KR102100197B1 - Continuous monitoring device of micro algae using flow cell - Google Patents

Abstract

The present invention is to obtain a high-resolution sample image by taking a sample taken from a river, a lake, etc., and flowing along a micro-channel, and to analyze a microalgae continuously by determining the type and amount by analyzing the shape of the micro-type shown in the sample image. It is about.
The microalgae continuous monitoring apparatus using a flow cell according to the present invention includes a flow cell in which a microchannel flows through a sample, an optical module device for photographing a sample flowing along the microchannel to generate a sample image, and various microalgae cultured purely It is characterized by comprising a reading device that detects one or more microalgae by comparing the image data with the database and the sample image stored in a large number of image data of the cultured species and the sample data.

Description

Continuous monitoring device of micro algae using flow cell

The present invention relates to a device for continuously monitoring microalgae contained in a sample, and more specifically, a sample taken from a river, a lake, etc. and flowing along a microchannel to secure a high-resolution sample image, and the sample image It relates to a continuous monitoring device for microalgae to determine the type and amount by analyzing the shape of the micro-type shown in.

In general, plants that do photosynthesis using chlorophyll among lower plants that are not systematically differentiated into roots, stems, leaves, etc. are called algae, and these algae are large algae such as seaweed and kelp that inhabit only at certain locations and 50㎛. It has the following size and can be divided into microalgae that live freely and live in water.

In particular, microalgae have various advantages such as excellent oil productivity, can produce various chemicals, and can purify soil and water contaminated with heavy metals, but adversely affect the environment, such as causing green algae and red tide. There is also a disadvantage that can affect the, it is necessary to accurately determine the type or amount of microalgae contained in samples taken from rivers, lakes, culture tanks, etc. to accurately understand the culture or diffusion status of microalgae.

In particular, it is very important to measure the population quickly and accurately, since harmful blue-green algae such as microcistis secrete toxic substances and odor-causing substances, and the concentration of the population becomes a standard for issuing algae warnings.

Conventional inventions related to the measurement of microalgae contained in the water include “wireless monitoring system of microalgae underwater” in Korean Patent Registration No. 10-0917030 and “portable microalgae detection device” in Korean Patent Registration No. 10-1683379. , Republic of Korea Patent Publication No. 10-2018-0003757 "algae monitoring system" and Republic of Korea Patent Publication No. 10-2018-0084076 "microalgae monitoring device and microalgae monitoring method" has been proposed and published.

In the Republic of Korea Patent Publication No. 10-0917030 "wireless monitoring system of microalgae underwater" the body is formed with a curved portion, a pressure sensor for detecting the pressure applied to the body, and a plurality of LEDs for irradiating light to the underwater microalgae And, it is configured to include an analysis unit for analyzing and monitoring the spectral distribution according to the depth of the underwater microalgae by analyzing the pressure and fluorescence, and an optoelectronic amplifier for detecting and amplifying the fluorescence generated by the underwater microalgae, The invention of a system capable of wirelessly monitoring the distribution and photosynthetic properties of aquatic microalgae according to depth has been proposed, and in the “Portable Microalgae Detection Device” of Korean Patent Registration No. 10-1683379, pre-processed DNA and fluorescent materials Phosphorus is connected to the first QD (Quantum dot), the first probe that binds to the first site of the DNA, and the second QD is connected to the first DNA A sample housing in which a sample solution including a second probe that binds to two sites is located, a light source device that irradiates light to the sample solution, and a plurality of photodiodes and sample solutions that receive photons emitted from the sample solution It has been proposed to include an analysis circuit for correcting or quantitatively analyzing microalgal DNA, and an invention has been proposed for an apparatus capable of effectively analyzing microalgal DNA with a small amount of sample solution.

In addition, in the “Algae Monitoring System” of Korean Patent Publication No. 10-2018-0003757, the measurement unit and the measurement unit that detect fluorescence by irradiating light after ultrasonic pretreatment with water containing algae are controlled, and the measurement unit is controlled. An invention has been proposed for a system configured to include a control unit for receiving data and a display unit for displaying the received data, and capable of wirelessly monitoring the distribution and photosynthetic properties of underwater microalgae according to the depth of the water. In the publication 10-2018-0084076, “A device for monitoring microalgae and a method for monitoring microalgae” includes a flow cell through which a fluid flows, an excitation light source for irradiating excitation light to the flow cell, and a fine excitation light. Fluorescence detector that detects autofluorescence from each algae lipid and scattered light from each microalgae An invention has been proposed that is configured to include a scattering light detector to detect, and is capable of observing lipids contained in microalgae in a simple, rapid and detailed manner.

However, the prior art as described above is an invention for grasping the distribution status of the microalgae contained in the water, and there is a problem that the type of the microalgae distributed in the water cannot be clearly determined. Therefore, there is a problem because the microalgae distribution state, DNA analysis, etc. are performed.

In addition, a known method in which a measuring instrument reads the type and amount of microalgae using a microscope requires physicochemical treatment time such as diluting and concentrating a sample, and classifying and counting the shape of microalgae by highly skilled experts. Since the process is necessary, it is difficult to immediately derive the results, and there is a problem in that the cost is increased as the number of readings increases. Therefore, the present invention relates to an apparatus for quickly reading the type and amount of microalgae by solving the above problems. This is required.

Republic of Korea Registered Patent Publication No. 10-0917030 (2009. 09. 04) Republic of Korea Registered Patent Publication No. 10-1683379 (2016. 11. 30) Republic of Korea Patent Publication No. 10-2018-0003757 (2018. 01. 10) Republic of Korea Patent Publication No. 10-2018-0084076 (2018. 07. 24)

Microalgae continuous monitoring apparatus using a flow cell according to the present invention is a technique proposed to solve the problems of the conventional invention as described above,

Physicochemical treatment time, such as dilution and concentration of samples, is required to detect microalgae by analyzing samples collected from rivers or lakes, and to read the type and amount of microalgae detected. As microalgae shape classification and counting process is required,

The purpose of this study is to present a solution to the problem, since it is difficult to immediately derive the result and there is a problem in that the cost is increased as the number of readings increases.

Microalgae continuous monitoring apparatus using a flow cell according to the present invention to achieve the above object,

A flow cell in which a micro-channel through which the sample flows is formed; An optical module device that photographs a sample flowing along the micro-channel to generate a sample image; A database in which a large number of image data of various cultured microalgae cultured are stored; And a detection and reading device that detects one or more microalgae by comparing the sample image with the image data, and reads the type and amount thereof. It provides a continuous monitoring device for microalgae using a flow cell, characterized in that comprises a.

Microalgae continuous monitoring apparatus using a flow cell according to the present invention,

By capturing the sample flowing along the microchannel, a high-resolution sample image is obtained, and by analyzing it, the type and amount of the detected microalgae are read, but by automating such a process, results can be quickly obtained and cost can be reduced. There was an effect.

1 is an overall configuration diagram of a microalgae continuous monitoring apparatus using a flow cell according to the present invention.
2 (a) and 2 (b) is a block diagram and an exemplary view showing a flow cell as a component of the continuous monitoring device for microalgae using a flow cell according to the present invention.
3 is a block diagram of an optical module device that is one component of the microalgae continuous monitoring device using a flow cell according to the present invention.
Figure 4 (a) to 4 (c) is an exemplary view showing a sample image generated by the optical module device.
Figure 5 is an exemplary view showing a variety of shapes that can be formed coelastrum.
6 is a configuration diagram of a detection and reading device that is a component of the continuous monitoring device for microalgae using a flow cell according to the present invention.
7 (a) and 7 (b) are exemplary views showing a state in which microalgae are detected from a sample image and the magnification of the detected microalgae is enlarged to adjust the magnification.
8 (a) and 8 (b) are exemplary views showing a state in which microalgae are detected from a sample image, and the magnification is adjusted by enlarging the shape of the detected microalgae.
9 (a) and 9 (b) are exemplary views showing a state in which microalgae are detected from a sample image, and a magnification is adjusted by enlarging the shape of the detected microalgae.

The present invention relates to an apparatus for continuously monitoring microalgae contained in a sample,

A flow cell 100 in which a microchannel 101 through which a sample flows is formed; An optical module device 110 for photographing a sample flowing along the micro-channel 101 to generate a sample image; A database 120 in which a large number of image data of various cultured microalgae cultured are stored; A detection and reading device 130 that detects one or more microalgae by reading the image and the image data, and reads the type and amount thereof; It relates to a microalgae continuous monitoring apparatus using a flow cell, characterized in that comprises a.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings.

First, as shown in FIG. 1, the apparatus for continuously monitoring microalgae using a flow cell according to the present invention is characterized in that it comprises a flow cell 100 in which a microchannel 101 through which a sample flows is formed.

The flow cell 100 is an experimental article configured to flow for a certain period of time by introducing samples taken from the outside, such as rivers, lakes, and culture tanks, or samples supplied in real time from the outside for the purpose of observation, analysis, experiment, or photographing. As shown in Figure 2 (a) and 2 (b), the sample is injected and the chemical injection is performed as needed, one or more sample inlet 102, and the microalgae in the sample is composed of aggregated individual observation If this is difficult, a mixing section in which a straight section and a curved section are repeatedly formed so that chemicals such as diluents or surfactants injected through the sample inlet 102 move to the left and right repeatedly with the sample and are effectively mixed ( 103), the width is expanded, all or part is formed transparently, including a shooting section 104 in which a moving sample is photographed One characterized in that, there is a position, the sample injection port (102) side such as the detailed construction can be configured differently depending on the system receiving the sample from the outside.

That is, the flow cell 100 may be configured to be capable of injecting samples taken from the outside into the sample inlet 102 while being placed indoors in a laboratory or the like, or placed in the outdoors such as a river, a lake, or a sample The inlet 102 may be directly connected to a river, a lake, or the like, and may be supplied with a sample from a river, a lake, etc., and a sample outlet or sample discharge for discharging a sample flowing along the micro-channel 101 to the outside Furnaces should be formed.

At this time, the sample injection port 102 may be replaced with a shape of a sample injection path composed of a tubular shape such as a hose.

The micro-channel 101 is a tube made of a transparent material having excellent optical properties to be suitable for observation and photographing from the outside to the inside, and flows into the sample inlet 102 so that the microalgae contained in the sample can be evenly distributed. The sample is repeatedly flowed and mixed in the linear and curved directions along the mixing section 103, and the shooting section 104 formed after the mixing section 103 is a sample of the sample by the following optical module device 110. The diameter or width may be formed to be longer than that of other parts of the micro-channel 101 to facilitate shooting.

Therefore, the photographing section 104 is formed to have a longer diameter or width than other parts of the micro-channel 101, thereby causing a bottleneck, thereby lowering the flow rate of the sample in the photographing section 104. The quality of the sample image generated by the optical module device 110 can be maintained above a certain level.

In addition, the flow cell 100 is configured to be capable of forming an inclination of a certain angle toward the sample discharge port or the sample discharge path from which the sample is discharged to the outside from the sample inlet 102 side where the sample flows, the sample collected or supplied Even if the amount of is small, it is possible to smooth the flow of the sample flowing along the micro-channel 101.

In addition, as shown in Figure 1, the microalgae continuous monitoring device using the flow cell 100 according to the present invention is an optical module device 110 for photographing a sample flowing along the micro-channel 101 to generate a sample image (110) It characterized in that it is configured to include.

The optical module device 110 detects the microalgae contained in the sample, and for the purpose of reading the type and amount of the detected microalgae, the moment of the sample flowing along the photographing section 104 of the microchannel 101 It is a device that shoots static still images.

Accordingly, as shown in FIG. 3, the optical module device 110 moves close to the flow cell 100 to perform an objective lens 111 capable of expanding a sample flowing along the microchannel 101 at a high magnification. And, by including a CCD (Charge coupled device) sensor 112, etc. capable of photographing a high-resolution still image, the detection and reading device 130 to clearly detect the microalgae contained in the sample flowing along the microchannel 101 The lighting device 113 for adjusting brightness can be added.

At this time, the sample introduced into the flow cell 100 is mixed in the mixing section 103 before entering the shooting section 104 so that the microalgae can be distributed evenly, but the mixing section 103 is performed only by such mixing action. Since the microalgae are not uniformly distributed throughout the sample passing through, the optical module device 110 photographs the sample two or more times to generate a plurality of sample images, compared to the measurement result by one sample image. Accurate and reliable measurement results can be obtained.

4 (a) to 4 (c) are exemplary views showing a plurality of sample images generated by the optical module device 110, and these sample images are provided to the following detection and reading device 130 to the sample The included microalgae are detected and the type and amount can be read at the same time, and the read sample image is matched with the exact name of the microalgae and stored in the following database 120 to quickly and accurately detect the microalgae later. It can be used as data for reading.

That is, the present invention detects microalgae and reads the type and amount by accumulating data such as sample images stored in the database 120 as the number of times of detecting the microalgae and reading the type and amount increases. An effect that can improve the speed and accuracy.

In addition, the microalgae continuous monitoring apparatus using the flow cell 100 according to the present invention, the flow cell 100 and the objective lens according to the size of the microalgae contained in the sample, the water level change of the sample in the microchannel 101 It may be configured to include a focal length control device for adjusting the distance between (111), the focal length control device is connected to the detection and reading device 130, the position of the objective lens 111 in a vertical distance By raising or lowering, the focus of the objective lens 111 can be adjusted for the purpose of improving the discriminative power regarding the shape of the microalgae.

According to an embodiment of the present invention, the focal length control device may schedule the optical module device 110 when the shape size of the microalgae included in the sample image generated by the optical module device 110 is so small that it is unreadable. By repeating the process of descending the distance to shorten the distance between the objective lens 111 and the sample so that a clear microalgae shape can be read, and when the shape of the microalgae included in the generated sample image is unclearly expressed By repeating the process of raising the optical module device 110 by a certain distance, the distance between the objective lens 111 and the sample is increased so that a clear microalgae shape can be read, and accordingly, the optical module device 110 is raised or lowered. May be determined according to the reason that causes the inability to judge when the microalgae are not readable by the following detection and reading device 130.

That is, when the size of the shape of the microalgae included in the sample image is small enough to be unreadable, the focus distance control device repeats the process of descending the optical module device 110 by a certain distance until it is readable. When the shape of the microalgae is unclear and cannot be read, the focal length control device repeats the process of raising the optical module device 110 by a certain distance until it is readable.

In addition, as shown in Figure 1, the microalgae continuous monitoring apparatus using a flow cell according to the present invention includes a database 120 in which a number of microalgae cultured species purely cultured and image data matching them are stored in a number It characterized in that the configuration.

The database 120 is a device for storing a plurality of image data including various shapes and characteristics of a single individual, along with the names and shapes of a single individual, as well as the names of each of the known microalgae. The image data is provided to the following detection and reading device 130 so that it can be contrasted with the sample image generated by the optical module device 110, and the sample image can be temporarily stored.

5 is an exemplary view showing various shapes in which a coelastrum, which is a kind of green algae, can be formed among various microalgae that can be stored in the database 120, and the nose is detected by the detection and reading device 130 as described above. Sample images read into the Elastroom can be added and stored.

In addition, a plurality of sample images in which the shape of the microalgae is blurred may be stored in the database 120, and the sample images may be detected and unreadable by the following detection and reading device 130. Can be used as

In addition, as shown in Figure 1, the microalgae continuous monitoring apparatus using a flow cell according to the present invention detects one or more microalgae by comparing the sample image and the image data, and detects the type and amount And a reading device 130.

The detection and reading device 130 receives the sample image from the optical module device 110 and enlarges the shape of the microalgae shown in the still image, and directly contrasts it with the shape of various microalgae included in the image data. As a device for detecting microalgae and reading the type and amount thereof, as shown in FIG. 6, a detector 131 for detecting microalgae from the sample image and an enlarged shape of the detected microalgae and adjusting magnification It is characterized in that it comprises a magnification adjustment unit 132 and a determination unit 133 for determining the type and amount of microalgae by respectively comparing the shape of the enlarged microalgae with the shape of various microalgae included in the image data. The extraction unit 134 extracts one or more features from the expanded microalgae shape, and includes the expanded microalgae features and the image data. It may be configured to further include a feature control unit 135 for contrasting the unique characteristics of the microalgae.

Therefore, the detection and reading device 130 can obtain the effect of determining the type and amount of microalgae in a manner through the contrast of the shape itself, and the type of microalgae in a manner through the contrast of the features shown in the shape And it is possible to obtain the effect of determining the amount, it is possible to obtain an effect that can improve the reliability of the type and amount of the microalgae determined by applying both methods.

In this case, the feature indicated in the shape refers to any one or more features common to the image data and the sample image according to the type of microalgae.

The following is an embodiment of the present invention in which the detection and reading device 130 detects microalgae contained in a sample and reads the type and amount thereof.

First, the optical module device 110 photographs a sample flowing along the micro channel 101 to generate various sample images as shown in FIGS. 2 (a) to 2 (c), and then detects and reads the device 130 The database 120 provides various pre-stored image data as shown in FIG. 5 to the detection and reading device 130.

Next, the detection and reading device 130 detects the microalgae by enlarging the microalgae included in the provided sample image and contrasting with the image data, and reads the type and amount of the detected microalgae, and reads the microalgae. The sample image of is stored in the database 120 with the name of the microalgae matched, and can be used as data for detecting and reading the microalgae later.

7 (a) and 7 (b), FIGS. 8 (a) and 8 (b) and FIGS. 9 (a) and 9 (b), the detection unit 131 is connected to the optical module device 110. The microalgae are detected from the generated sample image, and the enlargement adjustment unit 132 is an exemplary view showing a state in which the magnification of the detected microalgae is adjusted and the magnification is adjusted. The microalgae shown in FIGS. 7 (b), 8 (b), and 9 (b) are respectively determined by comparing the shape of the microalgae with the shapes of various microalgae included in the image data, respectively, It is obvious that the amount is judged differently.

In the same manner as above, the detection and reading device 130 can read the type and amount of all microalgae included in the sample image to calculate the concentration of the microalgae, divide the sample image into a plurality of zones and simultaneously each zone By reading the type and amount of microalgae included in the results can be quickly derived, and the color of the microalgae read out of all the microalgae shown in the sample image can be sequentially converted to prevent duplicate reading.

The embodiments introduced above are provided as examples, so that the technical spirit of the present invention can be sufficiently transmitted to a person having ordinary knowledge in the technical field to which the present invention pertains, and the present invention is directed to the embodiments described above. It is not limited and may be embodied in other forms.

In order to clearly describe the present invention, parts not related to the description are omitted in the drawings, and in the drawings, the width, length, and thickness of components may be exaggerated or reduced for convenience.

In addition, the same reference numbers throughout the specification indicate the same components.

100: flow cell
101: micro flow path 102: sample inlet
103: mixed section 104: shooting section
110: optical module device
111: objective lens 112: CCD sensor
113: lighting device
120: database
130: detection and reading device
131: detection unit 132: enlargement adjustment unit
133: judgment unit 134: extraction unit
135: feature contrast

Claims (6)

A flow cell 100 in which a microchannel 101 through which a sample flows is formed;
An optical module device 110 for photographing a sample flowing along the micro-channel 101 to generate a sample image;
A database 120 in which a large number of image data of various cultured microalgae cultured are stored;
A detection and reading device 130 that detects one or more microalgae by reading the sample image and the image data, and reads the type and amount thereof; Consisting of, including
The flow cell 100,
One or more sample injection port 102 to which the sample is introduced;
A mixing section 103 in which a straight section and a curved section are repeatedly formed and samples are repeatedly moved and mixed to the left and right;
A photographing section 104 in which a width is expanded and a moving sample is photographed in whole or in part; It comprises,
The optical module device 110,
An objective lens 111 capable of expanding the sample flowing along the micro-channel 101 at a high magnification;
Charge coupled device (CCD) sensor 112 capable of photographing a high-resolution still image; It comprises,
The shooting section 104,
Compared to other parts of the micro-channel 101, the diameter or width is formed longer,
The flow cell 100,
Continuous monitoring device for microalgae using a flow cell, characterized in that it is configured to form an inclination of a certain angle toward the sample discharge port or the sample discharge path from which the sample is discharged from the sample injection port 102 side where the sample flows.
delete delete According to claim 1,
Microalgae continuous monitoring device using the flow cell,
A focal length control device that adjusts the distance between the flow cell 100 in which the microchannel 101 is formed and the objective lens 111 according to the size of the microalgae included in the sample, the water level change in the microchannel 101, etc. Continuous device for monitoring microalgae using a flow cell, characterized in that it comprises a (not shown).
According to claim 1,
The detection and reading device 130,
A detection unit 131 for detecting microalgae from the sample image;
An enlargement adjustment unit 132 for enlarging the detected microalgae shape and adjusting the magnification;
A determination unit 133 for determining the type and amount of the microalgae by respectively comparing the shape of the enlarged microalgae with the shape of various microalgae included in the image data; Continuous monitoring device for microalgae using a flow cell, characterized in that comprises a.
The method of claim 5,
The detection and reading device 130,
An extraction unit 134 for extracting one or more features from the enlarged shape of the microalgae;
A feature matching unit (135) for comparing the characteristics of the enlarged microalgae with the characteristics of the microalgae included in the image data; Continuous monitoring device for microalgae using a flow cell, characterized in that further comprises a.

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