JP2000338030A - Method and apparatus for counting blue-green algae, algae and fine particle - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a method and apparatus capable of counting separately the whole of blue-green algae and algae and the whole of fine particles so as to be capable of smoothing process control at a time of an increase in respective measured values while monitoring the number and density of the whole of the blue-green algae and algae and the whole of fine particles because a problem such as the obstruction of flocculation, the clogging of a filter, offensive smell and taste or the outflow of algae to a water purifying basin is generated by the generation of algae, in particular, blue-green algae in raw water of a water purifying plant. SOLUTION: The detection of the whole of blue-green algae by the fluorescence generated from phycocyanin contained in blue-green algae, the detection of the whole of algae by the fluorescence generated from chlorophyll a and the detection of the whole of fine particles containing algae by a conventional light scattering or cutting-off system are performed and, by combining these three systems, an apparatus for counting the whole of blue-green algae and algae and the whole of fine particles separately can be produced.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method and an apparatus for counting the number concentration of whole fine particles, the number concentration of whole algae and the number concentration of cyanobacteria in a sample water.

[0002]

2. Description of the Related Art In the water supply field, it is necessary to control the turbidity of the water at the outlet of a filtration pond and the concentration of fine particles as a measure against pathogenic microorganisms such as cryptosporidium. Demand for turbidimeters and particle counters is increasing.

A high-sensitivity turbidity meter of a particle counting type and a fine particle counting type described in Japanese Patent Application No. 9-54612, “Method and Apparatus for Measuring Turbidity,” filed by the present inventors, can be used in a sample water. It has a function of counting fine particles, and the measuring methods of these devices include a light scattering method and a light blocking method.

[0004] In the light blocking method, when a light beam is irradiated from a light source toward sample water, a light beam passing through the sample water is converted into an electric signal by a photoelectric conversion element installed on the opposite side of the light source through the sample water. Has a mechanism to convert. When light is blocked by fine particles in the sample water, the voltage of the electric signal drops, so that a pulse is observed in the electric signal every time the fine particle passes through the light beam. Particle counting by the light blocking method is performed by counting this pulse with a counter. Since the pulse peak value corresponds to the particle size, if a threshold value corresponding to each particle size is provided, the particle number concentration for each particle size category can be measured.

On the other hand, in the light scattering method, when a light beam is irradiated from a light source, side scattered light is collected by a lens system installed at a certain angle with respect to an optical axis connecting the light source and the sample. Later, the light is directly excluded by the side scattered light method, which has a mechanism to convert to an electric signal by the photoelectric conversion element, and the beam stop installed on the optical axis opposite to the light source through the sample, and then forward by the lens system. There is a forward scattered light method having a mechanism of collecting scattered light and converting the scattered light into an electric signal by a photoelectric conversion element. Both, the observation area defined by the pinhole and the light receiving area of the photoelectric conversion element, by counting the pulse signal observed by the photoelectric conversion element every time the fine particles pass by the counter,
Perform particle counting. If a threshold value corresponding to each particle size is provided in the same manner as in the light blocking method, the concentration of the number of fine particles in each particle size category can be measured.

The light blocking method has a disadvantage that the sensitivity to particles having a particle size of 1 μm or less is small, but it can measure a wide range of particles ranging from tens to hundreds of μm, and the optical system is relatively inexpensive. It has the feature that it can be designed to.

[0007] The side scattered light method of the light scattering method requires a light source having a higher output and an expensive light receiving system as compared with the light blocking method, but can measure particles of submicron or less. is there.

In the forward scattered light method among the light scattering methods, the optical axis connecting the light source and the sample coincides with the optical axis of the light receiving system, and stray light is easily generated from the boundary between the flow cell and the sample other than the observation area. Therefore, the measurable particles are limited to about submicron, but an optical system which is inexpensive and easy to adjust as compared with the side scattering method can be used.

Recently, there has been a growing need to monitor algae generated in large quantities in water purification plants. -128235, "Method and apparatus for counting algae and fine particles". In this application, a fluorescence condensing optical system for counting algae by detecting a fluorescent pulse due to chlorophyll a or phycocyanin in algae, a scattered light condensing optical system for counting the whole fine particles, and a transmitted light receiving optical system And algae and other fine particles are counted separately.

[0010]

In a water treatment plant,
A large amount of algae is generated in raw water, which may cause problems such as aggregation inhibition, filtration blockage, off-flavor, and outflow of algae to a water purification pond. When a large amount of algae is generated, measures such as changing the chlorine injection rate and the coagulant injection rate and stopping water intake are taken. At this time, it is better to monitor the number concentration of algae and immediately take measures such as changing the treatment method when the algae increase, rather than taking measures after a serious problem such as filtration blockage occurs. In addition, process control can be performed smoothly.

The most suitable method for monitoring algae is to count organisms such as algae while distinguishing them from other fine particles. However, in the above-described ordinary fine particle counter or fine particle counting type high-sensitivity turbidity meter, a sample is used. Although it is possible to count fine particles in water, it has no function to determine whether the fine particles are inorganic substances such as sand or living things such as algae. However, by applying the method described in Japanese Patent Application No. 11-128235, it is possible to distinguish and count algae and other fine particles.

However, among the algal disorders in the water purification process, the disorders caused by blue-green algae are particularly serious problems. Even among the same algae, cyanobacteria and other algae have different methods of treating algal disorders such as different chlorine injection points. Therefore, if blue-green algae and other algae can be distinguished and counted and the number concentration of both can be monitored, more precise process control can be performed when algae are generated. Japanese Patent Application Hei 11
According to the algae counting method described in -128235, since blue-green algae and other algae cannot be counted separately, there is a problem that this process control cannot be performed.

[0013] The present invention provides a method and apparatus capable of distinguishing and counting cyanobacteria, whole algae and whole fine particles in a sample water in order to solve the above-mentioned problem of distinguishing cyanobacteria from other algae. Is to do.

[0014]

In order to achieve the above object, the present invention provides (a) detection of cyanobacteria by fluorescence generated from phycocyanin contained in cyanobacteria, and (b) chlorophyll a Detecting the whole algae by the fluorescence generated from, and (c) detecting the whole fine particles including the algae by the conventional light scattering method and light blocking method,
By combining the three methods (a), (b), and (c), the cyanobacteria, the whole algae, and the whole fine particles are counted separately.

As a more specific configuration, (a) and (b) show a method of irradiating a light beam from a laser toward a sample water containing algae flowing in a flow cell, and generating algae generated when the light beam passes through the light beam. And a fluorescence pulse counting unit for detecting a fluorescence pulse due to phycocyanin or chlorophyll a therein.

(C) is a scattered light focusing optical system for detecting scattered light pulses generated when fine particles (including algae or other fine particles) pass through the light beam, and a scattered light pulse counting unit. A transmitted light receiving optical system and a transmitted light for detecting a dimming pulse of the transmitted light generated when the particles pass through the light beam (including algae or other fine particles) and the light beam is cut off; This is a pulse counting unit.

The optical system and the counting section are combined to form an apparatus. In the fluorescence condensing optical systems for (a) and (b), scattered light from inorganic substances is blocked by a filter, and the number of fluorescent pulses specific to the entire algae and the number of fluorescent pulses specific to the cyanobacteria are counted. Kind can be detected.

In the scattered light collecting optical system for (c), the conventional light scattering system and the transmitted light receiving optical system are equivalent to the optical system used in the light blocking type particle counter, and the whole particles are counted. It has no function to distinguish algae from other fine particles.

Here, there are four types of combinations of the above optical systems depending on the purpose. (1) When counting the whole algae and the cyanobacteria, the fluorescence condensing optical system is used, and (2) the algae are used. The fluorescence condensing optical system and the scattered light condensing optical system are used for simultaneous counting of microparticles and submicron particles. (3) The fluorescence condensing optical system and transmitted light are used for simultaneous counting of algae and fine particles of several microns or more. The light receiving optical system is configured using (4) a fluorescence condensing optical system, a scattered light condensing optical system, and a transmitted light receiving optical system when simultaneously counting algae and fine particles ranging from submicron to several microns or more. .

The details of each optical system and the counting unit will be described below. However, the scattered light collecting optical system and the transmitted light receiving optical system, that is, the light scattering system and the light blocking system are conventional technologies, and will not be described. Omitted, the characteristics of algae fluorescence and the method of counting fluorescence pulses are described.

The wavelength of the fluorescence generated when the algae is irradiated with excitation light is approximately 650 nm for phycocyanin contained in cyanobacteria as shown in FIG. 1 and about 650 nm for chlorophyll a as shown in FIG. There is a peak around 680 nm.

In general, the excitation spectrum of phycocyanin has a peak over a wide range of about 500 to 640 nm, and has a maximum at about 610 nm. On the other hand, the wavelength of the excitation light of chlorophyll a is 430 nm in order to suppress the fluorescence other than chlorophyll a as much as possible. However, the actual excitation spectrum of chlorophyll a is wide and has a peak near 670 nm. Are known.

Since algae have the above-mentioned fluorescent properties,
The fluorescence optical system captures only the fluorescence from algae,
It is necessary to install a color filter that transmits only light of about 650 nm or more, or an interference filter that transmits light near 650 to 750 nm.

In order to observe a fluorescent pulse having a large peak value, it is suitable that the excitation light source has a longer wavelength in the range of 500 to 640 nm. So 635nm
The use of the semiconductor laser is more advantageous than the use of an excitation light source having a wavelength of 600 nm or less from the viewpoint of miniaturization and cost reduction of the entire apparatus. However, when an excitation wavelength of 600 nm or more is used, the excitation light may pass through the filter of the fluorescence optical system, making it impossible to distinguish between the fluorescence of the algae and the scattered light of other fine particles. It is necessary to use either an interference filter or a method for shifting the fluorescence detection wavelength of phycocyanin to a wavelength longer than 650 nm.

FIG. 3 is a schematic diagram of a pulse signal observed by each light detecting means when a sample water containing algae is irradiated with a light beam from the excitation light source using the above-described excitation light source and filter.

From this figure, it can be seen that (a) the phycocyanin fluorescence pulse is for cyanobacteria, (b) the chlorophyll a fluorescence pulse is for the entire algae, and (c) the scattered light pulse or light blocking pulse is for the entire microparticles containing algae. It turns out that it can be observed. In other words, in terms of the type of fine particles, in other words, fine particles other than algae are scattered light pulses and light-blocking pulses, and algae other than blue-green algae are scattered light pulses, light-blocking pulses and chlorophyll-a fluorescent pulses. (Scattered light, light blocking, chlorophyll a fluorescence, and phycocyanin fluorescence pulses).

The phycocyanin fluorescence pulse, chlorophyll-a fluorescence pulse, scattered light pulse and light blocking pulse thus observed are counted by the following method. Phycocyanin, which is generated each time an algae passes through the light beam,
Alternatively, the fluorescence of chlorophyll a is condensed by the respective fluorescence condensing optical systems after the scattered light and the direct light beam are excluded by the above-mentioned filter, and is converted into an electric signal by the photoelectric converter. This electric signal is counted by the fluorescence pulse counting unit, and the total number of alga and cyanobacteria in the sample water is calculated and output.

The scattered light generated every time the algae or other fine particles pass through the light beam is collected by a scattered light collecting optical system, and converted into an electric signal by a photoelectric converter. The electric signal is counted by the scattered light pulse counting unit, and the number concentration of the fine particles including algae in the sample water is calculated and output.

The transmitted light, which is dimmed every time algae or other fine particles pass through the light beam, is converted into an electric signal by a photoelectric converter. This electric signal is counted by the transmitted light pulse counting unit, and the number concentration of the fine particles including algae in the sample water is calculated and output.

The peak values of the fluorescent pulse by the algae, the scattered light pulse by the algae or other fine particles, and the transmitted light pulse increase according to the size of the algae or other fine particles. Measured by a peak hold circuit, compared with a predetermined threshold group corresponding to the particle size classification of cyanobacteria, or the whole algae, or the whole fine particles, and count for each particle size class. The number of comparators can be prepared, threshold values corresponding to each particle size section are provided, and the number of comparators is compared with each pulse signal to count for each particle size section.

[0031]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below based on two embodiments. [Example 1] An optical system for detecting forward scattered light pulses, transmitted light pulses, and forward fluorescent light pulses and a counting unit were provided in order to separately count cyanobacteria, whole algae, and whole fine particles in sample water. FIG. 4 shows an example of a device configuration diagram, and FIG. 5 shows an example of a block diagram of a transmitted light pulse counting unit using a peak hold circuit.

In FIG. 4, the forward scattered light pulse, the transmitted light pulse and the forward fluorescent light pulse are counted, and the number concentration of the whole fine particles including algae, the number concentration of the whole algae, and the number concentration of the cyanobacteria are distinguished. FIG. 2 shows a configuration diagram of an apparatus for measuring.

In this apparatus, by counting forward scattered light pulses due to fine particles, the number concentration of fine particles including algae having a particle size on the order of submicrons is determined. Measures the number concentration of fine particles including algae of several microns or more, and counts the number of forward fluorescence pulses by the whole algae and blue algae, thereby measuring the number concentration of the whole algae and blue algae, respectively.

1. Counting of transmitted light pulse: A light beam 1a emitted from a laser 1 that emits a light beam having a wavelength of 640 nm or less with an Ar laser, a He-Ne laser, an LD-pumped solid-state laser, a semiconductor laser, or the like is collected by a focusing optical system 2. In a direction perpendicular to the direction in which the sample water 4 flows, the shape is flat, and in the direction in which the sample water flows, light is condensed. After the light beam is attenuated by the filter 6 embedded in the perforated plano-convex lens 5, the light beam is converted into an electric signal 7 a by the photodiode 7. The electric signal 7a becomes a pulse signal every time the algae 8 or other fine particles pass through the observation region in the light beam.
The pulse signal due to the light blocking of the fine particles is transmitted to the transmitted light pulse counting unit 9 to measure the number concentration 9a of the fine particles in the sample water.
Is output for each particle size classification. In the light blocking method, it is not possible to measure fine particles on the order of submicrons, so the particle size is set within a range of several microns or more.

FIG. 5 is a block diagram of a transmitted light pulse counting unit using a peak hold circuit for counting transmitted light pulses. In the transmitted light pulse counting unit 9 in this figure, first, an electric signal 7 a from the photodiode 7 is amplified by the preamplifier 10 and the main amplifier 11. An output electric signal of the main amplifier 11 is input to a low-pass filter (hereinafter, referred to as LPF) 13 and an LPF 12 in parallel. The LPF 13 has a higher cutoff frequency than the transmitted light pulse signal in the output electric signal of the main amplifier 11, and the output electric signal is a transmitted light pulse signal from which high-frequency noise has been removed. On the other hand, LPF12 is LP
It has a cutoff frequency and a smoothing circuit sufficiently lower than F13, and by smoothing this low-frequency output electric signal, an average value, that is, an output electric signal of a DC component due to stray light or the like is obtained. By subtracting the output electric signal of the LPF 12 from the electric signal of the LPF 13 by the differential amplifying unit 14, a DC component due to stray light or the like is subtracted from these two outputs,
The peak value of the transmitted light pulse signal as a measured value is obtained in the next peak hold circuit 15.

Next, the peak value measured by the peak hold circuit 15 every time a pulse signal is generated by passing a fine particle through the observation region in the light beam is calculated by the arithmetic circuit 16
Is compared with a threshold value category corresponding to the particle size category determined in advance, and counting is performed for each particle size category. The value counted for each particle size classification is output from the display output circuit 17 as the number concentration 9a of the whole fine particles in the sample water.

2. Counting scattered light pulses: with the device of FIG.
The forward scattered light 18 or forward fluorescence 19 generated when algae or other fine particles pass through the light beam 1a,
A parallel light flux is formed by the plano-convex lens 5 and the achromatic lens 20 having a hole at the center, and light of the wavelength of the light beam 1a is reflected, and only the forward scattered light 18 is reflected by the dichroic mirror 21 that transmits the other light. The light is reflected at a right angle and enters the achromatic lens 22. Next, the forward scattered light 18 reflected at right angles is condensed by the achromatic lens 22, and after the stray light is removed by the pinhole 23, the light is converted into an electric signal 24 a by the photodiode 24. The electric signal 24a is transmitted from a scattered light fluorescence condensing optical system constituted by the plano-convex lens 5, the achromatic lens 20, and the dichroic mirror 21 and a forward scattered light condensing optical system constituted by the achromatic lens 22, and the pinhole 23. Each time an algae or other fine particles pass through the observation area, a pulse signal is generated. The pulse signal due to the forward scattering of the fine particles is output by the scattered light pulse counting unit 25 having the same function as that of the transmitted light pulse counting unit 9 as the number concentration measurement value 25a of the fine particles in the sample water for each particle size classification. . The particle size classification is set mainly in the submicron range that cannot be measured by the light blocking method.

3. Fluorescence pulse counting: The forward fluorescence 19, which is generated when the algae 8 passes through the light beam 1a and is made into a parallel light flux by the perforated plano-convex lens 5 and the achromatic lens 20, passes through the dichroic mirror 21,
Next, although the fluorescent light having a peak wavelength of 680 nm is transmitted, the fluorescent light having a peak wavelength of 650 nm is separated into fluorescent light of chlorophyll a and phycocyanin by a dichroic mirror 26 which reflects at right angles.
And 28, and pinholes 29 and 30
After the stray light is removed, the photomultiplier tubes 31 and 32 convert the light into electric signals 31a and 32a.
The electric signal 31a is composed of a scattered light fluorescence condensing optical system composed of a plano-convex lens 5, an achromatic lens 20, and a dichroic mirror 21, and a chlorophyll a fluorescence collection composed of a dichroic mirror 26, an achromatic lens 27, and a pinhole 29. Each time an algae passes through the observation area of the optical optical system, it becomes a pulse signal. Further, the electric signal 32a is provided by a scattered light fluorescence condensing optical system including a plano-convex lens 5, an achromatic lens 20, and a dichroic mirror 21,
Dichroic mirror 26, achromatic lens 2
8. Each time a cyanobacterium passes through the observation area of the phycocyanin fluorescence condensing optical system constituted by the pinhole 30, it becomes a pulse signal. The fluorescence pulse signal is supplied to the chlorophyll-a fluorescence pulse counting unit 33 and the phycocyanin fluorescence pulse counting unit 34 having the same function as the transmitted light pulse counting unit 9 to measure the number concentration 33a of the whole algae in the sample water and the cyanobacteria. Is output for each particle size classification. [Example 2] An optical system for detecting forward scattered light pulses, transmitted light pulses, and side fluorescent pulses, and a counting unit were provided in order to distinguish and count cyanobacteria, whole algae, and whole fine particles in sample water. FIG. 6 shows an example of a device configuration diagram, and FIG. 7 shows an example of a block diagram of a transmitted light pulse counting unit using a plurality of comparators.

In FIG. 6, the forward scattered light pulse, transmitted light pulse, and side fluorescence pulse are counted, and the number concentration of the whole fine particles including algae, the number concentration of the whole algae, and the number concentration of the blue algae are classified. FIG. 2 shows a configuration diagram of an apparatus for separately measuring.

In this apparatus, by counting forward scattered light pulses due to fine particles, the number concentration of fine particles including algae having a particle size on the order of submicrons is determined. Measure the number concentration of fine particles including algae of several microns or more, and count the number of side fluorescence pulses by the whole algae and blue algae, thereby measuring the number concentration of the whole algae and blue algae, respectively.

1. Counting of transmitted light pulse: A light beam 1a emitted from a laser 1 that emits a light beam having a wavelength of 640 nm or less with an Ar laser, a He-Ne laser, an LD-pumped solid-state laser, a semiconductor laser, or the like is collected by a focusing optical system 2. In a direction perpendicular to the direction in which the sample water 4 flows, the shape is flat, and in the direction in which the sample water flows, light is condensed. After the light beam is attenuated by the filter 6 embedded in the perforated plano-convex lens 5, the light beam is converted into an electric signal 7 a by the photodiode 7. The electric signal 7a becomes a pulse signal every time the algae 8 or other fine particles pass through the observation region in the light beam.
The pulse signal due to the light blocking of the fine particles is converted by the transmitted light pulse counting unit 35 into a measured number concentration 3 of the fine particles in the sample water.
5a is output for each particle size category. In the light blocking method, it is not possible to measure fine particles on the order of submicrons, so the particle size is set within a range of several microns or more.

FIG. 7 is a block diagram of a transmitted light pulse counting unit using a plurality of comparators to count transmitted light pulses. The transmitted light pulse counting unit 35 in FIG.
First, the electric signal 7 a from the photodiode 7 is amplified by the preamplifier 10 and the main amplifier 11. The output electric signal of the main amplifier 11 is input to the LPF 13 and the LPF 12 in parallel. LPF13 is the main amplifier 1
1 has a higher cutoff frequency than the transmitted light pulse signal in the output electric signal, and the output electric signal is a transmitted light pulse signal from which high-frequency noise has been removed. On the other hand, the LPF 12 has a cutoff frequency and a smoothing circuit that are sufficiently lower than the LPF 13, and by smoothing the low-frequency output electric signal, an average value, that is, an output electric signal of a DC component due to stray light or the like is obtained. These two outputs are LPF
By subtracting the output electric signal of the LPF 12 from the output electric signal of the LPF 12 by the differential amplifier 14, a DC component due to stray light or the like is subtracted, and the next plurality of comparators 36 to 3
8 is input.

Next, in each of the plurality of comparators 36 to 38, the threshold value of each comparator is set to a voltage corresponding to each particle size section, the pulse signal is binarized, and a threshold corresponding to each particle size section is set. The arithmetic circuit 39 counts pulses having a peak value equal to or greater than the value. The value counted for each particle size division is output from the display output circuit 17 as the number concentration 35a of the whole fine particles in the sample water, as in the first embodiment. Here, the number of comparators is three, that is, there are three types of particle size divisions. However, the number of comparators can be changed according to the number of necessary particle size divisions.

2. Counting scattered light pulses: with the device of FIG.
Forward scattered light 18 generated when algae or other fine particles pass through the light beam 1a is condensed by the plano-convex lens 5, achromatic lens 20, and achromatic lens 22 having a hole at the center, and the pinhole is formed. After the stray light is eliminated by 23, the light is converted into an electric signal 24a by the photodiode 24. Electrical signal 24
a is a plano-convex lens 5, an achromatic lens 20, a dichroic mirror 21, an achromatic lens 2
2. Each time an algae or other fine particles pass through the observation area of the forward scattered light collecting optical system constituted by the pinhole 23, a pulse signal is generated. The pulse signal due to the forward scattering of the fine particles is output by the scattered light pulse counting unit 40 having the same function as the transmitted light pulse counting unit 35 as the number concentration measurement value 40a of the fine particles in the sample water for each particle size classification. .
The particle size classification is set mainly in the submicron range that cannot be measured by the light blocking method.

3. Fluorescence pulse counting: Among the side fluorescences generated when the algae 8 pass through the light beam 1a, the fluorescence having a peak wavelength of 685 nm is transmitted, but the peak wavelength is 650n.
The fluorescence 42 of chlorophyll a is separated by the interference filter 41 that blocks the fluorescence of m. Next, the fluorescence 42 of the chlorophyll a is condensed by the plano-convex lens 43 and the achromatic lenses 44 and 45, and after the stray light is eliminated by the pinhole 46, is converted into an electric signal 47 a by the photomultiplier 47. The electric signal 47a becomes a pulse signal every time algae pass through the observation region of the fluorescence condensing optical system including the interference filter 41, the plano-convex lens 43, and the achromatic lenses 44 and 45. The pulse signal of the fluorescence 42 of chlorophyll-a is converted by the chlorophyll-a fluorescence pulse counting unit 48 having the same function as that of the transmitted light pulse counting unit 35 into the number concentration measurement value 48a of the whole algae in the sample water for each particle size classification. Is output.

Of the side fluorescence generated when the algae 8 pass through the light beam 1a, the fluorescence having a peak wavelength of 650 nm is transmitted, but the fluorescence 50 of phycocyanin is separated by an interference filter 49 which blocks the fluorescence having a peak wavelength of 680 nm. Is done. Next, the phycocyanin fluorescence 50 is condensed by the plano-convex lens 51, the achromatic lenses 52 and 53, and after the stray light is removed by the pinhole 54, it is converted into the electric signal 55a by the photomultiplier 55. The electric signal 55a becomes a pulse signal every time cyanobacteria pass through the observation region of the fluorescence condensing optical system including the interference filter 49, the plano-convex lens 51, and the achromatic lenses 52 and 53. The pulse signal of the phycocyanin fluorescence 50 is output by the phycocyanin fluorescence pulse counting section 56 having the same function as that of the transmitted light pulse counting section 35 to the number concentration measurement 56a of cyanobacteria in the sample water for each particle size classification. You.

Further, the transmitted light pulse counting section 3 of this embodiment
5, the scattered light pulse counting unit 40, the chlorophyll a fluorescence pulse counting unit 48, and the phycocyanin fluorescence pulse counting unit 56 are replaced by the transmitted light pulse counting unit 9, the scattered light pulse counting unit 25, and the chlorophyll a fluorescence pulse counting unit 33 of the first embodiment. , And a phycocyanin fluorescence pulse counting unit 34.

[0048]

The present invention relates to an apparatus for counting the number concentration of the whole fine particles, the number concentration of the whole algae, and the number concentration of the whole blue-green algae in the sample water, and irradiates the sample water flowing in the flow cell with a light beam. Fluorescence receiving optical system and fluorescence pulse counting unit for detecting the phycocyanin in cyanobacteria generated when passing through the light beam, or chlorophyll a in the algae, and the fluorescence pulse counting unit, the whole algae in the sample water and cyanobacteria It is possible to measure the number concentration.

Further, the above-mentioned fluorescence focusing optical system and fluorescence pulse counting unit, and a scattered light focusing optical system for detecting a scattered light pulse generated when algae or other fine particles pass through the light beam, A transmitted light receiving optical system for detecting a dimming pulse of transmitted light generated when algae or other fine particles pass through the light beam and the light beam is cut off, a scattered light pulse counting unit, and a transmitted light pulse counting. The combination of the parts enables the algae and other fine particles to be distinguished and the number concentration to be measured.

[Brief description of the drawings]

FIG. 1 shows a fluorescence spectrum of phycocyanin specifically contained in cyanobacteria.

FIG. 2 is a diagram showing a fluorescence spectrum of chlorophyll a contained in algae in general.

FIG. 3 is a schematic diagram of a pulse signal observed by each light detection unit when a sample water containing algae is irradiated with a light beam from an excitation light source.

FIG. 4 counts forward scattered light pulse, transmitted light pulse, and forward fluorescent light pulse, and counts the number concentration of the whole fine particles including algae;
Configuration diagram of the device for distinguishing and measuring the number concentration of whole algae and the number concentration of cyanobacteria

FIG. 5 is a block diagram of a transmitted light pulse counting unit that uses a peak hold circuit to count transmitted light pulses.

FIG. 6 counts forward scattered light pulses, transmitted light pulses, and side fluorescence pulses to determine the number concentration of the whole fine particles including algae;
Configuration diagram of the device for distinguishing and measuring the number concentration of whole algae and the number concentration of cyanobacteria

FIG. 7 is a block diagram of a transmitted light pulse counting unit using a plurality of comparators to count transmitted light pulses;

[Explanation of symbols]

 1: laser 1a: light beam 2: focusing optics 3: flow cell 4: sample water 5: plano-convex lens 6: filter 7: photodiode 7a: electric signal 8: algae 9: transmitted light pulse counting section 9a: particle count concentration Measured value 10: Preamplifier 11: Main amplifier 12, 13: Low pass filter (LPF) 14: Differential amplification 15: Peak hold circuit 16: Operation circuit 17: Display output circuit 18: Forward scattered light 19: Forward fluorescence 20: Achromatic Lens 21: Dichroic mirror 22: Achromatic lens 23: Pinhole 24: Photodiode 24a: Electric signal 25: Scattered light pulse counting unit 26: Dichroic mirror 27, 28: Achromatic lens 29, 30: Pinhole 31, 32: Photomultiplier tube 3 3: chlorophyll a fluorescence pulse counting unit 34: phycocyanin fluorescence pulse counting unit 35: transmitted light pulse counting unit 35a: measured value of particle number concentration 36 to 38: comparator 39: arithmetic circuit 40: scattered light pulse counting unit 40a: particle number concentration Measured value 41: Interference filter 42: Fluorescence of chlorophyll a 43: Plano-convex lens 44, 45: Achromatic lens 46: Pinhole 47: Photomultiplier tube 47a: Electric signal 48: Chlorophyll a fluorescence pulse counter 48a: Number concentration measurement Value 49: Interference filter 50: Phycocyanin fluorescence 51: Plano-convex lens 52, 53: Achromatic lens 54: Pinhole 55: Photomultiplier tube 55a: Electric signal 56: Chlorophyll a fluorescence pulse counting section 56a: Number concentration measurement value

Continuation of the front page (72) Inventor Tokio Oto 1-1, Tanabe Nitta, Kawasaki-ku, Kawasaki, Kanagawa Prefecture Inside Fuji Electric Co., Ltd. (72) Inventor Kenji Harada 1-1, Tanabe Nitta, Kawasaki-ku, Kawasaki-ku, Kanagawa Prefecture Inside Fuji Electric Co., Ltd. (72) Inventor Akinori Sasaki 1-1-1, Tanabe-Nitta, Kawasaki-ku, Kawasaki-city, Kanagawa Prefecture F-term (reference) 2G043 AA03 BA16 CA03 CA06 EA01 EA13 EA14 GA04 GB01 GB03 HA01 HA09 JA03 KA05 KA09 LA02

Claims (7)

[Claims] 1. A light beam emitted from a light source is condensed by a condensing optical system, and when the light beam is irradiated toward a flow cell through which sample water containing fine particles flows, only scattered light having the same wavelength as the irradiation light is condensed. Then, the scattered light pulse signal is detected by photoelectric conversion, the transmitted light pulse signal is detected by the dimmed irradiation light by photoelectric conversion, and only the light of chlorophyll a contained in algae having a fluorescence wavelength of 680 nm is collected. Fluorescence pulse signal of chlorophyll a is detected by conversion, only the light of phycocyanin fluorescence wavelength 650 nm contained only in cyanobacteria is collected, and the fluorescence pulse signal of phycocyanin is detected by photoelectric conversion. Are separately counted, the count value of the scattered light pulse signal and the transmitted light pulse signal is defined as the total number of fine particles, and the count value of the chlorophyll a fluorescent pulse is determined as the total number of algae. And, cyanobacteria the count value of the phycocyanin fluorescence pulse and outputs as a blue-green algae total number counting method of algae and particulates. 2. The light emitted from the light source according to claim 1 is 640.
A method for counting cyanobacteria, algae and fine particles, wherein the wavelength is not more than nm. 3. The counting of the light detection pulse signals according to claim 1, wherein the peak value of each pulse signal is detected, and discrimination counting for recording a count value for each predetermined peak value is performed. A method for counting cyanobacteria, algae and fine particles. 4. A light beam emitted from a light source is condensed by a condensing optical system, and when irradiated toward a flow cell through which sample water containing fine particles flows, only scattered light having the same wavelength as the irradiated light is transmitted or transmitted. Scattered light detecting means for reflecting, condensing and photoelectrically converting, transmitted light detecting means for photoelectrically converting the reduced irradiation light, and fluorescence wavelength 68 of chlorophyll a contained in algae
A chlorophyll-a fluorescence detecting means for transmitting and reflecting only light of 0 nm and condensing and photoelectrically converting the light; and a phycocyanin for transmitting and reflecting only light having a fluorescence wavelength of 650 nm of phycocyanin contained only in cyanobacteria and condensing and photoelectrically converting the light. 2. The apparatus according to claim 1, further comprising: fluorescence detection means; and discrimination counting means for separately counting pulse signals from the four light detection means.
A total number of fine particles, a total number of algae, and a total number of blue-green algae are output by the counting method of (1). 5. An apparatus for counting cyanobacteria, algae and fine particles, wherein the light source according to claim 4 is a light source having a wavelength of 640 nm or less. 6. The discrimination counting means according to claim 4, further comprising discrimination counting means for detecting a peak value of each pulse signal and recording a count value for each predetermined peak value section. Blue-green algae, which is characterized by performing discrimination counting by the method of
Algae and particulate counting equipment. 7. An apparatus for counting blue-green algae, algae and fine particles, wherein the whole particle, the whole algae, and the size classification of blue-green algae are output from the counted values of the pulse signals according to wave heights.