WO2023008580A1 - Dispositif et procédé de mesure de particules fines - Google Patents

Dispositif et procédé de mesure de particules fines Download PDF

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Publication number
WO2023008580A1
WO2023008580A1 PCT/JP2022/029394 JP2022029394W WO2023008580A1 WO 2023008580 A1 WO2023008580 A1 WO 2023008580A1 JP 2022029394 W JP2022029394 W JP 2022029394W WO 2023008580 A1 WO2023008580 A1 WO 2023008580A1
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Prior art keywords
light
channel
signal
measuring device
particle measuring
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PCT/JP2022/029394
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English (en)
Japanese (ja)
Inventor
和真 馬渡
慶之 津山
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国立大学法人 東京大学
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Publication of WO2023008580A1 publication Critical patent/WO2023008580A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/02—Investigating particle size or size distribution
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • 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
    • G01N21/47—Scattering, i.e. diffuse reflection
    • 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
    • G01N21/47—Scattering, i.e. diffuse reflection
    • G01N21/49—Scattering, i.e. diffuse reflection within a body or fluid
    • G01N21/53—Scattering, i.e. diffuse reflection within a body or fluid within a flowing fluid, e.g. smoke
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N37/00—Details not covered by any other group of this subclass

Definitions

  • the present invention relates to a particle measuring device and a particle measuring method for measuring the properties of particles supplied to a channel.
  • a method of increasing sensitivity using an interference optical system has also been developed.
  • optical interference detection is realized by splitting a laser into two, irradiating one of them onto a sample, adjusting the phase of the other, and superimposing them again.
  • Such a method inevitably complicates the optical system and complicates the optical adjustment, and is difficult to be compatible with other measurement methods.
  • simultaneous measurement of fluorescence and scattered light is often used.
  • the present invention has been made in view of the above background art, and provides a particle measurement apparatus and method that enable easy and highly sensitive measurement of scattered light from particles by utilizing the optical diffraction phenomenon of a flow path. intended to provide
  • a particle measuring apparatus includes an illumination device that illuminates a flow channel and a detection device that detects light from the flow channel, and detects scattered light from particles passing through the flow channel and the flow. Interference with diffracted light from the channel is measured using a detector to obtain information about particles passing through the channel.
  • the interference between the scattered light from the particles passing through the flow path and the diffracted light from the flow path is measured using a detection device.
  • a simple and highly sensitive measurement of scattered light from particles is possible without the need for a complicated optical system that splits and recombines the light.
  • the signal derived from the particles passing through the channel is detected in a predetermined frequency range from the signal output of the detection device obtained while the particles are flowing in the channel illuminated by the illumination device.
  • signals originating from particles passing through the channel can be selectively detected with high sensitivity, and information regarding the properties of the particles passing through the channel can be accurately detected.
  • a signal processing device is further provided for extracting a component corresponding to a predetermined frequency range from the detection signal by the detection device.
  • the predetermined frequency range corresponds to the reciprocal of the passage time of particles through the channel. In this case, it is possible to perform measurement that reflects the state of appearance of the particles with reference to the area observed by the optical system.
  • the predetermined frequency range is set in consideration of the flow velocity of particles in the channel. In this case, it becomes possible to perform measurement corresponding to the particle flow velocity.
  • the predetermined frequency range is 1-3 kHz.
  • the signal processing device extracts the frequency and intensity of peak signals from the detection signal by the detection device and performs data processing. In this case, the flow rate number and size distribution of particles passing through the channel can be evaluated.
  • the illumination device has a plurality of laser light sources with different wavelengths, and the signal processing device evaluates wavelength characteristics of detection signals from the detection device.
  • the signal processing device evaluates wavelength characteristics of detection signals from the detection device.
  • particles of different sizes and compositions can be distinguished.
  • an excitation device that irradiates a channel with excitation light modulated by a chopper, a probe device that irradiates the channel with probe light, and diffraction of the probe light by the channel a photothermal conversion spectroscopic device including a measuring device for measuring;
  • optical diffraction type photothermal conversion spectroscopic measurement becomes possible.
  • optical diffraction scattered light measurement and optical diffraction photothermal conversion spectroscopic measurement can be performed in parallel, and a single scattered light measurement device can also be operated as a photothermal conversion spectroscopic measurement device.
  • data processing is performed by extracting the intensity and width of the peak signal from the detection signal by the detection device and the measurement output by the measurement device of the photothermal conversion spectroscopic device.
  • particles of different sizes and compositions can be distinguished.
  • the cross-sectional size of the channel is 0.1 ⁇ or more and 10 ⁇ or less, where ⁇ is the wavelength of the light that illuminates the channel.
  • the cross-sectional size of the channel means the length of the long side when the channel has a rectangular cross section, for example, and the diameter when the channel has a circular cross section, for example.
  • a particle measurement method measures interference between scattered light from particles passing through an illuminated channel and diffracted light from the channel, and obtains information about the particles passing through the channel.
  • the interference between the scattered light from the particles passing through the illuminated channel and the diffracted light from the channel is measured.
  • a simple and highly sensitive measurement of scattered light from particles becomes possible without the need for a complex optical system for resynthesis.
  • FIG. 1 is a block diagram of a particle measuring device according to a first embodiment
  • FIG. FIG. 3 is a conceptual perspective view for explaining the principle of detecting diffracted light and scattered light generated in the vicinity of a microchip corresponding to a channel
  • FIG. 3A shows the time variation of the detection signal corresponding to the signal light
  • FIG. 3B shows the time variation of the extraction signal obtained by extracting only specific high frequency components from the detection signal.
  • FIG. 2 is a conceptual diagram illustrating the structure of a microchip that receives fluid supply
  • FIG. 5A is a chart for explaining the S/N ratio or signal count number at the measurement frequency
  • FIG. 5B is a chart for explaining the relationship between the slit position and the signal count number.
  • FIG. 4 shows specific measurement results obtained by the particle measuring device of the second embodiment. It is a block diagram explaining the structure of the microparticles
  • FIG. 1 is a functional block diagram of the entire particle measuring device 100 according to this embodiment.
  • the microparticle measuring apparatus 100 includes a light source device 20 that outputs illumination light IL for illuminating a channel 12 formed in the microchip 10, and an observation light OL that supplies the illumination light IL to the microchip 10 and emits the observation light OL from the microchip 10.
  • an observation optical system 30 that takes in the observation light OL from the observation optical system 30, a light receiving device 40 that takes in scattered light and the like from the observation light OL from the observation optical system 30; a lock-in amplifier 51; and a control device 80 for overall control of the operation of each part of the device.
  • the light source device 20 and the observation optical system 30, which is closer to the light source device 20 than the microchip 10 are collectively referred to as an illumination device 101.
  • the two sides are collectively referred to as the detection device 102 .
  • the light source device 20 includes a light source 21 and a beam expander 22.
  • the type of light source 21 related to the light emission principle of the light emitting element and the like is not limited.
  • the light source device 20 is preferably composed of a laser light source with high coherency from the viewpoint of causing interference between scattered light and diffracted light from the observation optical system 30 .
  • a laser light source a solid (including semiconductor) laser, gas laser, liquid laser, or the like can be used.
  • the illumination light IL output from the light source device 20 is not limited in wavelength as long as it forms scattered light by the fluid SL passing through the channel 12 of the microchip 10 or the detection target object SM.
  • External light visible light, ultraviolet light, or the like can be used.
  • a Nd:YAG laser second harmonic: 532 nm
  • a solid-state laser fundamental wavelength: 488 nm
  • the beam expander 22 expands the beam diameter of the illumination light IL emitted from the light source 21 and can make the illumination light IL incident on the observation optical system 30 suitable for the observation optical system 30 .
  • the beam expander 22 can be of a variable type that can change the expansion ratio of the beam diameter.
  • the observation optical system 30 has an objective lens 31 and a condenser lens 32 .
  • the objective lens 31 converges the illumination light IL expanded in diameter through the beam expander 22 to form a beam spot of desired size in the observation area A1 including the channel 12 of the microchip 10 .
  • the beam spot can have a size close to the diffraction limit, and can be set to a size of about 1 ⁇ m or less, for example.
  • the condenser lens 32 collimates the observation light OL that has passed through the microchip 10 .
  • the NA of the objective lens 31 was set to 0.45 and the NA of the condensing lens 32 was set to 0.90, but they are not limited to these.
  • the objective lens 31 and condenser lens 32 can be of variable focal length type or interchangeable type to adjust the spot size.
  • the light receiving device 40 is a part of the detection device 102 and includes a transmitted light blocker 41 and a photodetector 42 .
  • the transmitted light blocker 41 is a slit plate having a slit 41a for selectively passing signal light LS, which is a component other than transmitted light or rectilinear light in the observation light OL that has passed through the microchip 10 .
  • the slit 41a is an aperture formed at a position a predetermined distance away from the optical axis AX of the observation optical system 30, and allows the signal light LS to pass therethrough.
  • the main body of the transmitted light blocking portion 41 that is, the periphery of the slit 41a selectively blocks straight light that passes through without being diffracted by the microchip 10.
  • FIG. 2 is a conceptual perspective view explaining extraction of the interfering light L3 by the transmitted light blocking part 41 and the like.
  • the condensing lens 32 (see FIG. 1) is omitted from the viewpoint of simplicity of expression.
  • the straight light L0 formed by the illumination light IL passing through the microchip 10 has its divergence angle restricted by the observation optical system 30 and is confined within the original optical path near the optical axis AX. , and does not reach the photodetector 42 .
  • part of the diffracted light L2 formed by the illumination light IL being diffracted at the interface between the channel 12 of the microchip 10 and its outside passes through the slit 41a of the transmitted light blocking part 41, and is detected by light detection. incident on the device 42 .
  • part of the scattered light L1 formed by the illumination light IL being scattered by the detection target object SM passing through the channel 12 of the microchip 10 also passes through the slit 41a of the transmitted light blocking part 41, Incident on detector 42 . That is, part of the scattered light L1 formed by the particulate detection target object SM passing through the channel 12 of the microchip 10 and part of the diffracted light L2 formed by the channel 12 of the microchip 10 are combined and interfere when passing through the flow path 12, and the interference light L3 between the scattered light L1 and the diffracted light L2 enters the photodetector 42 as the signal light LS. It should be noted that the intensity and output timing of the scattered light L1 change due to the shape, size, etc. of the detection target object SM passing through the flow path 12 .
  • the diffracted light L2 has different output directions and intensity distributions depending on the difference in refractive index between the inside and outside of the channel 12 of the microchip 10 and the size of the channel.
  • the cross-sectional size of the channel 12 is 0.1 ⁇ or more and 10 ⁇ or less, where ⁇ is the wavelength of the illumination light IL that illuminates the channel 12 .
  • ⁇ is the wavelength of the illumination light IL that illuminates the channel 12 .
  • the cross section size is preferably set within the range of 53 nm to 5.3 ⁇ m.
  • the cross-sectional size of the flow channel 12 is set to 10 ⁇ or less, the flow channel size, that is, the cross-sectional size of the flow channel 12, can be made equal to or smaller than the spot size of the illumination light IL. It can be said that the reliability of the intensity of the signal value is also increased through the focus or spot. If the cross-sectional size of the flow channel 12 is less than 0.1 ⁇ , the illumination light IL becomes smaller than the diffraction limit. The signal value obtained from diffracted light and scattered light also decreases.
  • the photodetector 42 of the light receiving device 40 only needs to have sensitivity in the wavelength range of the observation light OL or the interference light L3, and includes, for example, a photodiode. Instead, a photoconductive cell, an image sensor, a phototube, a photomultiplier tube, or the like can be used.
  • a signal output from the photodetector 42 (a signal corresponding to the signal light LS) is input to the lock-in amplifier 51 .
  • a lock-in amplifier 51 extracts, from the signal output of the photodetector 42 , a predetermined frequency component that reflects periodic variations in scattering caused by particles passing through the flow path 12 .
  • the predetermined frequency range to be extracted by the lock-in amplifier 51 corresponds to the reciprocal of the passage time of particles through the channel.
  • the predetermined frequency range of interest is determined based on the size of the observation area A1 or the beam spot of the illumination light IL and the flow velocity of the fluid SL passing through the channel 12 .
  • the predetermined frequency range set by the lock-in amplifier 51 is, for example, 1 to 3 kHz.
  • the predetermined frequency range set by the lock-in amplifier 51 is determined based on a standard value corresponding to the product of the reciprocal of the pot size of the illumination light IL and the flow velocity of the fluid SL. For example, when the spot size of the illumination light IL is 1 ⁇ m and the flow velocity of the fluid SL is several mm/s, the standard value of the predetermined frequency range set by the lock-in amplifier 51 is 2 to 3 kHz. However, the predetermined frequency range set by the lock-in amplifier 51 is not limited to a theoretical standard value, and is adjusted or modified to a value that enhances sensitivity in consideration of other measurement conditions.
  • FIG. 3A and 3B are diagrams for explaining extraction of scattered signal components by the lock-in amplifier 51.
  • FIG. 3A shows the time change of the detection signal corresponding to the signal light LS
  • FIG. 3B shows the time change of the extraction signal obtained by extracting only specific high frequency components from the detection signal of FIG. 3A.
  • ED represents the amplitude of the diffracted light L2 incident on the photodetector 42 as an electromagnetic wave
  • ES represents the amplitude of the scattered light L1 incident on the photodetector 42 as an electromagnetic wave.
  • 2 corresponds to the detected intensity of the diffracted light L2
  • 2 corresponds to the detected intensity of the scattered light L1
  • 2ED ⁇ ES ⁇ cos ⁇ It corresponds to the detected intensity of the interference light L3 formed by interference.
  • the detected intensity of the diffracted light L2 is extremely high compared to the detected intensity of the scattered light L1.
  • 2 caused only by diffraction is extremely large compared to the interference light L3, and the detected intensity of the interference light L3 is buried in the detected intensity of
  • a predetermined frequency component determined based on the size of the observation area by the illumination light IL and the flow velocity of the fluid SL passing through the flow path 12 from the detection signal of FIG. 3A, only the interference light L3 A corresponding detection signal can be retrieved (see FIG. 3B).
  • a plurality of peak signals indicate instantaneously generated detection signals for each particle, indicating that the interference light L3 can be detected.
  • the fluid supply device 60 has a reservoir 61 that stores fluid SL containing fine particles, and a pressure controller 62 that adjusts the air pressure supplied to the reservoir 61 .
  • FIG. 4 is a conceptual diagram explaining the structure of the microchip 10 that receives the supply of the fluid SL from the reservoir 61.
  • the microchip 10 has a flat plate shape and is formed, for example, by microfabrication of quartz glass, Pyrex (registered trademark), other glass, transparent plastic, or the like.
  • the microchip 10 includes, as the channel 12, a nanochannel 12a extending to a region corresponding to the observation region A1, and a pair of microchannels 12c and 12d formed so as to be connected to both ends of the nanochannel 12a. have.
  • One microchannel 12c is formed with a supply port 14a connected to the reservoir 61 shown in FIG. 1, and the other microchannel 12d is formed with a discharge port 14b.
  • the cross-sectional size of the nanochannel 12a is desirably set to 0.1 ⁇ or more and 10 ⁇ or less as described above.
  • the control device 80 manages the operating states of the light source device 20, the lock-in amplifier 51, the fluid supply device 60, and the like. Specifically, the controller 80 controls the light source 21 to output stable illumination light IL. The control device 80 controls the pressure controller 62 to adjust the flow velocity of the fluid SL passing through the channel 12 of the microchip 10 . The control device 80 appropriately operates the lock-in amplifier 51 and receives, as a digital signal, a detection signal obtained by extracting a component corresponding to the interference light L3 from the output signal. The control device 80 and the lock-in amplifier 51 function as a signal processing device 50 that extracts a component corresponding to a predetermined frequency component from the signal detected by the light receiving device 40 .
  • the signal processing device 50 or the control device 80 performs data processing by extracting the frequency and intensity of peak signals from the signals detected by the light receiving device 40 (that is, the signal extracted by the lock-in amplifier 51) constituting the detection device 102.
  • the flow rate number and size distribution of the fine particles passing through the channel 12 can be evaluated. That is, the frequency of the peak signals corresponds to the flow rate number of particles passing through the channel 12 and the intensity of the peak signals gives information about the size of the particles passing through the channel 12 .
  • the predetermined frequency set in the lock-in amplifier 51 is fixed, it is also possible to make the predetermined frequency set in the lock-in amplifier 51 variable and measure the frequency spectrum characteristics of the peak signal. In this case, the spot size of the illumination light IL and the flow velocity of the fluid SL passing through the channel 12 are increased in degree of freedom, enabling measurement under various conditions.
  • FIGS. 5A and 5B are charts for explaining specific measurement examples.
  • FIG. 5A shows the relationship between the frequency extracted by the lock-in amplifier 51 and the detected output, etc.
  • FIG. 5B shows the relationship between the slit position of the transmitted light blocking section 41 and the count number of fine particles.
  • a relatively high S/N ratio can be achieved at 1-3 kHz.
  • the measurement was performed with the spot size of the illumination light IL set to about 1 ⁇ m and the flow velocity of the fluid SL set to about 0.2 mm/s. Considering noise, 2 kHz is optimal.
  • FIG. 5A shows the relationship between the frequency extracted by the lock-in amplifier 51 and the detected output, etc.
  • FIG. 5B shows the relationship between the slit position of the transmitted light blocking section 41 and the count number of fine particles.
  • a relatively high S/N ratio can be achieved at 1-3 kHz.
  • the measurement was performed with the spot size of the illumination light IL set to about 1 ⁇ m and the flow velocity of the fluid
  • 10 mm on the horizontal axis is the position of the optical axis AX, and 0 mm on the horizontal axis corresponds to a position 15 mm away from the optical axis AX.
  • fine particles are detected when the slit position is in the range of 5.5 to 9.0 mm away from the optical axis AX. That is, when the slit position is less than 5.5 mm from the optical axis AX, the detection of the interfering light L3 is hindered by the straight light L0, and a sufficient detection signal cannot be obtained. Also, when the slit position exceeds 9.0 mm from the optical axis AX, the incident amount of the interference light L3 decreases, and a sufficient detection signal cannot be obtained.
  • the interference between the scattered light L1 from the particles passing through the flow path 12 and the diffracted light L2 from the flow path 12 is measured using the detection device 102.
  • a complicated optical system for splitting a coherent light source into two optical paths and recombining them is not required, and the scattered light L1 from fine particles can be easily and highly sensitively measured.
  • a particle measuring device and the like according to the second embodiment of the present invention will be described below.
  • the particle measuring apparatus of the second embodiment is obtained by partially changing the wavelength used in the particle measuring apparatus of the first embodiment, and the description of common parts will be omitted.
  • light source device 20 includes a first light source 21a and a second light source 21b, and both light sources 21a and 21b output illumination lights IL1 and IL2 having different wavelengths.
  • the first wavelength of the illumination light IL1 is, for example, 488 nm
  • the second wavelength of the illumination light IL2 is, for example, 532 nm.
  • the illumination light IL1 from the first light source 21a is reflected by the mirror 25a, passes through the dichroic mirror 25b, and enters the beam expander 22.
  • Illumination light IL 2 from second light source 21 b is reflected by dichroic mirror 25 b and enters beam expander 22 .
  • the observation optical system 30 receives the illumination lights IL1 and IL2 having different wavelengths in combination.
  • the objective lens 31 and the condenser lens 32 that constitute the observation optical system 30 can be of an achromatic type corresponding to the wavelengths of the illumination lights IL1 and IL2.
  • the light receiving device 40 includes a transmitted light blocking portion 41, a first photodetector 42a, and a second photodetector 42b.
  • the wavelength component corresponding to the illumination light IL1 (corresponding to the signal light LS1) is reflected by the dichroic mirror 45a, passes through a lens or the like, and enters the first photodetector 42a.
  • the wavelength component corresponding to the illumination light IL2 (corresponding to the signal light LS2) passes through the dichroic mirror 45a, is reflected by the mirror 45b, passes through the lens or the like, and becomes the second light. Incident on the detector 42b.
  • the signal output (signal corresponding to LS1) of the first photodetector 42a is input to the first lock-in amplifier 51a, and the signal output (signal light) of the second photodetector 42b is input to the second lock-in amplifier 51b.
  • LS2 is input.
  • the fluid SL passing through the channel 12 of the microchip 10 is measured with two wavelengths.
  • FIG. 7 shows specific measurement results by the particle measuring device 100 of the second embodiment.
  • the horizontal axis represents the intensity of the scattering signal obtained by processing the output signal of the first photodetector 42a by the first lock-in amplifier 51a when the illumination light IL1 has a wavelength of 488 nm
  • the vertical axis represents the intensity of the illumination light.
  • IL2 has a wavelength of 532 nm
  • the intensity of the scattered signal obtained by processing the output signal of the second photodetector 42b by the second lock-in amplifier 51b is shown.
  • the channel 12 of the microchip 10 had a rectangular cross section, a width of 1.2 ⁇ m and a depth of 550 nm.
  • the fluid SL supplied to the microchip 10 contained gold particles with a diameter of 40 nm, gold particles with a diameter of 60 nm, silver particles with a diameter of 40 nm, and silver particles with a diameter of 60 nm as detection target objects SM.
  • the two-dimensional distribution area differs depending on the type and size of the fine particles. That is, by performing data processing based on scattered signals corresponding to a plurality of wavelengths from the object to be detected SM, it becomes possible to identify fine particles having different sizes and compositions.
  • a 90% correct answer rate can be achieved by performing experimental machine learning for distinguishing when two types of fine particles are mixed, and experimental machine learning is performed for distinguishing when four types of fine particles are mixed. By doing so, we were able to achieve an accuracy rate of 70%.
  • the measurement is performed using the illumination lights IL1 and IL2 with two different wavelengths.
  • the light L1 can be detected for each wavelength.
  • the illumination device 101 has a plurality of light sources 21a and 21b with different wavelengths, and the signal processing device 50 evaluates the wavelength characteristics of the detection signal from the detection device 102. Therefore, information can be obtained that distinguishes between particles of different sizes and compositions.
  • a particle measuring apparatus and the like according to the third embodiment of the present invention will be described below.
  • the particle measuring device of the third embodiment is obtained by adding a photothermal conversion spectroscopic device to the particle measuring device of the first embodiment, and the description of the common parts will be omitted.
  • the particle measuring device 200 of the third embodiment is an application of the photothermal conversion spectroscopic device 300 .
  • the photothermal conversion spectroscopic device 300 operates on the principle described in Japanese Patent Application Laid-Open No. 2020-204594, and has the same structure as that described in Japanese Patent Application Laid-Open No. 2020-204594.
  • the photothermal conversion spectrometer 300 performs optical diffraction type photothermal conversion spectrometry, and originally evaluates fine particles passing through the flow path 12 by measuring diffracted light. , can also be operated as the particle measuring apparatus 100 shown in FIG.
  • the microparticle measuring apparatus 200 includes an excitation device 71 that irradiates the channel 12 of the microchip 10 with the modulated excitation light LE, a probe device 72 that irradiates the channel 12 with the probe light LP, and a probe by the channel 12. and a measuring device 73 for measuring the diffraction of the light LP.
  • the excitation device 71 can be turned on and off at required timing.
  • probe device 72 includes probe light source 327 , beam expander 322 and objective lens 331 .
  • the excitation device 71 also includes an excitation light source 328 , a chopper 29 , a beam expander 322 and an objective lens 331 .
  • the measurement device 73 includes a condenser lens 332 , a transmitted light blocker 341 , an interference filter 46 a , a lens 46 b , a color filter 46 c , a photodetector 342 and lock-in amplifiers 351 and 352 .
  • the excitation light LE enters the objective lens 331 through the chopper 29 and the dichroic mirror 26b in a state where the intensity periodically increases and decreases.
  • the probe light LP enters the objective lens 331 via the mirror 26a and the dichroic mirror 26b without passing through the chopper.
  • the excitation light LE that has entered the channel 12 or observation area A1 of the microchip 10 through the objective lens 331 heats the channel 12, causing a refractive index change in the fluid inside the channel 12 and its surroundings.
  • the probe light LP that has entered the flow path 12 of the microchip 10 or the observation area A1 through the objective lens 331 travels straight through the observation area A1 or is diffracted in the observation area A1 and emitted, and the observation light emitted from the observation area A1.
  • the OL enters the transmitted light blocking portion 341 of the light receiving device 340 via the condenser lens 332 .
  • the interference light L4 resulting from diffraction that has passed through the slit 41a of the transmitted light blocking portion 341 enters the photodetector 342 via the interference filter 46a, the lens 46b, and the color filter 46c.
  • the interference filter 46a and the color filter 46c selectively transmit the interference light L4 and block the excitation light LE.
  • a signal output from the photodetector 342 is input to the lock-in amplifiers 351 and 352 .
  • One lock-in amplifier 351 extracts a signal component corresponding to the modulation frequency of the intensity modulation by the chopper 29 from the signal output of the photodetector 342 .
  • the interference light L4 from the observation area A1 of the microchip 10 can be selectively detected with high accuracy, and the absorption by the fine particles passing through the flow path 12 can be evaluated.
  • the particle measuring device 200 functions as a device that performs optical diffraction photothermal conversion spectroscopic measurement, and such a particle measuring device 200 is called a photothermal conversion spectroscopic device 300 .
  • the other lock-in amplifier 352 is operated at a frequency different from the synchronous frequency of the lock-in amplifier 351 . Specifically, the lock-in amplifier 352 extracts a predetermined frequency component determined based on the size of the observation area by the illumination light IL and the flow velocity of the fluid SL passing through the channel 12. It can also function in the same way as the scattered light measurement device 100 shown in FIG. In this case, the particle measuring device 200 is called the scattered light measuring device 100 .
  • FIG. 9 shows specific measurement results by the particle measuring device 200 of the third embodiment.
  • the horizontal axis indicates the light absorption based on the diffracted light obtained from the output signals of the photodetector 342 and the lock-in amplifier 351 operating as the photothermal conversion spectroscopic device 300
  • the vertical axis indicates the scattered light measuring device 100 operating.
  • 2 shows the intensity of scattered light obtained from the output signals of the photodetector 42 and the lock-in amplifier 352.
  • the fluid SL supplied to the microchip 10 contained gold particles with a diameter of 20 nm, gold particles with a diameter of 40 nm, silver particles with a diameter of 40 nm, and silver particles with a diameter of 60 nm as detection target objects SM.
  • the two-dimensional distribution area differs depending on the type and size of the fine particles. In other words, it is possible to identify fine particles having different sizes and compositions based on scattered signals from the object SM to be detected.
  • FIGS. 10A to 10D are the results of examining the frequency conditions under which the absorption signal and the light scattering signal can be distinguished by changing the modulation frequency of the excitation light. For this reason, we plotted and compared the signals when the excitation light is on (light absorption + light scattering) and when the excitation light is off (light scattering only), and investigated the conditions under which the two can be separated.
  • the horizontal axis indicates a signal having the same frequency as the modulation frequency of the excitation light extracted by the lock-in amplifier 351 (light absorption signal + mixed light scattering signal).
  • the modulation frequency of the excitation light is 2 to 4 kHz
  • the measurement frequency of the scattered light is about 1 kHz. It is desirable that the modulation frequency of the excitation light is relatively high. have understood.
  • the particle measuring device 200 may incorporate the scattered light measuring device 100 and the photothermal conversion spectroscopic device 300 independently.
  • the particle measuring device according to the embodiment has been described above, the particle measuring device according to the present invention is not limited to the above.
  • it can be combined with other various measurement methods such as a fluorescence measurement method in which fluorescence is measured with a fluorescent label.
  • the data processing in the signal processing device 50 or the control device 80 is performed not only for (1) identifying fine particles having different sizes and compositions based on scattering signals corresponding to a plurality of wavelengths, but also for (2) identifying peak signals in the scattering signals.
  • (3) light absorption based on diffracted light obtained from the output signal of the photothermal conversion spectrometer 300 and light scattering obtained from the output signal of the scattered light measurement device 100 Machine learning can also be used when identifying fine particles of different sizes and compositions based on this.

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Abstract

Le problème décrit par la présente invention est de fournir un dispositif et un procédé de mesure de particules fines qui permettent de mesurer facilement et avec une grande précision la lumière diffusée par les particules fines, en utilisant un effet de diffraction de la lumière. La solution selon l'invention porte sur un dispositif de mesure de particules fines 100 qui comprend : un dispositif d'illumination 101 qui illumine un trajet d'écoulement 12 ; et un dispositif de détection 102 qui détecte la lumière provenant du trajet d'écoulement 12. Le dispositif de détection 102 est utilisé pour mesurer l'interférence entre la lumière diffusée L1 provenant des particules traversant le trajet d'écoulement 12 et la lumière diffractée L2 provenant du trajet d'écoulement 12, et des informations relatives aux particules traversant le trajet d'écoulement 12 sont ainsi acquises.
PCT/JP2022/029394 2021-07-30 2022-07-29 Dispositif et procédé de mesure de particules fines WO2023008580A1 (fr)

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JP2021126164A JP2023020673A (ja) 2021-07-30 2021-07-30 微粒子計測装置及び微粒子計測方法

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990005310A2 (fr) * 1988-11-11 1990-05-17 Public Health Laboratory Service Board Mesure optique de vitesse
US20090290156A1 (en) * 2008-05-21 2009-11-26 The Board Of Trustee Of The University Of Illinois Spatial light interference microscopy and fourier transform light scattering for cell and tissue characterization
JP2020204594A (ja) * 2019-06-19 2020-12-24 国立大学法人 東京大学 光熱変換分光装置及び微量検体検出方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990005310A2 (fr) * 1988-11-11 1990-05-17 Public Health Laboratory Service Board Mesure optique de vitesse
US20090290156A1 (en) * 2008-05-21 2009-11-26 The Board Of Trustee Of The University Of Illinois Spatial light interference microscopy and fourier transform light scattering for cell and tissue characterization
JP2020204594A (ja) * 2019-06-19 2020-12-24 国立大学法人 東京大学 光熱変換分光装置及び微量検体検出方法

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