KR20150025700A - Method for examining microbe having fluorescence with range of specific wavelength - Google Patents

Abstract

The present invention relates to a method for determining the number of microbe individuals included in a sample by using an optical device, which comprises: a light source unit for supplying light to the sample including microbes having fluorescence reflecting the light at a range of specific wavelength range; and a detection unit including an imaging acquiring unit for producing images by collecting the light reflected from the microbes, and an image processing unit for processing the produced images. The present invention relates to a method for examining microbes having fluorescence at the range of the specific wavelength, which comprises the steps of: (a) the image acquiring unit collecting the light reflected from the microbes included in a sample, which radiate light at the range of the specific wavelength; (b) the image acquiring unit producing the image of the collected light; (c) the image processing unit displaying the image produced by the image acquiring unit on a display, analyzing each pixel constituting the display on which the image is displayed to classify a plurality of pixels of continuously emitting light among pixels included in the display as one individual, and extracting the individuals of which the average brightness against the pixels constituting the individual among the individuals corresponds to a range of predetermined brightness, as an active individual; and (d) the image processing unit calculating the number of the active individuals included in the display. The present invention can provide a method for examining microbes having fluorescence at the range of the specific wavelength, which can precisely count the number of active microbe individuals among the microbes having fluorescence at the range of the specific wavelength included in a sample.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a method for testing microorganisms having a fluorescence characteristic at a specific wavelength,

[0001] The present invention relates to an inspection method for detecting the number of microorganisms contained in a sample, and more particularly, to an inspection method for detecting the number of microorganisms in an active microorganism having a fluorescence characteristic at a specific wavelength included in a sample, The present invention relates to a method of inspecting microorganisms having fluorescence characteristics at specific wavelengths.

The present invention relates to a microorganism testing method.

Particularly, in the case of a microorganism such as plankton having a fluorescence characteristic for a blue-based light having a wavelength of 380 nm to 480 nm, identification of the population thereof is not limited to specific understanding and study of a green or red tide phenomenon, It is considered very important for the protection of marine ecosystems.

As a measure to prevent the destruction of marine ecosystems due to the discharge of ship ballast water contained in ships returning from one country to another, if a ship departing from a certain area discharges ballast water to anchor in another area, Treatment of plankton contained in ballast water that may threaten ecosystems should be undertaken first.

In particular, the International Maritime Organization (IMO), in February 2004, called for '' ballast water for ships and control and management of sediments '' in order to prevent ecological and economic damage that could be caused by ballast water movement and to preserve biodiversity. International Convention '.

Specifically, there are two main ways in which ship ballast water can be used to meet the standards of an adopted convention. The first is an in-line process, and the second is an in-tank process. The pipe treatment method is a method of treating the ballast water at the time of inflow and outflow, and the tank treatment method is a method of treating the ballast water at sea after the inflow of the ballast water. Currently, international ballast water treatment technologies are pipeline treatment methods. The tank treatment method can be smaller than the pipeline treatment method, but the treatment speed must be set in consideration of the sailing time. have.

In both of the above methods, the number of plankton contained in the ballast water is identified and a suitable treatment method can be selected, and a final inspection of the ballast water discharged before discharge of the treated ballast water is performed, It should be checked whether the plankton, etc., which is problematic in the ship ballast water discharged, has been treated according to the standards.

However, up to now, there has not been provided a professional inspection method for detecting microbial populations such as plankton contained in ballast water.

Conventional microorganism inspection methods such as plankton contained in ship equilibrium water have only taken the method of judging the image of the photographed sample by the expert and judging the number of plankton contained in the sample, This conventional method has a problem that it is very difficult to accurately calculate the number of microorganisms such as plankton contained in the sample.

In addition, it is difficult to distinguish microorganisms including existing plankton from various foreign substances, and it is also difficult to distinguish between active microorganisms and inactive microorganisms, which makes it difficult to grasp the exact number of microorganisms.

The background art of the present invention is disclosed in Korean Patent Laid-Open Publication No. 10-1995-7001601 and the like, but a solution to the above-mentioned problem is not presented.

It is an object of the present invention, which has been devised to solve the above-mentioned problems, to provide a method and an apparatus for collecting light reflected by a microorganism having a fluorescence characteristic for a specific wavelength to generate an image thereof, (C) classifying a plurality of pixels that emit light continuously and classifying the plurality of pixels into one object, extracting only objects corresponding to an average value of pixel brightness of the objects within a predetermined range as an active object, and And (d) calculating the number of active objects. The microorganism having fluorescence characteristic for a specific wavelength, which enables to accurately grasp only the number of active microorganisms among the microorganisms having fluorescence characteristic at a specific wavelength included in the sample, To provide an inspection method.

In addition, by further including classifying the plurality of pixels that continuously emit light by introducing the boundary detection processing method in step (c) into one object, it is possible to reduce the number of microbes contained in the sample to a specific wavelength The present invention also provides a method for testing microorganisms having fluorescence properties.

The method further includes the steps of (c-1) to (c-3) categorizing the plurality of pixels that are continuously emitted within the predetermined number of pixels in step (c) The present invention provides a method for inspecting a microorganism having a fluorescence characteristic for a specific wavelength, which enables more accurate identification of the microorganism population by a filtering process for various foreign substances.

According to an aspect of the present invention for achieving the above object, the present invention provides a light source comprising: a light source unit for supplying light to a sample containing fluorescent microorganisms reflecting only light of a specific wavelength range; A method for determining the number of objects of a microorganism included in the sample using an optical device including an image acquisition unit for collecting an image and an image processing unit for processing the generated image, Collecting light from a microorganism contained in the sample, wherein the image acquiring unit radiates light in a specific wavelength range; (b) the image acquiring unit generating an image for the collected light; (c) The image processing unit displays the image generated by the image obtaining unit on the screen, analyzes each pixel constituting the image displayed on the screen, and detects a plurality of pixels that continuously emit light among the pixels included on the screen Extracting only an object having an average value of brightness of pixels constituting the object corresponding to a predetermined brightness range among the objects as an active object; (d) the image processing unit calculating the number of active objects included in the screen; .

In this case, the step (c) may further include: (c-1) performing the edge detection processing on the generated image by the image processing unit; (c-2) the image processing unit scans the image subjected to the boundary detection processing in the step (c-1), and detects a plurality of continuous light beams included in each boundary line with respect to each of the boundary lines included in the boundary- Classifying pixels into one object; (c-3) The image processing unit determines whether the average value of the brightness of all the pixels constituting the object is included in the predetermined brightness range for each of the objects extracted in the step (c-2) Extracting only an object included in the brightness range as an active object; And a control unit.

In the step (c), the image processing unit may extract only the number of pixels constituting the one active object among the extracted active objects within a predetermined number of ranges as a target active object, The total number of the active objects included in the screen calculated by the image processing unit is a total number of the target active objects.

In the step (c), the image processing unit classifies the plurality of consecutively emitting pixels into one object, and consecutively consecutively consecutively arranges one or more pixels Only a plurality of pixels emitting light within a preset contrast ratio range are determined as one object.

As described above, according to the present invention, it is possible to provide a method for inspecting a microorganism having a fluorescence characteristic for a specific wavelength, which can precisely grasp only the number of active microorganisms among the microorganisms having fluorescence characteristics for a specific wavelength included in the sample .

In addition, according to the present invention, it is possible to provide a method for inspecting a microorganism having a fluorescence characteristic for a specific wavelength capable of accurately grasping the microorganisms contained in the sample.

In addition, according to the present invention, it is possible to provide a method for inspecting microorganisms having a fluorescence characteristic for a specific wavelength capable of accurately grasping the number of microorganisms by filtering the various foreign substances contained in the sample.

BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is a schematic block diagram of an optical apparatus for performing a microorganism inspection method having fluorescence characteristics for a specific wavelength according to the present invention. FIG.
2 is a perspective view showing an internal configuration of the optical device.
3 is an explanatory view showing the structure of the optical device.
4 is a block diagram of the detection unit.
5 and 6 are flow charts of a method for testing microorganisms having fluorescence characteristics for a specific wavelength according to the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention and the manner of achieving them will become apparent with reference to the embodiments described in detail below with reference to the accompanying drawings.

The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. To fully disclose the scope of the invention to those skilled in the art, and the invention is only defined by the scope of the claims. Like reference numerals refer to like elements throughout the specification.

Hereinafter, the present invention will be described with reference to the drawings for explaining a microorganism inspection method having fluorescence characteristics for a specific wavelength according to embodiments of the present invention.

FIG. 1 is a schematic block diagram of an optical apparatus for performing a microorganism inspection method having fluorescence characteristics for a specific wavelength according to the present invention, FIG. 2 is a perspective view for showing an internal configuration of the optical apparatus, Is an explanatory diagram showing the structure of the optical device, and Fig. 4 is a block diagram of the detector.

5 and 6 are flowcharts of a microorganism inspection method having fluorescence characteristics for a specific wavelength according to the present invention.

A method for inspecting a microorganism having a fluorescence characteristic at a specific wavelength according to a preferred embodiment of the present invention includes a light source unit 150 for supplying light to a sample containing fluorescent microorganisms reflecting only light of a specific wavelength range, An optical apparatus 100 including an image obtaining unit 121 for collecting the reflected light and generating an image and a detecting unit 120 including an image processing unit 122 for processing the generated image, To a method for determining the number of objects of an included microorganism.

First, the configuration of the optical device 100 used in the microorganism inspection method having the fluorescence characteristic for a specific wavelength according to the present invention will be described.

1 to 3, the optical device 100 includes a light source unit 150, an optical filter unit 110, a ground unit 190, and a detection unit 120.

The light source unit 150 is a means for performing a function of a light source used in an optical microscope or the like and is a means for emitting most visible light or light of a specific wavelength range.

When the light of the entire visible light region is emitted from the light source unit 150, the optical filter unit 110 may filter the light of the wavelength range of the light of the emitted visible light region, And an excitation filter 111 to be transmitted to the platform 190 side.

At this time, the light source unit 150 may be a light source that emits only light of a specific wavelength range that the microorganism exhibits fluorescence characteristics. In this case, the excitation filter described above may be excluded.

Examples of microorganisms exhibiting fluorescence characteristics with respect to the light of the specific wavelength range include plankton containing chlorophyll and the like.

The plankton is a microorganism having a fluorescence characteristic of emitting red light having a wavelength of 620 nm to 780 nm. Hereinafter, the plankton will be described as an example.

The platform 190 is preferably a supporting stand on which samples such as ship equilibrium water or seawater are placed, and the sample is preferably placed on the platform in a state where the ship equilibrium water or seawater is dried.

The detector 120 is a means for collecting the reflected light of the sample placed on the platform 190 and reflecting the light of the light source unit 150 and detecting the image of the reflected light, and includes a camera used for the optical microscope do.

In this case, it is preferable that the camera used in the detection unit 120 is capable of observing objects of at least 20 mu m, and preferably has a resolution of 1250 * 1250 pixels or more.

That is, the detector 120 is a means for collecting the light reflected from the sample placed on the platform 190 and detecting the image of the reflected light. The light emitted from the light source 150 is received by the detector 120, And is disposed at a position opposite to the platform 190 to detect an image of the plankton contained in the sample.

The optical filter unit 110 is a means for transmitting the light provided from the light source unit 150 to the sample side disposed on the platform 190 and transmitting the light reflected by the sample to the detection unit 120 side.

The light from the light source unit 150 is transmitted to the light source unit 150 through the optical filter unit 110 without interfering with the external light between the light source unit 150, the optical filter unit 110, the detection unit 120, 190 to the sample side and to allow the light reflected from the sample to travel through the optical filter unit 110 to the detection unit 120 without loss.

The optical filter unit 110 includes an excitation filter 111 and a dichroic mirror 112.

The excitation filter 111 is a filter for passing only the light of the wavelength of 380 nm to 480 nm among the light emitted from the light source unit 150.

At this time, if the microorganism is configured to emit light of a specific wavelength showing fluorescence characteristic in the light source unit, the excitation filter may be excluded.

The dual filter 112 is a means for transmitting the light of the blue series which has passed through the excitation filter 111 to the sample side disposed on the ground and for transmitting the light reflected from the sample to the detection unit 120 side .

The platform 190 and the detection unit 120 are disposed on a straight line and the light source unit 150 is disposed in a direction intersecting a straight line connecting the detection unit 120 and the platform 190.

At this time, the optical filter 112 reflects the blue-based light, which is the light passing through the excitation filter 111, to the sample side on the flooring 190, and only the red- Refracted or refracted) and converged on the detection unit 120. The optical filter shown in Fig.

That is, the excitation filter 111 passes only the blue light having a wavelength of 380 nm to 480 nm, the double filter 112 refracts the blue light to the sample side of the platform 190, Red light having a wavelength of 620 nm to 780 nm, which is the light reflected by the light source, passes through the detection unit 120 as it travels.

That is, the dual filter 112 is an optical filter that reflects light of 380 nm to 480 nm and passes light of 620 nm to 780 nm as it is.

That is, the optical filter unit 110 is disposed between the light source unit 150 and the detection unit 120 so that the excitation filter 111 and the dual filter 112 can perform the functions described above.

The optical filter unit 110 is formed with side surfaces opposed to the light source unit 150, the platform 190, and the detection unit 120, and each of the side surfaces is provided with a light passage ) Is formed.

At this time, the excitation filter 111 is coupled to a light path of a side surface of the optical box facing the light source unit 150, and the double filter 112 reflects light, 190 so that the light reflected by the plankton contained in the sample passes through the optical part 120 side.

At this time, it is preferable that the side surface of the optical box facing the detection unit 120 further includes an emission filter 113.

The emission filter 113 is an optical filter for passing only light having a wavelength of 600 nm or more, and is a filter for minimizing the transmission of light corresponding to the noise to the detection unit side.

And a tube-shaped light guide member 130 connecting the light source unit 150 and the excitation filter coupled to the optical box so that the light emitted from the light source unit 150 is concentrated on the excitation filter coupled to the optical box .

The light guide member 130 functions to allow the light of the light source unit 150 to pass through the excitation filter 111 of the optical filter unit 110 without interference with external light.

The optical filter unit 110 may further include a separate light guide member for connecting the optical filter unit 110 to the platform 190 and the detection unit 120.

The detection unit 120 includes an image acquisition unit 121 for sensing light reflected by the plankton included in the sample and generating an image of plankton contained in the sample, And an image processor 121 for analyzing each pixel to determine the number of plankton contained in the sample.

That is, the light of the light source unit 150 is transmitted to the microorganisms included in the sample that emits only light of a specific wavelength range of the ground through the optical filter unit 110, Is condensed by the image acquisition unit 121 of the detection unit 120 through the optical filter unit 110 to generate an image of the microorganism, and the image processing unit 122 processes the generated image Thereby calculating the number of microorganisms contained in the sample.

Hereinafter, a method for inspecting microorganisms having fluorescence characteristics for a specific wavelength according to the present invention will be described.

The method for inspecting a microorganism having fluorescence characteristic for a specific wavelength according to the present invention is characterized in that the image acquiring unit 121 collects light from microorganisms included in the sample which radiate light in a specific wavelength range (b) generating an image of the collected light by the image obtaining unit 121; and (b) displaying the image generated by the image obtaining unit 121 on the screen And analyzing each pixel constituting a screen on which the image is displayed to classify a plurality of pixels that continuously emit light among the pixels included in the screen into one object, (C) extracting only an object whose average value of brightness corresponds to a predetermined brightness range to an active object, and calculating the number of active objects included in the image by the image processing unit And a step (d) to.

That is, the method for inspecting microorganisms having fluorescence characteristics for a specific wavelength according to the present invention is characterized in that the image obtaining unit 121 irradiates light emitted from a microorganism included in the sample, which radiates light in a specific wavelength range, (Step (a)) of collecting the collected light, and the step (b) of generating the image of the collected light by the image acquiring unit.

For example, when the microorganism is phytoplankton, when blue light having a wavelength of 380 nm to 480 nm and passed through the excitation filter 111 is reflected to the sample side by the double filter 112, light having a wavelength of 380 nm to 480 nm Through the photosynthetic action of the chloroplast contained in the phytoplankton, etc., the energy level is changed to the light of the wavelength of 620 nm to 780 nm and is radiated. At this time, the double filter 112 passes the emitted light having a wavelength of 620 nm to 780 nm directly to the detection unit, thereby generating an image of the phytoplankton contained in the sample to be inspected in the image acquisition unit of the detection unit.

Then, the image processing unit 122 displays the image generated by the image obtaining unit on the screen, analyzes each pixel constituting the image displayed on the screen, and analyzes a plurality of Pixels are classified into one object (step (c)).

For example, when there are two pixels (first object), five pixels (second object), twelve pixels (third object) and twenty pixels (fourth object) It is classified as having four objects on the screen.

At this time, only an object having an average value of brightness for the pixels constituting the object among the objects corresponding to a predetermined brightness range is extracted as an active object (step (c)).

That is, in the above example, the intensity range for the brightness of light for the microorganism being actually activated, that is, the microorganism being actually active as life is preset to the image processing unit as a range of, for example, 10 lux (lux) to 20 lux When the brightness range is set from 10 lux to 20 lux, the average brightness of the pixels constituting the first object is 8 lux, the average brightness of the pixels constituting the second object is 10 lux, If the average brightness of the pixels constituting the third object is 16 lux and the average brightness of the pixels constituting the fourth object is 25 lux, the image processing unit 122 may calculate the average brightness of the second object and the fourth object, Only 3 objects are extracted as active objects.

That is, two active objects are treated as being present on the screen.

In this case, the predetermined range of the brightness or intensity of the predetermined light may be determined by using various units for the intensity or brightness of the existing light such as lumen (luminous flux) or lux (illumination) or candela (cd) It can be set.

The detector may be configured to digitize the image of the collected light so that the brightness of each pixel constituting the image has a value between 256, which is regarded as the brightness of 0 to the brightest light, And a predetermined range of brightness or intensity of the predetermined light may be preset to a range of the appropriate one of the values between 0 and 256. [

Finally, the image processing unit 122 calculates the number of active objects included in the screen (step (d)).

That is, as described above, in the above example, the screen for the sample contains two active objects.

It is needless to say that the manner in which the image processing unit 122 in step (c) distinguishes a plurality of consecutively emitting pixels can be easily extracted from the information on the emitting pixels.

At this time, in the present invention, the image processing unit 122 may perform edge discrimination processing for the image to perform discrimination for consecutively emitting pixels.

The edge, or contour, is a feature that represents the boundary of a region in an image.

In this case, the boundary detection is a method of obtaining a pixel corresponding to a contour line as a discontinuous point of the image brightness at the boundary (edge, edge), and a conventional boundary detection processing method can be applied.

That is, the step (c) includes a step c-1 in which the image processing unit 122 performs edge detection processing on the generated image, and a step c-1 of scanning the boundary detection processed image in the step (C-2) categorizing a plurality of pixels, which are included in each boundary line included in the boundary detection processed image, into a single object, Extracting only the objects included in the predetermined brightness range as an active object (step < RTI ID = 0.0 > c -3) step.

By classifying an object using the above-described boundary detection method, an object included in the image can be more accurately extracted.

In step (c), the image processor 122 may extract only the extracted active objects that are included in the predetermined number of pixels constituting the one active object as the target active objects.

This is to exclude floats and the like other than plankton which should be actually grasped by previously setting a range of the number of consecutive pixels corresponding to the size of the target plankton actually in question.

In the above example, when the microorganism as the actual problem has a range of 4 to 13 pixels, the first object and the fourth object are excluded from the microorganisms that are actually a problem, and only the second object and the third object Will be extracted as the target active object.

In this case, the total number of the active objects included in the screen calculated by the image processing unit 122 in the step (d) may be the total number of the target active objects included in the screen.

In the step (c), the image processing unit classifies the plurality of consecutively emitting pixels into one object, and consecutively consecutively consecutively arranges one or more pixels Only a plurality of pixels emitting light within a preset contrast ratio range can be determined as one object.

In other words, even when 10 pixels are consecutively emitted, two of the pixels emit light in such a manner as to deviate from a predetermined contrast ratio with all adjacent pixels, and only eight pixels excluding the two pixels constitute one object .

This is for accurately extracting the number of pixels constituting one object and extracting the target active object accurately.

According to the method of the present invention for detecting a microorganism having fluorescence characteristic at a specific wavelength, it is possible to accurately grasp the number of microorganisms from a sample containing microorganisms having fluorescence characteristics of light of a specific wavelength, such as plankton In addition, the microorganism can be selectively screened for the active microorganisms.

Hereinafter, a microorganism testing apparatus having fluorescence characteristics for a specific wavelength according to the present invention will be described.

An apparatus for inspecting a microorganism having a fluorescence characteristic for a specific wavelength includes: a light source part (150) for emitting light; A platform 190 in which a sample containing microorganisms having fluorescence characteristics for a specific wavelength to be inspected is disposed; The light source unit 150 receives light emitted from the light source unit 150 and collects the light reflected by the microorganisms contained in the sample to collect the microorganisms included in the sample, A detection unit 120; And the light emitted from the light source unit 150 is transmitted to the sample side disposed on the flooring 190 and the light reflected by the sample is transmitted to the detection unit 120 side, An excitation filter 111 for passing only light of a wavelength of 380 nm to 480 nm and having a wavelength of 380 nm to 480 nm and a reflector for reflecting the light passed through the excitation filter to the sample side of the platform 190, An optical filter unit 110 including an optical filter 112 for passing only red light of 620 nm to 780 nm toward the detection unit 120; .

At this time, the optical filter unit 110 has side surfaces opposed to the light source unit 150, the platform 190, and the detection unit 120, respectively, and each side has an optical path through which light passes, Wherein the excitation filter (111) is coupled to a light path on a side of the optical path of the optical box opposite to the light source part (150), the optical filter (112) Is reflected by the sample side of the sample (190), and is allowed to pass through the optical part (120) through the light reflected by the plankton contained in the sample.

In addition, a tube-shaped light guide member 130 connecting the light source unit 150 and the excitation filter coupled to the optical box so that the light emitted from the light source unit 150 is concentrated on the excitation filter coupled to the optical box And further comprising:

The detecting unit 120 may include an image obtaining unit 121 for detecting light reflected by the plankton contained in the sample and generating an image of the microorganism included in the sample and displaying the image on a screen; The method comprising the steps of: analyzing each pixel included in a screen on which the generated image is displayed to determine a number of the microorganisms included in the sample, and determining a plurality of pixels that continuously emit light among the pixels included in the screen as one object An image processor (122) for calculating a total number of the objects included in the screen and determining the total number of target microorganisms included in the screen; And a control unit.

In addition, the image processing unit 122 may recognize only the object corresponding to the number of consecutively emitting pixels within the preset number of the objects as a target microorganism to be actually inspected, And the total number of the target microorganisms included in the screen is determined by calculating the total number of the target microorganisms.

The image processing unit 122 may exclude the target microorganisms when the light intensity average value of all the pixels constituting one object among the objects identified as the target microorganisms is out of the preset light intensity range do.

In addition, the image processing unit 122 processes each of a plurality of consecutively emitting pixels perceived by the one object such that each of the plurality of pixels consecutively emits light within a predetermined contrast ratio and adjacent pixels.

It will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive. The scope of the present invention is defined by the appended claims rather than the foregoing detailed description, and all changes or modifications derived from the meaning and scope of the claims and the equivalents thereof are included in the scope of the present invention Should be interpreted.

100: Optical device applied to a microorganism inspection method having fluorescence characteristic for a specific wavelength according to the present invention
110: optical filter unit 111: excitation filter
112: Optical filter 120: Detector
121: Image acquiring unit 121:
130: light guide member 141, 142, 143: support member
150: Light source

Claims (4)

An image obtaining unit for obtaining an image by collecting the light reflected from the microorganism and an image processing unit for processing the generated image, A method for determining the number of objects of a microorganism contained in the sample using an optical device including a detection unit including a detection unit,
(a) collecting light from a microorganism included in the sample, wherein the image acquiring unit radiates light in a specific wavelength range;
(b) the image acquiring unit generating an image for the collected light;
(c) The image processing unit displays the image generated by the image obtaining unit on the screen, analyzes each pixel constituting the image displayed on the screen, and detects a plurality of pixels that continuously emit light among the pixels included on the screen Extracting only an object having an average value of brightness of pixels constituting the object corresponding to a predetermined brightness range among the objects as an active object;
(d) the image processing unit calculating the number of active objects included in the screen; Wherein the microorganism has a fluorescence characteristic for a specific wavelength.
The method of claim 1, wherein the step (c)
(c-1) the image processing unit performs edge detection processing on the generated image;
(c-2) the image processing unit scans the image subjected to the boundary detection processing in the step (c-1), and detects a plurality of continuous light beams included in each boundary line with respect to each of the boundary lines included in the boundary- Classifying pixels into one object;
(c-3) The image processing unit determines whether the average value of the brightness of all the pixels constituting the object is included in the predetermined brightness range for each of the objects extracted in the step (c-2) Extracting only an object included in the brightness range as an active object; Wherein the microorganism has a fluorescence characteristic with respect to a specific wavelength.
The method according to claim 1,
In the step (c), the image processing unit,
Extracting only the extracted active objects that include the number of pixels constituting the one active object within a predetermined number of ranges as a target active object,
Wherein the total number of the active objects included in the screen calculated by the image processing unit in the step (d) is the total number of the target active objects.
The method according to claim 1,
In the step (c), the image processing unit,
The method according to claim 1, wherein the plurality of consecutive light emitting pixels are classified into one object, and the plurality of pixels emitting light within a predetermined contrast ratio range continuously with one or more pixels among all adjacent pixels The microorganism having the fluorescence characteristic with respect to a specific wavelength.

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