JP2014042463A - Method of testing microorganism and device thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a device of testing microorganisms which can measure the amount of microorganisms in ballast water easily, rapidly and accurately, by using a batch-type measurement cell.SOLUTION: A device 1 of testing microorganisms for measuring the amount of microorganisms in a sample solution is equipped with; agitating and mixing means 7 for agitating and mixing a sample solution in which a sample and a fluorescent dye reagent are added in a batch-type sample container 5 formed of a translucent material; an excitation light source 10 equipped with a light source which irradiates a surface to be irradiated on the sample container 5 with excitation light while the sample solution 5 is agitated by the agitating and mixing means 7; light receiving means 14 for detecting fluorescent light excited by the excitation light from the excitation light source 10; and control means 23 which converts the light detected by the light receiving means 14 into electric signals to detect the number of light, and calculates the amount of microorganisms included in the sample in the sample container 5 from the number of light.

Description

  The present invention relates to a microorganism testing method and apparatus, and more particularly to a microorganism testing method and apparatus suitable for detecting microorganisms such as plankton that are contained in ballast water and are alive.

In order to stabilize the ship, the ship that is not loaded with cargo travels with ballast water and discharges the ballast water in the sea area where the luggage is loaded.
Ballast water is usually discharged into a sea area different from the sea area on which it is mounted. Therefore, problems such as the destruction of ecosystems by transporting microbes such as plankton and bacteria contained in the ballast water to sea areas other than their original habitats. There is a risk of causing it.

  In order to deal with such problems, international rules regarding the regulation of ballast water have been formulated, and the “International Convention for the Regulation and Management of Ship Ballast Water and Sediment (Ballast Water Management Convention)” has been adopted. Yes.

The “Guidelines for Ballast Water Sampling (G2)” related to the above Ballast Water Management Convention is based on the “Ballast Water Emission Standard (D-2)”. For example, for a microorganism having a minimum size of 50 μm or more (hereinafter referred to as “L size organism”), the allowable number of individuals is 10 / m ³ or less, and a microorganism having a minimum size of 10 μm or more and less than 50 μm (hereinafter referred to as “S size organism”). “)” Is defined as 10 pieces / mL or less according to the minimum size of the microorganism.

  To date, as a method for confirming whether or not the above discharge standards are satisfied when discharging the above ballast water, the seawater pumped up by the water pump is passed through the flow cell and image measurement is performed (for example, patents) Literature 1), known is a microorganism testing apparatus such as a device that counts microorganisms by passing seawater pumped by a water pump through filter units with different openings and causing the microorganisms on the filter to emit light (for example, Patent Document 2). ing.

According to the microorganism testing apparatus described in Patent Document 1, a staining unit that stains a living organism having living cells present in the sample while flowing a liquid sample, and the organism that flows the stained sample. The concentration unit for concentrating so as to increase the concentration of the individual, the individual measurement unit for acquiring image information of the individual including the organism in the concentrated specimen, and the image information of the individual output from the individual measurement unit And a control means for measuring a living thing.
As a result, the process of staining organisms in the liquid of the specimen, the process of concentrating organisms in the liquid, the process of acquiring information on organisms in the liquid, etc. can be performed by the flow method. The waiting time until a part of a sample that has completed one process proceeds to the next process can be greatly reduced or reduced to zero, and stable organisms can be prevented in order to prevent deterioration of the staining state during the waiting time. There is an advantage that life and death information can be acquired.

  However, in the microorganism testing apparatus described in Patent Document 1, seawater pumped up by a water pump is sequentially passed through various processes, and there is a problem that the apparatus becomes large and the manufacturing cost is high. And although it is made to pass water sequentially to various processes and waiting time is shortened, there exists a problem that it takes at least several hours to complete a measurement.

Moreover, the microorganism testing apparatus described in Patent Document 2 includes a step of passing seawater through a filter unit in which three types of filters having different openings are arranged in series, and a microorganism that is collected and survives in the filter. A process of generating any one of coloration, luminescence, and fluorescence due to the above, and a process for detecting the coloration, luminescence, and fluorescence and counting the number of microorganisms in the ballast water or seawater by image analysis. It is a feature.
As a result, it is possible to realize capture of microorganisms for each step size, and as a result, there is an advantage that it is possible to quickly measure whether or not the allowable residual standard regulated by the standard for each size is satisfied.

  However, even in the microorganism testing apparatus described in Patent Document 2, the seawater pumped up by the water pump is sequentially passed through various processes in the same manner as Patent Document 1, and the apparatus becomes large and the manufacturing cost increases. There was a point.

JP 2009-85898 A JP2007-135582

  In view of the above problems, the present invention provides a microorganism testing method and apparatus capable of measuring the amount of microorganisms in ballast water simply, in a short time, and with high accuracy by using a batch type measurement cell. Providing is a technical issue.

  In order to solve the above problems, the present invention is a microorganism testing apparatus for measuring the amount of microorganisms in a sample solution, wherein a sample and a fluorescent staining reagent are placed in a batch-type sample container formed of a light-transmitting material. A stirring and mixing unit that stirs and mixes the sample solution by adding a light source, and an excitation light source that includes a light source that irradiates the irradiated surface of the sample container while stirring the sample solution by the stirring and mixing unit; Light receiving means for detecting the light emitted by the excitation light from the excitation light source, and detecting the number of emitted light by converting the light detected by the light receiving means into an electrical signal, and the sample in the sample container from the number of emitted light And a control means for calculating the amount of microorganisms contained in the product.

  The invention according to claim 2 is arranged such that the excitation light source is irradiated with excitation light orthogonal to the surface to be irradiated of the sample container, while the light receiving means is excitation light of the excitation light source. A light receiving surface for receiving fluorescent light emission at an angle orthogonal to is arranged.

  The invention described in claim 3 is characterized in that a slit for regulating an observation range is provided between the light receiving means and the sample container.

  Furthermore, the invention according to claim 4 further comprises a parallel light converting means for converting the diffused light from the light source into parallel light that is uniformly irradiated at the same angle toward one surface between the excitation light source and the sample container. It is provided.

  The invention according to claim 5 is characterized in that the parallel light converting means is formed by drilling a threaded hole having a predetermined diameter in a flat plate having a predetermined thickness.

  The invention according to claim 6 is characterized in that the parallel light converting means is formed of a convex lens.

  The invention according to claim 7 is a method for inspecting microorganisms for measuring the amount of microorganisms in a sample solution, and stirring and mixing the sample solution in which a fluorescent staining reagent is added to the sample in a batch type sample container. A stirring and mixing step, an excitation step of irradiating the irradiated surface of the sample container with excitation light while stirring the sample solution, a light receiving step of counting fluorescence of microorganisms that emit fluorescence by the excitation light, and the light receiving step. A microorganism number estimation step of calculating the amount of microorganisms contained in the sample in the sample container from the detected number of luminescence.

  According to the invention of claim 1, after adding the sample and the fluorescent staining reagent to the batch type sample container, the sample solution is stirred and mixed by the stirring and mixing means, and then the sample solution is stirred while the sample solution is stirred. The excitation light is incident on the irradiated surface of the sample container, and the fluorescent light emission of the microorganisms is received by the light receiving means, so that the microorganisms emit light brightly in a very short time compared to the measurement without standing and stirring. In addition, the amount of microorganisms in the ballast water can be measured easily and in a short time. Since the apparatus of the present invention is a batch type, the apparatus can be miniaturized and the manufacturing cost can be reduced.

  According to a second aspect of the present invention, the excitation light source is disposed so that excitation light orthogonal to the surface to be irradiated of the sample container is incident, while the light receiving means has its light receiving surface. Is arranged so that the fluorescence emission is received at an angle orthogonal to the excitation light of the excitation light source, the excitation light from the excitation light source does not directly enter the light receiving surface of the light receiving means, and the fluorescence emission (For example, as shown in FIG. 2, the light emitting portion having a conventional width of 20 to 30 mm is narrowed to have a width M (3 mm) and the thickness portion is thin). The difference in the amount of light between the background and the fluorescence emission of the microorganism becomes very clear, and the detection accuracy of the fluorescence emission of the microorganism is improved.

  According to the invention of claim 3, since the slit for regulating the observation range is provided between the light receiving means and the sample container, the area of background fluorescent light emission that becomes noise is narrowed. The ratio of the fluorescence emission signal of the microorganism to the background fluorescence emission is improved, and the detection accuracy of the fluorescence emission of the microorganism is improved.

  According to a fourth aspect of the invention, there is provided parallel light conversion means for converting the diffused light from the light source into parallel light that is uniformly irradiated at the same angle toward one surface between the excitation light source and the sample container. Because it is provided, the spread of the excitation light from the excitation light source is suppressed and the irradiated surface of the sample container is irradiated with parallel light, so that the thickness of the background fluorescence emission is reduced, and the background fluorescence emission The ratio of the fluorescence emission signal of the microorganism to that of the microorganism is improved, and the detection accuracy of the fluorescence emission of the microorganism is improved.

  According to the invention described in claim 5, since the parallel light converting means is formed by drilling a threaded hole having a predetermined diameter in a flat plate having a predetermined thickness, the excitation light from the excitation light source is made of an inexpensive material. However, the angle is corrected by the threaded hole, and the directivity angle of the light emitted from the threaded hole can be narrowed. For this reason, since the thickness portion of the background fluorescent light emission becomes thin, the ratio of the fluorescent light emission signal of the microorganism to the background fluorescent light emission is improved, and the detection accuracy of the fluorescent light emission of the microorganism is improved.

  According to the invention described in claim 6, since the parallel light converting means is formed of a convex lens, the directivity angle of the excitation light from the excitation light source can be narrowed by an inexpensive material. For this reason, the thickness portion of the background fluorescence emission is reduced, the ratio of the fluorescence emission signal of the microorganism to the background fluorescence emission is improved, and the detection accuracy of the fluorescence emission of the microorganism is improved.

  According to the seventh aspect of the present invention, there is provided a microorganism testing method for measuring the amount of microorganisms in a sample solution, wherein the sample solution is stirred and mixed with a fluorescent staining reagent added to the sample in a batch type sample container. An agitation and mixing step, an excitation step of irradiating the irradiated surface of the sample container with excitation light while stirring the sample solution, a light receiving step of counting fluorescence of microorganisms that emit fluorescence by the excitation light, and the light reception And the number of microorganisms estimation step for calculating the amount of microorganisms contained in the sample in the sample container from the number of luminescence detected in the process. The microorganisms emit light brightly in a short time, and the amount of microorganisms in the ballast water can be measured easily and in a short time. In addition, since the thickness of the fluorescent light emission is reduced, the light amount difference between the background and the fluorescent light emission of the microorganism becomes extremely clear, and the detection accuracy of the fluorescent light emission of the microorganism can be improved.

It is a perspective view showing the whole microbe inspection device concerning one embodiment of the present invention. It is a general | schematic plane sectional view of the measurement part which concerns on one Embodiment of this invention. 1 is a block diagram showing the overall configuration of a microorganism testing apparatus according to an embodiment of the present invention. It is a flowchart which shows the measurement flow of the microbe inspection apparatus which concerns on one Embodiment of this invention. It is a schematic sectional drawing which shows one Embodiment of a parallel light conversion means. It is an effect | action figure which shows that an observation surface narrows with the presence or absence of a slit. It is a graph which shows the correlation with the number of microorganisms and the light reception count of a photomultiplier tube (PMT). It is a graph which shows the test of whether detection is possible by the life and death of microorganisms.

  A mode for carrying out the present invention will be described with reference to the drawings. FIG. 1 is a perspective view showing an entire microorganism testing apparatus according to an embodiment of the present invention, FIG. 2 is a schematic plan sectional view of a measuring unit according to an embodiment of the present invention, and FIG. It is a block diagram which shows the whole structure of the microbe inspection apparatus which concerns on one Embodiment.

  As shown in FIGS. 1 and 2, an inspection apparatus 1 according to the present invention includes a main body 2 that incorporates a control mechanism such as a CPU board and performs information processing work such as measurement results and statistical processing work, and the main body 2 A display unit 4 formed of a liquid crystal panel or the like for displaying the measurement results, and a transparent material that transmits light (for example, glass or A main part is a measurement unit 6 that accommodates a batch-type sample container 5 formed of quartz, acrylic resin, or the like and optically counts the number of microorganisms in the sample solution S. Reference numeral 7 denotes a rotor for agitating the sample solution S accommodated in the sample container 4. The rotor 7 is accommodated in the sample container 5 together with the sample solution S and the luminescent reagent, and the sample container 5 is accommodated in the sample container 5. When accommodated in the measuring unit 6, the magnetic stirrer 27 built in the measuring unit 5 is rotationally driven. Thereby, the number of microorganisms in the sample solution S can be counted while stirring and mixing the sample solution S composed of the sample and the luminescent reagent in the sample container 5 at a predetermined temperature, and the sample solution S can be measured without being stirred. Compared with, the microorganisms emit light brightly in a very short time, and the amount of microorganisms in the ballast water can be measured easily and in a short time.

The dimensions of the inspection apparatus 1 shown in FIG. 1 are 300 mm in width, 300 mm in depth, 100 mm in height, and about 2 to 4 kg in weight, and are accommodated in a handheld trunk (not shown) or the like. It can be carried anywhere and can be measured inside a ship or measured outdoors.
The batch-type sample container 5 formed of a transparent material that transmits light is formed in a prismatic shape having a bottom surface of 50 mm × 50 mm and a height of 60 mm, and has an internal volume of 100 ml (milliliter when the water level is 40 mm). ) Is set. The sample container 5 is not limited to such a prism shape, and may be cylindrical or cubic as long as the internal volume can be secured about 100 ml (milliliter).

  As shown in FIGS. 1, 2, and 3, the measurement unit 6 includes a sample container storage unit 9 that stores and holds the sample container 5, and a light source unit 13 that emits excitation light toward the sample container 5. And a light receiving unit 19 for observing microorganisms that are stained in the luminescent reagent by the excitation light emitted from the light source unit 13 and drift in the sample container 5. The light receiving unit 19 is electrically connected to a CPU substrate 23 that counts the number of microorganisms in the sample solution S and performs information processing operations such as measurement results and statistical processing operations.

The sample container storage unit 9 is formed by holding plates 8a and 8b surrounding at least two surfaces of the sample container 5, and stores and holds the sample container 5 so as not to block light irradiation from the light source unit 13. It is.
Then, as shown in FIG. 2, the light source unit 13 is arranged so that excitation light from the normal line AP is incident on the irradiated surface G of the sample container 5. The light source unit 13 includes an LED light source 10 disposed in the vicinity of the sample container storage unit 9 and a parallel light conversion unit 11 (LED is a light source) that is disposed in front of the LED light source 10 and converts diffused light into parallel light. Since the light is diffused and irradiated in a random direction, the sample container 5 is irradiated with excitation light consisting of parallel light that is uniformly irradiated with light rays at the same angle toward one surface) and slit-shaped parallel light. And an excitation light band-pass filter 12.

  FIG. 5 is a schematic sectional view showing an embodiment of the parallel light converting means 11. In the example shown in FIG. 5A, the parallel light converting means 11 is formed by drilling a threaded hole 32 having a predetermined diameter in a flat plate 31 having a predetermined thickness, and the thickness of the flat plate 31 is adjusted to the optical path length. L and the hole diameter of the threaded hole are appropriately set. Thereby, the scattered light of the incident angle θ irradiated from the LED light source 10 is converted into parallel light when passing through the threaded hole 32. In the example shown in FIG. 5A, the optimum condition of θ and L is determined by the S / N ratio test. For example, if M3 (outer diameter of screw hole) × 0.5 (pitch), θ Was 9.5 ° and L was 15 mm.

  The parallel light conversion means 11 shown in FIG. 5B is provided with a convex lens 33 on the front surface of the LED light source 10, and the scattered light emitted from the LED light source 10 passes through the convex lens 33 and is emitted to the outside. In this case, the light is converted into parallel light.

  Note that the light source unit 13 of the present embodiment uses the LED light source 10 as a light source. However, the parallel light that can be irradiated with parallel light is not limited to the LED light source 10 as long as it can excite the fluorescent substance contained in the microorganism. An LED light source, a laser light source, and a light bulb can also be employed. Needless to say, when a parallel light LED or a laser light source capable of irradiating parallel light is employed, the above-mentioned parallel light conversion means 11 is not necessary.

  As shown in FIG. 2, the light receiving unit 19 is provided such that the light receiving surface F is arranged at an angle orthogonal to the excitation light by the normal line AP from the light source unit 13. The light-receiving unit 19 is a photomultiplier tube arranged and configured to receive fluorescence with an optical axis orthogonal to the parallel light irradiated with excitation light from the LED light source 10 toward the sample container 5 ( PMT) 14, a fluorescent bandpass filter 15 disposed in front of the photomultiplier tube (PMT) 14, a condensing lens 16 disposed in front of the fluorescent bandpass filter 15, and the condensing A slit 17 disposed on the front surface of the lens 16 and a gap between the slit 17 and the sample container 5 are used to excite a fluorescent substance contained in microorganisms, thereby condensing and imaging the emitted fluorescence. And a relay lens 18.

  The slit 17 between the photomultiplier tube (PMT) 14 and the sample container 5 narrows the observation surface in a slit shape. That is, as shown in FIG. 6, (a) in the state without slits, the background where the light receiving surface F is formed in a circle is monitored, while (b) in the state with slits, the light receiving surface F is shaded. The background formed by the vertically long slits except for is monitored. Therefore, as a result of the light receiving area of the light receiving surface F being narrowed as shown in (b), the area of background fluorescent light emission, which becomes noise, is also narrowed. This improves the detection accuracy of the fluorescence emission of microorganisms.

  Although the light receiving unit 19 uses the photomultiplier tube (PMT) 14 as the light receiving sensor, the light receiving unit 19 is not limited to this, and is not limited to this, but a silicon photodiode (SiPD) or an avalanche photodiode (APD). As in the case of a photomultiplier tube (PMT), various types of photodetectors that can detect the emission of a fluorescent substance contained in a microorganism can be employed.

  Furthermore, with reference to FIG. 3, the electrical control structure of the inspection apparatus 1 of this embodiment is demonstrated. In the center of the housing 20 forming the main body 2, an output signal converted from light to electricity by the photomultiplier tube (PMT) 14 is supplied with power from an AC power source 21 or a secondary battery 22. A CPU board 23 is provided for performing analysis, determining whether or not it is within an arbitrary luminance range, pulse counting an arbitrary luminance signal, and controlling on / off of the LED light source 10. . An AC / DC converter 24 is interposed between the AC power source 21 and the CPU board 23.

  The CPU substrate 23 is electrically connected to the photomultiplier tube (PMT) 14, the LED light source 10, a RAM 25 serving as a read / write storage unit, and a ROM 26 serving as a read-only storage unit. Further, the power button 3a, the measurement start button 3b, the external output button 3c, and the setting button 3d of the operation unit 3 shown in FIG. 1 are electrically connected. Then, on / off switching control is performed by pressing the power button 3a, measurement is started by pressing the measurement start button 3b, and data is transferred to an external printer or personal computer by pressing the external output button 3c. By pressing the setting button 3d, the measurement type is switched (L size microorganism measurement or S size microorganism measurement is switched), the judgment reference setting is changed, the threshold value setting is changed, and the measurement is performed. The time setting can be changed.

  In addition, the CPU board 23 includes a magnetic stirrer 27 that rotates the rotor 7 by magnetic force, a display unit 4 formed of a liquid crystal panel, a cooling fan 28 for a control device such as the CPU board 23, and RS-232C. An external output terminal 29 is connected.

  FIG. 4 is a flowchart showing a measurement flow, and the operation of the above configuration will be described with reference to FIGS.

  First, the operator uses a pipette or the like to collect 100 ml (milliliter) as a sample from ballast water at a temperature of about 20 ° C. and put it into the sample container 5 (step 1 in FIG. 4). Next, a fluorescent staining reagent is added into the sample container 5 (step 2 in FIG. 4). As this fluorescent staining reagent, generally known calcein AM (Calcein-AM, manufactured by Promocell GMBH, Germany), FDA or the like can be used. Calcein AM tends to stain phytoplankton, while FDA tends to stain zooplankton. Therefore, staining with a staining reagent is mixed with calcein AM and FDA. When dyeing is performed with the reagent, it is possible to shorten the dyeing time of the reagent and halve the time required for dyeing. Then, after the operator puts the rotor 7 into the sample container 5, the operator accommodates it in the measurement unit 6 of the inspection apparatus 1 and attaches the lid 30 of the measurement unit 6 to complete the measurement preparation. Here, when the power button 3a is pressed, the rotor 7 is rotated by driving the magnetic stirrer 27 built in the measuring unit 6, and the sample solution S is stirred (step 3 in FIG. 4). .

  Next, when the operator depresses the measurement start button 3b of the operation unit, the LED light source 10 is turned on after a predetermined time, and the light transmitted through the excitation light band-pass filter 12 is irradiated to the sample container 5. At this time, for example, light having a wavelength of 450 nm to 490 nm is irradiated as wavelength characteristics, and the specimen (microorganism) in the sample container 5 emits fluorescence (step 4 in FIG. 4). This fluorescence passes through the fluorescence band-pass filter 15 and is detected by the photomultiplier tube (PMT) 14 (step 5 in FIG. 4).

  The photomultiplier tube (PMT) 14 converts light energy into electrical energy by utilizing the photoelectric effect, and has a current amplification function, and can detect fluorescence emission with high sensitivity. The detected electrical signal is sent to the CPU substrate 23, and the received light waveform exceeding a certain threshold value is counted (step 6 in FIG. 4).

  Further, the CPU board 23 estimates the number of microorganisms present in 100 ml (milliliter) of water in the sample container 5 from the received light wave count value, and displays on the display unit 4 whether or not the drainage standard is satisfied. Yes (step of FIG. 4).

  Examples of the present invention will be described below. First, a confirmation test of the inspection accuracy of the microorganism inspection apparatus of the above embodiment was performed.

  The correlation between the microbial population and the photomultiplier tube (PMT) light reception count was investigated. S-type scallop worms (minimum size of about 100 μm = L size organisms) are accommodated in a plurality of sample containers 5 (100 mL capacity) by 5 individuals, 10 individuals, 50 individuals, 100 individuals, 1000 individuals, respectively. Staining was performed with a fluorescent staining reagent FDA (concentration 0.01 [millimol / liter]). As a result, the number of waveform counts increased according to the number of microorganisms accommodated, and five samples of 5, 10, 50 and 100 responded linearly (FIG. 7 (a)). FIG. 7 (b)). Therefore, the number of microorganisms present in 100 mL of ballast water can be estimated from the obtained waveform count.

  A test was conducted as to whether or not detection was possible due to the life and death of microorganisms (see FIGS. 8A to 8D). S-type rotifer (minimum size of about 100 μm = L size organism) that has been killed by heat (heated at 60 ° C. for 30 minutes) with a fluorescent staining reagent FDA (concentration 0.01 [millimol / liter]) Stain. Three samples were prepared 1 hour after the killing treatment, 20 hours after the killing treatment, and 5 days after the killing treatment. As a result, no waveform with a certain threshold value or more was found in any of the killed samples, and it was possible to discriminate between samples that contained microorganisms without killing and samples that did not contain microorganisms. Is possible.

As described above, according to the present embodiment, after adding the sample and the fluorescent staining reagent to the batch type sample container 5, the sample solution is stirred and mixed by the stirring and mixing means 7, and then the sample solution is stirred. However, since the excitation light is incident on the irradiated surface of the sample container, and the fluorescence emission of the microorganisms is received by the light receiving means, the microorganisms can be measured in a very short time as compared with those measured by standing without stirring. Emits bright light, and the amount of microorganisms in the ballast water can be measured easily and in a short time. And since the apparatus of this embodiment is a batch type, it becomes possible to reduce the size of the apparatus and to reduce the manufacturing cost.
In addition, since the light receiving means 14 has the light receiving surface F disposed at an angle orthogonal to the excitation light, the excitation light from the excitation light source 10 does not directly enter the light receiving surface F of the light receiving means 14, The difference in the amount of light between the background and the fluorescence emission of the microorganism becomes very clear, and there is an extremely remarkable action and effect that the detection accuracy of the fluorescence emission of the microorganism is improved.

  The present invention can be applied to a microorganism testing apparatus for confirming whether or not the discharge standard is satisfied when discharging ballast water.

DESCRIPTION OF SYMBOLS 1 Inspection apparatus 2 Main body part 3 Operation part 4 Display part 5 Sample container 6 Measuring part 7 Rotor 8 Holding plate 9 Sample container accommodating part 10 LED light source 11 Parallel light conversion means 12 Excitation light band pass filter 13 Light source part 14 Photoelectron increase Double pipe (PMT)
DESCRIPTION OF SYMBOLS 15 Fluorescence band pass filter 16 Condensing lens 17 Slit 18 Relay lens 19 Light-receiving part 20 Case 21 AC power supply 22 Secondary battery 23 CPU board 24 AC / DC converter 25 RAM
26 ROM
27 Magnetic Stirrer 28 Fan 29 External Output Terminal 30 Lid 31 Flat Plate 32 Screw Cutting Hole 33 Cylindrical Lens

Claims (7)

A microorganism testing apparatus for measuring the amount of microorganisms in a sample solution,
A stirring and mixing means for stirring and mixing the sample solution by adding the sample and the fluorescent staining reagent to a batch type sample container formed of a light transmitting material;
An excitation light source comprising a light source for irradiating the irradiated surface of the sample container with excitation light while stirring the sample solution by the stirring and mixing means;
A light receiving means for detecting light emitted by the excitation light from the excitation light source;
Control means for detecting the number of emitted light by converting the light detected by the light receiving means into an electrical signal, and calculating the amount of microorganisms contained in the sample in the sample container from the number of emitted light. Microbe inspection device.   The excitation light source is arranged so that excitation light orthogonal to the irradiated surface of the sample container is incident, while the light receiving means has an angle orthogonal to the excitation light of the excitation light source. The microbe inspection apparatus according to claim 1, wherein the microbe inspection apparatus is arranged to receive fluorescent light emission.   The microorganism testing apparatus according to claim 1 or 2, wherein a slit for regulating an observation range is provided between the light receiving means and the sample container.   The parallel light conversion means for converting the diffused light from the light source into parallel light that is uniformly irradiated at the same angle toward one surface is provided between the excitation light source and the sample container. The microorganism testing apparatus according to any one of the above.   5. The microorganism testing apparatus according to claim 4, wherein the parallel light converting means is formed by drilling a threaded hole having a predetermined diameter in a flat plate having a predetermined thickness.   The microbe inspection apparatus according to claim 4, wherein the parallel light converting means is formed of a convex lens. A method for testing microorganisms for measuring the amount of microorganisms in a sample solution,
A stirring and mixing step of stirring and mixing a sample solution in which a fluorescent staining reagent is added to the sample in a batch type sample container;
An excitation step of irradiating the irradiated surface of the sample container with excitation light while stirring the sample solution;
A light receiving step for counting the fluorescence of the microorganisms that emit fluorescence by the excitation light;
And a microorganism number estimation step of calculating the amount of microorganisms contained in the sample in the sample container from the number of luminescence detected in the light receiving step.

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