KR102606708B1 - The Preprocessing Apparatus for Apparatus for Continuously Monitoring Image of Microalgae - Google Patents

Abstract

The present invention relates to a preprocessing device for continuous measurement of microalgae images. More specifically, when a microalgae sample is injected to continuously measure high-resolution microalgae images, a preprocessing step is automatically performed on the microalgae sample. This relates to a preprocessing device for continuous measurement of microalgae images that can stably capture multiple microalgae images.

Description

Preprocessing Apparatus for Continuously Monitoring Image of Microalgae}

The present invention relates to a preprocessing device for continuous measurement of microalgae images. More specifically, when a microalgae sample is injected to continuously measure high-resolution microalgae images, a preprocessing step is automatically performed on the microalgae sample. This relates to a preprocessing device for continuous measurement of microalgae images that can stably capture multiple microalgae images.

In water ecosystems such as large-scale dams, rivers, and lakes in Korea and other countries, environmental destruction is caused mainly by the occurrence of green and red tides, and this causes great difficulties in water management. Therefore, various methods for removing algae are continuously being studied, and preventive measures are also being taken to fundamentally block or suppress the occurrence of algae.

However, prior to research into these preventive measures or bird removal methods, a bird monitoring system must be established to identify in advance where birds occur. In addition, the dominant species of these bird groups may vary depending on the region, environment, and season, and in order to properly manage and monitor the water quality and environment of lakes and rivers, it is important to secure a database for each of these bird groups.

Accordingly, conventional bird monitoring methods include a method of irradiating light with multiple wavelengths to birds and analyzing the wavelengths generated by the birds, a method of analyzing DNA extracted from birds, and a method of directly imaging birds using a microscope, etc. There are ways to take pictures, etc. However, in order to classify species for each group of birds and secure a database for the birds, DNA analysis or image taking methods are desirable, but are difficult to implement in small-scale research institutes or private companies due to time and cost issues. there is.

Meanwhile, research has recently been conducted on technology to learn images and classify objects using artificial intelligence algorithms, and image classification through deep learning has the advantage of being able to analyze a large amount of images with high accuracy. , this requires a sufficient amount of learning, and it is generally known that at least 2000 images are needed for learning to classify one object.

Therefore, in order to classify images through deep learning for samples collected from rivers or water sources, a preprocessing process to remove flocs, which are aggregates of microalgae present in the sample, is essential, and as thousands of high-resolution microalgae images must be measured, a high-resolution process is required. There is a need for inventions related to preprocessing devices with high performance and efficiency.

The present invention relates to a preprocessing device for continuous measurement of microalgae images. More specifically, when a microalgae sample is injected to continuously measure high-resolution microalgae images, a preprocessing step is automatically performed on the microalgae sample. The purpose is to provide a preprocessing device for continuous measurement of microalgae images that can stably capture multiple microalgae images.

In order to solve the above problems, the present invention is a preprocessing device for a continuous microalgae image measurement device, comprising: a dilution unit that supplies a dilution solvent to a microalgae sample to control turbidity of the microalgae; An ultrasonic disperser for dispersing the microalgae by radiating ultrasonic waves to the microalgae sample at an ultrasonic intensity for each type of microalgae for pretreatment of the microalgae sample, wherein the dilution unit includes a casing; an internal partition dividing the casing into an upper space and a lower space; a diluted solvent storage tank disposed in the lower space of the casing and storing a diluted solvent; a sample tank disposed in the upper space of the casing and receiving microalgae samples from a sample storage tank; a first diluting tank disposed in the upper space of the casing and receiving a microalgae sample from the sample tank and receiving a dilution solvent from the dilution solvent storage tank to dilute the concentration of the microalgae sample; a second dilution tank disposed in the upper space of the casing and receiving a microalgae sample from the first dilution tank and receiving a dilution solvent from the dilution solvent storage tank to dilute the concentration of the microalgae sample; and a pump unit including a plurality of pumps that perform fluid movement between the dilution solvent storage tank, the sample tank, the first dilution tank, the second dilution tank, and the ultrasonic disperser. A microalgae image comprising a. Provides preprocessing equipment for continuous measurement devices.

In one embodiment of the present invention, the first dilution tank and the second dilution tank each include a dilution chamber in which the microalgae sample and the dilution solvent are mixed; A turbidity meter attached to the dilution chamber and measuring turbidity of the microalgae sample contained in the dilution chamber; A stirring motor installed at the lower end of the dilution chamber; a first magnet located between the stirring motor and the dilution chamber, connected to the stirring motor, and rotating when the stirring motor is activated; It may include a second magnet located at a lower portion inside the dilution chamber and rotating in response to the first magnet when the first magnet rotates.

In one embodiment of the present invention, the ultrasonic disperser includes an ultrasonic dispersion chamber that receives a microalgae sample from a second dilution tank by the pump unit; An ultrasonic irradiation device that irradiates ultrasonic waves to the microalgae sample supplied to the ultrasonic dispersion chamber; And it may include a temperature sensor that measures the temperature of the microalgae sample supplied to the ultrasonic dispersion chamber.

In one embodiment of the present invention, the sample tank includes a first turbidity meter capable of measuring turbidity, the first dilution tank includes a second turbidity meter capable of measuring turbidity, and the second dilution tank measures turbidity. It includes a third turbidity meter, wherein the pump unit includes a first dilution pump for moving the dilution solvent from the dilution solvent storage tank to the first dilution tank, and a second dilution pump for moving the dilution solvent from the dilution solvent storage tank to the second dilution tank. 2 It includes a dilution pump, and the pretreatment device controls the operation of the first dilution pump and the second dilution pump based on turbidity information sensed by the first turbidity meter, the second turbidity meter, and the third turbidity meter. It further includes a transfer pump control unit, wherein the transfer pump control unit sets a first target turbidity of the first dilution tank and a second target turbidity of the second dilution tank based on the first turbidity information sensed by the first turbidity meter. Setting stage; A first dilution pump control step of controlling the first dilution pump so that the second turbidity information sensed by the second turbidity meter approaches the first target turbidity; and a second dilution pump control step of controlling the second dilution pump so that the third turbidity information sensed by the third turbidity meter approaches the second target turbidity.

In one embodiment of the present invention, the sample tank includes a first turbidity meter capable of measuring turbidity, the pump unit includes a first dilution pump for moving the dilution solvent from the dilution solvent storage tank to the first dilution tank, and the dilution It includes a second dilution pump that moves the dilution solvent from the solvent storage tank to the second dilution tank, and the pretreatment device is based on the turbidity information sensed by the first turbidity meter, and the first dilution pump and the second dilution pump. and a transfer pump control unit that controls the operation, wherein the transfer pump control unit determines the first target turbidity of the first dilution tank and the second target turbidity of the second dilution tank based on the first turbidity information sensed by the first turbidity meter. Setting target turbidity setting step; A first dilution pump control step of controlling the first dilution pump based on the first target turbidity; and a second dilution pump control step of controlling the second dilution pump based on the second target turbidity.

In one embodiment of the present invention, the ultrasonic disperser includes a calibration step of deriving an energy unit corresponding to an input current or a device drive value for the input energy unit; and a device driving step of driving the ultrasonic disperser using the device driving value derived in the calibration step.

In one embodiment of the present invention, the device driving step is to irradiate ultrasonic waves to the microalgae sample for a preset time at ultrasonic intensity according to the device driving value derived in the calibration step to produce flocs, which are microalgae aggregates. It can be dispersed.

According to one embodiment of the present invention, the turbidity of the microalgae sample in the dilution chamber can be measured through a turbidity meter attached to the dilution chamber, and the dilution process can be performed based on this, thereby reducing the time required for the dilution process and accurately It can be effective in performing a sufficient amount of dilution.

According to one embodiment of the present invention, by performing a double dilution process, a high-multiplier dilution process can be performed with a small amount of dilution solvent, thereby reducing the space and cost problems of the conventional pretreatment device. .

According to one embodiment of the present invention, by using a modular pretreatment device, it is easy to replace the microalgae sample and dilution solvent, and cleaning and replacing the dilution chamber is simple, which has the effect of reducing maintenance costs. .

According to one embodiment of the present invention, a plurality of pumps provided in the pretreatment device can be automatically operated by the transfer pump control unit, so that the microalgae sample is pretreated by simply injecting the microalgae sample into the microalgae storage tank. Because the process can be performed, the efficiency of the pretreatment process can be increased, and because it is performed automatically, it can have the effect of reducing contamination that may occur with the sample.

According to one embodiment of the present invention, the pretreatment method using an ultrasonic disperser can increase the single cell recovery rate and solve the efficiency and time problems compared to the conventional pretreatment method using chemicals and heat treatment.

According to an embodiment of the present invention, the intensity of the ultrasonic disperser can be standardized through a calibration step, which has the effect of implementing cross-validation between laboratories.

Figure 1 schematically shows a device for continuously measuring microalgae images according to an embodiment of the present invention.
Figure 2 shows a microalgae image according to a preprocessing method according to an embodiment of the present invention.
Figure 3 shows microalgae images according to the intensity of an ultrasonic disperser that irradiates ultrasonic waves to a microalgae sample according to an embodiment of the present invention.
Figure 4 schematically shows the steps performed in the calibration step according to an embodiment of the present invention.
Figure 5 shows a graph for calculating relationship information between device drive values and energy units in the calibration step according to an embodiment of the present invention.
Figure 6 shows an actual photograph and design drawing of a flow cell according to an embodiment of the present invention.
Figure 7 schematically shows the structure of a flow path in a flow cell according to an embodiment of the present invention.
Figure 8 shows microalgae images according to the height of the flow cell flow path according to an embodiment of the present invention.
Figure 9 shows an image of microalgae according to the shape of a flow path in a flow cell according to an embodiment of the present invention.
Figure 10 schematically shows the structure of an inverted microscope according to an embodiment of the present invention.
Figure 11 schematically shows the structure and fluid flow of a pretreatment device according to an embodiment of the present invention.
Figure 12 schematically shows the structures of the first and second dilution tanks according to an embodiment of the present invention.
Figure 13 schematically shows a dilution process performed in a dilution unit according to an embodiment of the present invention.
Figure 14 schematically shows the steps of the dilution process performed in the dilution unit according to an embodiment of the present invention.
Figure 15 schematically shows the steps of the dilution process performed in the dilution unit according to another embodiment of the present invention.
Figure 16 schematically shows the structure of an ultrasonic disperser according to an embodiment of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS Various embodiments and/or aspects are now disclosed with reference to the drawings. In the following description, for purposes of explanation, numerous specific details are set forth to facilitate a general understanding of one or more aspects. However, it will be appreciated by those skilled in the art that this aspect(s) may be practiced without these specific details. The following description and accompanying drawings set forth in detail certain example aspects of one or more aspects. However, these aspects are illustrative and some of the various methods in the principles of the various aspects may be utilized, and the written description is intended to encompass all such aspects and their equivalents.

Additionally, various aspects and features may be presented by a system that may include multiple devices, components and/or modules, etc. It is also understood that various systems may include additional devices, components and/or modules, etc. and/or may not include all of the devices, components, modules, etc. discussed in connection with the drawings. It must be understood and recognized.

As used herein, “embodiments,” “examples,” “aspects,” “examples,” etc. may not be construed to mean that any aspect or design described is better or advantageous over other aspects or designs. . The terms '~part', 'component', 'module', 'system', 'interface', etc. used below generally refer to computer-related entities, such as hardware, hardware, etc. A combination of and software, it can mean software.

Additionally, the terms “comprise” and/or “comprising” mean that the feature and/or element is present, but exclude the presence or addition of one or more other features, elements and/or groups thereof. It should be understood as not doing so.

Additionally, terms including ordinal numbers, such as first, second, etc., may be used to describe various components, but the components are not limited by the terms. The above terms are used only for the purpose of distinguishing one component from another. For example, a first component may be named a second component, and similarly, the second component may also be named a first component without departing from the scope of the present invention. The term and/or includes any of a plurality of related stated items or a combination of a plurality of related stated items.

In addition, in the embodiments of the present invention, unless otherwise defined, all terms used herein, including technical or scientific terms, are generally understood by those skilled in the art to which the present invention pertains. It has the same meaning as Terms defined in commonly used dictionaries are interpreted as having meanings consistent with their meanings in the context of related technologies.

It should be, and unless clearly defined in the embodiments of the present invention, it should not be interpreted in an ideal or excessively formal sense.

1. Microalgae image continuous measurement device

Before explaining the preprocessing device for the continuous microalgae image measurement device of the present invention, the microalgae image continuous measurement device will be described.

Figure 1 schematically shows a device for continuously measuring microalgae images according to an embodiment of the present invention.

As shown in Figure 1, it is a continuous microalgae image measurement device, including a filter unit 100 that removes contaminants contained in the microalgae sample; A dilution unit 110 that supplies a solvent to the microalgae sample to control turbidity of the microalgae; An ultrasonic disperser 120 that disperses the microalgae by irradiating ultrasonic waves to the microalgae sample at an ultrasonic intensity for each type of microalgae for preprocessing of the microalgae sample; A flow cell 130 through which the microalgae sample to which ultrasonic waves are applied by the ultrasonic disperser 120 flows; A microscope that continuously photographs the microalgae sample inside the flow cell 130; And a transfer pump 140 that transfers the microalgae sample from the filter unit 100 to the flow cell 130. It provides a continuous microalgae image measurement device including a.

Meanwhile, the dilution unit 110 includes a turbidity meter for measuring turbidity of the microalgae sample; A dilution solvent capable of diluting the microalgae sample based on the turbidity measured by the turbidity meter; And a dilution chamber for diluting the microalgae sample using the dilution solvent, wherein the dilution solvent includes distilled water and may optionally further include a chemical agent for fixation of the microalgae.

Specifically, in the filter unit 100, the first step among the preprocessing steps to disperse flocs, which are microalgae aggregates present in the microalgae sample, is performed, and the microalgae sample is filtered using a microsieve filter. The microsieve filter is a mesh-type filter with a pore size of tens to thousands of μm and removes contaminants present in the microalgae sample. Unlike algae cultured species, samples obtained from water sources necessarily require a contaminant removal process and a dilution process. The microsieve filter can be used to remove foreign substances depending on the characteristics of the algae or to adjust the pore size so that only algae smaller than the target size can flow into the flow cell 130.

According to one embodiment of the present invention, a microsieve filter with a pore size of 1000 ㎛ can be used to prevent foreign substances from entering the flow cell 130, and a microsieve filter with a pore size of 50 ㎛ can be used to prevent microalgae colonies. Can be used to distribute to objects. According to another embodiment of the present invention, even when a microsieve filter with a small pore size is used, the object dispersion effect may not be clearly displayed, and an additional preprocessing step may be performed to compensate for this. The additional pretreatment steps include a dilution step and an ultrasonic dispersion step, and details about these will be described later.

In the dilution unit 110, the second step among the pretreatment steps for dispersing the floc in the microalgae sample is performed. After measuring the turbidity of the microalgae sample, a dilution solvent is used based on the measured turbidity to dilute the microalgae sample. Samples can be diluted.

The turbidity of the microalgae sample can be measured by a turbidity meter provided in the dilution unit 110. Specifically, the turbidity meter can measure the turbidity of the microalgae sample by irradiating light to the microalgae sample and comparing the degree to which the light transmitted through the sample is reflected or scattered by dispersed particles with a standard solution.

Based on the turbidity of the microalgae sample measured through the turbidity meter, the dilution unit 110 can adjust the amount of dilution solvent injected into the dilution chamber provided. As an embodiment of the present invention, it is preferable to inject the dilution solvent into the dilution chamber so that the turbidity of the microalgae sample is less than 200 NTU, and distilled water is used as the dilution solvent.

Meanwhile, as an embodiment of the present invention, a chemical agent may be optionally included in the diluting solvent to fix algae in the microalgae sample. Formaldehyde, etc. may be used as the chemical agent, and as an embodiment of the present invention, a chemical agent such as KOH (potassium hydroxide) or NaOH (sodium hydroxide) may be additionally used to disperse the floc in the microalgae sample. .

As another embodiment of the present invention, the dilution solvent can be automatically injected into the dilution chamber through an automated system without operator intervention. As shown in FIG. 1, the dilution solvent is contained in a separate dilution solvent reservoir and can be injected into the dilution chamber through the transfer pump 140. Through the automated system described above, contamination of the microalgae sample can be prevented and the effect of increasing the efficiency of the measurement method can be expected.

The ultrasonic disperser 120 is a device used in the last step of the preprocessing step to disperse flocs in the microalgae sample by irradiating ultrasonic waves to the microalgae sample.

Algae dispersion pretreatment techniques include chemical treatment techniques and heat treatment techniques in addition to techniques using ultrasonic waves. However, the present invention adopted the ultrasonic technique, which showed excellent results in terms of dispersion time and single cell recovery rate in single cell recovery rate experiments conducted independently. . A detailed comparison between the preprocessing technique using ultrasonic waves and the preprocessing technique without using ultrasonic waves will be described later.

Meanwhile, by performing a calibration step in the ultrasonic disperser 120, the energy unit corresponding to the injected microalgae sample or the device drive value for the input energy unit can be derived. In other words, the optimal ultrasonic intensity corresponding to the microalgae sample can be derived to maximize the dispersion effect of microalgae, and since the energy unit is a standardized value, cross-verification with other laboratories can be implemented. Details about the calibration step will be described later.

The flow cell 130 includes a plurality of thin flow paths, through which the microalgae sample that has undergone a pretreatment process can flow, and by photographing the microalgae sample flowing through the flow path through a microscope, the corresponding microalgae sample can flow. Microalgae images can be obtained for algae samples. As an embodiment of the present invention, the width of the passage is preferably set to 50 ㎛ to 1000 ㎛ in order to capture images of microalgae of various sizes. Details about the flow cell 130 will be described later.

The transfer pump 140 is located between the filter unit 100 and the dilution unit 110 and between the dilution solvent and the dilution chamber to help transfer the microalgae sample or the dilution solvent. The transfer pump 140 may use a micro pump to precisely control the amount of the microalgae sample flowing into the flow cell 130. The flow rate and flow rate of the microalgae sample transported through the micropump can be adjusted depending on the size and shape of the flow path within the flow cell 130. The micro pump refers to a fluid device capable of transporting a minute flow rate of 1 μl or less per minute.

Additionally, the operating state of the transfer pump 140 can be controlled through pump control software. Through the pump control software, it is possible to control the flowing amount of the microalgae sample or dilution solvent, control the operation of the syringe, and control the continuous operation of the pump. Once the transfer pump 140 is set up through the pump control software, it can automatically transfer the set amount of microalgae sample and dilution solvent to the dilution chamber, ultrasonic disperser 120, and flow cell 130, This can have the effect of increasing efficiency in measuring the microalgae images.

As described above, in one embodiment of the present invention, the microalgae image continuous measurement device automatically performs a preprocessing step when only the microalgae sample is initially injected, and then the sample is transferred to the flow cell 130, and the sample is transferred to the flow cell 130. The image of the microalgae sample can be automatically measured in the flow cell 130. An example of the device for continuously measuring microalgae images that is performed automatically is as follows.

When the microalgae sample is injected into the microalgae image continuous measurement device, the microalgae sample is transferred to the filter unit 100 by the transfer pump 140, and impurities and foreign substances present in the microalgae sample are removed. .

The microalgae sample that has passed through the filter unit 100 is automatically transferred to the dilution chamber of the dilution unit 110 by the transfer pump 140, and the turbidity meter automatically measures the microalgae sample in the dilution chamber. Measure turbidity. The dilution unit 110 automatically calculates the amount of dilution solvent required to dilute the microalgae sample based on the measured turbidity, and based on the calculated amount of dilution solvent, the microalgae sample is preset. Dilute below turbidity.

The microalgae sample that has passed through the dilution unit 110 is transferred to the ultrasonic disperser 120 by the transfer pump 140, and the ultrasonic disperser 120 is based on the preset ultrasonic intensity and ultrasonic irradiation time. Ultrasonic waves are irradiated to the microalgae sample transferred to the ultrasonic disperser 120.

The microalgae sample that has completed the preprocessing step in the ultrasonic disperser 120 is transferred to the flow cell 130 by the transfer pump 140, and images of the microalgae sample are continuously captured in the flow cell 130. You can shoot.

Movement of the microalgae sample by the transfer pump 140 operates automatically according to preset values.

Figure 2 shows a microalgae image according to a preprocessing method according to an embodiment of the present invention.

Schematically, Figure 2 (a) shows a microalgae image of a microalgae sample using a chemical treatment technique among the algae dispersion pretreatment techniques, and Figure 2 (b) shows microalgae images using an ultrasonic irradiation technique among the algae dispersion pretreatment techniques. An image of microalgae in the sample is shown.

Specifically, algae spraying pretreatment techniques can be largely divided into single pretreatment techniques and complex pretreatment techniques. The single pretreatment techniques include chemical treatment techniques, heat treatment techniques, and ultrasonic techniques, and the complex pretreatment technique is a technique that uses two or more of the above single pretreatment techniques in combination. In general, in the case of complex preprocessing techniques, the dispersion effect increases compared to a single preprocessing technique, but time and cost may also increase.

For the present invention, a plurality of experiments were performed to derive the optimal pretreatment technique, and in Figure 2, as an example of the present invention, the single cell recovery rate of the pretreatment technique using ultrasound and the pretreatment technique without ultrasound are shown. An image showing the difference is shown. The algae used in the experiment corresponding to Figure 2 is Chlorella vulgaris, and the multiple experiments were conducted on a plurality of algae including Chlorella vulgaris, but this is not shown separately.

Figure 2 (a) is an example of the present invention, a microscope image of Chlorella vulgaris using a chemical treatment technique, and in this experiment, a 0.01M sodium hydroxide solution was used. As shown in Figure 2 (a), areas 6 and 8 appear to be relatively well dispersed, but in areas 19 and 21, flocs that are not properly dispersed exist, as shown in the red circle. You can check that.

Figure 2 (b) is an example of the present invention, a microscope image of Chlorella vulgaris using an ultrasonic technique. In this experiment, ultrasonic waves were irradiated for 20 seconds at an ultrasonic intensity of 200 kJ/L. As shown in Figure 2 (b), it can be seen that algae are evenly distributed in all areas as a result of using the ultrasonic technique.

Based on the above, the present invention uses an ultrasonic treatment technique with high dispersion effect. However, depending on the type of microalgae, the intensity of ultrasonic waves and the ultrasonic irradiation time at which the dispersion effect on the microalgae is most effective are different. To compensate for this, the present invention determines the optimal ultrasonic intensity according to the type of microalgae through the calibration step. and ultrasonic irradiation time can be derived.

Figure 3 shows a microalgae image according to the intensity of the ultrasonic disperser 120 that irradiates ultrasonic waves to a microalgae sample according to an embodiment of the present invention.

Schematically, Figure 3 (a) shows a microalgae image when the microalgae sample was irradiated with an ultrasonic intensity of 0.004 W/ml. Figure 3(b) shows a microalgae image when the microalgae sample was irradiated with an ultrasonic intensity of 0.08 W/ml. Figure 3(c) shows a microalgae image when the microalgae sample was irradiated with an ultrasonic intensity of 0.3 W/ml.

Specifically, referring to FIG. 3 as an embodiment of the present invention, it can be seen that the more ultrasonic waves are irradiated to the microalgae sample with a strong intensity of ultrasonic waves, the better the algae dispersion effect appears. In the case of Figure 3 (c), a single The cell recovery rate was measured to be approximately 90%. As another embodiment of the present invention, when a complex pretreatment technique using a chemical treatment technique and an ultrasonic treatment technique is used simultaneously, the single cell recovery rate may be measured higher than that of the ultrasonic treatment technique. However, even when only the ultrasonic treatment technique is used, the single cell recovery rate is high. Since this has been measured, in the present invention, considering cost and time issues, a single pretreatment technique using only ultrasonic treatment is used.

Figure 4 schematically shows the steps performed in the calibration step according to an embodiment of the present invention.

As shown in FIG. 4, the ultrasonic disperser 120 includes a calibration step of deriving an energy unit corresponding to an input current or a device drive value for the input energy unit; A device driving step of driving the ultrasonic disperser 120 with the device driving value derived in the calibration step may be performed, and the calibration step includes driving the ultrasonic disperser 120 with the first device driving value to use the reference medium. A first measurement step (S100.1) of applying ultrasonic waves to and measuring first change information of the temperature of the reference medium; A second measurement step (S100.2) of driving the ultrasonic disperser 120 with a second device driving value to apply ultrasonic waves to a reference medium and measuring second change information of the temperature of the reference medium; A first derivation step (S110.1) of deriving a first energy unit for the first device drive value based on the first change information; A second derivation step (S110.2) of deriving a second energy unit for the second device drive value based on the second change information; and the first device drive value and the second device drive value. A step of deriving relationship information between the device drive value and the energy unit based on regression information including the first energy unit and the second energy unit, and based on the relationship information between the device drive value and the energy unit. The ultrasonic intensity for each type of microalgae can be derived.

Specifically, the calibration step is a step of standardizing the device drive value of the ultrasonic disperser 120 through a calorimetric method. In the method of using the conventional ultrasonic disperser 120, even if the ultrasonic disperser 120 is operated with the same device driving value, the result value varies depending on the laboratory environment, equipment used, and processing container, etc., resulting in the same Even if the samples were used in the same experimental method, the results were different, which caused a problem of low reliability of the experiment. In order to solve this problem, the present invention performs a calibration step to enable more rigorous comparison through standardized power no matter which laboratory the experiment is conducted in, and to increase the reliability of the experiment through cross-validation (Round Robin Test) between laboratories. It can have an improving effect. In addition, if the intensity of the ultrasonic waves applied to the microalgae sample is too strong, the algae cells may be destroyed, so by performing the calibration step, the device can maximize the dispersion effect of the microalgae depending on the type of microalgae. You can find the driving value. The reference medium, as an example of the present invention, may be a microalgae sample diluted by the dilution unit 110.

Figure 5 shows a graph for calculating relationship information between device drive values and energy units in the calibration step according to an embodiment of the present invention.

As shown in FIG. 5, the ultrasonic disperser 120 performs a device driving step of driving the ultrasonic disperser 120 with the device driving value derived in the calibration step, and in the device driving step, the calibration Depending on the device drive value derived in the step, ultrasonic waves can be irradiated to the microalgae sample for a preset time at ultrasonic intensity to disperse floc, which is an aggregate of microalgae.

Specifically, with reference to Figure 4, in the calibration step, a measurement step (S100.1 to S100.N, S100) and a derivation step (S110.1 to S110.N, S110) are performed, and the measurement step ( S100) is sequentially performed from the first measurement step (S100.1) to the Nth measurement step (S100.N), and the derivation step (S110) is performed corresponding to the measurement step. In the measurement step, the device drive value is set and the ultrasonic disperser 120 is driven with the device drive value. The ultrasonic disperser 120 applies ultrasonic waves to the reference medium, and the ultrasonic wave is applied according to the irradiation time of the reference medium to which the ultrasonic waves are applied. Record change information about the changed temperature of the reference medium.

As an embodiment of the present invention, Figures 5 (a) and (b) show graphs for the measurement step (S100), and Figures 5 (a) and (b) show results performed in different laboratories, respectively. is shown in a graph.

Referring to Figure 5(a), the first device drive value was set to 10% and the temperature change according to the ultrasonic irradiation time was recorded. After completing the first measurement step (S100.1), the second measurement step (S100.2) was performed by setting the second device drive value to 20%, and the sixth device drive value was sequentially set to 100%. Up to 6 measurement steps (S100.6) were performed.

In the case of Figure 5(b), the experiment was conducted twice with one device drive value and the temperature change according to the ultrasonic irradiation time was recorded. In Figure 5(b), the first change information was obtained by setting the first device drive value to 20%, and sequentially in the fifth measurement step (S100.5), the fifth device drive value was set to 100%. , Obtain the fifth change information. After obtaining the fifth change information, the sixth device drive value is set to 20% again using the same reference medium as the reference medium used in the first measurement step (S100.1) to the fifth measurement step (S100.5). Then, the 6th change information is acquired, and in the 10th measurement step (S100.10), the 10th device drive value is set to 100% and the 10th change information is acquired.

In the derivation step (S110), change information obtained through the measurement step (S100); And the mass and specific heat information of the reference medium according to the initial conditions of the experiment; the device drive value and energy unit ( ) relationship information can be derived. In the derivation step (S110), the energy unit applied to the reference medium can be obtained by substituting the three condition information into Equation (1) below.

- 식 (1)

- Equation (1)

The above equation (1) can be derived by modifying the relationship between heat quantity and specific heat. The unit of the amount of heat is J (Joule), and dividing J by time (s) is W (Watt). Therefore, the equation (1) can be obtained by dividing both sides of the relationship between the amount of heat and specific heat by unit time.

As an embodiment of the present invention, Figures 5 (c) and (d) show graphs for the derivation step (S110) corresponding to Figures 5 (a) and (b), respectively.

Looking at Figures 5 (c) and (d), each blue dot represents the device drive value. Set the value as the energy unit derived based on the change information when the ultrasonic disperser 120 is driven with the device drive value. It was recorded as a value.

The relationship between the device drive value and the energy unit can be derived through the relationship information between the device drive value and the energy unit. Specifically, with reference to FIG. 4, the regression information may correspond to a relational expression between the device drive value and energy, and for the relational expression, a coefficient of determination ( ), the accuracy of the regression information can be determined.

As an embodiment of the present invention, referring to FIG. 5, it can be further confirmed that the coefficient of determination increases when the number of measurement steps (S100) in the calibration step is increased. That is, as the value of N in FIG. 4 increases, an accurate correlation equation between the device drive value and the energy unit can be obtained. Specifically, as shown in Figures 5 (b) and (d), the level of the device drive value can be subdivided into n levels and then the value of N can be increased through repeated measurements. Here, 'N = n ‘Number of repetitions’. As another embodiment of the present invention, the value of N may be increased by further subdividing the level of the device drive value, that is, by increasing the value of n. Here, 'N = n'.

However, in the case of Figure 5 (c), the coefficient of determination is 0.9981 when the N value is 5, and in the case of Figure 5 (d), the result can be confirmed that the coefficient of determination is 0.9997 when the N value is 10, so the measurement step In (S100), it is desirable to set the value of n to 4 or more, and considering efficiency aspects, it is more desirable to set the value of n to 4 to 6.

In this way, if the ultrasonic intensity applied to the sample is converted into an energy level through the calibration step and the experiment is conducted, the experiment can be conducted and recorded at a standardized energy level without affecting the laboratory environment.

In addition, when preprocessing actual algae, the energy delivered to the microalgae sample can be quantified and standardized, so the optimal ultrasonic intensity that can maximize the dispersion effect is determined depending on the type of microalgae or the source of the microalgae sample. By applying the optimal ultrasound intensity derived in a laboratory environment to the continuous microalgae image measurement device, it is possible to achieve the effect of stably acquiring high-resolution microalgae images.

Meanwhile, as an embodiment of the present invention, as data information on the optimal ultrasonic intensity for each laboratory is accumulated, additional characteristics of microalgae can be found.

FIG. 6 shows an actual photograph and design diagram of the flow cell 130 according to an embodiment of the present invention, and FIG. 7 schematically shows the structure of a flow path within the flow cell 130 according to an embodiment of the present invention. .

As shown in FIGS. 6 and 7, the flow cell 130 includes one or more flow paths, and each of the one or more flow paths includes an inlet flow path 131 formed in a vertical direction, and a horizontal inlet flow path 131 in the inlet flow path 131. It includes a middle flow path 132 extending in a direction, and an outlet flow path 133 extending vertically from the middle flow path 132, and the inner surface of the middle flow path 132 is designed to alleviate the problem of residual algae adhesion. It is coated with nanomaterials.

Schematically, (a) in FIG. 6 is an actual photograph of an actual flow cell 130 with different flow path widths, and (b) in FIG. 6 is a photograph of the flow cell 130 shown in (a) of FIG. 6. It shows the design diagram. Figure 7 shows the structure of a flow path located inside the flow cell 130.

Specifically, as shown in FIG. 6, one flow cell 130 includes two or more flow paths. Each flow path has a different width, so a technical feature is that a single flow cell 130 can photograph microalgae samples flowing through the flow path at different magnifications. This technical feature allows for stable measurement depending on the size of the microalgae. Image shooting is possible. In addition, as shown in Figure 7, when the microalgae sample flows into each flow path through the inlet flow path 131 formed in the vertical direction, an intermediate flow path 132 extending horizontally from the inlet flow path 131 is formed. It is operated in such a way that the microalgae sample flows through it and the microalgae sample is discharged through the outlet passage 133 extending vertically from the middle passage 132. The inlet passage 131 and the outlet passage 133 are opened in the same direction.

Referring to FIG. 7, the PDMS chip, which is the part containing the flow cell 130, uses PDMS material, and the lower part of the PDMS chip is made of slide glass. The PDMS material is a colorless polymer material similar to silicon, and has excellent moldability, so it is used in various fields such as biology, medicine, pharmacy, materials engineering, and mechanical engineering. In order to compensate for the disadvantage of poor durability of the PDMS material, it can be manufactured with a slide glass attached to the bottom of the PDMS chip. As an embodiment of the present invention, the PDMS chip can be manufactured to a height of 4.5 mm, and is preferably manufactured to have a width of 63 mm and a height of 20.5 mm, and may be larger or larger depending on the laboratory environment and the size of the flow path. It can also be manufactured in small form.

Additionally, the inner surface of the channel is coated with nanomaterial. When the microalgae sample passes through the flow cell 130 at low speed, a problem may occur where the microalgae adhere to the inner surface of the flow path and block the flow path. Therefore, as described above, the remaining algae are removed by coating the inner surface of the flow path. This alleviates the adhesion problem and allows the microalgae sample to easily pass through the flow cell 130.

Figure 8 shows microalgae images according to the height of the flow cell flow path according to an embodiment of the present invention.

Specifically, the height of the flow path refers to the height of the middle flow path 132, referring to FIG. 7. The height of the flow path is an important setting factor in taking microalgae images. As shown in FIG. 8, when the height of the flow path is higher than a certain height, the amount of light transmitted through the microalgae sample decreases, and the amount of light transmitted through the microalgae sample decreases. Difficulties may arise. However, if the height of the flow path is lower than a certain height, the microalgae sample may not flow smoothly within the flow path, which may cause the flow path to become clogged. In order to solve the problem, the flow rate within the flow cell 130 is increased. The resolution of microalgae images may be lowered.

Referring to Figure 8, as an example of the present invention, an image taken by passing a microalgae sample through a flow path with a height of 20 ㎛, 50 ㎛, and 80 ㎛ is shown, and the level that can be recognized when the height of the flow path is 20 ㎛ It can be confirmed that the focus is secured. Based on the plurality of microalgae images shown in FIG. 8, the height of the flow path is preferably set to less than 50 ㎛, and more preferably, the height of the flow path can be set to 20 ㎛ to 30 ㎛. The problem of the microalgae sample not flowing smoothly inside the flow cell due to the low height of the channel described above can be solved by adjusting the width of the channel.

Figure 9 shows an image of microalgae according to the shape of the flow path within the flow cell 130 according to an embodiment of the present invention.

As shown in FIG. 9, the intermediate passage 132 includes an inlet-side intermediate passage 132.1 having a first width; a middle side middle flow path (132.2) extending from the inlet side middle flow path (132.1) and having a second width; and an outlet-side intermediate passageway (132.3) extending from the middle-side intermediate passageway (132.2) and having a third width, and the second width may be larger than the first width and the third width.

Schematically, Figure 9 (a) shows a straight-shaped intermediate channel 132 used in conventional experiments as an embodiment of the present invention; And a microscope image when using the intermediate flow path 132; Figure 9(b) shows an embodiment of the present invention, which has a first to third width and is of the form used in the present invention. middle flow path (132); and a microscope image when the intermediate flow path 132 is used.

Specifically, the middle passage 132 is a part that photographs microalgae samples flowing within the flow cell 130 through a microscope. As shown in (a) of FIG. 9, the straight-shaped intermediate channel 132 used in conventional experiments is easy to manufacture, costs less, and has advantages in terms of maintenance, so it was widely used. However, in order to secure high-resolution microalgae images, it is preferable that the microalgae sample flows at low speed in the middle passage 132 in the flow cell 130, and in order to secure focus, it is preferable that the height of the middle passage 132 is low. do. On the other hand, in the straight-shaped middle flow path 132 shown in (a) of Figure 9, when the height of the flow path is manufactured low and the microalgae sample flows at low speed, the flow of algae does not occur smoothly, and the flow of algae does not occur smoothly. For this purpose, if the flow cell 130 is used to flow the microalgae sample at high speed or has a channel height higher than a certain level, referring to FIG. 8, difficulties arise in stably obtaining high-resolution images.

In order to solve the above-described problem, in the present invention, an intermediate flow passage 132 of the form shown in (b) of FIG. 9 was manufactured. Republic of Korea Patent No. 10-2100197 also used a flow cell 130 with a different width of the flow path of the imaged area, but in the registered patent, the process of injecting chemicals into the flow cell 130 was performed. , Compared to the present invention, the size of the flow cell 130 is large and the structure is complex. Due to these characteristics, the registered patent implements the function of capturing microalgae images at various magnifications through two or more channels with various widths in one flow cell 130, which is a technical feature of the present invention. It has its difficulties. In addition, the flow cell 130 of the registered patent has a complex structure, is expensive to manufacture, is unsuitable for obtaining high-resolution images, and is difficult to maintain. However, as shown in (b) of FIG. 9, this The flow cell 130 with a flow path used in the invention improves the above-mentioned problems, and enables efficient microalgae imaging.

Figure 10 schematically shows the structure of a microscope 200 according to an embodiment of the present invention.

As shown in Figure 10, in one embodiment of the present invention, the microscope 200 includes an illumination unit 210; A stage unit 220 disposed below the lighting unit 210 and having a through hole formed therein; and a lens unit 230 located below the stage unit 220 and capable of moving up and down, wherein the flow cell 130 is seated on the upper surface of the stage unit 220, and the lens unit 230 may be introduced into the through hole.

Specifically, as shown in Figure 10, the present invention uses an inverted microscope. Microscopes can be largely divided into upright microscopes and inverted microscopes. The upright microscope has an objective lens located above the stage 220, and the inverted microscope has an objective lens located below the stage 220. It has a positional shape.

The upright microscope has an advantage in observing slide samples because the lighting unit 210 is located at the bottom of the stage unit 220 and the objective lens is located at the top of the stage unit 220, and the inverted microscope has the lighting unit 210 It is located at the top of the stage part 220, and the objective lens is located at the bottom of the stage part 220, so it is suitable for photographing the sample contained in the culture medium.

Meanwhile, it is desirable to use an objective lens with high magnification and high resolution in the lens unit 230, and in the present invention, an objective lens with a maximum magnification of 40x is used. Through this, it is possible to take images of microalgae flowing in the flow path inside the flow cell 130 mounted on the stage unit 220, and as an embodiment of the present invention, images of microalgae with a size of less than 10 ㎛ can be captured by using the objective lens. Measurement is also possible.

The lighting unit 210 is located at the top of the stage unit 220 and irradiates light downward from the top of the inverted microscope. When the light emitted from the lighting unit 210 passes through the flow cell 130, the lens unit 230 photographs the inside of the flow cell 130 that receives the light from the lighting unit 210. In detail, the lens unit 230 can photograph the middle passage 132 located inside the flow cell 130. More specifically, with reference to FIG. 9, the second width of the middle passage 132 is The branch takes pictures of microalgae samples flowing in the middle channel (132.2).

Meanwhile, as the magnification of the objective lens used in the lens unit 230 increases, the working distance may become shorter, making it difficult to focus. That is, when the upright microscope is used in the present invention, because the flow path is located at the bottom of the PDMS chip of the flow cell 130, when a high magnification lens is used, the thickness of the flow cell 130 determines the working distance. Since the problem of overshooting occurs, the present invention uses the inverted microscope in consideration of its structural and functional features. The working distance means the vertical distance between the objective lens surface and the slide glass when in focus. Therefore, by using the inverted microscope, a higher magnification objective lens can be used than when using an upright microscope, and through this, microalgae images can be taken stably.

The stage unit 220 is located in the middle part of the inverted microscope, that is, between the lighting unit 210 and the lens unit 230, and is a part on which the flow cell 130 is placed. As shown in FIG. 10, two flow cells 130 can be placed on the stage unit 220, and a lens unit 230 and a camera capable of photographing each flow cell 130 are provided on the stage unit ( 220) Be sure to install each at the bottom. The lens unit 230 may be provided with objective lenses of different magnifications, and each objective lens is connected to the camera, and the camera is connected to a computer, so that the microalgae image captured through the camera is The images are collected using an image analysis program installed on the computer. In addition, as an embodiment of the present invention, the two flow cells 130 may include a plurality of passages with different widths inside, and different magnifications for one microalgae sample through the plurality of passages. It can be effective in obtaining images of microalgae.

The stage unit 220 includes a through hole therein, and the flow cell 130 is seated at the top of the through hole. The slide glass of the flow cell 130 is seated to face the lens unit 230, and the flow cell 130 is held by the stage unit (220) through a fixing device (not shown) included in the stage unit 220. 220). Through the fixing device, small movements caused by the transfer pump 140 can be controlled, making it possible to obtain clear images of microalgae. As an embodiment of the present invention, the middle side intermediate passage 132.2 inside the flow cell 130 may be installed to be located at the exact center of the through hole.

The lens unit 230 can install two or more objective lenses of one microscope 200, and each objective lens can use a different magnification, so that birds of different sizes can be photographed simultaneously, or the same For large-sized algae, you can photograph them broadly so that many algae cells can be seen through a single low-magnification objective lens, or you can photograph each of multiple algae cells in detail using a high-magnification objective lens. Additionally, the lens unit 230 can move up and down to focus, and the lens unit 230 can be inserted into the through hole to focus.

2. Preprocessing device for continuous microalgae image measurement device

As described above, the continuous microalgae image measurement device can be largely divided into a preprocessing stage and a measurement stage. In particular, in order to capture thousands of high-resolution microalgae images in the measurement step, a technology or device that can reduce time and increase efficiency compared to the conventional pre-processing step is required.

Meanwhile, by efficiently performing the conventional preprocessing step through the preprocessing device provided in the present invention, the effect of being able to capture microalgae images faster and more stably than before through the continuous microalgae image measurement device can be expected.

Below, the preprocessing device for the continuous microalgae image measurement device will be described in detail.

Figure 11 schematically shows the structure and fluid flow of a pretreatment device according to an embodiment of the present invention.

As shown in Figure 11, it is a preprocessing device for a continuous microalgae image measurement device, including a dilution unit 110 that supplies a dilution solvent to a microalgae sample to control the turbidity of the microalgae; It includes an ultrasonic disperser 120 that disperses the microalgae by irradiating ultrasonic waves to the microalgae sample at an ultrasonic intensity for each type of microalgae for pretreatment of the microalgae sample, and the dilution unit 110 includes a casing 1000. ); An internal partition 1100 that divides the casing 1000 into an upper space and a lower space; A diluted solvent storage tank (300) disposed in the lower space of the casing (1000) and storing a diluted solvent; A sample tank (110.1) disposed in the upper space of the casing (1000) and receiving microalgae samples from the sample storage tank (310); A first dilution tank is disposed in the upper space of the casing 1000 and receives a microalgae sample from the sample tank 110.1, and receives a dilution solvent from the dilution solvent storage tank 300 to dilute the concentration of the microalgae sample. (110.2); A second tank is disposed in the upper space of the casing 1000 and receives the microalgae sample from the first dilution tank 110.2, and receives the dilution solvent from the dilution solvent storage tank 300 to dilute the concentration of the microalgae sample. Dilution tank (110.3); and a plurality of fluids that perform fluid movement between the dilution solvent storage tank 300, the sample tank 110.1, the first dilution tank 110.2, the second dilution tank 110.3, and the ultrasonic disperser 120. It may include a pump unit (141 to 146) including a pump.

Specifically, as an embodiment of the present invention, the pretreatment device includes a casing 1000 in the form of a hexahedral frame, and the shape of the casing 1000 is not limited to the above embodiment. The inside of the casing 1000 can be divided into an upper space and a lower space through an internal partition 1100, and the upper space includes the sample tank 110.1, the first dilution tank 110.2, and the second dilution tank ( 110.3), the ultrasonic disperser 120, and the pump units 141 to 146 may be provided. Meanwhile, as shown in FIG. 11, the transfer pump control unit 400 and the ultrasonic disperser control unit 410 are disposed in the upper space of the casing 1000, but the transfer pump control unit 400 and the ultrasonic disperser control unit ( The installation area of 410) is not limited to the upper space of the casing 1000. A dilution solvent storage tank 300 capable of containing a dilution solvent and a sample storage tank 310 capable of containing the microalgae sample may be disposed in the lower space of the casing 1000. As an embodiment of the present invention, the lower space includes a wastewater storage tank (not shown) capable of containing wastewater generated during the pretreatment process; and a washing water storage tank (not shown) for cleaning the pretreatment device after the pretreatment process is completed.

The pump units 141 to 146 include a first sample pump 141, a second sample pump 142, a third sample pump 143, a fourth sample pump 144, a first dilution pump 145, and a second dilution pump 146, wherein each pump includes the sample tank 110.1, the first dilution tank 110.2, the second dilution tank 110.3, the ultrasonic disperser 120, and the sample tank 110.1. Fluid can be moved between the diluted solvent storage tank 300 and the sample storage tank 310 by connecting through a hose.

The first sample pump 141 is a transfer pump that moves the microalgae sample from the sample storage tank 310 to the sample tank 110.1, and the second sample pump 142 is a transfer pump of the sample tank 110.1. It is a transfer pump that moves the microalgae sample to the first dilution tank (110.2), and the third sample pump (143) transfers the microalgae sample from the first dilution tank (110.2) to the second dilution tank (110.3). ), and the fourth sample pump 144 is a pump that moves the microalgae sample from the second dilution tank 110.3 to the ultrasonic disperser 120.

The first dilution pump 145 is a transfer pump that moves the dilution solvent from the dilution solvent storage tank 300 to the first dilution tank 110.2, and the second dilution pump 146 is a transfer pump that moves the dilution solvent from the dilution solvent storage tank 300 to the first dilution tank 110.2. ) is a transfer pump that moves the dilution solvent to the second dilution tank (110.3).

The sample tank 110.1 can receive the microalgae sample by the first sample pump 141, and can evenly disperse the microalgae sample using a stirring motor included in the sample tank 110.1. there is. Details about the stirring motor will be described later.

In the first dilution tank (110.2), the microalgae sample supplied from the sample tank (110.1); And the dilution solvent supplied from the dilution solvent storage tank 300 is stirred to lower the turbidity of the microalgae sample. The amount of dilution solvent supplied to the first dilution tank (110.2) can be set based on the turbidity of the microalgae sample in the sample tank (110.1).

In the second dilution tank (110.3), the microalgae sample supplied from the first dilution tank (110.2); And the dilution solvent supplied from the dilution solvent storage tank 300 is stirred to lower the turbidity of the microalgae sample. The amount of dilution solvent supplied to the second dilution tank (110.3) can be set based on the turbidity of the microalgae sample in the sample tank (110.1).

When compared to the conventional one-step dilution process by diluting the microalgae sample in the sample storage tank 310 in two steps through the first dilution tank 110.2 and the second dilution tank 110.3, Even if the dilution ratio is the same, less dilution solvent can be used, which has the effect of increasing the efficiency of the dilution process. In addition, there is an advantage in that the volume of the pretreatment device can be reduced by reducing the size of the first dilution tank (110.2) and the second dilution tank (110.3).

The ultrasonic disperser 120 can receive microalgae samples from the second dilution tank 110.3 by the fourth sample pump 144, and the microalgae samples are supplied using ultrasonic waves within the ultrasonic disperser 120. Flocs in the microalgae sample can be removed through the technique. A detailed description of the ultrasonic use technique is described in detail in the description of FIGS. 2 and 3.

The first to fourth sample pumps (141 to 144) and the first to second dilution pumps (145 to 146) are operated by commands from the transfer pump control unit 400, and the ultrasonic disperser 120 uses the ultrasonic waves. It is operated by commands from the disperser control unit 410. In the ultrasonic disperser control unit, referring to FIGS. 4 and 5, the ultrasonic disperser can be operated by setting the device drive value, and by performing the calibration step, the energy unit corresponding to the device drive value can be obtained. .

Figure 12 schematically shows the structure of the first dilution tank 110.2 and the second dilution tank 110.3 according to an embodiment of the present invention.

As shown in Figure 12, the first dilution tank (110.2) and the second dilution tank (110.3) each include a dilution chamber (111) in which the microalgae sample and the dilution solvent are mixed; A turbidity meter attached to the dilution chamber 111 and measuring the turbidity of the microalgae sample contained in the dilution chamber 111; A stirring motor 114 installed at the lower end of the dilution chamber 111; A first magnet 115 located between the stirring motor 114 and the dilution chamber 111, connected to the stirring motor 114, and rotating when the stirring motor 114 is operated; It may include a second magnet 116 that is located at the inner lower end of the dilution chamber 111 and rotates in response to the first magnet 115 when the first magnet 115 rotates.

Schematically, Figure 12 (a) briefly shows the structure of the first to second dilution tanks (110.2 to 110.3). Figure 12(b) briefly shows the upper surface of the dilution chamber 111. Figure 12(c) briefly shows a cross section of the first to second dilution tanks 110.2 to 110.3.

Specifically, as shown in (a) of Figure 12, in the dilution chamber 111, the microalgae sample and the dilution solvent are stirred to dilute the microalgae sample. A light emitting sensor 112 and a light receiving sensor 113 are attached to the outer side of the dilution chamber 111 at positions facing each other, and the light emitting sensor 112 and the light receiving sensor 113 are included in the turbidity meter. Additionally, the stirring motor 114 and the first magnet 115 are disposed at the bottom of the dilution chamber 111. The stirring motor 114 and the first magnet 115 are physically connected, and when the stirring motor 114 is operated, the first magnet 115 rotates around the motor shaft of the stirring motor 114. . Meanwhile, the first magnet 115 and the dilution chamber 111 are not in contact with each other.

As shown in (b) of FIG. 12, the upper surface of the dilution chamber 111 includes three holes. The three holes each include a sample inlet (111.1) through which microalgae samples flow into the dilution chamber (111); A dilution solvent inlet (111.2) through which the dilution solvent flows into the dilution chamber (111); And a sample outlet (111.3) through which the diluted microalgae sample is discharged from the dilution chamber (111).

The sample inlet 111.1 of the first dilution tank 110.2 is connected to the second sample pump 142 to receive a microalgae sample from the sample tank 110.1, and the first dilution tank 110.2 ), the dilution solvent inlet (111.2) is connected to the first dilution pump (145) and can receive the dilution solvent from the dilution solvent storage tank (300). In addition, the sample outlet (111.3) of the first dilution tank (110.2) is connected to the third sample pump to dilute the microalgae sample in the dilution chamber (111) of the first dilution tank (110.2) into the second dilution. It can be supplied as crude (110.3).

The sample inlet 111.1 of the second dilution tank 110.3 is connected to the third sample pump 143 to receive a microalgae sample from the first dilution tank 110.2, and the second dilution tank 110.3 is connected to the sample inlet 111.1. The diluted solvent inlet (111.2) at (110.3) is connected to the second diluted pump (146) and can receive the diluted solvent from the diluted solvent storage tank (300). In addition, the sample outlet 111.3 of the second dilution tank 110.3 is connected to the fourth sample pump to distribute the microalgae sample in the dilution chamber 111 of the second dilution tank 110.3 to the ultrasonic disperser ( 120).

As shown in (c) of FIG. 12, a second magnet 116 that can rotate corresponding to the first magnet 115 is provided inside the dilution chamber 111, an embodiment of the present invention. As such, the first magnet 115 and the second magnet 116 may have a disc-shaped shape. The first magnet 115 exists outside the dilution chamber 111 and does not contact the lower surface of the dilution chamber 111. Additionally, the second magnet 116 exists inside the dilution chamber 111 and does not contact the lower surface of the dilution chamber 111.

The first magnet 115 is connected to the stirring motor 114, and when the stirring motor 114 operates, the first magnet 115 also rotates according to the speed of the motor, and the second magnet 116 ) rotates corresponding to the rotational movement of the first magnet 115. The second magnet 116 rotates within the dilution chamber 111 so that the microalgae sample within the dilution chamber 111 and the dilution solvent are well mixed.

As another embodiment of the present invention, the stirring efficiency in the dilution chamber 111 can be increased by modifying the shape of the second magnet 116. For example, by changing the shape of the upper end of the second magnet 116, when the second magnet 116 rotates within the dilution chamber 111, a vortex is generated within the dilution chamber 111. It can be created. When creating a vortex within the dilution chamber 111, the stirring efficiency within the dilution chamber 111 can be increased, and the microalgae sample in the dilution chamber 111 sinks and accumulates at the bottom of the dilution chamber 111. phenomenon can be reduced. As a method of modifying the shape of the second magnet 116, there is a method of modifying the shape of the second magnet 116 itself, or attaching an attachment such as a propeller shape to the upper end of the second magnet 116. .

Meanwhile, as described above, the sample tank 110.1, similar to the structure of the first to second dilution tanks 110.2 to 110.3 shown in FIG. 12, includes a sample chamber (not shown) containing a sample; Turbidimeter including a light emitting sensor and a light receiving sensor; A stirring motor 114 disposed at the bottom of the sample chamber; A first magnet 115 connected to the stirring motor; and a second magnet 116 located inside the sample chamber and capable of rotating in response to the rotational movement of the first magnet.

However, the sample tank (110.1), unlike the first to second dilution tanks (110.2 to 110.3), has only a sample tank inlet (not shown) and a sample tank outlet (not shown) on the upper surface of the sample chamber. There is no inlet through which the diluting solvent present in the first to second diluting tanks (110.2 to 110.3) flows.

The sample tank inlet of the sample tank (110.1) is connected to the first sample pump 141 to receive microalgae samples from the sample storage tank 310, and the sample tank outlet of the sample tank (110.1) is connected to the first sample pump 141. , It is connected to the second sample pump 142 and can supply the microalgae sample in the sample chamber to the first dilution tank (110.2).

As an embodiment of the present invention, by modifying the shape of the second magnet 116 as described above in the description of (c) of FIG. 12, a vortex can be generated in the sample chamber, and the vortex can be generated. By doing so, the dispersion effect of the microalgae sample introduced into the sample chamber can be increased.

Figure 13 schematically shows the dilution process performed in the dilution unit 110 according to an embodiment of the present invention.

Schematically, the solid arrow shown in FIG. 13 indicates that the transfer pump control unit 400 receives each turbidity information from the sample tank (110.1), the first dilution tank (110.2), and the second dilution tank (110.3). It shows a process for receiving input and controlling the first dilution pump 145 and the second dilution pump 146, and the dotted arrow shown in FIG. 13 indicates the moving direction of the dilution solvent.

Specifically, the sample tank (110.1) includes a first turbidity meter capable of measuring the turbidity of the sample tank (110.1), and the first dilution tank (110.2) includes a turbidity meter of the first dilution tank (110.2). It includes a second turbidity meter capable of measuring , and the second dilution tank 110.3 includes a third turbidity meter capable of measuring the turbidity of the second dilution tank 110.3.

The first to third turbidity meters each include first to third light emitting sensors (not shown); and first to third light receiving sensors (113.1 to 113.3), wherein the first to third light receiving sensors (113.1 to 113.3) each include the sample tank (110.1) and the first dilution tank (110.2). ), and after measuring the turbidity of the second dilution tank (110.3), information on the measured turbidity can be transmitted to the transfer pump control unit 400.

The transfer pump control unit 400 controls the operation of the first dilution pump 145 and the second dilution pump 146 based on the turbidity of the sample tank 110.1 received from the first light receiving sensor 113.1. Control. In one embodiment of the present invention, the transfer pump control unit 400 detects turbidity from the second light-receiving sensor 113.2 and the third light-receiving sensor 113.3 along with the turbidity information of the sample tank 110.1, as necessary. By receiving information, the operation of the first dilution pump 145 and the second dilution pump 146 can be controlled.

The first dilution pump 145 and the second dilution pump 146 pump the dilution solvent from the dilution solvent storage tank 300 based on the command received from the transfer pump control unit 400, and the first dilution pump 145 The dilution pump 145 supplies the dilution solvent to the first dilution tank 110.2, and the second dilution pump 146 supplies the dilution solvent to the second dilution tank 110.3.

Figure 14 schematically shows the steps of the dilution process performed in the dilution unit 110 according to an embodiment of the present invention.

As shown in FIG. 14, the sample tank 110.1 includes a first turbidity meter capable of measuring turbidity, and the first dilution tank 110.2 includes a second turbidity meter capable of measuring turbidity, The second dilution tank (110.3) includes a third turbidity meter capable of measuring turbidity, and the pump units (141 to 146) dilute the dilution solvent of the dilution solvent storage tank (300) into the first dilution tank (110.2). It includes a first dilution pump 145 that moves the dilution solvent from the dilution solvent storage tank 300 to the second dilution tank 110.3, and the pretreatment device includes the above. A transfer pump control unit 400 that controls the operation of the first dilution pump 145 and the second dilution pump 146 based on the turbidity information sensed by the first turbidity meter, the second turbidity meter, and the third turbidity meter. It further includes, wherein the transfer pump control unit 400 determines the first target turbidity of the first dilution tank (110.2) and the first target turbidity of the second dilution tank (110.3) based on the first turbidity information sensed by the first turbidity meter. 2Target turbidity setting step to set target turbidity; A first dilution pump control step of controlling the first dilution pump 145 so that the second turbidity information sensed by the second turbidity meter approaches the first target turbidity; and a second dilution pump control step of controlling the second dilution pump 146 so that the third turbidity information sensed by the third turbidity meter approaches the second target turbidity.

Specifically, Figure 14 shows one embodiment of Figure 13. One embodiment of the present invention is described as follows.

When the microalgae sample is injected into the sample storage tank 310 of the pretreatment device, the microalgae sample in the sample storage tank 310 is injected into the sample tank 110.1 by the first sample pump 141.

The first turbidity meter can measure the turbidity of the microalgae sample injected into the sample tank 110.1 (S200). The first turbidity meter irradiates light through the first light emitting sensor 112 to the microalgae sample injected into the sample tank 110.1. When the light irradiated through the first light-emitting sensor 112 passes through the microalgae sample and is detected by the first light-receiving sensor 113, the first light-receiving sensor 113 analyzes the detected light and The turbidity of algae samples can be measured. The second and third turbidity meters can also measure the turbidity of a microalgae sample in the same way as the first turbidity meter as described above.

The transfer pump control unit 400 receives the first turbidity information measured in the sample tank 110.1 from the first light receiving sensor 113. The transfer pump control unit 400 performs a target turbidity setting step (S210) of setting the first target turbidity and the second target turbidity based on the first turbidity information.

Meanwhile, the microalgae sample whose turbidity has been measured in the sample tank (110.1) is supplied to the first dilution tank (110.2) by the second sample pump (142). The turbidity of the microalgae sample supplied to the first dilution tank (110.2) is measured by the second turbidity meter (S220). The transfer pump control unit 400 receives the second turbidity information calculated by the second turbidity meter from the second light receiving sensor 113, and receives the second turbidity information until the second turbidity information approaches the first target turbidity. The first dilution pump control step (S230) of operating the first dilution pump (145) is performed.

When the second turbidity information reaches the first target turbidity, the transfer pump control unit 400 stops operation of the first dilution pump 145 and operates the third sample pump 143 to The microalgae sample diluted in the dilution tank (110.2) is supplied to the second dilution tank (110.3).

The turbidity of the microalgae sample supplied to the second dilution tank (110.3) is measured by the third turbidity meter (S240). The transfer pump control unit 400 receives the third turbidity information calculated by the third turbidity meter from the third light receiving sensor 113, and receives the third turbidity information until the third turbidity information approaches the second target turbidity. 2The second dilution pump control step (S250) of operating the dilution pump 146 is performed.

When the third turbidity information reaches the second target turbidity, the transfer pump control unit 400 stops operation of the second dilution pump 146 and operates the fourth sample pump 144 to The microalgae sample diluted in the dilution tank (110.3) is supplied to the ultrasonic disperser (120).

Figure 15 schematically shows the steps of the dilution process performed in the dilution unit 110 according to another embodiment of the present invention.

As shown in FIG. 15, the sample tank (110.1) includes a first turbidity meter capable of measuring turbidity, and the pump unit transfers the dilution solvent of the dilution solvent storage tank (300) into the first dilution tank (110.2). It includes a first dilution pump 145 that moves the dilution solvent from the dilution solvent storage tank 300 to the second dilution tank 110.3, and the pretreatment device includes the above. Based on the turbidity information sensed by the first turbidity meter, it further includes a transfer pump control unit 400 that controls the operation of the first dilution pump 145 and the second dilution pump 146, and the transfer pump control unit ( 400) is a target turbidity setting step of setting the first target turbidity of the first dilution tank (110.2) and the second target turbidity of the second dilution tank (110.3) based on the first turbidity information sensed by the first turbidity meter. ; A first dilution pump control step of controlling the first dilution pump 145 based on the first target turbidity; and a second dilution pump control step of controlling the second dilution pump 146 based on the second target turbidity.

Specifically, Figure 15 shows another embodiment of Figure 13. Another embodiment of the present invention is described as follows.

When the microalgae sample is injected into the sample storage tank 310 of the pretreatment device, the microalgae sample in the sample storage tank 310 is injected into the sample tank 110.1 by the first sample pump 141. When the microalgae sample flows into the sample tank (110.1), the first turbidity meter measures the turbidity of the microalgae sample (S300) to calculate first turbidity information, and the first light receiving sensor 113 measures the turbidity of the microalgae sample (S300). 1Turbidity information is transmitted to the transfer pump control unit 400.

The transfer pump control unit 400 performs a target turbidity setting step of setting the first target turbidity and the second target turbidity (S310) based on the first turbidity information. In the target setting step, the amount of dilution solvent supplied to the first dilution tank (110.2) to achieve the first target turbidity in consideration of the turbidity and amount of the microalgae sample in the sample tank (110.1); And the amount of dilution solvent supplied to the second dilution tank (110.3) to achieve the second target turbidity; can be calculated. After calculating the amount of the dilution solvent, the transfer pump control unit 400 operates the second sample pump 142 to move the microalgae sample from the sample tank 110.1 to the first dilution tank 110.2. I order it.

When the microalgae sample is supplied to the first dilution tank (110.2), the transfer pump control unit 400 controls the first dilution pump 145 based on the first target turbidity. Step (S320) is performed, and according to this, the first dilution pump 145 supplies the dilution solvent to the first dilution tank 110.2 by the calculated amount of the dilution solvent supplied to the first dilution tank 110.2. supply. The first dilution pump 145 supplies the dilution solvent to the first dilution tank 110.2 based on the command. At this time, the transfer pump control unit 400 waits for a preset time so that the microalgae sample can be well diluted, and then operates the third sample pump 143 to dilute the diluted microalgae sample to the second dilution. Move to Joe (110.3).

When the diluted microalgae sample is supplied to the second dilution tank (110.3), the transfer pump control unit 400 controls the second dilution pump 146 based on the second target turbidity. A pump control step (S330) is performed, and according to this, the second dilution pump 146 dilutes the dilution into the second dilution tank (110.3) by the calculated amount of dilution solvent supplied to the second dilution tank (110.3). Supply solvent. At this time, the transfer pump control unit 400 waits for a preset time so that the microalgae sample can be well diluted, and then operates the fourth sample pump 144 to transfer the diluted microalgae sample to the ultrasonic disperser ( 120).

The preset time should be set to a time during which the dilution solvent and the microalgae sample can be sufficiently mixed by the second magnet 116 in the dilution chamber 111, and may vary depending on the type of the microalgae sample. there is.

Referring to the description of FIGS. 14 and 15, the differences between the two embodiments are summarized. In the embodiment of the description of FIG. 14, the first turbidity information measured by the first turbidity meter in the transfer pump control unit 400 Based on this, the first target turbidity and the second target turbidity are set, and the turbidity of the first dilution tank (110.2) and the second dilution tank (110.3) is set to the first target turbidity and the second target turbidity, respectively. Supply diluting solvent until it is reached. That is, in the embodiment described in FIG. 14, the sample tank 110.1, the first dilution tank 110.2, and the second dilution tank 110.3 all require turbidity meters.

On the other hand, in the embodiment in the description of FIG. 15, after the transfer pump control unit 400 sets the first target turbidity and the second target turbidity based on the first turbidity information measured by the first turbidity meter, the Without receiving turbidity information from the first dilution tank (110.2) and the second dilution tank (110.3), the amount of dilution solvent required for the first target turbidity and the second target turbidity is added to the first dilution tank (110.3). 110.2) and the second dilution tank 110.3, the turbidity meter is needed only for the sample tank 110.1, and the turbidity meter is not needed for the first to second dilution tanks 110.2 to 110.3.

That is, in the embodiment mentioned in the description of FIG. 14, the transfer pump control unit 400 receives turbidity information in real time from each of the first dilution tank 110.2 and the second dilution tank 110.3. It can have the effect of performing a dilution process that can accurately reach the target turbidity, and in the embodiment mentioned in the description of FIG. 15, the process performed in the dilution process can be simplified, so the dilution process can be performed. It can have the effect of speeding up the speed.

Figure 16 schematically shows the structure of an ultrasonic disperser 120 according to an embodiment of the present invention.

As shown in FIG. 16, the ultrasonic disperser 120 includes an ultrasonic dispersion chamber 121 that receives microalgae samples from the second dilution tank 110.3 by the pump units 141 to 146; An ultrasonic irradiation device (122) that irradiates ultrasonic waves to the microalgae sample supplied to the ultrasonic dispersion chamber (121); and a temperature sensor 123 that measures the temperature of the microalgae sample supplied to the ultrasonic dispersion chamber 121. A calibration step of deriving a device drive value for an energy unit; and a device driving step of driving the ultrasonic disperser 120 using the device driving value derived in the calibration step. In addition, in the device driving step, flocs, which are aggregates of microalgae, can be dispersed by irradiating ultrasonic waves to the microalgae sample for a preset time at ultrasonic intensity according to the device driving value derived in the calibration step.

Specifically, the ultrasonic disperser 120 can remove flocs present in the microalgae sample by irradiating ultrasonic waves to the microalgae sample supplied from the second dilution tank 110.3. Meanwhile, in the ultrasonic disperser 120, the calibration step can be performed before performing the preprocessing step for the microalgae sample. The calibration step is performed as shown in FIG. 4.

The microalgae sample is supplied from the second dilution tank (110.3) to the ultrasonic dispersion chamber (121). When the microalgae sample is supplied to the ultrasonic dispersion chamber 121, referring to FIG. 11, the ultrasonic disperser control unit 410 drives the ultrasonic irradiation device 122 with the first device driving value. After driving the ultrasonic irradiation device 122 with the first device drive value, the temperature sensor 123 installed inside the ultrasonic dispersion chamber 121 calculates first change information corresponding to the first device drive value. and transmits it to the ultrasonic disperser control unit 410. The ultrasonic disperser control unit 410 may derive the first energy unit for the first device driving value based on the first change information.

After completing the first measurement step (S100.1) for calculating the first change information and the first derivation step (S110.1) for deriving the first energy unit, the ultrasonic disperser control unit 410 is connected to the second device. The ultrasonic device is driven using the driving value. After driving the ultrasonic irradiation device 122 with the second device drive value, the temperature sensor 123 calculates second change information corresponding to the second device drive value and sends it to the ultrasonic disperser control unit 410. Deliver. The ultrasonic disperser control unit 410 may derive the second energy unit for the second device driving value based on the second change information. Meanwhile, in order to precisely measure temperature changes, the ultrasonic dispersion chamber 121 is preferably located inside a space surrounded by an insulating material.

The ultrasonic disperser control unit 410 determines the device drive value and the energy unit based on regression information including the first device drive value, the second device drive value, the first energy unit, and the second energy unit. The calibration step that can derive relationship information can be performed. As described above in the description of FIG. 5, the regression information may correspond to the relationship between the device drive value and the energy unit.

Based on the energy unit information obtained through the calibration step, the device drive value in the laboratory that can achieve the greatest dispersion effect for the microalgae sample can be calculated. In addition, as an embodiment of the present invention, since the energy unit information is standardized information, the device drive value and ultrasonic irradiation time with the highest dispersion effect for the corresponding algae can be calculated through cross-verification between laboratories.

Using the device drive value calculated through the calibration step, ultrasonic waves are irradiated to the microalgae sample supplied to the ultrasonic dispersion chamber 121 using the device drive value. Referring to FIG. 1, the ultrasonicated microalgae sample passes through the flow cell 130, and an image of the microalgae sample can be measured.

According to one embodiment of the present invention, the turbidity of the microalgae sample in the dilution chamber 111 can be measured through a turbidity meter attached to the dilution chamber 111, and the dilution process can be performed based on this, thereby reducing the cost required for the dilution process. It can be effective in reducing time and performing dilution exactly as needed.

According to one embodiment of the present invention, by performing a double dilution process, a high-multiplier dilution process can be performed with a small amount of dilution solvent, thereby reducing the space and cost problems of the conventional pretreatment device. .

According to one embodiment of the present invention, by using a modular pretreatment device, it is easy to replace the microalgae sample and dilution solvent, and cleaning and replacing the dilution chamber 111 is simple, which has the effect of reducing maintenance costs. It can be performed.

According to one embodiment of the present invention, a plurality of pumps provided in the pretreatment device can be automatically operated by the transfer pump control unit 400, so that the microalgae sample is simply injected into the microalgae storage tank. The pretreatment process can be performed on the sample, thereby increasing the efficiency of the pretreatment process, and because it is performed automatically, it can have the effect of reducing contamination that may occur with the sample.

According to one embodiment of the present invention, the pretreatment method using the ultrasonic disperser 120 can increase the single cell recovery rate and solve the efficiency and time problems compared to the conventional pretreatment method using chemicals and heat treatment. .

According to an embodiment of the present invention, the intensity of the ultrasonic disperser can be standardized through a calibration step, which has the effect of implementing cross-validation between laboratories.

The present invention has been described with reference to the embodiments shown in the drawings, but these are merely illustrative, and those skilled in the art will understand that various modifications and other equivalent embodiments are possible therefrom. For example, the described techniques are performed in a different order than the described method, and/or components of the described device, structure, device, circuit, etc. are combined or combined in a different form than the described method, or other components are used. Alternatively, appropriate results may be achieved even if substituted or substituted by an equivalent. Therefore, the true technical protection scope of the present invention should be determined by the technical spirit of the attached registration claims.

Claims (7)

delete As a preprocessing device for a continuous measurement device for microalgae images,
A dilution unit that supplies a dilution solvent to the microalgae sample to control turbidity of the microalgae;
An ultrasonic disperser for dispersing the microalgae by irradiating ultrasonic waves to the microalgae sample at an ultrasonic intensity for each type of microalgae for pretreatment of the microalgae sample,
The dilution part,
casing;
an internal partition dividing the casing into an upper space and a lower space;
a diluted solvent storage tank disposed in the lower space of the casing and storing a diluted solvent;
a sample tank disposed in the upper space of the casing and receiving microalgae samples from a sample storage tank;
a first diluting tank disposed in the upper space of the casing and receiving a microalgae sample from the sample tank and receiving a dilution solvent from the dilution solvent storage tank to dilute the concentration of the microalgae sample;
a second dilution tank disposed in the upper space of the casing and receiving a microalgae sample from the first dilution tank and receiving a dilution solvent from the dilution solvent storage tank to dilute the concentration of the microalgae sample; and
It includes a pump unit including a plurality of pumps that perform fluid movement between the dilution solvent storage tank, the sample tank, the first dilution tank, the second dilution tank, and the ultrasonic disperser,
The ultrasonic disperser,
A calibration step of deriving an energy unit corresponding to the input tidal current or a device drive value for the input energy unit; and
Performing a device driving step of driving the ultrasonic disperser with the device driving value derived from the calibration step,
The device operation step is,
According to the device drive value derived in the calibration step, ultrasonic waves are irradiated to the microalgae sample for a preset time at ultrasonic intensity to disperse flocs, which are microalgae aggregates,
The first and second dilution tanks are, respectively,
A dilution chamber in which the microalgae sample and the dilution solvent are mixed;
A turbidity meter attached to the dilution chamber and measuring turbidity of the microalgae sample contained in the dilution chamber;
A stirring motor installed at the lower end of the dilution chamber;
a first magnet located between the stirring motor and the dilution chamber, connected to the stirring motor, and rotating when the stirring motor is activated;
A preprocessing device for a continuous measurement device for microalgae images, including a second magnet located at the lower end inside the dilution chamber and rotating in response to the first magnet when the first magnet rotates.
As a preprocessing device for a continuous measurement device for microalgae images,
A dilution unit that supplies a dilution solvent to the microalgae sample to control turbidity of the microalgae;
An ultrasonic disperser for dispersing the microalgae by irradiating ultrasonic waves to the microalgae sample at an ultrasonic intensity for each type of microalgae for pretreatment of the microalgae sample,
The dilution part,
casing;
an internal partition dividing the casing into an upper space and a lower space;
a diluted solvent storage tank disposed in the lower space of the casing and storing a diluted solvent;
a sample tank disposed in the upper space of the casing and receiving microalgae samples from a sample storage tank;
a first diluting tank disposed in the upper space of the casing and receiving a microalgae sample from the sample tank and receiving a dilution solvent from the dilution solvent storage tank to dilute the concentration of the microalgae sample;
a second dilution tank disposed in the upper space of the casing and receiving a microalgae sample from the first dilution tank and receiving a dilution solvent from the dilution solvent storage tank to dilute the concentration of the microalgae sample; and
It includes a pump unit including a plurality of pumps that perform fluid movement between the dilution solvent storage tank, the sample tank, the first dilution tank, the second dilution tank, and the ultrasonic disperser,
The ultrasonic disperser,
A calibration step of deriving an energy unit corresponding to the input tidal current or a device drive value for the input energy unit; and
Performing a device driving step of driving the ultrasonic disperser with the device driving value derived from the calibration step,
The ultrasonic disperser,
an ultrasonic dispersion chamber that receives microalgae samples from the second dilution tank by the pump unit;
An ultrasonic irradiation device that irradiates ultrasonic waves to the microalgae sample supplied to the ultrasonic dispersion chamber; and
A preprocessing device for a continuous microalgae image measurement device comprising a temperature sensor that measures the temperature of the microalgae sample supplied to the ultrasonic dispersion chamber.
As a preprocessing device for a continuous measurement device for microalgae images,
A dilution unit that supplies a dilution solvent to the microalgae sample to control turbidity of the microalgae;
An ultrasonic disperser for dispersing the microalgae by irradiating ultrasonic waves to the microalgae sample at an ultrasonic intensity for each type of microalgae for pretreatment of the microalgae sample,
The dilution part,
casing;
an internal partition dividing the casing into an upper space and a lower space;
a diluted solvent storage tank disposed in the lower space of the casing and storing a diluted solvent;
a sample tank disposed in the upper space of the casing and receiving microalgae samples from a sample storage tank;
a first diluting tank disposed in the upper space of the casing and receiving a microalgae sample from the sample tank and receiving a dilution solvent from the dilution solvent storage tank to dilute the concentration of the microalgae sample;
a second dilution tank disposed in the upper space of the casing and receiving a microalgae sample from the first dilution tank and receiving a dilution solvent from the dilution solvent storage tank to dilute the concentration of the microalgae sample; and
It includes a pump unit including a plurality of pumps that perform fluid movement between the dilution solvent storage tank, the sample tank, the first dilution tank, the second dilution tank, and the ultrasonic disperser,
The ultrasonic disperser,
A calibration step of deriving an energy unit corresponding to the input tidal current or a device drive value for the input energy unit; and
Performing a device driving step of driving the ultrasonic disperser with the device driving value derived from the calibration step,
The sample tank includes a first turbidity meter capable of measuring turbidity,
The first dilution tank includes a second turbidity meter capable of measuring turbidity,
The second dilution tank includes a third turbidity meter capable of measuring turbidity,
The pump unit includes a first dilution pump for moving the dilution solvent in the dilution solvent storage tank to the first dilution tank, and a second dilution pump for moving the dilution solvent in the dilution solvent storage tank to the second dilution tank,
The pretreatment device further includes a transfer pump control unit that controls the operation of the first dilution pump and the second dilution pump based on turbidity information sensed by the first turbidity meter, the second turbidity meter, and the third turbidity meter, ,
The transfer pump control unit,
A target turbidity setting step of setting a first target turbidity of the first dilution tank and a second target turbidity of the second dilution tank based on the first turbidity information sensed by the first turbidity meter;
A first dilution pump control step of controlling the first dilution pump so that the second turbidity information sensed by the second turbidity meter approaches the first target turbidity; and
A preprocessing device for a continuous microalgae image measurement device that performs a second dilution pump control step of controlling the second dilution pump so that the third turbidity information sensed by the third turbidity meter approaches the second target turbidity.
As a preprocessing device for a continuous measurement device for microalgae images,
A dilution unit that supplies a dilution solvent to the microalgae sample to control turbidity of the microalgae;
An ultrasonic disperser for dispersing the microalgae by irradiating ultrasonic waves to the microalgae sample at an ultrasonic intensity for each type of microalgae for pretreatment of the microalgae sample,
The dilution part,
casing;
an internal partition dividing the casing into an upper space and a lower space;
a diluted solvent storage tank disposed in the lower space of the casing and storing a diluted solvent;
a sample tank disposed in the upper space of the casing and receiving microalgae samples from a sample storage tank;
a first diluting tank disposed in the upper space of the casing and receiving a microalgae sample from the sample tank and receiving a dilution solvent from the dilution solvent storage tank to dilute the concentration of the microalgae sample;
a second dilution tank disposed in the upper space of the casing and receiving a microalgae sample from the first dilution tank and receiving a dilution solvent from the dilution solvent storage tank to dilute the concentration of the microalgae sample; and
It includes a pump unit including a plurality of pumps that perform fluid movement between the dilution solvent storage tank, the sample tank, the first dilution tank, the second dilution tank, and the ultrasonic disperser,
The ultrasonic disperser,
A calibration step of deriving an energy unit corresponding to the input tidal current or a device drive value for the input energy unit; and
Performing a device driving step of driving the ultrasonic disperser with the device driving value derived from the calibration step,
The sample tank includes a first turbidity meter capable of measuring turbidity,
The pump unit includes a first dilution pump for moving the dilution solvent in the dilution solvent storage tank to the first dilution tank, and a second dilution pump for moving the dilution solvent in the dilution solvent storage tank to the second dilution tank,
The pretreatment device further includes a transfer pump control unit that controls the operation of the first dilution pump and the second dilution pump based on the turbidity information sensed by the first turbidity meter,
The transfer pump control unit,
A target turbidity setting step of setting a first target turbidity of the first dilution tank and a second target turbidity of the second dilution tank based on the first turbidity information sensed by the first turbidity meter;
A first dilution pump control step of controlling the first dilution pump based on the first target turbidity; and
A preprocessing device for a continuous microalgae image measurement device that performs a second dilution pump control step of controlling the second dilution pump based on the second target turbidity. delete delete

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