2019131376 A1 WO 2019131376A1 JP 2018046769 W JP2018046769 W JP 2018046769W WO 2019131376 A1 WO2019131376 A1 WO 2019131376A1
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WIPO (PCT)
Prior art keywords
thin film
sample
organic
electron beam
infrared
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PCT/JP2018/046769
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English (en)
Japanese (ja)
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小椋　俊彦
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国立研究開発法人産業技術総合研究所
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Publication of WO2019131376A1 publication Critical patent/WO2019131376A1/fr

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    • 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
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • 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
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
    • G01N21/3563—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing solids; Preparation of samples therefor
    • 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
    • G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides

Definitions

  • the present invention relates to an observation method and an observation system for obtaining information reflecting the composition of an organic sample, and more particularly to an observation method and an observation system obtaining information reflecting the composition distribution of an organic sample from infrared absorption.
  • An optical microscope is widely used for surface observation of various samples.
  • optical microscopes using infrared rays having a longer wavelength or the like have been proposed (for example, Patent Document 1).
  • an infrared (IR) microscope has also been proposed which condenses infrared light onto an observation sample with an aperture or the like, detects transmitted light or reflected light with a semiconductor detector, and gives qualitative analysis or quantitative analysis of minute parts.
  • Patent Document 2 discloses an observation system in which a Fourier transform infrared spectrophotometer (FTIR) is combined with an infrared microscope that performs microspectroscopy in an infrared wavelength region.
  • FTIR Fourier transform infrared spectrophotometer
  • the irradiation position of the infrared light flux is determined based on the visible light image of the area including the analysis position on the sample surface, and this is irradiated to the area of about 15 ⁇ m square to perform microspectroscopic light There is.
  • Patent Document 3 and Non-patent Document 1 soft X-rays generated by irradiating a thin metal thin film with an electron beam are irradiated to an observation sample and soft X-rays transmitted are detected, and organisms containing moisture are detected. Describes how to provide an internal view of the sample.
  • soft X-rays it is possible to observe a water-containing observation sample (a biological sample or a sample in a solution) as it is, and since the wavelength is shorter than visible light, high resolution observation beyond an optical microscope It is said that it is possible.
  • Patent Document 4 and Non-Patent Document 2 as in the above-mentioned Patent Document 3 and Non-Patent Document 1, the biological sample in the aqueous solution is not subjected to staining treatment by generating potential fluctuation using an electron beam irradiation apparatus. Describes what can be observed with high contrast.
  • a heavy metal thin film formed on the upper surface of the insulating thin film having pressure resistance is irradiated with an electron beam to form a local potential change, and the potential fluctuation causes an attenuation state when passing through the observation sample as an image It observes.
  • the dielectric constant of water is as high as about 80 and the potential change is well transmitted
  • the dielectric constant of the biological sample is as low as about 2 to 3 to inhibit the transmission of the potential change, so observation with high contrast can be obtained It is
  • the image observation is performed without staining processing of bacteria, virus, protein or protein complex in water or aqueous solution, and as it is alive It becomes possible. Also in this case, even if it is attempted to incorporate an infrared microscope for composition analysis, it is difficult to guide the optical path to the observation sample because there is a plate to be irradiated that receives electron beam irradiation.
  • the present invention has been made in view of the above situation, and the object of the present invention is an observation method for obtaining information reflecting the composition from infrared absorption of an organic sample with high sensitivity while being simple. And providing an observation system.
  • an aqueous solution containing an organic substance sample is interposed between opposing main surfaces of a pair of first and second insulating thin films, and the pair of opposing surfaces of the first insulating thin film
  • the conductive thin film provided on the outer main surface is irradiated with an electron beam to be locally heated, and the intensity of infrared rays transmitted through the second insulating thin film is measured by an infrared sensor as information reflecting the composition of the organic substance sample It is characterized by
  • the electron beam may be scan-irradiated along the conductive thin film. According to this invention, it is possible to obtain information reflecting the composition from the infrared absorption of the organic substance sample with high sensitivity while being simple.
  • the infrared sensors are arranged in an array to be opposed to the second insulating thin film, and each detection signal from this is calculated based on the mutual position with the irradiation position of the electron beam to the conductive thin film.
  • Processing may be performed to obtain three-dimensional information including the composition distribution of the organic sample. According to this invention, it is possible to obtain three-dimensional information including the composition distribution of the organic substance sample with high sensitivity while being simple.
  • the infrared sensor may be provided with filters having different transmission wavelengths to obtain an absorbance spectrum. According to this invention, it is possible to obtain information reflecting the composition from the infrared absorption of the organic substance sample with high sensitivity while being simple.
  • the conductive thin film may be characterized by being made of a single substance of manganese, tungsten, tantalum, gold, platinum, silver, copper, iron, titanium, osmium, or an alloy containing the same.
  • the first and second insulating thin films may be made of a silicon nitride thin film, a silicon oxide thin film, or a polyimide thin film. According to this invention, it is possible to obtain information reflecting the composition from the infrared absorption of the organic matter sample with high sensitivity while being simple.
  • a pair of first and second insulating thin films capable of interposing an aqueous solution containing an organic substance sample between opposing main surfaces of the organic substance sample;
  • An electron beam irradiated portion including a conductive thin film provided on an outer major surface forming a pair with the opposite major surface and capable of locally heating by irradiating an electron beam thereto, and the second as information reflecting the composition of the organic sample
  • an infrared sensor for measuring the intensity of infrared rays transmitted through the insulating thin film.
  • the apparatus may include an irradiation control unit which scans and irradiates the electron beam along the conductive thin film. According to this invention, it is possible to obtain information reflecting the composition from the infrared absorption of the organic matter sample with high sensitivity while being simple.
  • the infrared sensors are arranged in an array to be opposed to the second insulating thin film, and each detection signal from this is calculated based on the mutual position with the irradiation position of the electron beam to the conductive thin film. It may be characterized by having an operation part which processes and obtains three-dimensional information including composition distribution of the organic matter sample. According to this invention, it is possible to obtain three-dimensional information reflecting the composition distribution from the infrared absorption of the organic substance sample with high sensitivity while being simple.
  • the conductive thin film may be characterized by being made of a single substance of manganese, tungsten, tantalum, gold, platinum, silver, copper, iron, titanium, osmium, or an alloy containing the same.
  • the first and second insulating thin films may be made of a silicon nitride thin film, a silicon oxide thin film, or a polyimide thin film. According to this invention, it is possible to obtain information reflecting the composition from the infrared absorption of the organic matter sample with high sensitivity while being simple.
  • the infrared sensor may include filters having different transmission wavelengths to obtain an absorbance spectrum. According to this invention, it is possible to obtain information reflecting the composition from the infrared absorption of the organic matter sample with high sensitivity while being simple.
  • the observation system 1 of the organic substance sample includes a sample chamber 2 which can be evacuated to a predetermined degree of vacuum, and is in communication with it and emits electrons from an electron source 30 near the top of the housing 3 above it.
  • the wire 31 is appropriately guided to a predetermined position of the observation holder 10 in the sample chamber 2 while passing through the diaphragm 32, and the conductive thin film 13 (see FIG. 2) to be described later is locally heated by the irradiated electron beam 31 to To generate an observation of the organic matter sample 18 (see FIG. 2).
  • the electron source 30 is a field emission type electron gun.
  • the traveling direction of the emitted electron beam 31 can be changed by the polarizing plate 33, and the electron beam 31 can be scanned and irradiated on the observation holder 10 (irradiation control unit).
  • the electron source 30 may be configured to be able to irradiate the electron beam 31 whose output changes in a pulse shape of a predetermined frequency onto the observation holder 10 by using the function generator 34.
  • a sample exchange chamber 41 is provided in the sample chamber 2 with an openable shutter 40 interposed therebetween, and a stage provided in the sample chamber 2 using the sample exchange rod 42 while maintaining the degree of vacuum in the sample chamber 2
  • the observation holder 10 is removable above 20.
  • a measuring unit A described later is provided on the insulating insulating casing 21 of the stage 20, and the signal from the measuring unit A can be extracted to the outside of the sample chamber 2.
  • the signal is amplified by the amplifier 23 (see FIG. 2) incorporated in the measurement unit A, guided to the frequency separation device 35, separated in frequency, and output to the composition analysis device 36.
  • the frequency separation device 35 receives the reference signal of the output change of the electron beam described above from the function generator 34.
  • a DC power supply 37 for the operation of the amplifier 23 and the like is connected to the measurement unit A.
  • the observation holder 10 includes an outer frame 11 having windows at the top and bottom, and a pair of first insulating thin film 12a and second insulating thin film 12b closing the upper and lower windows from the inside.
  • the insulating thin film 12a closing the upper window holds the aqueous solution 18b containing the organic sample 18 from the upper side by the lower side (opposing main surface) from the upper side, and the conductive thin film 13 is laminated on the upper side (outside main surface) Ru.
  • the insulating thin film 12 b closing the lower window holds the aqueous solution 18 b containing the organic sample 18 from the lower side by the upper side surface (opposing main surface).
  • the main surfaces of the insulating thin films 12a and 12b are opposed to each other, and the organic sample 18 and the aqueous solution 18b are interposed between each other.
  • the insulating thin films 12a and 12b are in contact with the inner surface of the observation holder 10 by an O-ring 17 and a packing not shown, respectively, and the inside is sealed against the vacuum outside the observation holder 10 to It can be held.
  • the insulating thin films 12a and 12b have a strength sufficient to withstand these pressure differences.
  • a thin silicon nitride thin film having high pressure resistance it is preferable to use a thin silicon oxide thin film having high pressure resistance.
  • a silicon oxide thin film, a polyimide thin film, and the like can also be suitably used.
  • the conductive thin film 13 may be provided on the outer major surface of the insulating thin film 12a and irradiated with the electron beam 31 to be locally heated, thereby generating an infrared ray by such heating.
  • the lower the thermal conductivity of the metal the higher the temperature of the region irradiated with the electron beam, and the larger the detection signal can be obtained. Therefore, a metal having a low thermal conductivity is more preferable.
  • a single element of manganese, tungsten, tantalum, gold, platinum, silver, copper, iron, titanium, osmium, an alloy containing the same, or the like can be used.
  • the measuring unit A of the stage 20 includes an infrared sensor 22 connected to the amplifier 23 to receive the infrared light transmitted through the insulating thin films 12 a and 12 b.
  • the amplifier 23 can amplify a detection signal based on the infrared intensity received by the infrared sensor 22 and output it to the frequency separation device 35 through the connector 24 as described above. Further, the amplifier 23 and the power supply 37 are connected via the connector 24.
  • the measuring unit A may further include a potential measurement terminal 25 capable of receiving a potential change due to the irradiation of the electron beam 31 to the conductive thin film 13. Thus, such potential changes can be transmitted through the insulating thin films 12a and 12b and can be received via the organic sample 18 and / or the aqueous solution 18b having different dielectric constants.
  • the observation system 1 to which the observation holder 10 is attached has an electron beam irradiation unit, and after evacuating the sample chamber 2 to a predetermined degree of vacuum, the electron beam 31 is emitted from the electron source 30.
  • the output of the electron beam 31 can be changed in a pulse shape by the control signal from the function generator 34.
  • the electron beam 31 may be chopped by the polarizing plate 33 to perform frequency modulation.
  • an element requiring frequency modulation such as a pyroelectric infrared sensor or a sensor based on thermoelectromotive force such as a thermopile can be used as the infrared sensor 22 and this contributes to the improvement of detection sensitivity and the reduction of noise.
  • the electron beam 31 irradiated toward the observation holder 10 is made incident on the conductive thin film 13 from the window of the observation holder 10 and absorbed, and the incident portion is locally heated.
  • Such local heating generates infrared light from the conductive thin film 13, and the insulating thin film 12 a, the organic sample 18 and the layer of the aqueous solution 18 b, and the insulating thin film 12 b are sequentially transmitted or partially absorbed, and infrared light intensity is measured by the infrared sensor 22. It is detected. That is, it is possible to obtain information reflecting the composition of the organic sample 18 based on the infrared absorption detected by the organic sample 18 and the like.
  • the infrared sensor 22 detects infrared rays at different detection angles.
  • the two-dimensional image is an inclined image in which the images directed to the respective detection angles are continuous.
  • a potential image of the organic substance sample 18 can be obtained also from the potential change received by the above-described potential measurement terminal 25, and combined with the infrared image, analysis of the organic substance sample 18 in more detail can also be performed.
  • infrared light is obtained by local heating at the irradiation position of the electron beam 31 irradiated to the conductive thin film 13, its generation source (light source) can be made very small, and high-definition information can be obtained by the infrared sensor 22. Can be obtained by Furthermore, there is no need for an optical system for focusing infrared light optically or a precise mechanism for optical scanning. That is, it is possible to obtain information with high sensitivity infrared light with a simple configuration.
  • the observation system 1 of the organic substance sample it is possible to obtain information reflecting the composition from the infrared absorption of the organic substance sample with high sensitivity while being simple.
  • the observation holder 10 a is formed by arranging a plurality of infrared sensors 22 in an array on the measurement unit A. Thereby, a plurality of infrared images can be obtained by one electron beam scan. That is, the above-described two-dimensional image obtained by scanning can be obtained by each of the infrared sensors 22.
  • the detection angle of infrared rays is calculated based on the mutual position of each infrared sensor 22 and the irradiation position of the electron beam 31, and three-dimensional information ( It is also possible to use calculation processing to obtain data) (calculation unit).
  • the infrared filter 22 a may be disposed on the upper surface of the infrared sensor 22. By making the transmission wavelengths different for each of the infrared filters 22a, it is possible to obtain absorbance spectra of different infrared wavelengths and to obtain two-dimensional images separately.
  • an infrared absorption spectrum can be calculated from the plurality of obtained two-dimensional images, and the composition of the organic sample 18 can be analyzed in more detail.
  • a biological sample such as a cell
  • proteins and oil components are contained inside.
  • proteins and oil components produce unique absorption spectra in the infrared region, it is possible to analyze the composition distribution in cells by obtaining the absorption spectra.
  • an electrode with a thin tip is brought close to the upper surface of the conductive thin film 13 and a voltage is applied to the conductive thin film 13 to generate a current. It can also be heated. For example, local heating can be made to be a minute spot of about 10 nm by setting the tip of the electrode to about 10 nm in diameter.
  • the observation holder 10 can be placed in the atmosphere. That is, a simple structure with low airtightness can be obtained for both the observation holder and the entire observation system of the organic substance sample.
  • observation system 10 observation holder 12a, 12b insulating thin film 13 conductive thin film 18 organic sample 18 b aqueous solution 31 electron beam

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  • Physics & Mathematics (AREA)
  • Chemical & Material Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Optics & Photonics (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)
  • Microscoopes, Condenser (AREA)
  • Analysing Materials By The Use Of Radiation (AREA)

Abstract

L'invention concerne un procédé et un système d'observation pour l'acquisition, avec une grande sensibilité et de façon très simple, d'informations reflétant la composition d'un échantillon organique à partir de son absorption infrarouge. Le système d'observation comprend : une paire de premier et second films minces isolants, entre les surfaces principales opposées desquels une solution aqueuse contenant un échantillon organique peut être interposée ; une unité d'émission de faisceau d'électrons qui émet un faisceau d'électrons en direction d'un film mince conducteur disposé sur la surface principale externe formant une paire avec la surface principale opposée du premier film mince isolant de façon à pouvoir chauffer localement le film mince conducteur ; et un capteur infrarouge qui mesure la distribution d'intensité des rayons infrarouges transmis à travers le second film isolant, en tant qu'information reflétant la composition de l'échantillon organique. Le procédé d'observation comprend : l'interposition d'une solution aqueuse contenant un échantillon organique entre les surfaces principales opposées d'une paire de premier et second films minces isolants ; l'émission d'un faisceau d'électrons en direction d'un film mince conducteur disposé sur la surface principale externe formant une paire avec la surface principale opposée du premier film mince isolant de façon à chauffer localement le film mince conducteur ; et la mesure de la distribution d'intensité des rayons infrarouges transmis à travers le second film isolant, en tant qu'information reflétant la composition de l'échantillon organique.
PCT/JP2018/046769 2017-12-28 2018-12-19 Procédé et système d'observation d'échantillon organique WO2019131376A1 (fr)

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JP2017-253657 2017-12-28
JP2017253657A JP2019120526A (ja) 2017-12-28 2017-12-28 有機物試料の観察方法及び観察システム

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JP7204114B2 (ja) * 2019-06-27 2023-01-16 株式会社大一商会 遊技機
JP7204120B2 (ja) * 2019-06-27 2023-01-16 株式会社大一商会 遊技機
JP7204117B2 (ja) * 2019-06-27 2023-01-16 株式会社大一商会 遊技機

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JP2015031515A (ja) * 2013-07-31 2015-02-16 独立行政法人物質・材料研究機構 近接場光学観察装置、試料含有環境セル作製方法、走査電子光学顕微鏡及び走査電子光学顕微鏡の使用方法
WO2016024503A1 (fr) * 2014-08-12 2016-02-18 株式会社村田製作所 Appareil de mesure de gaz, substrat multi-dispositif, procédés de fabrication associés, et procédés de fabrication de source de lumière infrarouge et de capteur infrarouge pyroélectrique
WO2017065135A1 (fr) * 2015-10-16 2017-04-20 国立研究開発法人産業技術総合研究所 Méthode d'observation de spécimen organique, et support d'observation ainsi que platine d'observation utilisée en son sein
WO2017154936A1 (fr) * 2016-03-09 2017-09-14 国立研究開発法人産業技術総合研究所 Microscope à constante diélectrique et procédé d'observation d'un spécimen organique

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JP2015031515A (ja) * 2013-07-31 2015-02-16 独立行政法人物質・材料研究機構 近接場光学観察装置、試料含有環境セル作製方法、走査電子光学顕微鏡及び走査電子光学顕微鏡の使用方法
WO2016024503A1 (fr) * 2014-08-12 2016-02-18 株式会社村田製作所 Appareil de mesure de gaz, substrat multi-dispositif, procédés de fabrication associés, et procédés de fabrication de source de lumière infrarouge et de capteur infrarouge pyroélectrique
WO2017065135A1 (fr) * 2015-10-16 2017-04-20 国立研究開発法人産業技術総合研究所 Méthode d'observation de spécimen organique, et support d'observation ainsi que platine d'observation utilisée en son sein
WO2017154936A1 (fr) * 2016-03-09 2017-09-14 国立研究開発法人産業技術総合研究所 Microscope à constante diélectrique et procédé d'observation d'un spécimen organique

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"Development of new visualizing technology of the liquid samples based on scanning electron microscopy", OLEOSCIENCE, vol. 15, no. 11, November 2015 (2015-11-01), pages 511 - 516 *
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