JPH0650287B2 - Methanogen measuring device - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a methanogenic bacterium measuring device which is one of microbial activity measuring devices, and more particularly to measuring the concentration or activity of methanogenic bacteria in fermentation processes, sewage treatment processes, etc. The present invention relates to a methanogen measuring device for measuring.

[Conventional technology]

A conventional device of this type is shown in FIG.

This device utilizes the fact that the methanogenic bacterium has a unique fluorescent substance F420. The principle is to identify the methanogenic bacterium by taking a fluorescent image and processing the image.

In FIG. 4, 8 is a subject containing microorganisms, 24 is a conduit for guiding the subject 8 to a measuring instrument, 25 is a plunger for holding and fixing the subject 8 during measurement, and 26 is a condenser 10. It is a cover glass for irradiating the subject 8 with the condensed excitation light and taking out the fluorescence emitted from the subject 8 to the outside of the conduit 24.

The subject 8 sent through the conduit 24 is clamped and fixed by the plunger 25 between the cover glass 26 and the plunger 25. At this time, it is emitted from the light source 2,
The excitation light, which is limited to a specific wavelength region by the optical filter 12 and is condensed by the condenser 10, irradiates the fixed subject 8 through the cover glass 26. The fixed subject 8 receives this excitation light and emits fluorescence.
This fluorescence is taken out of the conduit 24 through the cover glass 26, and is excluded by the optical filter 16 except for the fluorescence wavelength region, and is limited to a specific wavelength. Then, after being magnified by the lens optical system 20 and the fluorescence image is photographed by the camera 21, it is sent to the image processing device 22 via the signal line 23.
The image processing device 22 performs image processing to eliminate the fluorescence image based on the foreign substances other than the methanogen, and then measures the concentration or activity of the methanogen from the fluorescence image based on the methanogen.

[Problems to be Solved by the Invention]

Since the conventional methanogen measuring device is configured as described above, various substances coexist in the system in which the methanogen is cultured, and fluorescence that cannot be distinguished from the methanogen is present in the system. It may include things to emit. Further, it is known that F420 is discharged to the outside of the bacterial cell. When a fluorescent image is taken in such a situation, a bright background appears, and sometimes image recognition may not be possible, making it impossible to measure the activity of methanogens from the fluorescence intensity of the fluorescent image. There was a problem that it would end up.

The present invention has been made to solve the above problems, it is possible to improve the image recognition rate by removing the background of the fluorescence image of the methanogen, and obtaining a clearer fluorescence image. Has an aim.

[Means for Solving the Problems]

The methanogenic bacterium measuring device according to the present invention filters a predetermined amount of a suspension containing a methanogenic bacterium, which is a sample, before irradiating with excitation light, and then rinses the medium with pure water or the like. The cleaning means is provided.

[Action]

In the methanogenic bacterium measuring apparatus of the present invention, since the means for previously washing the suspension containing the methanogenic bacterium as the test object is provided at the stage before the irradiation of the excitation light, the methanogenic bacterium contained in the medium is generated. Fluorescent substances other than bacteria are removed,
The background of the fluorescent image can be removed.

〔Example〕

 An embodiment of the present invention will be described below with reference to the drawings.

FIG. 1 is a diagram showing the configuration of a methanogenic bacterium measuring apparatus of the present invention, in which 1 is a fermenter, 2 is a light source, 8 is a suspension containing a microorganism, and 12 is excitation light. An optical filter for limiting the wavelength region, 10 is a condenser lens, 1
6 is an optical filter for limiting the fluorescence wavelength region, 20 is a photographing lens, 21 is a video camera, 23 is a signal line, 2
2 is an image processing device, 24 is a conduit for guiding the subject 8 in the fermenter 1 to the measuring section, 26 is a cover glass, 25 is a plunger, 35 is a filter for filtration, 27 is a sluice valve, 3
6 is a flexible hose connecting the solid-liquid separator 31 and the plunger 25, 32 is a suction pump, 33 is a discharge valve, 3
0 is a drive device having a rack and pinion mechanism for moving the plunger 25 up and down, 28 is a cover glass 26,
A nozzle for injecting water for removing deposits on the filter 35, 37 is an opening for introducing a cleaning liquid or air into the space partitioned by the partition valve 27, which flows through the cleaning liquid tank 40 and the three-way valve 38. Has been done. The other side of the three-way valve is open to the air. 34 is a sluice valve 2
7, a suction pump 32, a drive device 30, a three-way valve 38, and the like, which are cleaning control devices.

Next, the operation will be described.

A signal from the cleaning control device 34 initializes each device as follows. Both gates 27 are opened and the pump 2
9, the three-way valve 38, the discharge valve 33, the waste liquid valve 39 are closed, and the plunger 25 is at the lowermost end (the upper surface of the filter 35 is the conduit 24).
Outside or inside).

Next, when the suspension 8 containing the methanogen is introduced into the measuring section from the fermenter 1 through the conduit 24, a signal from the cleaning control device causes the upstream sluice valve 27, and then the downstream sluice valve. The valve 27 is closed. In this way, the subject is confined in the space surrounded by the cover glass 26, the gate valves 27, and the plunger 25.

Next, the suction pump 32 is started by a signal from the cleaning control device 34, and the three-way valve 38 is connected so that the opening 37 is connected to the atmosphere.
Works. Then, the sample is sucked by the filter 35 at the upper end of the plunger 25, and the filtrate is collected in the solid-liquid separator 31. A suitable membrane filter is a membrane filter having a representative pore diameter of 0.22 micron, and methanogens are fixed on the upper surface of the membrane. The volume of the confined subject and the area of the filter 35 are determined by the bacterial concentration. Then, the opening part 3 is generated by a signal from the cleaning control device 34.
The three-way valve 38 operates so that the No. 7 and the cleaning liquid tank 40 are connected. In this case also, since the suction pump 32 is operating, the medium on the filter 35 is washed. When the cleaning is completed, the signal from the cleaning control device 34 closes the three-way valve 38 and stops the suction pump 32.

Next, the drive device 30 is actuated by the signal of the cleaning control device 34, and the surface of the filter 35 is about 1 below the cover glass 26.
The plunger 25 is pushed up until it reaches 00 microns. FIG. 2 shows a structural diagram centering on the movable part at this time. After that, image capturing and image processing are performed as in the conventional example, and methanogens are measured.

When the image capturing is completed, a signal from the cleaning control device 34 is output, the pump 29 operates to clean the filter 35 and the cover glass 26, and at the same time, the waste liquid valve 39 is opened and the cleaning liquid is discarded. Since the preparation for the next measurement is completed as described above, when the measurement start signal is output, the cleaning control device 34 is initialized again and the above process is repeated.

Next, the effect will be described.

FIG. 3 is a fluorescent image photograph of a methanogenic bacterium taken by the methanogenic bacterium measuring device of the present invention, and FIG. 5 is a fluorescent image photograph of a methanogenic bacterium taken by a conventional methanogenic bacterium measuring device.

As can be seen by comparing FIG. 3 and FIG. 5, in the present invention, a means for previously washing the suspension containing the methanogenic bacterium, which is the analyte, is provided in the stage before the irradiation of the excitation light. , The background of the fluorescence image is completely removed.
In addition, the contrast of the fluorescence image is also improved, so that the image recognition rate of the methanogen is improved.

〔The invention's effect〕

As described above, according to the methanogenic bacterium measuring apparatus according to the present invention, since the means for preliminarily washing the suspension containing the methanogenic bacterium as the test object is provided at the stage before the irradiation of the excitation light, The background of the fluorescence image is completely removed, the fluorescence image of the methanogen can be made clearer, and image processing such as binarization processing becomes significantly easier.
It has the effect of improving the image recognition rate of methanogens.

[Brief description of drawings]

FIG. 1 is a block diagram of a methanogen measuring device according to an embodiment of the present invention, and FIG. 2 mainly shows a movable part at a stage where a plunger of the methanogen measuring device of FIG. FIG. 3 is a diagram showing a fluorescent image of methanogenic bacteria photographed by the methanogen measuring device of the present invention, and FIG. 4 is a block diagram of a conventional methanogenic measuring device, FIG. FIG. 4 is a diagram showing a fluorescent image of a methanogenic bacterium taken by a conventional methanogen measuring device. 1 is a fermenter, 2 is a light source, 8 is a suspension, 10 is a light collector, 1
2, 16 are optical filters, 20 is a lens optical system, 21 is a video camera, 22 is an image processing device, 23 is a signal line, 24
Is a conduit, 25 is a plunger, 26 is a cover glass, 2
7 is a sluice valve, 28 is a nozzle, 29 is a pump, 30 is a drive device, 31 is a solid-liquid separator, 32 is a suction pump, 33 is a discharge valve, 34 is a cleaning control device, 35 is a filter, 36 is a flexible pipe, 37 is a gas-liquid inlet, 38 is a three-way valve, 39 is a waste liquid valve, and 40 is a cleaning liquid tank. The same reference numerals in the drawings indicate the same or corresponding parts.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Kenki Ozawa 8-1-1 Tsukaguchi Honcho, Amagasaki City, Hyogo Sanryo Electric Co., Ltd. Applied Equipment Research Laboratory (56) Reference JP-A-62-174636 (JP, A) ) JP 62-269045 (JP, A)

Claims (1)

[Claims] 1. A conduit for flowing an analyte containing a methanogenic bacterium, an irradiation means for irradiating the analyte with excitation light in a predetermined wavelength range, and an analyte containing the methanogenic bacterium is excited with the excitation light. A means for acquiring fluorescence in a predetermined wavelength range out of fluorescence emitted by irradiation of light as a fluorescence image, and an image processing circuit for image-processing the fluorescence image to eliminate fluorescence based on substances other than methanogens. A methanogenic bacterium measuring apparatus for measuring the concentration or activity of a methanogenic bacterium, comprising a means for previously washing a sample before irradiating the excitation light. .