Calibration method and calibration device for biological aerosol monitor

Abstract

The invention discloses a calibration method and a calibration device for a biological aerosol monitor, a physical calibration method is combined with a biological verification method, and the calibration method comprises the following steps: S1, calibrating flow parameters of biological aerogel by using a flow meter; s2, putting polystyrene microspheres and polystyrene microspheres with specific fluorescence into the calibration cabin, and uniformly stirring and mixing; s3, connecting a connector on the left side of the calibration cabin with a TSI particle size spectrometer, a particle counter and a calibrated monitor respectively, and carrying out second-step calibration on the monitor by monitoring the particle size and the particle concentration of aerosol particles in the cabin; s4, biological verification: carrying out calibration verification on the biological sampling rate of the biological aerosol sampler; and S5, after the calibration is completed, monitoring the particle concentration in the calibration cabin by adopting a TSI particle size spectrometer until the cleanliness in the cabin meets the requirement. According to the invention, a physical calibration method and a biological verification method can be combined, a uniform, stable and satisfactory aerosol environment is ensured to be provided for calibration work, and the calibration result is more accurate.

Description

Technical Field

[0001] The present invention relates to the technical field of agricultural automation. Specifically, it relates to a calibration method and a management method for a bioaerosol monitor, which can achieve precise irrigation and fertilization of crops and comprehensive management. Background Art

[0002] Bioaerosol refers to an important component of atmospheric aerosol, which is particles directly derived from biological organisms, such as viruses, bacteria, fungi, pollen, cells, or plant debris, animal tissues, etc. However, when the bioaerosol contains pathogenic microorganisms such as bacteria, viruses, allergenic pollen, mold spores, and parasite eggs, the biological characteristics of the aerosol have undergone a fundamental change. Exactly speaking, it should be called pathogenic bioaerosol. Bioaerosol has the characteristics of large pollution area, strong infectivity, and long influence time, and has always threatened human safety. Therefore, it highlights the importance of monitoring, early warning, detection, and identification of harmful bioaerosols.

[0003] Traditional bioaerosol detection mainly relies on on-site sampling, cultivation, analysis, and identification. Its detection process is long, the speed is slow, and contact sampling detection poses a biosafety threat to the safety of the detection personnel themselves. At the end of the 20th century, with the development of laser technology, optoelectronic device manufacturing technology, and spectral detection technology, the technologies based on laser-induced fluorescence technology and light scattering technology have become the main technologies for current bioaerosol monitoring.

[0004] Microbial particles such as bacteria, viruses, and pollen contain various fluorescent substances, such as amino acids and coenzymes. Among them, amino acids exist in all biological organisms and are the basic units that make up protein peptides, while coenzymes, as biomarkers of living organisms, exist in biological organisms with strong metabolism. For organisms with no obvious metabolic activity, the coenzyme content is very low. These organic substances will generate intrinsic fluorescence under the excitation of light with a specific wavelength, which is an important condition for distinguishing biological properties. Since some inorganic minerals also emit fluorescence under ultraviolet light excitation, the particle size range of the collected particles is measured by light scattering technology to exclude interfering substances. By continuously detecting the fluorescence characteristics of biological particles with a specific particle size, a sudden increase in the concentration of biological particles in the air can be found. When the device detects that the increase rate of the concentration of biological particles in the air exceeds the threshold, an alarm will be triggered.

[0005] As a front-end early warning induction device for small portable active biological particles, the working principle of a bioaerosol monitor is as follows: after air pump sampling, it measures the particle size by light scattering method and uses fluorescence spectroscopy to identify the biological and non-biological of aerosol particles to achieve the monitoring of biological particles emitting fluorescence signals in the environment. Therefore, physical parameters such as the excitation light wavelength, fluorescence emission wavelength, and quantum yield provide characteristic information of fluorescent substances and are the basis for qualitative and quantitative analysis of the detected bioaerosol particles.

[0006] Domestic bioaerosol monitors often produce false alarms during actual use. On the one hand, because there are a large number of substances in nature that can produce fluorescence, such as polycyclic aromatic hydrocarbon compounds and natural organic matter, the wavelength ranges emitted by these substances cover or partially overlap with coenzymes, resulting in interference to the bioaerosol monitor and false alarms. On the other hand, when users use bioaerosol monitors, they usually think that the lower the biological alarm threshold, the more sensitive it is, resulting in false alarms due to too low threshold settings. Therefore, it is necessary to establish a scientific evaluation technology to evaluate the performance of bioaerosol monitors, and at the same time provide a reference for setting working parameters according to the monitor performance. However, there are no standard substances and calibration methods for calibrating bioaerosol monitors in China.

[0007] In 2020, the Beijing Institute of Metrology and Testing Science proposed: Select fluorescent reagents with different emission wavelengths, and prepare fluorescent microsphere reference materials that can generate fluorescence by themselves through the SI-ATRP method; combine with a static chamber calibration device, and generate aerosols with different concentrations to calibrate the fluorescence particle counting efficiency of the bioaerosol monitor.

[0008] Beijing Frog Minghuaqing Environmental Protection Technology Co., Ltd. proposed a gas and bioaerosol calibration system and method: Generate bioaerosols into the calibration chamber, use a six-stage sieve-hole air impact sampler to sample into the culture medium, use PBS as a negative control, and calibrate the monitor before leaving the factory by the method of culture counting.

[0009] There are three problems in the existing technology:

[0010] 1) The Beijing Institute of Metrology and Testing Science selects fluorescent reagents with different emission wavelengths and prepares fluorescent microsphere reference materials that can generate fluorescence by themselves through the SI-ATRP method. Since the fluorescence intensity of the fluorescent reagent is much higher than the intrinsic fluorescence intensity of microorganisms, the prepared fluorescent microspheres cannot characterize the fluorescence light intensity of biological characteristics.

[0011] 2) Beijing Frog Minghuaqing Environmental Protection Technology Co., Ltd. uses a method of generating bioaerosols with bacterial solutions and calibrating the monitor by culture counting. Due to the influence of factors such as the activity of biological particles, the generation method, the temperature and humidity in the chamber, and the sampling method in the chamber, the sampling efficiency of bioaerosols varies greatly, and it is difficult to quantitatively characterize the performance parameters of the monitor. At the same time, the biological method is prone to environmental safety risks, requiring laboratories to be equipped with corresponding biological safety protection measures, and operators must also have corresponding biotechnology sample operation and culture technologies.

[0012] 3) The Beijing Institute of Metrology and Measurement Sciences characterized the performance of the calibration device by calibrating the stability, uniformity, and response time of the aerosol in the calibration chamber. A light-scattering particulate matter sensor was placed at the sampling position and the center point in the chamber to monitor the uniformity and stability of the aerosol concentration in the chamber. By observing the changes in the intake air flow rate and the particle concentration in the chamber, the response time of the calibration chamber was verified. This method did not verify the performance of the generation system.

[0013] The present invention aims to solve the problems described above. One object of the present invention is to provide a calibration method and calibration device for a bioaerosol monitor applicable to light scattering and laser-induced fluorescence techniques, and another object is to establish a performance evaluation method for a bioaerosol calibration chamber. Summary of the Invention

[0014] In view of this, the present invention provides a calibration method and calibration device for a bioaerosol monitor applicable to light scattering and laser-induced fluorescence techniques, and an automatic sprinkler fertilization detection management system and management method that can accurately meet the growth requirements of crops.

[0015] To achieve the above object, the present invention adopts the following technical solutions:

[0016] A calibration method for a bioaerosol monitor, which adopts a physical calibration method combined with a biological verification method, includes the following steps:

[0017] S1: Perform the first-step calibration of the flow parameters of the bioaerogel using a flow meter.

[0018] S2: Generate an aerosol into the calibration chamber using polystyrene microspheres and polystyrene microspheres with specific fluorescence through an aerosol generation system. Control the concentration of the aerosol in the calibration chamber by adjusting the generation flow rate and generation time, and stir and mix evenly with a calibration chamber fan.

[0019] S3: Connect the left connection port of the calibration chamber to a TSI particle size spectrometer, a particle counter, and the monitor to be calibrated respectively. Perform the second-step calibration of the monitor by monitoring the particle size and particle concentration of the aerosol in the chamber.

[0020] S4: Biological verification. Atomize the Bacillus subtilis reference material to generate an aerosol with a concentration of 2*10 6 cfu / L. The Andersen sampler and the aerosol monitor sample simultaneously. Compare the results of the Andersen sampling with the results of the aerosol monitor using the colony counting method to calibrate and verify the biological sampling rate of the bioaerosol sampler.

[0021] S5: After calibration is completed, turn on the calibration chamber purification system, and use a TSI particle size spectrometer to monitor the particle concentration in the calibration chamber until the cleanliness of the chamber meets the requirements.

[0022] Further, in step S2, the preparation of the specific fluorescent polystyrene microspheres is as follows: The polystyrene microspheres are ultrasonically dispersed to be uniformly dispersed in an ethanol solvent to form a stable dispersion liquid, and the ultrasonic time is 10 to 30 minutes;

[0023] Transfer the polystyrene microsphere dispersion liquid into a device with an electromagnetic stirring device, protect it with nitrogen filling, control the reaction temperature through a constant temperature water bath, and set the initial reaction temperature at 25 to 35 °C; slowly dropwise add the riboflavin stock solution into the polystyrene microsphere dispersion liquid, control the dropping speed at 1 to 2 drops per second, and at the same time start stirring, adjust the stirring speed to 300 to 600 rpm to make riboflavin fully contact with the polystyrene microspheres, and the reaction time is 1 to 3 hours;

[0024] Under the excitation of the excitation light, by measuring the intrinsic fluorescence intensity of Bacillus subtilis and Phi-X174 phage, and the fluorescence intensity of the fluorescent microspheres, and adjusting the fluorescence intensity of the fluorescent microspheres by controlling the amount of riboflavin, so that the light intensity of the prepared fluorescent microspheres is close to the intrinsic fluorescence light intensity of the microorganisms, and adopt the absolute measurement method, use a scanning electron microscope to calibrate the labeled fluorescent particles, and use spectral analysis to test the fluorescence characteristics of the labeled fluorescent particles;

[0025] Reaction termination and post-treatment: When it is determined that the same intrinsic fluorescence intensity of the microorganisms as that of Bacillus subtilis and Phi-X174 phage is reached, add a terminator to terminate the reaction, and the dosage is 1 to 2 times the molar amount of riboflavin. After adding the terminator, continue to stir the reaction for 10 to 15 minutes to completely terminate the reaction; then, centrifuge the reaction product at a speed of 8000 to 12000 rpm for 10 to 20 minutes to separate, collect the precipitate of polystyrene microspheres grafted with riboflavin; wash the precipitate with deionized water repeatedly for 3 to 5 times to remove the ungrafted riboflavin and other impurities, and finally obtain a suspension of 1 μm and 3 μm fluorescent microspheres.

[0026] Further, the terminator is ethanolamine.

[0027] Further, in step S3, the second calibration includes calibration of the indication error of particle size measurement, calibration of the indication error of particle concentration, and calibration of the counting deviation of fluorescent particles.

[0028] Further, the calibration of the fluorescence particle counting deviation is to dilute the fluorescence microsphere suspensions of 1 μm and 3 μm by 5 to 10 times respectively. After shaking well, monodisperse fluorescence microsphere aerosol is generated through the calibration system, and the aerosol particle concentration is controlled at 45,000 - 55,000 particles / L. Put it into and turn on the bioaerosol monitor. After normal operation for 5 minutes, start recording the particle size measurement results of the bioaerosol monitor. Record the data every 10 s, and record 3 groups in total. Calculate the particle size measurement indication error ΔD according to Equation (1):

[0029]

[0030] In the formula: is the average value of three measurements of the bioaerosol monitor, μm;

[0031] D s is the standard value of the fluorescence microsphere particle size, μm.

[0032] Further, the calibration of the particle concentration indication error is to dilute the fluorescence microsphere suspensions of 1 μm and 3 μm by 5 to 10 times respectively. After shaking well, monodisperse fluorescence microsphere aerosol is generated through the calibration system, and the aerosol particle concentration is controlled at 45,000 - 55,000 particles / L. Turn on the particle counter and the bioaerosol monitor respectively. After normal operation for 5 minutes, record the particle concentration measurement results of the particle counter and the bioaerosol monitor every 10 s. Continuously record 10 groups, and calculate the particle concentration indication error γ according to Equations (2) - (4): d :

[0033]

[0034] In the formula: C si is the particle concentration value measured by the particle counter for the i-th time, particles / L;

[0035] C di is the particle concentration value measured by the bioaerosol monitor for the i-th time, particles / L.

[0036] Further, the calibration of the fluorescence particle counting deviation is to dilute the fluorescence microsphere suspensions of 1 μm and 3 μm by 5 to 10 times respectively. After shaking well, turn on the bioaerosol monitor and the AGI sampler respectively, sample for 5 minutes, and record the fluorescence particle concentration and the total particle concentration measured by the bioaerosol monitor; add the collected liquid of the AGI sampler to the flow cytometer, and measure the fluorescence particle concentration and the total particle concentration in the collected liquid; calculate the fluorescence particle counting deviation according to Equation (5):

[0037]

[0038] In the formula: C M is the number of fluorescence particles measured by the bioaerosol monitor, particles;

[0039] C MT is the total number of particles measured by the bioaerosol monitor, in pieces;

[0040] C S is the number of fluorescent particles in the aerosol sample solution, in pieces;

[0041] C ST is the total number of particles in the aerosol sample solution, in pieces.

[0042] A calibration device for a bioaerosol monitor, the calibration device of the bioaerosol monitor is calibrated by the above-mentioned calibration method of the bioaerosol monitor, and at least includes an aerosol generation and sampling module, a constant temperature and humidity module, a stirring and mixing module, a disinfection module, a purification module and a bioaerosol monitor;

[0043] Further, the aerosol generation and sampling module includes a pump, a generator, an AGI sampler, a TSI particle size spectrometer, a particle counter and an Andersen impact sampler; one end of the pump is connected to the generator, and the other end is connected to the cabin of the bioaerosol monitor; the AGI sampler, the TSI particle size spectrometer, the particle counter and the Andersen impact sampler are all connected to the cabin of the bioaerosol monitor;

[0044] Further, the stirring and mixing module: the stirring and mixing module is arranged at the top and bottom of the cabin of the bioaerosol monitor;

[0045] Further, the disinfection module includes a disinfectant solution box and a peristaltic pump, one end of the peristaltic pump is connected to the disinfectant solution box, and the other end is connected to the generator;

[0046] The purification module is arranged at the top and bottom of the cabin of the bioaerosol monitor, and the gas in the cabin of the bioaerosol monitor is discharged through the high-efficiency filter at the bottom of the side wall, and the supplemented gas enters the cabin through the high-efficiency filter at the top for circulating purification.

[0047] Further, it also includes a constant temperature and humidity module, and the constant temperature and humidity module is arranged in the cabin of the bioaerosol monitor.

[0048] The constant temperature and humidity module equipped in the cabin of the present invention can control the temperature and humidity in the cabin to obtain a stable experimental environment.

[0049] Further, the stirring and mixing module includes at least one large fan arranged on the top of the cabin of the bioaerosol monitor and at least two small fans arranged on the bottom of the cabin of the bioaerosol monitor.

[0050] The bilge small fan of the present invention is combined with the cabin top large fan. Vortices are formed among the small fans and dispersed by the large fan, which can effectively stir and mix biological aerosols.

[0051] The fluorescence value of the fluorescent polystyrene microspheres prepared by the present invention is close to the intrinsic fluorescence of microorganisms. It can realize the use of physical calibration instead of biological sampling and monitoring methods, which can not only avoid the environmental biosafety risks in the process of biological sampling and monitoring, but also avoid the differential influence of factors such as the activity of biological particles, the generation efficiency of biological aerosols and the sampling efficiency on the calibration results. The "fluorescent particle counting deviation" calibration is proposed in the calibration method, which can not only evaluate the fluorescent particle counting efficiency of the biological aerosol monitor, but also verify the anti-interference ability of the fluorescent monitoring system of the monitor. An integrated evaluation method for the working performance of the aerosol calibration chamber is established, which can provide a basis for setting the working parameters of the calibration chamber to ensure a uniform, stable and compliant aerosol environment for the calibration work. Brief Description of the Drawings

[0052] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0053] Figure 1 Technical roadmap for calibrating a biological aerosol monitor;

[0054] Figure 2 Technical route for preparing fluorescent polystyrene microspheres;

[0055] Figure 3 Technical roadmap for evaluating the performance of the calibration system. Detailed Embodiments

[0056] The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the drawings in the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, rather than all embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.

[0057] This embodiment provides a calibration method and a management method for a biological aerosol monitor.

[0058] Such as Figure 1As shown: The calibration method of the bioaerosol monitor adopts a physical calibration method combined with a biological verification method. The physical calibration method is to generate an aerosol of polystyrene microspheres with specific fluorescence into the calibration chamber through an aerosol generation system. The concentration of the aerosol in the calibration chamber is controlled by adjusting the generation flow rate and generation time. The aerosol in the calibration chamber is stirred and mixed evenly by a large fan at the top of the calibration chamber and 4 small fans at the bottom to ensure the uniformity inside the chamber. The left connection port of the calibration chamber is connected to a TSI particle size spectrometer, a particle counter, and the monitor to be calibrated respectively. The physical calibration of the monitor is achieved by monitoring the particle size and particle concentration of the aerosol in the chamber. The specific calibration method is as follows:

[0059] 1) Indication error of particle size measurement. Dilute the suspension of fluorescent microspheres with diameters of 1μm and 3μm, shake well, and generate a monodisperse fluorescent microsphere aerosol through the calibration system. Control the aerosol particle concentration between (45000 - 55000) particles / L or within the range specified by the manufacturer. Turn on the bioaerosol monitor, and after normal operation for 5 minutes, start recording the particle size measurement results of the bioaerosol monitor. Record data every 10 seconds, and record a total of 3 groups. Calculate the indication error ΔD of particle size measurement according to Equation (1):

[0060]

[0061] In the formula: is the average value of the three measurements of the bioaerosol monitor, μm;

[0062] D s is the standard value of the fluorescent microsphere particle size, μm.

[0063] 2) Indication error of particle concentration. Dilute the suspension of fluorescent microspheres with diameters of 1μm and 3μm respectively, shake well, and generate a monodisperse fluorescent microsphere aerosol through the calibration system. Control the aerosol particle concentration between (45000 - 55000) particles / L. Turn on the particle counter and the bioaerosol monitor respectively. After normal operation for 5 minutes, record the particle concentration measurement results of the particle counter and the bioaerosol monitor every 10 seconds. Continuously record 10 groups, and calculate the indication error γ of particle concentration according to Equations (2) - (4): d :

[0064]

[0065] In the formula: C si is the particle concentration value measured by the particle counter for the i - th time, particles / L;

[0066] C di is the particle concentration value measured by the bioaerosol monitor for the i - th time, particles / L;

[0067] 3) Fluorescence particle counting deviation. Dilute the 1 μm and 3 μm fluorescence microsphere suspensions and non-fluorescent microsphere suspensions after rinsing in equal proportions. Generate monodisperse aerosol from the mixed solution through the calibration system. Turn on the bioaerosol monitor and the AGI sampler simultaneously, and sample for 5 minutes. Record the fluorescence particle concentration and total particle concentration measured by the bioaerosol monitor; Add the collected liquid of the AGI sampler to the flow cytometer to measure the fluorescence particle concentration and total particle concentration in the collected liquid; Calculate the fluorescence particle counting deviation according to Equation (5).

[0068]

[0069] In the formula: C M is the number of fluorescence particles measured by the bioaerosol monitor, in pieces;

[0070] C MT is the total number of particles measured by the bioaerosol monitor, in pieces;

[0071] C S is the number of fluorescence particles in the aerosol sample solution, in pieces;

[0072] C ST is the total number of particles in the aerosol sample solution, in pieces.

[0073] After calibration is completed, turn on the calibration chamber purification module, and use a TSI particle size spectrometer to monitor the particle concentration in the calibration chamber. When the particle concentration in the calibration chamber is not greater than the ISO 5 level requirement specified in GB / T 25915.1-2010, the cleanliness of the chamber meets the requirements.

[0074] As Figure 2 shown: The preparation of the polystyrene microspheres with specific fluorescence is as follows: Ultrasonically disperse the polystyrene microspheres to make them uniformly dispersed in an ethanol solvent to form a stable dispersion. The ultrasonic time is 10 to 30 minutes;

[0075] Transfer the polystyrene microsphere dispersion to a container with an electromagnetic stirring device, protect it with nitrogen, control the reaction temperature through a constant temperature water bath, and set the initial reaction temperature at 25 to 35 °C; Slowly drop the riboflavin stock solution into the polystyrene microsphere dispersion, control the dropping speed at 1 to 2 drops per second, and turn on the stirring at the same time. Adjust the stirring speed to 300 to 600 rpm to make riboflavin fully contact with the polystyrene microspheres. The reaction time is 1 to 3 hours;

[0076] Under the excitation of the excitation light, by measuring the intrinsic fluorescence intensity of Bacillus subtilis and Phi-X174 phage, and the fluorescence intensity of the fluorescent microspheres, and by controlling the amount of riboflavin to adjust the fluorescence intensity of the fluorescent microspheres, so that the light intensity of the prepared fluorescent microspheres is close to the intrinsic fluorescence light intensity of the microorganisms, and the absolute measurement method is used, and a scanning electron microscope is used to calibrate the labeled fluorescent particles, and the fluorescence characteristics of the labeled fluorescent particles are tested by spectroscopic analysis;

[0077] Reaction termination and post-treatment: When it is determined that the same intrinsic fluorescence intensity of the microorganisms in Bacillus subtilis and Phi-X174 phage is reached, a terminator is added to terminate the reaction, and the dosage is 1 to 2 times the molar amount of riboflavin. After adding the terminator, the reaction is continuously stirred for 10 to 15 minutes to completely terminate the reaction; then, the reaction product is separated by high-speed centrifugation at a rotation speed of 8000 to 12000 rpm for 10 to 20 minutes, and the precipitate of polystyrene microspheres grafted with riboflavin is collected; the precipitate is washed repeatedly with deionized water 3 to 5 times to remove ungrafted riboflavin and other impurities, and finally a suspension of 1μm and 3μm fluorescent microspheres is obtained.

[0078] Further, the terminator is ethanolamine.

[0079] The calibration device consists of an aerosol generation and sampling module, a constant temperature and humidity module, a stirring and mixing module, a disinfection module, and a purification module.

[0080] 1) Generation and sampling module: The generation and sampling module includes a generation module and a sampling module, including a pump, a generator, an AGI sampler, a TSI particle size spectrometer, a particle counter, and an Andersen impact sampler; the pump provides power to the generator or is externally connected to a Collisin generator through the left connection port for aerosol generation, and the pump can be connected to an AGI sampler, a TSI particle size spectrometer, a particle counter, an Andersen impact sampler, and an aerosol monitor through the sampling port or the left external interface for aerosol collection;

[0081] 2) Constant temperature and humidity module: The constant temperature and humidity module equipped in the chamber can control the temperature and humidity in the chamber to obtain a stable experimental environment;

[0082] 3) Stirring and mixing module: Aerosol generation occurs in the chamber, and it can be stirred and mixed by a large fan at the top and 4 small fans at the bottom to ensure the uniformity in the chamber;

[0083] 4) Disinfection module: After the test, the disinfectant is supplied by a peristaltic pump, and the generator generates the disinfectant into the chamber to disinfect the microorganisms in the chamber; the top ultraviolet lamp can also be turned on for disinfection in the chamber;

[0084] 5) Purification module: Before and after the start and end of the experiment, the gas in the chamber is purified. The gas in the chamber is discharged through the high-efficiency filter at the bottom of the side wall, and the supplemented gas enters the chamber through the high-efficiency filter at the top, and is purified by circulation accordingly.

[0085] As Figure 3 described, the present invention can evaluate the performance of the bioaerosol calibration chamber, specifically including:

[0086] Performance evaluation of the generation module. More specifically, generate known-sized particles, and use a TSI to measure their particle size distribution. If the geometric standard deviation of the particle size distribution < 3%, it is determined to be monodisperse; generate particle concentration repeatability. Under repeated conditions, generate known-sized particles, and use a TSI to measure their particle concentration values, with a total of 6 groups measured. Calculate its repeatability using Bessel's formula, and the repeatability is not greater than 5%.

[0087] Performance evaluation of the sampling module. More specifically, flow deviation: calibrate the sampling flow of the flowmeter in the calibration chamber, set the sampling time to 5 minutes, and compare it with the set flow of the calibration chamber.

[0088] Performance evaluation of the stirring and mixing performance. More specifically, calibrate the aerosol distribution uniformity in the chamber. Generate known-sized particles, evenly distribute 6 measurement points in the sampling plane, use a TSI particle size spectrometer to measure the particle concentration values at the 6 measurement points respectively, and calculate the aerosol distribution uniformity in the calibration chamber using the range method, and the uniformity is not greater than 5%.

[0089] Stability of the aerosol distribution in the calibration chamber per unit time. Generate known-sized particles, continuously sample for 10 minutes using a TSI particle size spectrometer, record a set of data every 1 minute, with a total of 10 groups recorded. Obtain the aerosol concentration stability in the calibration chamber using the range method, which is not greater than 5%

[0090] Performance evaluation of the purification module. More specifically, self-purification time: generate a specified concentration of aerosol, turn on the calibration self-purification mode, and use a TSI to monitor the aerosol concentration in the calibration chamber in real time. It is the time when the particle concentration value meets the requirements of cleanliness level IS05.

[0091] Calibration fluorescent polystyrene microspheres. Select riboflavin and use the chemical grafting method to achieve the fluorescence of polystyrene microspheres with specific particle sizes. And according to three excitation light excitation conditions, by measuring the intrinsic fluorescence intensities of two representative microorganisms (Bacillus subtilis and Phi-X174 phage) in different growth cycles, at four concentrations and in two forms (liquid and aerosol), control the amount of riboflavin to adjust the fluorescence intensity of the fluorescent microspheres, so that the light intensity of the prepared fluorescent microspheres is as close as possible to the intrinsic fluorescence light intensity of the microorganisms. Use the absolute measurement method, use a scanning electron microscope to determine the value of the labeled fluorescent particles, and use spectral analysis to test the fluorescence characteristics of the labeled fluorescent particles.

[0092] Under excitation by 405 nm excitation light on the same optical detection platform, the fluorescence spectra of the fluorescent microsphere solution after labeling, commercially available fluorescent microspheres, and two representative microbial bacterial solutions were compared. It was found that the fluorescence intensity of the fluorescent microspheres provided by the present invention was significantly weaker than that of the commercially available fluorescent microspheres; diluting the fluorescent microsphere solution provided by the present invention by 10 times could characterize the fluorescence intensity of the phage bacterial solution at the exponential phase of 2.0×10 6 pfu / mL.

[0093] The performance evaluation method of the calibration device established by the present invention is to atomize polystyrene microspheres with known particle sizes to generate aerosol into the calibration chamber. By measuring its particle size distribution and particle concentration, the performance of the generation and sampling module, mixing and homogenization module, and purification module of the calibration chamber is verified. The performance of the generation and sampling module of the calibration chamber is verified with a flow meter, and the performance of the constant temperature and humidity module of the calibration chamber during operation is verified with a temperature and humidity meter, so as to comprehensively evaluate the working performance of the calibration device.

[0094] The innovation points of the present invention: 1) Calibrate the fluorescence particle counting deviation of the bioaerosol monitor by physical methods. The specific method is: atomize the equally diluted fluorescent microsphere suspension and non-fluorescent microsphere suspension to generate monodisperse aerosol into the calibration chamber. After mixing evenly, turn on the bioaerosol monitor and AGI sampler respectively, and record the fluorescence particle concentration and total particle concentration measured by the bioaerosol monitor; add the AGI sampler collection solution to the flow cytometer, measure the fluorescence particle concentration and total particle concentration during sampling, and use the deviation between the ratio of the fluorescence particle counting result and the total particle counting result of the bioaerosol monitor and the ratio of the actual fluorescence particle number and total particle number contained in the generated aerosol to obtain the fluorescence particle counting deviation of the bioaerosol monitor.

[0095] 2) Use polystyrene microspheres to generate aerosol through the generation module of the calibration chamber. Verify the monodispersity, aerosol concentration repeatability, aerosol concentration uniformity, and aerosol concentration stability of the calibration chamber generation module by measuring the aerodynamic particle size and particle concentration of the emitted aerosol particles with a TSI particle size spectrometer.

[0096] 3) Prepare polystyrene fluorescent microspheres with specific particle sizes by the epoxy method. Measure the biological intrinsic fluorescence spectra of Bacillus subtilis and phages in different growth cycles and at four gradient concentrations in the bacterial solution state and aerosol state respectively, and adjust the fluorescence value of the fluorescent microspheres to prepare standard microspheres close to the biological intrinsic fluorescence of microorganisms.

[0097] Example 1

[0098] Preparation work

[0099] Calibration device preparation:

[0100] Ensure that the calibration chamber is intact, with good sealing performance and smooth inner walls. Check the aerosol generation and sampling module to ensure that the pump in the chamber is tightly connected to the generator, and the connecting pipelines of the sampling pump to each external device (AGI sampler, TSI particle size spectrometer, particle counter, Andersen impact sampler, and aerosol monitor) are firm and unblocked. Confirm that the temperature sensor and humidity sensor of the constant temperature and humidity module are working properly, the temperature control and humidity control modules are in good condition, and the heat exchanger, ultrasonic atomization or condensation dehumidification equipment can operate normally. The one large fan at the top and four small fans at the bottom of the stirring and mixing module can rotate normally, and the blades are not damaged. The peristaltic pump, generator, and top ultraviolet lamp of the disinfection module can all work normally, the flexible pipeline has no leakage, and the disinfectant storage tank has enough disinfectant. The high-efficiency filters at the air inlet and outlet of the purification module are installed correctly and not damaged.

[0101] Preparation of fluorescent polystyrene microspheres for calibration:

[0102] In the spectroscopy laboratory, use a high-precision spectrometer to measure Bacillus subtilis and Phi-X174 phage. Select the excitation light wavelengths of 350nm, 450nm, and 550nm respectively, and measure the intrinsic fluorescence intensities of the two microorganisms at four concentration levels of 10 3 、10 4 、10 5 、10 6 CFU / mL in the logarithmic growth phase, stationary phase, and decline phase of the three growth cycles, and in both liquid and aerosol states. The sensitivity of the photodetector of the spectrometer reaches 0.01mV, and the resolution of the grating spectroscopic element is 0.1nm. Accurate data is obtained and stored through a complex signal processing algorithm.

[0103] In the chemical reaction kettle, according to the above measurement data, accurately weigh riboflavin using an electronic balance with a precision of 0.0001g, and slowly drip the riboflavin solution into the polystyrene microsphere dispersion with particle sizes of 1μm and 3μm using a pipette, while stirring with a magnetic stirrer at a speed of 300r / min. During the reaction process, the temperature is controlled at 25°C ± 0.5°C, and the pH value is controlled at 7.0 ± 0.1. Real-time monitoring is carried out through the temperature sensor and pH sensor, and the heating device and the addition amount of the acid-base regulator are adjusted.

[0104] After preparation, take a small amount of the fluorescent particle sample and place it under a scanning electron microscope with a magnification of 50,000 times and an image acquisition pixel of 10 million to determine the particle size and morphology. Then place the fluorescent particles in a spectral analyzer, irradiate them with 488 nm excitation light, collect the fluorescence spectrum, and analyze their fluorescence characteristics to ensure compliance with the calibration requirements. Record parameters such as the riboflavin addition amount during the preparation process (e.g., 0.5 mg for 1 μm microspheres and 1.0 mg for 3 μm microspheres), reaction time (3 hours), reaction temperature, etc., to form a quality traceability file.

[0105] Calibration of the indicated error in particle size measurement

[0106] Accurately pipette 5 mL each of the 1 μm and 3 μm fluorescent microsphere suspensions from the storage container using a calibrated pipette, transfer them to a clean beaker, add an appropriate amount of solvent, and use a vortex oscillator to shake well at a speed of 2000 r / min for 5 minutes to fully disperse the microspheres.

[0107] Slowly inject the diluted and shaken fluorescent microsphere suspension into the feed inlet of the aerosol generation device of the calibration device. The calibration device uses microfluidic chip technology and ultrasonic vibration dispersion technology to generate monodisperse fluorescent microsphere aerosol under the condition that the cabin pump provides stable energy for the generator. The particle concentration is monitored in real time by a high-precision concentration control device based on the principle of laser scattering and stabilized in the range of (45000 - 55000) particles / L.

[0108] Turn on the bioaerosol monitor, and the instrument starts to preheat. After 5 minutes, it enters a stable operating state. At this time, the connected computer data acquisition system automatically starts, records the particle size measurement data at 10-second intervals, and records a total of 3 groups of data. For example, the measured values are 1.02 μm, 1.03 μm, 0.98 μm (for 1 μm microspheres) and 3.05 μm, 3.03 μm, 3.02 μm (for 3 μm microspheres). Calculate the indicated error in particle size measurement according to the formula. The average measured value of 1 μm microspheres is 1.01 μm, the standard value is 1 μm, and the error is (1.01 - 1) / 1×100% = 1%; the average measured value of 3 μm microspheres is 3.03 μm, the standard value is 3 μm, and the error is (3.03 - 3) / 3×100% = 1%.

[0109] Calibration of the indicated error in particle concentration

[0110] On the ultra-clean workbench, perform dilution operations on the 1 μm and 3 μm fluorescent microsphere suspensions respectively. Weigh 0.1 g of the microsphere suspension using an electronic balance with a precision of 0.0001 g, accurately pipette 9.9 mL of solvent using a pipette, and shake well with a vortex oscillator.

[0111] The diluted solution is introduced into the calibration device, and monodisperse fluorescent microsphere aerosol is generated through high-pressure gas injection and Venturi tube atomization technology. The supporting concentration monitoring and regulation component controls the particle concentration at (45000 - 55000) particles / L.

[0112] The particle counter and the bioaerosol monitor are turned on simultaneously. After 5 minutes, the data acquisition line transmits the measurement signals of the two to the central data processing unit. The particle concentration measurement results measured by the particle counter and the bioaerosol monitor are recorded every 10 seconds, and 10 groups are continuously recorded. For example, the particle concentration values measured by the particle counter are successively 48000 particles / L, 47500 particles / L, 48200 particles / L, etc. (1μm microspheres), and the values measured by the bioaerosol monitor are successively 46500 particles / L, 47000 particles / L, 47800 particles / L, etc. (1μm microspheres). Calculate the indication error of the particle concentration according to the formula. The calculated indication error of the particle concentration of 1μm microspheres is (assuming the calculation result is) 2%, and similarly calculate the indication error of the particle concentration of 3μm microspheres (assuming the result is) 1.5%.

[0113] Calibration of fluorescence particle counting deviation

[0114] The 1μm and 3μm fluorescent microsphere suspensions and the non-fluorescent microsphere suspension are diluted in a ratio of 1:1 with the assistance of a high-precision balance. Use a magnetic stirrer to stir at a speed of 500 r / min for 10 minutes to ensure uniform mixing.

[0115] The diluted mixture is converted into monodisperse aerosol through the piezoelectric ceramic-driven microspray device of the calibration device. At the same time, the bioaerosol monitor and the AGI sampler are turned on. The sampling ports of the two are opposite and the distance is set to 10 cm, and synchronous sampling is carried out for 5 minutes. During the sampling period, the bioaerosol monitor real-time displays the fluorescence particle concentration and the total particle concentration, and the operator records. For example, the fluorescence particle concentration is 2000 particles / mL and the total particle concentration is 5000 particles / mL (1μm microspheres).

[0116] After the sampling is completed, carefully inject the collected liquid of the AGI sampler into the sample chamber of the flow cytometer. The flow cytometer uses the 488nm laser excitation, fluorescence detection and signal amplification functions to accurately measure the fluorescence particle concentration and the total particle concentration in the collected liquid. Assume that the measured fluorescence particle concentration is 1800 particles / mL and the total particle concentration is 4800 particles / mL (1μm microspheres). Calculate the fluorescence particle counting deviation according to the formula. The fluorescence particle counting deviation of 1μm microspheres is (assuming the calculation result is) 10%, and similarly calculate the fluorescence particle counting deviation of 3μm microspheres (assuming the result is) 8%.

[0117] Purify the calibration chamber after calibration

[0118] Press the start button of the calibration chamber purification module, the ventilation duct valve switches to the purification mode, and the air circulation pump runs at high speed. The TSI particle size spectrometer is connected to the gas in the chamber through a sampling tube to monitor the particle concentration in real time. When the monitored particle concentration is not greater than the ISO 5 standard specified in GB / T 25915.1-2010 (assumed to be 1000 particles / m 3 )), the purification control system emits a prompt tone, and the control panel displays "The cleanliness in the chamber meets the standard", completes the purification operation, and prepares for the next round of calibration or other operations.

[0119] Example 2

[0120] Preparation work

[0121] Calibration device preparation: The same as the calibration device preparation steps in Example 1 to ensure that all systems and equipment are working properly.

[0122] Preparation of fluorescent polystyrene microspheres for calibration:

[0123] In the spectral measurement step, select excitation light wavelengths of 360 nm, 460 nm, and 560 nm, and measure Bacillus subtilis and Phi-X174 phage at different growth cycles, concentration levels, and two morphological forms. The spectrometer parameters are the same as in Example 1, and the data is acquired and stored.

[0124] In the chemical synthesis reaction, according to the measurement data, adjust the riboflavin addition amount (0.6 mg for 1μm microspheres and 1.2 mg for 3μm microspheres), and control other reaction conditions such as the temperature at 26°C ± 0.5°C, the pH value at 7.2 ± 0.1, the stirring speed at 350 r / min, and the reaction time at 3.5 hours.

[0125] The quality inspection steps are similar to those in Example 1. The fluorescent particles are detected by a scanning electron microscope and a spectral analyzer to ensure that the quality meets the requirements, and the preparation parameters are recorded to form a file.

[0126] Calibration of the indication error of particle size measurement

[0127] Absorb 6 mL of each of the 1μm and 3μm fluorescent microsphere suspensions, add different solvents for dilution, and shake well with a vortex oscillator for 6 minutes.

[0128] Inject into the feed inlet of the aerosol generating device to generate monodisperse fluorescent microsphere aerosol, and regulate the particle concentration to be (45000 - 55000) particles / L. After turning on the bioaerosol monitor and preheating for 5 minutes, it enters stable operation, and record 3 groups of particle size measurement data. For example, the measured values of 1μm microspheres are 1.01μm, 1.04μm, and 0.99μm, and the measured values of 3μm microspheres are 3.04μm, 3.02μm, and 3.06μm. Calculate the indicated error of the particle size measurement. The error of the 1μm microsphere is (1.013 - 1) / 1×100% = 1.3%, and the error of the 3μm microsphere is (3.04 - 3) / 3×100% = 1.33%.

[0129] Calibration of the indicated error of particle concentration

[0130] Dilute the 1μm and 3μm fluorescent microsphere suspensions. Weigh 0.12g of the microsphere suspension and transfer 9.88mL of the solvent, then shake well.

[0131] Generate aerosol and control the concentration. Turn on the particle counter and the bioaerosol monitor, and record 10 groups of particle concentration measurement results after 5 minutes. For example, the particle concentration values of 1μm microspheres measured by the particle counter are 47800 particles / L, 48500 particles / L, etc., and the values measured by the bioaerosol monitor are 46800 particles / L, 47300 particles / L, etc. Calculate the indicated error of the particle concentration. The error of the 1μm microsphere is (assuming the calculation result is) 2.2%, and the error of the 3μm microsphere is (assuming the result is) 1.8%.

[0132] Calibration of the fluorescence particle counting deviation

[0133] Dilute the fluorescent microsphere and non-fluorescent microsphere suspensions in a ratio of 1:1.2 and stir with a magnetic stirrer for 12 minutes.

[0134] After converting to aerosol, turn on the bioaerosol monitor and the AGI sampler. Set the sampling port distance to 12cm and sample synchronously for 5 minutes, and record the data displayed by the monitor. For example, the fluorescence particle concentration is 2200 particles / mL, and the total particle concentration is 5500 particles / mL (1μm microspheres).

[0135] Inject the sampling liquid into the flow cytometer for measurement. Assume that the measured fluorescence particle concentration is 1900 particles / mL and the total particle concentration is 5000 particles / mL (1μm microspheres). Calculate the fluorescence particle counting deviation. The deviation of the 1μm microsphere is (assuming the calculation result is) 13.6%, and similarly calculate the deviation of the 3μm microsphere (assuming the result is) 10.5%.

[0136] Purify the calibration chamber after calibration

[0137] Start the purification module. When the particle concentration monitored by the TSI particle size spectrometer meets the standard (not greater than the ISO Class 5 standard), the purification is completed, and it is determined that the cleanliness of the chamber meets the standard, and subsequent operations can be carried out.

[0138] As can be seen from the above embodiments, the calibration method and device of the bioaerosol monitor can effectively calibrate the monitor, ensure the accuracy and reliability of its measurement, and achieve good calibration effects under different operating conditions and parameter settings. At the same time, the preparation process of the fluorescent polystyrene microspheres for calibration is strict and traceable, providing high-quality reference materials for the calibration work.

[0139] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present embodiments. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present embodiments.

[0140] Therefore, the present embodiments will not be limited to these embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for calibrating a bioaerosol monitor, characterized in that: The physical calibration method is combined with the biological verification method, including the following steps: S1: Use the flow meter to calibrate the flow parameters of the bioaerogel in the first step; S2: Use polystyrene microspheres and polystyrene microspheres with specific fluorescence to generate aerosols into the calibration chamber through an aerosol generation system, control the concentration of aerosols in the calibration chamber by adjusting the generation flow rate and generation time, and stir and mix them by the calibration chamber fan; S3: The connection ports on the left side of the calibration cabin are connected to the TSI particle size spectrometer, particle counter and calibrated monitor respectively, and the monitor is calibrated in the second step by monitoring the aerosol particle size and particle concentration in the cabin; S4: Biological verification: atomize the Bacillus subtilis standard substance to a concentration of 2*10 6 cfu / L aerosol, the Anderson sampler and the aerosol monitor sampled at the same time, and the results of the Anderson sampling were compared with those of the aerosol monitor using the colony counting method to calibrate and verify the biological sampling rate of the bioaerosol sampler; S5: After the calibration is completed, turn on the calibration cabin purification module and use the TSI particle size spectrometer to monitor the particle concentration in the calibration cabin until the cleanliness in the cabin meets the requirements.

2. The calibration method of a bioaerosol monitor according to claim 1, characterized in that: In step S2, the specific fluorescent polystyrene microspheres are prepared by: dispersing the polystyrene microspheres uniformly in an ethanol solvent by ultrasonic dispersion to form a stable dispersion, wherein the ultrasonic time is 10 to 30 minutes; The polystyrene microsphere dispersion is transferred to an electromagnetic stirring device, filled with nitrogen for protection, and the reaction temperature is controlled by a constant temperature water bath, and the initial reaction temperature is set at 25-35°C; the riboflavin stock solution is slowly added to the polystyrene microsphere dispersion, and the drop speed is controlled at 1-2 drops / second, and stirring is started at the same time, and the stirring speed is adjusted to 300-600rpm to make the riboflavin and polystyrene microspheres fully contact, and the reaction time is 1-3 hours; Under the condition of excitation light, the intrinsic fluorescence intensity of microorganisms of Bacillus subtilis and Phi-X174 bacteriophage was measured, and the fluorescence intensity of fluorescent microspheres was adjusted by controlling the amount of riboflavin, so that the light intensity of the prepared fluorescent microspheres was close to the intrinsic fluorescence intensity of microorganisms. The absolute measurement method and scanning electron microscope were used to determine the value of the labeled fluorescent particles, and the fluorescence characteristics of the labeled fluorescent particles were tested by spectral analysis. Reaction termination and post-treatment: When it is determined that the intrinsic fluorescence intensity of the microorganism is the same as that of Bacillus subtilis and Phi-X174 bacteriophage, a terminator is added to terminate the reaction in an amount of 1 to 2 times the molar amount of riboflavin. After adding the terminator, the reaction is continued to be stirred for 10 to 15 minutes to completely terminate the reaction; then, the reaction product is separated by high-speed centrifugation at a speed of 8000 to 12000 rpm and a centrifugation time of 10 to 20 minutes, and the polystyrene microsphere precipitate grafted with riboflavin is collected; the precipitate is repeatedly washed with deionized water for 3 to 5 times to remove ungrafted riboflavin and other impurities, and finally a 1μm and 3μm fluorescent microsphere suspension is obtained.

3. The calibration method of a bioaerosol monitor according to claim 2, characterized in that: The terminator is ethanolamine.

4. The calibration method of a bioaerosol monitor according to claim 3, characterized in that: In step S3, the second step calibration includes particle size measurement indication error calibration, particle concentration indication error calibration, and fluorescent particle counting deviation calibration.

5. The calibration method of a bioaerosol monitor according to claim 4, characterized in that: The fluorescent particle counting deviation calibration is to dilute the 1μm and 3μm fluorescent microsphere suspensions by 5 to 10 times, shake them thoroughly, and generate monodisperse fluorescent microsphere aerosols through the calibration system. The aerosol particle concentration is controlled at 45,000 to 55,000 particles / L. The bioaerosol monitor is placed and turned on. After 5 minutes of normal operation, the bioaerosol monitor particle size measurement results are recorded. The data are recorded every 10 seconds, and a total of 3 groups are recorded. The particle size measurement indication error ΔD is calculated according to formula (1): Where: is the average of three measurements by the bioaerosol monitor, μm; D s is the standard value of fluorescent microsphere particle size, μm.

6. The calibration method of a bioaerosol monitor according to claim 4, characterized in that: The particle concentration indication error calibration is to dilute the 1 μm and 3 μm fluorescent microsphere suspensions by 5 to 10 times, shake them thoroughly, generate monodisperse fluorescent microsphere aerosols through the calibration system, and control the aerosol particle concentration at 45,000 to 55,000 particles / L. The particle counter and the bioaerosol monitor are turned on respectively. After 5 minutes of normal operation, the particle concentration measurement results of the particle counter and the bioaerosol monitor are recorded every 10 seconds. Ten groups are recorded continuously, and the particle concentration indication error γ is calculated according to formulas (2) to (4): d : Where: C si is the particle concentration value measured by the particle counter for the i-th time, particles / L; C di is the particle concentration value measured by the bioaerosol monitor for the i-th time, particles / L.

7. The calibration method of a bioaerosol monitor according to claim 4, characterized in that: The fluorescent particle count deviation calibration is to dilute the 1 μm and 3 μm fluorescent microsphere suspensions by 5 to 10 times, shake them thoroughly, start the bioaerosol monitor and AGI sampler, sample for 5 minutes, record the fluorescent particle concentration and total particle concentration measured by the biosol monitor; add the AGI sampler collection liquid to the flow cytometer, measure the fluorescent particle concentration and total particle concentration in the collection liquid; calculate the fluorescent particle count deviation according to formula (5): Where: C M is the number of fluorescent particles measured by the bioaerosol monitor, pieces; C MT The total number of particles measured by the bioaerosol monitor, pcs; C S is the number of fluorescent particles in the aerosol sample solution, pieces; C ST is the total number of particles in the aerosol sample solution, pieces.

8. A calibration device for a bioaerosol monitor, characterized in that: The calibration device of the bioaerosol monitor is calibrated by the calibration method of the bioaerosol monitor based on the principle of laser-induced biological intrinsic fluorescence and light scattering according to any one of claims 1 to 7, and at least includes an aerosol generation and sampling module, a constant temperature and humidity module, a stirring and mixing module, a disinfection module, a purification module and a bioaerosol monitor; The aerosol generation and sampling module includes a pump, a generator, an AGI sampler, a TSI particle size spectrometer, a particle counter and an Anderson impact sampler; one end of the pump is connected to the generator, and the other end is connected to the bioaerosol monitor cabin; the AGI sampler, the TSI particle size spectrometer, the particle counter and the Anderson impact sampler are all connected to the bioaerosol monitor cabin; The stirring and mixing module: The stirring and mixing module is arranged at the top and bottom of the bioaerosol monitor cabin; The disinfection module includes a disinfectant box and a peristaltic pump, one end of the peristaltic pump is connected to the disinfectant box, and the other end is connected to the generator; The purification module is arranged at the top and bottom of the bioaerosol monitor cabin. The gas in the bioaerosol monitor cabin is discharged through the high efficiency filter at the bottom of the side wall, and the supplementary gas enters the cabin through the high efficiency filter at the top for circulation and purification.

9. The calibration device for a bioaerosol monitor according to claim 8, characterized in that: It also includes a constant temperature and humidity module, which is arranged in the cabin of the bioaerosol monitor.

10. The calibration device for a bioaerosol monitor according to claim 8, characterized in that: The stirring and mixing module includes at least one large fan arranged on the top of the bioaerosol monitor cabin and at least two small fans arranged on the bottom of the bioaerosol monitor cabin.

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