CN103940709A - Real-time microbial particle counter - Google Patents
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Abstract
The invention discloses a real-time microbial particle counter comprising a light path, an air channel intersected with the light path, and a signal processing system connected with the light path, wherein the light path comprises a lighting light path for irradiating tested particles and a collecting light path arranged along the advancing direction of the lighting light path; the collecting light path also comprises a relay system for separating the light path and separately detecting separated light paths; the air channel is used for sampling the tested particles; the signal processing system is used for analyzing and processing a signal, and comprises a fluorescent preamplifier and a scattered light preamplifier. The concentration of microbial particles in air is monitored in real time by taking laser-induced fluorescence detection as the principle, the particle sizes and the biological attributes of the tested particles are judged by detecting the intensities of scattered light and fluorescence emitted by exciting light, and the particles in a sampling airflow are counted, so that the automatic detection on planktonic bacteria microorganisms is achieved, and the real-time microbial particle counter is simple and convenient to operate, strong in detection instantaneity, high in sensitivity and high in accuracy.
Description
Technical field
The invention belongs to suspension microorganism detection field, particularly a kind of microorganism corpuscular counter in real time.
Background technology
The microorganisms such as infectiousness germ or virus are to be all present in atmosphere with particulate state, in the time that the concentration of infectiousness germ in atmosphere or virus exceedes a certain threshold value, will threaten to the mankind and vegeto-animal health.Meanwhile, because microorganism can produce various hypopus, can in air, survive for a long time, and by air dielectric diffusion and transmission, thereby cause the outburst of various infectious diseases and spread, cause serious harm.So be badly in need of detecting in real time technology and the equipment of microorganism particle in surrounding environment.
Microorganism particle contains multiple organic molecule, and wherein main fluorescent material is tryptophane, nicotinamide adenine dinucleotide (NADH), lactochrome and pyridinedicarboxylic acid (DPA) etc.These metabolins of biosome can send intrinsic fluorescence under ultraviolet light or royal purple irradiation, and wherein, tryptophane has three absorption peaks, is respectively 220nm, 280nm and 288nm; Nicotinamide adenine dinucleotide (NADH) has two absorption peaks, is respectively 260mn and 340nm; Lactochrome has three absorption peaks, is respectively 240nm, 370nm and 450nm; Pyridinedicarboxylic acid (DPA) is mainly present in bacterial spore inwall, and absorption bands is about 320～420nm, and absorption peak is about 400nm.The contained composition of different microorganism particles is often different, as often contained above-mentioned each organic molecular species in bacterium, fungi, spore, and generally only contains aminoacid ingredient in virus, toxin.So different microorganism particles present different light absorption and the characteristics of luminescence.
The detection of microbes in air particle is not only widely used in the evaluation of the indoor air quality such as medical industry toilet, hospital clean operation department, biological products toilet, and can be effectively for biological weapons early warning in military affairs and anti-terrorism field.
Existing microorganism detection method is mainly the bacterium sampling cultivation that swims: collected and be suspended in airborne active microorganism particle on the nutrient culture media of double dish by borne bacteria sampler, under suitable condition, allow its breeding count to visible bacterium colony, judge the viable microbial number in clean environment with the clump count in double dish, and evaluate the biomone cleanliness factor of toilet's (or clean area) with this.
There is following shortcoming in above-mentioned existing method:
(1) testing procedure complexity, flow process is loaded down with trivial details.
Each test all will be passed through the steps such as sterilization, sampling, cultivation, counting, all needs personnel to operate in test process.Before test, need to possess plurality of devices and the equipment such as borne bacteria sampler, double dish, nutrient culture media, constant incubator, autoclave sterilizer, magnifier.
(2) sense cycle is long, does not have real-time.
The bacterium that swims sampling cultivation generally needs several days time just can obtain testing result, and sense cycle is longer, does not have real-time.Therefore the method cannot be applied to the application higher to requirement of real-time, as biological weapons early warning.
(3) rate of failing to report is higher.
The bacterium that swims sampling cultivation is counted after microorganism is cultivated, but the different growth of microorganism cycles is not identical yet, is therefore difficult to ensure the quantity of all microorganisms to add up completely.In incubation time, some microorganism may not yet breed count enable degree, and some microorganism may be dead.In addition, exist and a large amount of can not cultivate microorganism in air, they cannot form bacterium colony on nutrient culture media, also cannot count by the bacterium sampling cultivation that swims.
Summary of the invention
Goal of the invention: the invention provides a kind of microorganism corpuscular counter in real time, the concentration of energy Real-Time Monitoring microbes in air particle.
Technical scheme: a kind of microorganism corpuscular counter in real time, comprises light path, the gas circuit crossing with light path, the signal processing system being connected with light path:
Light path, comprises illumination path for irradiating tested particle and along the collection light path arranging in illumination path working direction, collects light path and also comprise for separating of light path and to separating the relay system of light path separate detection;
Gas circuit, for the tested particle of sampling;
Signal processing system, for analyzing and processing signals, comprises fluorescence prime amplifier (31) and scattered light prime amplifier (36).
Wherein gas circuit, for sampling gas circuit, contains admission piece, outlet nozzle and vacuum air pump.Admission piece and outlet nozzle lay respectively at the both sides in the irradiation region of illumination path.Under the driving of vacuum air pump, be sampled gas and enter from admission piece, the particle in gas, is discharged by outlet nozzle one by one by irradiation region afterwards with certain speed.
Between described illumination path and relay system, be also provided with successively curved mirror, the first light trapping, non-spherical lens, the focus of curved mirror is positioned at described light path and the gas circuit light sensitive area forming that crosses.
Described curved mirror is parabola collecting lens, and the focus of described parabola collecting lens is positioned at the center of light sensitive area.
Described relay system comprises the collimating mirror, dichronic mirror and the phosphor collection light path that in light path working direction, set gradually along collecting, and the below of dichronic mirror reflection direction is provided with scattered light and collects light path.
Described relay system also comprises the second light trapping for absorbing unnecessary scattered light, and the second light trapping is arranged on the top of dichronic mirror reflection direction, relative with scattered light collection light path.
The incident angle of described dichronic mirror is 30 °～60 °.
Described phosphor collection light path comprises the fluorescent optical filter, fluorescence focus lamp, fluorescence diaphragm and the fluorescent probe that in light path working direction, set gradually along collecting, and fluorescence diaphragm is positioned at the focal plane place of fluorescence focus lamp, and fluorescent probe is connected with fluorescence prime amplifier.
Described scattered light is collected light path and is comprised the scattered light focus lamp, scattered light diaphragm and the detector for scattered light that set gradually along dichronic mirror reflection direction downwards, scattered light diaphragm is positioned at the focal plane place of scattered light focus lamp, and detector for scattered light is connected with scattered light prime amplifier.
Before relay system, scattered light light path and fluorescence light path are common light paths, by being arranged on after the dichronic mirror in relay system, scattered light and fluorescence are separated, after scattered light reflection turnover, converge at through scattered light focus lamp on the photosurface of detector for scattered light, fluorescence converges on the test surface of fluorescent probe through fluorescence focus lamp after directly seeing through dichronic mirror and fluorescent optical filter again.
Described relay system also comprises the first diaphragm, and the first diaphragm is positioned at described collimating mirror and non-spherical lens confocal and locates.
Described signal processing system also comprises the modular converter, processor, buffer and the host computer that connect in turn, and the input end of modular converter is connected with the output terminal of scattered light prime amplifier with the output terminal of fluorescence prime amplifier respectively.Signal processing system is for analyzing and process the original burst signal of detector for scattered light and fluorescent probe output.The original burst signal of detector for scattered light and fluorescent probe output is low current signal, after scattered light prime amplifier and the processing of fluorescence prime amplifier, all become analog voltage signal respectively, then enter modular converter simultaneously, digital signal after conversion is admitted to processor and extracts peak value of pulse, pulse amplitude is sent to host computer after buffer, host computer paired pulses amplitude is analyzed and is processed, and finally calculates the quantity of particle and the quantity of biomone in each particle diameter section.
Described illumination path comprises the semiconductor laser, illumination collimating mirror and the cylindrical mirror that set gradually, described light sensitive area is the light area of focus of cylindrical mirror and the intersecting area of gas circuit, the plane that described gas circuit forms perpendicular to the focal line of cylindrical mirror and the optical axis of illumination path.The laser beam that semiconductor laser sends, after over-illumination collimating mirror collimation, is become elongated hot spot by cylindrical mirror one-dimensional focusing, for irradiating tested particle.The irradiation region that the focus place of cylindrical mirror forms and the intersecting area of sampling air flow are called light sensitive area, and the intersection point of the optical axis of illumination path and cylindrical mirror focal line is the center of light sensitive area.The optical axis three conllinear of the optical axis of illumination path, the central axis of parabola collecting lens and fluorescence light path, the focus of parabola collecting lens and the center superposition of light sensitive area, the optical axis of scattered light light path and the optical axis of fluorescence light path after reflection turnover are vertical.The central axis of sampling air flow is through the center of light sensitive area.
Described semiconductor laser is that wavelength is the laser diode of 350nm～410nm.Can effectively excite most biosomes such as bacterium, fungi, gemma of fluorescent materials such as containing nicotinamide adenine dinucleotide (NADH), lactochrome or pyridinedicarboxylic acid (DPA) to produce fluorescence.
Principle of work: semiconductor laser gives off laser beam, after illumination collimating mirror, in the direction perpendicular to illumination path optical axis, form the laser beam that xsect is rectangle, the long limit of this laser beam is identical with sampling air flow central axis, then along the long side direction of rectangular light beam by cylindrical mirror one-dimensional focusing to light sensitive area, enter the first light trapping and be absorbed through the laser beam of light sensitive area.Under the driving of air pump, particle to be measured enters admission piece with sampling air flow, behind light sensitive area, discharges from outlet nozzle.Each particle to be measured, in through light sensitive area, can produce the proportional scattered light signal of intensity and its particle size, if biomone, meanwhile also can produce the fluorescence signal of some strength.Particle to be measured is just positioned near the focus of parabola collecting lens by laser beam irradiation time, therefore, the optical axis direction that the scattered light that it sends and fluorescence are parallel to illumination path after the reflection of parabola collecting lens penetrates, and then after non-spherical lens focusing and collimating mirror collimation, arrives dichronic mirror successively.Dichronic mirror is high anti-and thoroughly high to fluorescence to scattered light, and therefore scattered light is reflected after dichronic mirror, then focuses on scattered light diaphragm through scattered light focus lamp, projects on detector for scattered light through after scattered light diaphragm.Fluorescence is converged at fluorescence diaphragm by fluorescence focus lamp directly see through fluorescent optical filter after dichronic mirror after, finally arrives on fluorescent probe, and the second light trapping is for absorbing unnecessary scattered light.Detector for scattered light transfers signals on scattered light prime amplifier, obtains the analog voltage pulse signal proportional with scattered light intensity; Fluorescent probe transfers signals on fluorescence prime amplifier, obtains the analog voltage pulse signal proportional with fluorescence intensity.Modular converter at least contains two-way ALT-CH alternate channel, can described two-way analog voltage pulse signal be converted to digital signal simultaneously.Digital signal after conversion is admitted to processor and extracts pulse amplitude, and pulse amplitude is sent to host computer after buffer, and thus, host computer obtains scattered light and the fluorescence intensity information of tested particle simultaneously.Host computer is got rid of the interference of pseudo-biomone according to the scattered light intensity information of tested particle, then calculate particle concentration and biomone concentration value in different-grain diameter interval.
Beneficial effect: scattered light and fluorescence intensity that the present invention sends under exciting light irradiates by detection single particle judge particle size and the biological attribute of tested particle, and the particle in sampling air flow is counted.Compared with prior art, tool of the present invention has the following advantages:
(1) robotization that can realize the bacteria microorganism concentration of swimming detects, easy and simple to handle.
Before each use, after tool housing is carried out to strict sterilization, after unlatching instrument, can carry out microorganism concn detection, after having detected, can obtain testing result by the host computer of instrument.The invention belongs to automatic detection instrument, require low to operating personnel's technical ability.In addition, the present invention is without adopting the consumptive materials such as sampling membrane, nutrient culture media, and therefore operation steps is more easy.
(2) detection sensitivity is high, real-time.
Sampling cultivation generally needs several days time just can obtain testing result, detects in real time and can carry out scene after the present invention's unlatching, and sense cycle is short, and real-time is good.The present invention may detect single biomone, even when airborne microorganism particle concentration is very low, still can be detected, therefore detection sensitivity of the present invention is high.
(3) accuracy in detection is high.
The present invention is taking laser-Induced Fluorescence Detection as principle, the nearly all microorganism particle including not educable microorganism can be detected.In addition, the present invention can get rid of according to the scattered light information of tested particle the interference of pseudo-biomone, thereby effectively reduces rate of false alarm.Therefore, accuracy in detection of the present invention is high.
Brief description of the drawings
Fig. 1 is illumination path and the schematic diagram of collecting light path in the present invention;
Fig. 2 is the schematic diagram of gas circuit and illumination path in the present invention;
Fig. 3 is the schematic diagram of signal processing system in the present invention;
Fig. 4 is the particle diameter coverage of variety classes particle in air;
Fig. 5 is the transmittance curve figure of fluorescent optical filter in collection light path of the present invention.
Embodiment
A kind of microorganism corpuscular counter in real time, comprises light path, gas circuit and signal processing system, and light path comprises illumination path, collects light path, and gas circuit is sampling gas circuit.
As shown in Figure 1, illumination path comprises semiconductor laser 101, illumination collimating mirror 102, cylindrical mirror 103, and the laser beam that semiconductor laser 101 sends, after over-illumination collimating mirror 102 collimations, is become elongated hot spot by cylindrical mirror 103 one-dimensional focusings.The intersecting area of the irradiation region that the focus place of cylindrical mirror 103 forms and the sampling air flow of gas circuit forms light sensitive area 118, and the intersection point of the optical axis of illumination path and cylindrical mirror focal line is 118 center, light sensitive area.
Collect light path and comprise that curved mirror is parabola collecting lens 104 along the curved mirror setting gradually in illumination path working direction, the first light trapping 105, non-spherical lens 106 and relay system.Relay system comprises collimating mirror 108, dichronic mirror 109, the second light trapping 110, fluorescent optical filter 111, fluorescence focus lamp 112, fluorescence diaphragm 113, fluorescent probe 114, scattered light focus lamp 115, scattered light diaphragm 116 and detector for scattered light 117.Fluorescent optical filter 111, fluorescence focus lamp 112, fluorescence diaphragm 113, fluorescent probe 114 form phosphor collection light path; Scattered light focus lamp 115, scattered light diaphragm 116 and detector for scattered light 117 form scattered light and collect light path.Dichronic mirror 109 is 45 ° of dichronic mirrors.Before relay system, scattered light light path and fluorescence light path are common light paths, by being arranged on after 45 ° of dichronic mirrors in relay system, scattered light and fluorescence are separated, after 90 ° of scattered light reflection turnovers, converge at through scattered light focus lamp 115 on the photosurface of detector for scattered light 117, fluorescence converges on the test surface of fluorescent probe 114 through fluorescence focus lamp 112 after directly seeing through dichronic mirror 109 and fluorescent optical filter 111 again, and the second light trapping 110 is for absorbing unnecessary scattered light.
The optical axis three conllinear of the optical axis of illumination path, the central axis of parabola collecting lens 104 and fluorescence light path, is designated as OO', and the focus of parabola collecting lens 104 overlaps with the center O of light sensitive area 118, the optical axis O of the scattered light light path after reflection turnover
1o
1' vertical with the optical axis OO' of fluorescence light path.
As shown in Figure 2, sampling gas circuit comprises admission piece 21, outlet nozzle 22 and vacuum air pump.Under the driving of vacuum air pump, be sampled gas and enter from admission piece 21, the particle in gas, is discharged by outlet nozzle 22 one by one by light sensitive area 118 afterwards with certain speed.The central axis O of sampling air flow
2o
2' through the center O of light sensitive area 118, and the plane forming perpendicular to the focal line of cylindrical mirror 103 and the optical axis OO' of illuminator.
As shown in Figure 3, the original burst signal that signal processing system is exported for analyzing and process detector for scattered light 117 and fluorescent probe 114.Signal processing system comprises fluorescence prime amplifier 31, scattered light prime amplifier 36, modular converter 32, processor 33, buffer 34 and host computer 35, modular converter 32 is A/D modular converter, processor 33 is FPGA processor, and buffer 34 is FIFO buffer; The original burst signal that detector for scattered light 117 and fluorescent probe 114 are exported is low current signal, after scattered light prime amplifier 36 and 31 processing of fluorescence prime amplifier, all become analog voltage signal respectively, then enter modular converter 32 simultaneously, digital signal after conversion is admitted to processor 33 and extracts peak value of pulse, pulse amplitude is sent to host computer 35 after buffer 34, and host computer 35 paired pulses amplitudes are analyzed and processed.Collect light path and comprise 3 diaphragm for eliminating stray light, the first diaphragm 107 is arranged at non-spherical lens 106 and collimating mirror 108 confocal and locates, fluorescence diaphragm 113 is arranged at the fluorescent probe 114 focal plane place of fluorescence focus lamp 112 before, and scattered light diaphragm 116 is arranged at the detector for scattered light 117 focal plane place of scattered light focus lamp 115 before.
Semiconductor laser 101 gives off laser beam, and after illumination collimating mirror 102, forms the laser beam that xsect is rectangle, the long limit of this laser beam and sampling air flow central axis O in the direction perpendicular to illumination path optical axis OO'
2o
2' identical, then along the long side direction of rectangular light beam by cylindrical mirror 103 one-dimensional focusings to light sensitive area 118, enter the first light trapping 105 and be absorbed through the laser beam of light sensitive area 118.Under the driving of vacuum air pump, particle to be measured enters admission piece 21 with sampling air flow, behind light sensitive area 118, discharges from outlet nozzle 22.Each particle to be measured, in through light sensitive area 118, can produce the proportional scattered light signal of intensity and its particle size, if biomone, also can produce the fluorescence signal of some strength.Particle to be measured is just positioned near the focus O of parabola collecting lens 104 by laser beam irradiation time, therefore, the optical axis OO' direction that the scattered light that it sends and fluorescence are parallel to illumination path after 104 reflections of parabola collecting lens penetrates, then after non-spherical lens 106 focusing and collimating mirror 108 collimations, arrive 45 ° of dichronic mirrors successively, the first diaphragm 107 is for eliminating parasitic light.
Dichronic mirror 109 is high anti-and thoroughly high to fluorescence to scattered light, and therefore scattered light, by 90 ° of reflections after dichronic mirror 109, then focuses on scattered light diaphragm 116 through scattered light focus lamp 115, projects on detector for scattered light 117 through after scattered light diaphragm 116.Fluorescence is converged at fluorescence diaphragm 113 by fluorescence focus lamp 112 directly see through fluorescent optical filter 111 after dichronic mirror 109 after, finally arrives on fluorescent probe 114.Detector for scattered light 117 transfers signals on scattered light prime amplifier 36, obtains the analog voltage pulse signal proportional with scattered light intensity; Fluorescent probe 114 transfers signals on fluorescence prime amplifier 31, obtains the analog voltage pulse signal proportional with fluorescence intensity.A/D module contains two-way A/D ALT-CH alternate channel, can described two-way analog voltage pulse signal be converted to digital signal simultaneously.Digital signal after conversion is admitted to FPGA processor and extracts pulse amplitude, and pulse amplitude is sent to host computer 35 after FIFO buffer, and thus, host computer 35 obtains scattered light and the fluorescence intensity information of tested particle.Host computer 35 is got rid of the interference of pseudo-biomone according to the scattered light intensity information of tested particle, then calculate particle concentration and abiotic particle concentration value in different-grain diameter interval.In air, contain the multiple particles such as dust, smog, bacterium, fungi, pollen, except bacterium, fungi, spore, pollen, cigarette particle etc. also can produce fluorescence under the exciting of 405nm wavelength light source, can cause interference to the detection of microorganism particle.As shown in Figure 4, the particle size range difference that different types of dust particle covers, can get rid of the interference of the anaphylactogens such as pollen and part cigarette particle accordingly.
It is the laser diode of 405nm that the present embodiment semiconductor laser 101 adopts wavelength, and power is about 70mW.Can effectively excite most biosomes such as bacterium, fungi, gemma of fluorescent materials such as containing nicotinamide adenine dinucleotide (NADH), lactochrome or pyridinedicarboxylic acid (DPA) to produce fluorescence.Illumination collimating mirror 102 is non-spherical lens, and focal length is 7.8mm.The laser beam that semiconductor laser 101 sends is about directional light after collimating mirror 102, and beam cross section is about the rectangle of 4mm × 2mm.Cylindrical mirror 103 is planoconvex lens, adopts optical glass K9, and focal length is 45.3mm, and the focal line size that laser beam forms at the center O place of light sensitive area 118 after cylindrical mirror 103 is about 2mm × 0.027mm.The solid angle that parabola collecting lens 104 is opened the center O of light sensitive area 118 is 2.204 π sterad.The first surface of non-spherical lens 106 is ellipsoid, and second is sphere, and focal length is 119.8mm.Fluorescence numerical aperture after non-spherical lens 106 focuses on is 0.292, and the object space numerical aperture of relay system is 0.292.The clear aperature of the first diaphragm 107 is 7mm × 1mm.Collimating mirror 108, fluorescence focus lamp 112 and the scattered light focus lamp 115 collected in light path adopt the cemented doublet with same structure parameter, focal length is 35.0mm, numerical aperture NA=0.329, its value is greater than the numerical aperture that focuses on rear fluorescence from non-spherical lens 106, therefore can not cause energy loss.Dichronic mirror 109 materials are K9 glass, and its zone of reflections centre wavelength is 405nm, and bandwidth is 380-420nm, and transmission bandgap wavelength scope is 430-650nm.The transmittance curve of fluorescent optical filter 111 as shown in Figure 5.The clear aperature of fluorescence diaphragm 113 and scattered light diaphragm 116 is Φ 6.0mm.Fluorescent probe 114, for having the side window type photomultiplier of multialkali photocathode material, have very high quantum efficiency, and typical gains is high in 200-800nm wave band.Detector for scattered light 117 is for having the silicon PIN type photodiode of high sensitivity and high response frequency, and spectral response range is 320-1100nm.
The light trapping seat that the first light trapping 105 has delustring line by heat absorbing glass and inwall forms.The profile of heat absorbing glass is the right cylinder with 45 ° of inclined-planes, and material adopts neutral gray glass, after 45 ° of polishing inclined planes, plates 405nm high transmittance film.Enter absorption glass from the laser beam overwhelming majority transmission of semiconductor laser 101 and be absorbed, the unabsorbed laser beam of minority is reflexed on the inwall of light trapping seat by 45 ° of inclined-planes, after multiple reflections, is almost absorbed completely by the delustring line of light trapping inwall.The second light trapping 110 adopts the inner light trapping with taper delustring line, for absorbing unnecessary scattered light.
The outlet internal diameter of admission piece 21 and the entrance internal diameter of outlet nozzle 22 are respectively Φ 1.1mm and Φ 1.5mm.Fluorescence prime amplifier 31 and scattered light prime amplifier 36 are all current/voltage-converted and the voltage amplifier circuits that form taking high-performance field effect cast dual operational amplifier as core.A/D module adopts the high-speed a/d sampling A/D chip that contains two-way A/D ALT-CH alternate channel, the signal that FPGA processor transmits A/D module extracts pulse amplitude after processing, because FPGA processor does not have memory function, need additional FIFO buffer as register, to meet the processing requirements of particulate in air concentration when higher.Last host computer 35 judges particle size and the biological attribute of tested particle according to the scattered light recording and fluorescence information simultaneously, and calculates particle concentration and abiotic particle concentration value in different-grain diameter interval.The minimum detection resolution of embodiment is 1 biomone, and air sampling flow is 1L/min.
Claims (12)
1. a real-time microorganism corpuscular counter, comprises light path, the gas circuit crossing with light path, the signal processing system being connected with light path, it is characterized in that:
Light path, comprises illumination path for irradiating tested particle and along the collection light path arranging in illumination path working direction, collects light path and also comprise for separating of light path and to separating the relay system of light path separate detection;
Gas circuit, for the tested particle of sampling;
Signal processing system, for analyzing and processing signals, comprises fluorescence prime amplifier (31) and scattered light prime amplifier (36).
2. real-time microorganism corpuscular counter as claimed in claim 1, it is characterized in that, between described illumination path and relay system, be also provided with successively curved mirror, the first light trapping (105), non-spherical lens (106), the focus of curved mirror is positioned at described light path and the gas circuit light sensitive area (118) forming that crosses.
3. real-time microorganism corpuscular counter as claimed in claim 2, it is characterized in that, described curved mirror is parabola collecting lens (104), and the focus of described parabola collecting lens (104) is positioned at the center of light sensitive area (118).
4. real-time microorganism corpuscular counter as claimed in claim 1, it is characterized in that, described relay system comprises the collimating mirror (108), dichronic mirror (109) and the phosphor collection light path that in light path working direction, set gradually along collecting, and the below of dichronic mirror (109) reflection direction is provided with scattered light and collects light path.
5. real-time microorganism corpuscular counter as claimed in claim 4, it is characterized in that, described relay system also comprises the second light trapping (110) for absorbing unnecessary scattered light, the second light trapping (110) is arranged on the top of dichronic mirror (109) reflection direction, relative with scattered light collection light path.
6. the real-time microorganism corpuscular counter as described in claim 4 or 5, is characterized in that, the incident angle of described dichronic mirror (109) is 30 °～60 °.
7. real-time microorganism corpuscular counter as claimed in claim 4, it is characterized in that, described phosphor collection light path comprises the fluorescent optical filter (111), fluorescence focus lamp (112), fluorescence diaphragm (113) and the fluorescent probe (114) that in light path working direction, set gradually along collecting, fluorescence diaphragm (113) is positioned at the focal plane place of fluorescence focus lamp (112), and fluorescent probe (114) is connected with fluorescence prime amplifier (31).
8. real-time microorganism corpuscular counter as claimed in claim 4, it is characterized in that, described scattered light is collected light path and is comprised the scattered light focus lamp (115), scattered light diaphragm (116) and the detector for scattered light (117) that set gradually along dichronic mirror reflection direction downwards, scattered light diaphragm (116) is positioned at the focal plane place of scattered light focus lamp (115), and detector for scattered light (117) is connected with scattered light prime amplifier (36).
9. real-time microorganism corpuscular counter as claimed in claim 4, it is characterized in that, described relay system also comprises the first diaphragm (107), and the first diaphragm (107) is positioned at described collimating mirror (108) and non-spherical lens (106) confocal and locates.
10. real-time microorganism corpuscular counter as claimed in claim 1, it is characterized in that, described signal processing system also comprises the modular converter (32), processor (33), buffer (34) and the host computer (35) that connect in turn, and the input end of modular converter (32) is connected with the output terminal of scattered light prime amplifier (36) with the output terminal of fluorescence prime amplifier (31) respectively.
11. real-time microorganism corpuscular counters as claimed in claim 1, it is characterized in that, described illumination path comprises the semiconductor laser (101), illumination collimating mirror (102) and the cylindrical mirror (103) that set gradually, described light sensitive area (118) is the light area that focuses on of cylindrical mirror (103) and the intersecting area of gas circuit, the plane that described gas circuit forms perpendicular to the focal line of cylindrical mirror (103) and the optical axis of illumination path.
12. real-time microorganism corpuscular counters as claimed in claim 11, is characterized in that the laser diode that described semiconductor laser (101) is 350nm～410nm for wavelength.
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