WO2024259548A1 - Appareil de surveillance d'aérosol biologique et procédé de surveillance - Google Patents

Appareil de surveillance d'aérosol biologique et procédé de surveillance Download PDF

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Publication number
WO2024259548A1
WO2024259548A1 PCT/CN2023/100945 CN2023100945W WO2024259548A1 WO 2024259548 A1 WO2024259548 A1 WO 2024259548A1 CN 2023100945 W CN2023100945 W CN 2023100945W WO 2024259548 A1 WO2024259548 A1 WO 2024259548A1
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WIPO (PCT)
Prior art keywords
fluorescence
scattered light
light
particles
optical path
Prior art date
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Pending
Application number
PCT/CN2023/100945
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English (en)
Chinese (zh)
Inventor
张佩
王光辉
陈斌
刘定高
杨晓佳
朱菁
黄惠杰
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Shanghai Leishen Optoelectronic Technology Co Ltd
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Shanghai Leishen Optoelectronic Technology Co Ltd
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Application filed by Shanghai Leishen Optoelectronic Technology Co Ltd filed Critical Shanghai Leishen Optoelectronic Technology Co Ltd
Priority to CN202380013938.4A priority Critical patent/CN118176411A/zh
Priority to PCT/CN2023/100945 priority patent/WO2024259548A1/fr
Publication of WO2024259548A1 publication Critical patent/WO2024259548A1/fr
Anticipated expiration legal-status Critical
Pending legal-status Critical Current

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Classifications

    • 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
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1434—Optical arrangements
    • 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
    • G01N15/149—Optical investigation techniques, e.g. flow cytometry specially adapted for sorting particles, e.g. by their size or optical properties
    • 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

Definitions

  • the invention relates to real-time monitoring of bioaerosol, in particular to a bioaerosol monitoring device and a monitoring method.
  • Excitation light induced intrinsic fluorescence detection technology has many advantages in the field of real-time monitoring of bioaerosols, such as fast speed, high sensitivity, no consumables, and non-invasiveness. Its technical principle is that bioaerosol particles contain organic molecules such as tryptophan, reduced coenzyme I (ie NADH) and riboflavin. These components can produce intrinsic fluorescence under the induction of ultraviolet light, while non-biological aerosol particles are generally difficult to produce intrinsic fluorescence. Based on this, it can be distinguished whether aerosol particles have biological properties.
  • cigarette particles, kaolin, and dust containing polycyclic aromatic hydrocarbons in airborne particles can also generate intrinsic fluorescence signals. Most existing technologies are unable to distinguish these interferents, which may cause false detection or false alarms of the instrument; Second, there are many types of bioaerosol particles. In practical applications, it is often necessary to monitor only a few or a dozen biological targets, while most existing technologies can only monitor the total number of fluorescent particles in general and cannot classify aerosol particles. For example, pollen or some harmless microbial species do not need to be monitored. Therefore, preliminary screening of biological particles is required to reduce the frequency and cost of subsequent biological detection.
  • Patent document CN108375530A discloses a real-time bioaerosol detection method and detection device, including an optical path, an air path intersecting with the optical path, and a signal processing system connected to the optical path.
  • the optical path includes a laser emission optical path for irradiating the measured particles, a scattered light collection optical path for receiving scattered light signals, and a fluorescence collection optical path for receiving fluorescence signals.
  • the air path is used to sample the measured particles, and the minimum detection resolution is one microbial particle.
  • the optical signal processing system is used to analyze and process signals, including a scattered light preamplifier and a fluorescence preamplifier, which can simultaneously monitor the concentration of microbial particles and non-microbial particles in the air.
  • Patent document CN103940709 discloses a real-time microbial particle counter, which determines the particle size and biological properties of the measured particles by detecting the scattered light and fluorescence intensity emitted by a single particle under the irradiation of excitation light.
  • the above two patent documents can only detect the peak value of scattered light pulse and fluorescence pulse, the microbial determination index is single, and the fluorescence bleaching information cannot be tested. Many interference substances are detected, and the false alarm rate of the instrument is high.
  • Patent document CN110411995A discloses a bioaerosol monitoring device and method based on intrinsic fluorescence bleaching characteristics, which detects fluorescence bleaching information and distinguishes the types of aerosol particles according to the fluorescence bleaching characteristics.
  • the instrument first enriches and samples multiple aerosol particles together, and then irradiates the collected particle group for a long time to obtain the fluorescence bleaching characteristics.
  • the disadvantages are: 1 It measures the cumulative fluorescence of multiple particles, and cannot obtain the scattered light, intrinsic fluorescence and fluorescence bleaching characteristics of a single aerosol particle. For samples containing multiple aerosol particles, it is impossible to accurately determine the test results; 2 It needs to enrich a certain number of aerosol particles before testing, the detection sensitivity is low, and it is not real-time monitoring, and the result determination has a certain lag.
  • the present invention provides a bioaerosol monitoring device and a monitoring method, which utilizes that the intrinsic fluorescence bleaching rates generated by organic molecules in bioaerosol particles are different under the irradiation of the same excitation light, such as riboflavin and other components bleaching faster, while NADH and other components bleach slower or almost no bleaching.
  • the same excitation light such as riboflavin and other components bleaching faster, while NADH and other components bleach slower or almost no bleaching.
  • the present invention provides a bioaerosol monitoring device, comprising an optical path, an air path intersecting with the optical path, and a signal processing system connected to the optical path;
  • the optical path comprises a laser irradiation optical path for irradiating measured particles, a scattered light collection optical path for receiving scattered light signals, and a fluorescence collection optical path for receiving fluorescence signals;
  • the air path is used to sample measured particles;
  • the signal processing system is used to analyze and process signals;
  • the intersection area of the laser irradiation optical path and the air path is a light sensitive area;
  • the device is characterized in that the focus of the excitation light source for irradiating measured particles is located before or after the light sensitive area on the optical axis of the laser irradiation optical path, so that the cross section of the excitation light spot perpendicular to the optical axis of the laser irradiation optical path in the light sensitive area is rectangular, and when a single aerosol particle passes through the light sensitive area, it
  • the laser irradiation optical path for irradiating the measured particles includes an excitation light source, the light emitted by the excitation light source is parallel to the X-axis direction and collimated by an aspherical mirror to form a parallel light beam, and then focused by a cylindrical mirror and passes through the light sensitive area to reach the light trap.
  • the dichroic mirror In the Y-axis direction of the light sensitive area, there are a curved reflector and a dichroic mirror in sequence, and the dichroic mirror is set at 45° to the Y-axis.
  • the dichroic mirror has high reflectivity to elastic scattered light and high transmittance to the fluorescent band.
  • the scattered light collecting optical path for receiving the scattered light signal comprises a focusing mirror, a scattering aperture and a scattered light receiver which are sequentially placed along the transmission direction of the elastic scattered light.
  • the fluorescence collection optical path for receiving the fluorescence signal comprises a fluorescence filter, a fluorescence focusing lens, a fluorescence diaphragm and a fluorescence receiver which are sequentially placed along the fluorescence transmission direction.
  • the air circuit is composed of an air inlet nozzle, an air outlet nozzle and an air pump, and the air inlet direction of the air circuit is parallel to the Z axis and passes through the light sensitive area.
  • the signal processing system includes a scattered light amplifying circuit, a scattered light AD conversion circuit, a fluorescence amplifying circuit, a fluorescence AD conversion circuit and an FPGA circuit;
  • the elastic scattered light pulse signal collected by the scattered light receiver is sequentially amplified by the scattered light amplifying circuit, converted into a digital signal by the scattered light AD conversion circuit and reaches the FPGA circuit;
  • the intrinsic fluorescence pulse signal collected by the fluorescence receiver is sequentially amplified by the fluorescence amplifying circuit, converted into a digital signal by the fluorescence AD conversion circuit and reaches the FPGA circuit, and the FPGA circuit is used to calculate the relative fluorescence intensity of the measured aerosol particles, the ratio of the scattering peak values and the intrinsic fluorescence bleaching rate.
  • the present invention also provides a method for monitoring bioaerosols using the above-mentioned bioaerosol monitoring device, comprising the following steps:
  • the peak value, relative fluorescence intensity and intrinsic fluorescence bleaching rate of the elastic scattered light pulse signal of the biological aerosol particles to be tested are calculated by the FPGA circuit, and the type of the aerosol particles to be tested is determined by comparing the landing area in the three-dimensional coordinate system.
  • the present invention has the following beneficial effects:
  • the focus of the excitation light is not located at the photosensitive area, but before or after the photosensitive area on the optical axis of the illumination light path, so that the cross section of the excitation light spot perpendicular to the X-axis in the photosensitive area is approximately rectangular, rather than a thin line.
  • the bleaching speed of the fluorescence signal is not the same, and the fluorescence pulse waveform will also be significantly different.
  • the fluorescence pulse waveform is approximately rectangular; for aerosol particles that only contain a small amount of bleaching components in the fluorescence component, the fluorescence pulse waveform is approximately trapezoidal; for aerosol particles whose fluorescent substances are mainly bleaching components, the fluorescence pulse waveform is approximately triangular. Since the airflow velocity and the size of the illumination beam are fixed, the time it takes for the aerosol particles to pass through the photosensitive area, that is, the duration of the fluorescence pulse signal, is basically fixed. The ratio of the area to the peak value of the fluorescence pulse signal of a single aerosol particle is calculated by FPGA, reflecting the fluorescence bleaching speed of the particle.
  • FIG1 is a schematic diagram of the optical path of an embodiment of a bioaerosol monitoring device of the present invention.
  • FIG. 2 is a schematic diagram of the illumination light path and the gas path in the embodiment of the bioaerosol monitoring device of the present invention.
  • FIG. 3 is a schematic diagram of a signal processing system of an embodiment of a bioaerosol monitoring device of the present invention.
  • FIG. 4 is a schematic diagram of the fluorescence signal pulse waveform of aerosol particles with different bleaching speeds according to the present invention.
  • Figure 1 is a schematic diagram of the optical path of an embodiment of the bioaerosol monitoring device of the present invention
  • Figure 2 is a schematic diagram of the illumination optical path and the gas path in the embodiment of the bioaerosol monitoring device of the present invention.
  • the bioaerosol monitoring device of this embodiment includes an optical path, a gas path and a signal processing system.
  • the optical path includes an excitation light source 101.
  • the excitation light beam emitted by the excitation light source 101 is collimated by an aspherical mirror 102 in parallel with the X-axis direction to form a parallel light beam, and then focused by a cylindrical mirror 103 and passes through a light sensitive area G to reach a light trap 105.
  • the focus M of the excitation light beam is located between the light sensitive area G and the light trap 105 on the optical axis of the illumination optical path.
  • the light sensitive area G is approximately rectangular in cross section perpendicular to the X-axis. In the Y-axis direction of the light sensitive area G, there are curved reflectors 10 4.
  • Photosensitive area G, dichroic mirror 106, the dichroic mirror 10 is set at 45 with the Y axis, the dichroic mirror 106 has high reflectivity to elastic scattered light and high transmittance to the fluorescent band, along the direction of elastic scattered light are scattered light focusing mirror 111, scattered light diaphragm 112, scattered light receiver 113, along the direction of fluorescence are fluorescent filter 107, fluorescent focusing mirror 108, fluorescent diaphragm 109, fluorescent receiver 110, the scattered light receiver 113 and the fluorescent receiver 110 are synchronously reached to the FPGA circuit (not shown) after passing through the signal amplification circuit and AD conversion circuit; the air path is composed of an air inlet nozzle 201, an exhaust nozzle 20 and an air pump, the air inlet direction of the air path is parallel to the Z axis and passes through the photosensitive area G; the intersection of the illumination light path and the air path channel is the photosensitive area G.
  • the light emitted by the excitation light source 101 is collimated by the aspheric mirror 102 to form a parallel beam, and then focused by the cylindrical mirror 103 and passes through the photosensitive area G to reach the light trap 105.
  • the focus of the excitation light is between the photosensitive area G and the light trap 105 on the optical axis of the illumination light path.
  • the photosensitive area is approximately rectangular in the cross section perpendicular to the X axis.
  • the elastic scattered light and intrinsic fluorescence (if any) generated by the aerosol particles passing through the photosensitive area after being excited are collected by the curved reflector 104 of the collection unit and then converted into parallel light to reach the dichroic mirror 106.
  • the dichroic mirror 106 has high reflection for elastic scattered light and high transparency for the fluorescence band. Therefore, the elastic scattered light is reflected by the dichroic mirror 106 to the focusing mirror 111 and then passes through the scattering aperture 112 to reach the scattered light receiver 113. The fluorescence is transmitted by the dichroic mirror 106 and passes through the fluorescence filter 107, the focusing mirror 108 and the fluorescence aperture 109 in sequence to reach the fluorescence receiver 110.
  • the scattered light pulse signal obtained by the scattered light receiver 113 reaches the FPGA circuit 303 after passing through the first signal amplification circuit 301 and the first AD conversion circuit 302, and the fluorescent pulse signal obtained by the fluorescent receiver 110 reaches the FPGA circuit 303 synchronously after passing through the second signal amplification circuit 304 and the second AD conversion circuit 305, as shown in FIG. 3 .
  • FIG2 is a schematic diagram of the relative positions of the illumination optical path and the gas path.
  • the gas path is composed of an air inlet nozzle 201, an exhaust nozzle 202, and an air pump.
  • the air inlet direction of the gas path is parallel to the Z axis and passes through the light sensitive area G.
  • the fluorescence pulse waveforms of different aerosol particles will also be different, as shown in Figure 4.
  • their fluorescence pulse waveform is approximately rectangular, as shown in Figure 4 (a);
  • their fluorescence pulse waveform is approximately trapezoidal, as shown in Figure 4 (b);
  • their fluorescence pulse waveform is approximately triangular, as shown in Figure 4 (c).
  • the monitoring method of the present invention is as follows:
  • the aerosol particles to be measured will emit elastic scattered light pulse signals, and some particles will also emit intrinsic fluorescence pulse signals at the same time. If the measured particle only has elastic scattered light signals, it is determined to be a non-biological particle. Otherwise, the elastic scattered light signal and intrinsic fluorescence pulse signal of the measured particle are simultaneously transferred to the FPGA circuit after passing through the weak signal amplification circuit and the AD conversion circuit.
  • the FGPA circuit obtains the peak value h0 of the scattered pulse signal of the measured aerosol particle, the area s1 of the fluorescence pulse signal, and the peak value h1 of the fluorescence pulse signal through calculation;
  • FPGA further calculates the relative fluorescence intensity of the aerosol particles being measured, that is, the ratio of the fluorescence peak to the scattering peak h1/h0, and the intrinsic fluorescence bleaching rate s1/h1.
  • step (1) and step (2) the scattered light peak value h0, relative fluorescence intensity h1/h0 and intrinsic fluorescence bleaching rate s1/h1 of various aerosol particles that can emit intrinsic fluorescence are tested, and then a three-dimensional coordinate system is established with these three detection values, the coordinates of different types of aerosol particles are marked, and the regions corresponding to the different types of aerosol particles are divided in the coordinate system.
  • the monitoring device detects and calculates the scattered light peak, relative fluorescence intensity and intrinsic fluorescence bleaching rate of the aerosol particles to be tested, thereby obtaining the landing point position of the particles to be tested in the three-dimensional coordinate system, and then determining the type of the aerosol particles to be tested according to the corresponding division area in step (3) corresponding to the landing point position.
  • the present invention collects characteristic information of different types of aerosol particles by analyzing the area, amplitude and other information of the intrinsic fluorescence pulse signal generated when aerosol particles pass through the photosensitive area under continuous irradiation, and combining it with the elastic scattering light pulse signal of the particles, thereby providing a basis for the preliminary classification of aerosol particles.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Dispersion Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)

Abstract

L'invention concerne un appareil de surveillance d'aérosol biologique comprenant un trajet de lumière, un trajet de gaz et un système de traitement de signal, le trajet de lumière comprenant un trajet de lumière d'irradiation laser conçu pour irradier des particules à tester, un trajet de lumière de collecte de lumière diffusée conçu pour recevoir des signaux de lumière diffusée, et un trajet de lumière de collecte de fluorescence conçu pour recevoir des signaux de fluorescence. Un foyer (M) d'une source de lumière d'excitation (101) conçue pour irradier lesdites particules est situé avant ou derrière une zone photosensible (G) sur un axe optique du trajet de lumière d'irradiation laser, de telle sorte que la section transversale d'un point lumineux d'excitation dans la zone photosensible (G) qui est perpendiculaire à l'axe optique du trajet de lumière d'irradiation laser est de forme rectangulaire et, lorsqu'une seule particule d'aérosol passe à travers la zone photosensible (G), cette seule particule d'aérosol est irradiée en continu par une lumière d'excitation pour obtenir une vitesse de détérioration de la fluorescence intrinsèque. Des informations, telles qu'une vitesse de détérioration de la fluorescence intrinsèque, une valeur de pic de lumière diffusée et une intensité de fluorescence relative de particules d'aérosol, sont intégrées, de telle sorte qu'une classification préliminaire de particules biologiques est réalisée, des alertes d'instrument provoquées par des microorganismes non cibles sont significativement réduites, et la capacité d'un instrument pour identifier les types d'aérosols biologiques est améliorée, ce qui permet de réduire la fréquence et le coût de détection biologique ultérieure.
PCT/CN2023/100945 2023-06-19 2023-06-19 Appareil de surveillance d'aérosol biologique et procédé de surveillance Pending WO2024259548A1 (fr)

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Application Number Priority Date Filing Date Title
CN202380013938.4A CN118176411A (zh) 2023-06-19 2023-06-19 生物气溶胶监测装置和监测方法
PCT/CN2023/100945 WO2024259548A1 (fr) 2023-06-19 2023-06-19 Appareil de surveillance d'aérosol biologique et procédé de surveillance

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PCT/CN2023/100945 WO2024259548A1 (fr) 2023-06-19 2023-06-19 Appareil de surveillance d'aérosol biologique et procédé de surveillance

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CN119510268B (zh) * 2025-01-20 2025-08-01 安徽科创中光科技股份有限公司 一种花粉性生物气溶胶浓度监测系统及监测方法

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