KR102555257B1 - Optofluidic bioluminescence detector and bioaerosol real-time detecting system having the same - Google Patents

Abstract

An optofluidic bioluminescence detector according to an embodiment of the present invention includes a body portion; a sample liquid injection unit formed in the body; a solution injection unit formed on one side of the sample solution injection unit; a fluid channel formed in the body so as to communicate with the sample solution injection unit and the solution injection unit; a light-emitting reaction liquid injection unit formed in the body to communicate with the fluid channel; a fluid outlet formed in the body to communicate with the fluid channel; and a detection unit formed in the body so as to communicate with the light emitting reaction solution injection unit, the fluid channel, and the fluid discharge unit.

Description

Optofluidic bioluminescence detector and bioaerosol real-time detection system having the same

The present invention relates to an optofluidic bioluminescence detector and a bioaerosol real-time detection system having the same, and more particularly, an optofluidic bioluminescence detector capable of real-time monitoring of bioaerosol with the naked eye by detecting bioaerosol in air in real time, and the same. Provided is a bioaerosol real-time detection system equipped with

Various microorganisms such as fungi, bacteria, and viruses are floating in the air, and airborne infection problems such as avian influenza, severe acute respiratory syndrome, influenza, Middle East respiratory syndrome, and new coronavirus are emerging. Therefore, the need for technology for detecting microorganisms floating in the air is gradually increasing.

The existing method of measuring airborne microorganisms is to collect microorganisms in a solid or liquid in which biological particles can proliferate, incubate them in an environment where appropriate temperature and humidity are maintained for 1 to 2 days, and then visually check the number of colonies (colony number). , bacterial count) and a method of measuring the number of microorganisms using a fluorescence microscope after staining the collected microorganisms separately.

However, the conventional method described above takes 1 to 2 days, has limitations in detecting airborne microorganisms in real time, and requires user's manual work such as a separate sampling process and pre-processing process. Therefore, existing methods have limitations in that they cannot develop a system for detecting airborne microorganisms in real time.

Therefore, it is necessary to develop a bioaerosol detection device capable of detecting bioaerosol such as biofine dust or airborne microorganisms in real time and visually displaying the result.

The present applicant has proposed the present invention in order to solve the above problems.

Korean Patent Registration No. 10-2200600 (2021.01.04.)

The present invention has been made to solve the above problems, and an optofluidic bioluminescence detector capable of real-time monitoring of bioaerosol with the naked eye by detecting bioaerosol containing biofine dust in the air in real time, and bioaerosol having the same in real time detection system is provided.

In addition, the present invention provides an optofluidic bioluminescence detector and a bioaerosol real-time detection system equipped with the optofluidic bioluminescence detector capable of rapid and accurate measurement in the field without requiring separate sampling and a series of manual operations.

An optofluidic bioluminescence detector according to an embodiment of the present invention for achieving the above object includes a body portion; a sample liquid injection unit formed in the body; a solution injection unit formed on one side of the sample solution injection unit; a fluid channel formed in the body so as to communicate with the sample solution injection unit and the solution injection unit; a light-emitting reaction liquid injection unit formed in the body to communicate with the fluid channel; a fluid outlet formed in the body to communicate with the fluid channel; and a detection unit formed in the body so as to communicate with the light emitting reaction solution injection unit, the fluid channel, and the fluid discharge unit.

A portion of the fluid channel positioned between the sample solution injection unit, the solution injection unit, and the luminescence reaction solution injection unit may be formed such that the flow direction of the fluid flowing inside the fluid channel is repeatedly changed or switched. .

The fluid channel may have the longest portion located between the sample solution injection unit, the solution injection unit, and the luminescent reaction solution injection unit.

The bioaerosol included in the sample solution injected from the sample solution injection unit and the lysis solution injected from the lysis solution injection unit are positioned between the sample solution injection unit, the lysis solution injection unit, and the luminescence reaction solution injection unit. While flowing through the fluid channel, the light emitting reaction liquid injected from the light emitting reaction liquid injection unit and the detection unit may flow into the light emitting reaction liquid injection unit in a state of being mixed and reacting with each other.

The detection unit may be thicker or larger than the diameter or size of the fluid channel.

A portion of the fluid channel in which the mixed solution of the bioaerosol and the dissolving solution and the luminescent reaction solution are mixed and flows may be formed such that the flow direction of the fluid is repeatedly changed or switched.

The detection unit located between the fluid discharge unit and the fluid channel in which the bioaerosol, the dissolution solution, and the luminescent reaction solution are mixed and flows may be formed in a shape in which a diameter or size increases or decreases more gently than that of the fluid channel. there is.

A temperature control unit may be included to control the temperature of a portion of the fluid channel positioned between the sample solution injection unit, the solution injection unit, and the luminescent reaction solution injection unit.

A light reflection unit may be provided on a lower surface of the detection unit.

On the other hand, the present invention, the inlet through which the air containing the bio-aerosol is introduced; a collection unit communicating with the suction unit to collect the bioaerosol; a water storage unit supplying water to the collecting unit; the optofluidic bioluminescence detector into which the bioaerosol collected by the collecting unit is injected together with water; a lysis solution storage unit supplying lysis solution to the optofluidic bioluminescence detector; a luminescence reaction solution storage unit supplying a luminescence reaction solution to the optofluidic bioluminescence detector; It is possible to provide a bioaerosol real-time detection system comprising a fluorescence measurement sensor for detecting light generated from the detection unit of the optofluidic bioluminescence detector.

The collecting unit may be provided as a wet cyclone.

Temperature control elements for controlling refrigerated storage temperatures of the dissolution solution and the luminescence reaction solution may be respectively formed in the solution storage unit and the light emitting reaction solution storage unit.

The optofluidic bioluminescence detector and the bioaerosol real-time detection system equipped with the same according to the present invention do not require a user to manually sample the bioaerosol in the air, and can easily and simply grasp the concentration of the bioaerosol in the air in real time.

In addition, since the optofluidic bioluminescence detector and bioaerosol real-time detection system equipped with the same according to the present invention are composed of one system rather than separate systems, sampling and bioluminescence can be continuously and automatically measured for a long period of time.

In addition, the optofluidic bioluminescence detector and bioaerosol real-time detection system equipped with the same according to the present invention can visually present the detection result to the user wirelessly or wired, and user convenience can be increased because remote control is possible. there is.

In addition, the optofluidic bioluminescence detector and bioaerosol real-time detection system equipped with the same according to the present invention can easily perform processes from bioaerosol dissolution to bioluminescence measurement through a fine fluid channel.

1 is a configuration diagram exemplarily showing the configuration of a bioaerosol real-time detection system according to an embodiment of the present invention.
Fig. 2 is a perspective view exemplarily showing the system according to Fig. 1;
FIG. 3 is a diagram schematically illustrating a concept of detecting bioaerosol in the system according to FIG. 1 .
FIG. 4 shows a first embodiment of an optofluidic bioluminescence detector for use in the system according to FIG. 1 ;
FIG. 5 is a diagram for explaining the operation of the optofluidic bioluminescence detector according to FIG. 4 .
FIG. 6 shows a second embodiment of an optofluidic bioluminescence detector for use in the system according to FIG. 1 ;
7 to 9 are graphs showing bioaerosol detection performance by the system according to FIG. 1 .

Hereinafter, the embodiments disclosed in this specification will be described in detail with reference to the accompanying drawings, but the same or similar reference numerals are given to the same or similar components, and overlapping descriptions thereof will be omitted. The suffix "part" for components used in the following description is given or used interchangeably in consideration of ease of writing the specification, and does not itself have a meaning or role distinct from each other. In addition, in describing the embodiments disclosed in this specification, if it is determined that a detailed description of a related known technology may obscure the gist of the embodiment disclosed in this specification, the detailed description thereof will be omitted. In addition, the accompanying drawings are only for easy understanding of the embodiments disclosed in this specification, the technical idea disclosed in this specification is not limited by the accompanying drawings, and all changes included in the spirit and technical scope of the present invention , it should be understood to include equivalents or substitutes.

Terms including ordinal numbers, such as first and second, may be used to describe various components, but the components are not limited by the terms. These terms are only used for the purpose of distinguishing one component from another.

It should be understood that when an element is referred to as being “connected” to another element, it may be directly connected to the other element, but other elements may exist in the middle.

Singular expressions include plural expressions unless the context clearly dictates otherwise.

In this application, terms such as "comprise" or "have" are intended to designate that there is a feature, number, step, operation, component, part, or combination thereof described in the specification, but one or more other features It should be understood that the presence or addition of numbers, steps, operations, components, parts, or combinations thereof is not precluded.

It is advised that the drawings are schematic and not drawn to scale. Relative dimensions and proportions of parts in the drawings are shown exaggerated or reduced in size for clarity and convenience in the drawings, and any dimensions are illustrative only and not limiting. And like structures, elements or parts appearing in two or more drawings, like reference numerals are used to indicate like features.

The embodiments of the present invention specifically represent ideal embodiments of the present invention. As a result, various modifications of the drawings are expected. Therefore, the embodiment is not limited to the specific shape of the illustrated area, and includes, for example, modification of the shape by manufacturing.

1 is a configuration diagram exemplarily showing the configuration of a bioaerosol real-time detection system according to an embodiment of the present invention, FIG. 2 is a perspective view exemplarily showing the system according to FIG. 1, and FIG. 3 is a system according to FIG. 1 Figure 4 schematically shows the concept of detecting bioaerosols, Figure 4 shows a first embodiment of an optofluidic bioluminescence detector used in the system according to Figure 1, Figure 5 is an optofluidic bioluminescence detector according to Figure 4 6 is a view showing a second embodiment of an optofluidic bioluminescence detector used in the system according to FIG. 1, and FIGS. 7 to 9 are bioaerosol detection by the system according to FIG. 1 Here is a graph showing the performance.

Referring to FIGS. 1 and 2 , a bioaerosol real-time detection system (10, hereinafter abbreviated as 'system') according to an embodiment of the present invention includes an inlet ( 120); a collection unit 110 communicating with the suction unit 120 to collect the bioaerosol; Water storage unit 160 for supplying water to the collecting unit 110; an optofluidic bioluminescence detector 100 into which the bioaerosol collected by the collecting unit 110 is injected together with water; a lysate storage unit 170 supplying the lysed solution to the optofluidic bioluminescence detector 100; a luminescence reaction liquid storage unit 176 supplying the luminescence reaction liquid to the optofluidic bioluminescence detector 100; and a fluorescence measuring sensor 180 for detecting light generated from the detector 106 (see FIG. 4) of the optofluidic bioluminescence detector 100.

The system 10 according to an embodiment of the present invention configured as described above collects bioaerosol in the air using the collecting unit 110 of the wet cyclone and injects it into the optofluidic bioluminescence detector 100 to measure fluorescence. A relative light unit (RLU) may be measured by the sensor 180 and the value may be displayed on a display in real time.

Hereinafter, the detection target is a concept including bioaerosols including airborne microorganisms such as viruses, bacteria, and fungi in the air, and aerosols such as fine dust, and the bioaerosol real-time detection system (10) according to an embodiment of the present invention. ) Means an object (material) that is liquefied and collected by the collecting unit 110.

In addition, in the present specification, bioaerosol is defined as a concept including biofine dust.

Referring to FIGS. 1 and 2 , air containing bioaerosol is introduced through the suction unit 120 of the system 10, and the suction unit 120 transfers the air containing the bioaerosol to the collecting unit 110. convey Here, the collecting unit 110 may be provided as a wet cyclone.

The collection unit 110 of the system 10 according to an embodiment of the present invention is the same as the "liquid collection device 110" described in Korean Patent Application No. 10-2020-0080338 for which the present applicant has filed a patent application. , repetitive explanations are omitted.

As disclosed in Korean Patent Application No. 10-2020-0080338, since the collecting unit 110 collects the bioaerosol in a wet method, water is injected into the collecting unit 110 along with the air containing the bioaerosol. To this end, the collecting unit 110 may be connected to the water storage unit 160 . At this time, the water (ultrapure water) stored in the water storage unit 160 may be transported to the collecting unit 110 through the fluid machine 140 . It is preferable to use a pump having an on/off function as the fluid machine 140 connecting the water storage unit 160 and the collecting unit 110. can be done

The air from which the bioaerosol is removed from the collecting unit 110 may be discharged to the outside by the air pump 130 . The air flow rate collected by the collecting unit 110 may be determined by the air pump 130 connected to the collecting unit 110 .

In the system 10 according to an embodiment of the present invention, the bioaerosol can be collected in a highly concentrated state in the collecting unit 110 . As such, since highly concentrated bioaerosol can be collected, even if a small amount of bioaerosol is supplied to the optofluidic bioluminescence detector 100 to be described later, the concentration of bioaerosol suspended in the air can be accurately detected.

The bioaerosol collected in the collecting unit 110 may be discharged from the collecting unit 110 together with water and then introduced into the optofluidic bioluminescence detector 100 . At this time, the fluid (water) including the bioaerosol may flow into the optofluidic bioluminescence detector 100 after passing through the fluid machine 151 .

The system 10 according to an embodiment of the present invention may detect bioaerosol in real time by including the optofluidic bioluminescence detector 100 . When the bioaerosol collected by the collecting unit 110 is injected into the optofluidic bioluminescence detector 100, it is mixed with a Lysis solution inside the microfluidic channel 104 to dissolve the bioaerosol. In the process, ATP (adenosine triphosphate) is extracted, and the extracted ATP is mixed with a luminescent reaction solution (Luciferin-Luciferase solution) to react and emit light.

Therefore, a dissolving solution and a luminescent reaction solution as well as water containing bioaerosol are introduced into the optofluidic bioluminescence detector 100 . To this end, the system 10 according to an embodiment of the present invention may include a solution storage unit 170 and a light emitting reaction solution storage unit 176 .

A solution for dissolving the bioaerosol may be stored in the dissolving solution storage unit 170 , and a light-emitting reaction solution that emits light by reacting with the dissolved bioaerosol may be stored in the light-emitting reaction solution storage unit 176 .

A lysis solution may be used as the lysate stored in the lysate storage unit 170 . Lysis solution is an enzyme solution capable of dissolving bioaerosol or destroying cell walls of bioaerosol. When lysed by the lysis solution or when the cell wall is destroyed, the bioaerosol releases ATP.

A luciferin-luciferase solution may be used as the luminescence reaction liquid stored in the luminescence reaction liquid storage unit 176 . When ATP reacts with the Luciferin-luciferase solution, ATP is converted to AMP and emits light. The luciferin-luciferase solution that reacts with ATP to emit light is also an enzyme fluid.

Since the lysis solution and the luciferin-luciferase solution used in the optofluidic bioluminescence detector 100 are enzyme solutions, they must be frozen to stabilize them. In the system 10 according to an embodiment of the present invention, the lifespan of the lysis solution and the luciferin-luciferase solution can be extended and used for a long time by storing the lysis solution and the luciferin-luciferase solution in a refrigerator without freezing them. To this end, temperature control elements 175 and 179 for controlling refrigerated storage temperatures of the dissolution solution and the luminescence reaction solution may be formed in the solution storage unit 170 and the light emitting reaction solution storage unit 176, respectively.

The temperature control elements 175 and 179 are preferably provided as thermoelectric elements capable of adjusting or controlling temperature by supplying electricity. The operation of the temperature control elements 175 and 179 may be controlled by the control unit 190 . The control unit 190 controls the amount of current supplied to the temperature control elements 175 and 179 so that the lysis solution and the luciferin-luciferase solution stored in the solution storage unit 170 and the luminescent reaction solution storage unit 176 are stored at about 4 ° C. It can be kept refrigerated.

On the other hand, since the lysis solution and the luciferin-luciferase solution are expensive solutions, it is advantageous in terms of economic feasibility to extend the lifespan and use them for a long time. In the system 10 according to an embodiment of the present invention, lifespan can be extended by refrigerating the lysis solution and the luciferin-luciferase solution, and the optofluidic bioluminescence detector 100 into which the lysis solution and the luciferin-luciferase solution are introduced Since the microfluidic channel 104 is formed, the bioaerosol can be detected by using a small amount of the lysis solution and the luciferin-luciferase solution by reducing the amount of the injected solution, and as a result, the lysis solution and the luciferin-luciferase solution can be used for a long period of time. Because of this, the economy can be improved.

In the system 10 according to an embodiment of the present invention, between the solution storage unit 170 and the optofluidic bioluminescence detector 100, the luminescence reaction solution storage unit 176 and the optofluidic bioluminescence detector 100 are connected. Between them may include fluid machines 151 and 155 respectively provided.

The operating state or operation of the fluid machines 151 and 155 is controlled by the control unit 190, and may serve as a peristaltic pump and a valve. That is, since the fluid machines 151 and 155 have an On/Off function, they can also serve as one-way valves.

The fluid machines 151 and 155 are preferably provided in the form of peristaltic pumps. The control unit 190 controls the operating state of the fluid machines 151 and 155 so that the solution and the luminescent reaction solution are injected into the optofluidic bioluminescence detector 100. flow rate can be adjusted. Since the fluid machines 151 and 155 can adjust or control the injection flow rate of the dissolving solution and the luminescent reaction solution, only a small amount of the expensive dissolving solution and the luminescent reaction solution are injected into the optofluidic bioluminescence detector 100 to obtain the dissolving solution and the luminescent reaction. The expiration date of the liquid can be extended, and the operating frequency of the optofluidic bioluminescence detector 100 can be controlled.

Here, the fluid machine 151 provided between the lysate storage unit 170 and the optofluidic bioluminescence detector 100 collects the fluid discharged from the collecting unit 110, that is, water containing the highly concentrated collected bioaerosol. An optofluidic bioluminescence detector 100 may be injected.

The bioaerosol injected into the optofluidic bioluminescence detector 100 emits light by the dissolving solution and the luminescent reaction solution, and the light is emitted by the fluorescence measuring sensor 180 disposed on the upper part of the optofluidic bioluminescence detector 100. can be measured

The fluorescence measurement sensor 180 measures the RLU (Relative Light Unit) and transmits the resultant value to the control unit 190, and the control unit 190 transmits the measurement value of the fluorescence measurement sensor 180 through the display unit 195. will inform the user. The control unit 190 and the display unit 195 may communicate by wire or wirelessly, and the display unit 195 may be provided in the form of a computer or user terminal, and may be remotely controlled.

The system 10 according to an embodiment of the present invention is provided with the optofluidic bioluminescence detector 100, so that processes from bioaerosol dissolution to bioluminescence measurement can be easily performed through a microfluidic channel, and sampling and Since bioluminescence is implemented in one system rather than a separate system, the concentration of bioaerosol in the air can be continuously and automatically measured for a long period of time.

Hereinafter, the optofluidic bioluminescence detector 100 will be described in detail with reference to the drawings.

3 shows a simplified concept of operation of a system 10 comprising an optofluidic bioluminescence detector 100 . Referring to FIG. 3 , air containing bioaerosol (BA) flows into the first inlet 112 of the collecting unit 110 and water flows into the second inlet 114 of the collecting unit 110. do.

Air and water including bioaerosol flow along the inner surface of the cyclone unit 111 of the collecting unit 110, and in this process, the bioaerosol is adsorbed to water and collected in a highly concentrated state. The air discharged through the outlet 115 and from which the bioaerosol has been removed is discharged through the second outlet 114 of the collecting unit 110 .

Water (HS) containing bioaerosol flows into the optofluidic bioluminescence detector 100, and then the cell wall is destroyed by the lysate to release ATP, and the ATP reacts with the luminescent reaction solution to emit light. (180) will measure the light. A fine fluid channel 104 is formed inside the optofluidic bioluminescence detector 100. Since the fluid channel 104 is finely formed, only a small amount of the bioaerosol, the solution, and the luminescent reaction solution pass through the fluid channel 104. It is possible to measure the concentration of bioaerosol even when it is injected.

4 and 5 a first embodiment of an optofluidic bioluminescence detector 100 is shown.

As shown in FIGS. 4 and 5 , the optofluidic bioluminescence detector 100 according to the first embodiment of the present invention includes a body portion 101; a sample liquid injection part 102 formed in the body part 101; a solution injection unit 103 formed on one side of the sample solution injection unit 102; a fluid channel 104 formed in the body 101 to communicate with the sample solution injection unit 102 and the solution injection unit 103; a light emitting reaction solution injection unit 105 formed in the body portion 101 to communicate with the fluid channel 104; a fluid outlet 109 formed in the body 101 to communicate with the fluid channel 104; and a detection unit 106 formed in the body 101 to communicate with the light emitting reaction liquid injection unit 105, the fluid channel 104, and the fluid discharge unit 109.

The body portion 101 of the optofluidic bioluminescence detector 100 is preferably formed of a transparent material so that a user can observe the inside with the naked eye. For example, it may be formed of a material such as a transparent polymer, polydimethylsiloxane (PDMS), or polymethylmethacrylate (PMMA). In some cases, only the detection unit 106 to detect light may be formed transparently, and the rest of the body portion 101 may be formed of metal such as copper.

The optofluidic bioluminescence detector 100 may be manufactured by injecting a material such as polymer or PDMS into a mold or mold in which a shape such as the microfluidic channel 104 is formed.

Referring to FIG. 4 , the body portion 101 preferably has a flat hexahedron shape so that a fine fluid channel 104 can be sufficiently formed therein, but is not necessarily limited to this shape.

The fine fluid channel 104 is formed inside the body portion 101, and four holes for injecting fluid or liquid flowing through the fluid channel 104 may be formed in the body portion 101. The four holes are a sample solution injection unit 102, a solution injection unit 103, a light emitting reaction solution injection unit 105, and a fluid discharge unit 109. The sample solution injection unit 102, the solution injection unit 103, the light emitting reaction solution injection unit 105, and the fluid discharge unit 109 connect the surface of the body 101 and the fluid channel 104. It is desirable to provide

The sample liquid discharged from the collecting unit 110 , that is, water containing highly concentrated bioaerosol may be injected into the sample liquid injection unit 102 . The control unit 190 may control the supply flow rate, supply time, and supply frequency of the sample liquid injected into the optofluidic bioluminescence detector 100 by controlling the operation of the fluid machine 151 provided as a peristaltic pump.

The sample liquid introduced through the sample liquid injection unit 102 flows through the fluid channel 104. The sample liquid injection unit 102 is connected to one end of the sample liquid channel 102a and the other end of the sample liquid channel 102a. may be connected to the fluid channel 104. Here, the sample liquid channel 102a is preferably provided in the same shape as the fluid channel 104.

A solution injection unit 103 may be formed on one side of the sample solution injection unit 102 . A lysate that dissolves the bioaerosol included in the sample liquid or destroys cell walls may be introduced into the lysate injection unit 103 . A lysis solution may be used as the lysis solution. The control unit 190 may control the supply flow rate, supply time, and supply frequency of the solution injected into the optofluidic bioluminescence detector 100 by controlling the operation of the fluid machine 151 provided as a peristaltic pump.

The lysis solution introduced through the lysis solution injection unit 103 flows through the fluid channel 104. The lysis solution injection unit 103 is connected to one end of the lysis solution channel 103a and the lysis solution channel ( The other end of 103a) may be connected to the fluid channel 104. Here, the solution channel 103a is preferably provided in the same shape as the fluid channel 104.

Meanwhile, the other end of the solution channel 103a is preferably connected to a portion where the other end of the sample solution channel 102a is connected to the fluid channel 104. That is, it is preferable that the sample solution channel 102a and the solution solution channel 103a are connected to the fluid channel 104 so that the sample solution and the solution solution flow together into the fluid channel 104 . In this case, the sample solution channel 102a, the solution channel 103a, and the fluid channel 104 may be integrally formed.

Since the upstream end of the fluid channel 104 is connected to the sample solution channel 102a and the solution channel 103a, the sample solution mixes with the solution flow while flowing through the fluid channel 104, so that the bioaerosol of the sample solution dissolves. It can react with liquid.

Of the fluid channel 104, the portion (CLR) positioned between the sample solution injection unit 102, the solution injection unit 103, and the luminescence reaction solution injection unit 105 is a source of fluid flowing inside the fluid channel 104. It can be configured so that the flow direction changes or switches repeatedly. Referring to FIG. 4 , a portion (CLR) of the fluid channel 104 positioned below the body portion 101 is formed in a zig-zag shape so that the flow direction of the fluid repeatedly changes. Among the fluid channels 104, the portion (CLR) formed in a zig-zag shape in which the flow direction of fluid repeatedly changes is a cell lysis region.

The part (CLR) of the fluid channel 104 in which the bioaerosol and the lysate are mixed and flows may be formed such that the flow direction of the fluid is repeatedly changed or switched.

Referring to FIG. 5, the optofluidic bioluminescence detector 100 according to an embodiment of the present invention needs to detect luminescence using ATP inside the cells of bioaerosol (BA). To this end, the cell walls of bioaerosol are destroyed. Alternatively, ATP must be extracted by cell lysis.

Cell walls of the bioaerosol must be destroyed before the light emitting reaction liquid injected from the light emitting reaction liquid injection unit 105 meets the bioaerosol so that sufficient ATP can react with the light emitting reaction liquid. Therefore, the longer the time when the lysis solution such as the Lysis solution is mixed with the bioaerosol is, the cell walls of the bioaerosol (BA) are destroyed or the cells are lysed so that ATP can be smoothly discharged.

The fluid channel 104 of the optofluidic bioluminescence detector 100 according to an embodiment of the present invention is positioned between the sample solution injection unit 102, the solution injection unit 103, and the luminescent reaction solution injection unit 109. The part to do can be formed the longest. That is, the cell lysis region (CLR) occupies the longest part of the fluid channel 104 . Since the fluid channel 104 of the cell lysis region (CLR) occupies the longest part, it is possible to secure the time required for the bioaerosol to be sufficiently mixed with the lysate while passing through the cell lysis region (CLR), and to obtain the concentration of the bioaerosol. ATP needed to measure can be released.

It takes about 3 minutes for the Lysis solution used as the dissolving solution to dissolve or destroy the cell walls of the bioaerosol. Therefore, the fluidic channel 104 is preferably formed so that it takes 3 minutes or longer for the bioaerosol and the lysate to pass through the cell lysis region (CLR). However, the time taken to pass through the entire cell lysis region (CLR) does not have to be about 3 minutes, and the time taken to pass through several channels located in the front of the cell lysis region (CLR) is formed to be about 3 minutes. also free

On the other hand, in order for the cell wall of the bioaerosol to react with a lysing solution such as a lysis solution to be melted or destroyed, the reaction temperature is preferably maintained constant. That is, in order to increase the ATP extraction efficiency, it is preferable to maintain a constant temperature of the cell lysis region (CLR) where the bioaerosol and the lysate are mixed. To this end, the optofluidic bioluminescence detector 100 may include a temperature controller 107 that controls the temperature of the cell lysis region CLR in the fluidic channel 104 . Operation of the temperature control unit 107 may be controlled by the control unit 190 .

The temperature control unit 107 is preferably disposed behind the cell lysis region (CLR), and it is preferable that the entire cell lysis region (CLR) is placed on the temperature control unit 107. The temperature control unit 107 may adjust the temperature so that the temperature of the cell lysis region (CLR) is maintained at room temperature or 4°C to 30°C. When the temperature of the cell lysis region (CLR) is lower than 4°C, the activity decreases, so it is preferable to adjust the temperature to 4°C or higher.

While passing through the cell lysis region (CLR) of the fluid channel 104, the ATP extracted from the bioaerosol meets the luminescence reaction liquid 109 injected from the luminescence reaction liquid injection unit 105.

A light emitting reaction liquid in which ATP reacts and emits light may be introduced into the light emitting reaction liquid injection unit 105 . A luciferin-luciferase solution may be used as the luminescence reaction solution. The control unit 190 may control the supply flow rate, supply time, and supply frequency of the light emitting reaction liquid injected into the optofluidic bioluminescence detector 100 by controlling the operation of the fluid machine 155 provided as a peristaltic pump.

The light-emitting reaction liquid (luciferin-luciferase solution) introduced through the light-emitting reaction liquid injection unit 105 flows through the fluid channel 104, and the light-emitting reaction liquid injection unit 105 connects one end and the other end of the light-emitting reaction liquid channel 105a. The other end of the luminescent reaction solution channel 105a may be connected to the fluid channel 104 at the outlet of the cell lysis region CLR. Here, it is preferable that the luminescence reaction liquid channel 105a is provided in the same shape as the fluid channel 104.

The luminescent reaction liquid flowing through the luminescent reaction solution channel 105a and the ATP of the bioaerosol passing through the cell lysis region (CLR) flow into the luminescence reaction channel 109a. One end of the luminescence reaction channel 109a may be connected to the other end of the luminescence reaction liquid channel 105a and the fluid channel 104 at the outlet of the cell lysis region CLR. The other end of the luminescence reaction channel 109a may be connected to the detector 106 .

In this way, the bioaerosol included in the sample solution injected from the sample solution injection unit 102 and the lysate injected from the solution injection unit 103 are injected into the sample solution injection unit 102, the solution injection unit 103 and The light emitting reaction liquid injected from the light emitting reaction liquid injection unit 105 and the detection unit 106 are introduced into the light emitting reaction liquid injection unit 105 while being mixed and reacted with each other while flowing through the fluid channel 104 (CLR) located between the light emitting reaction liquid injection unit 105. It can be.

While flowing through the luminescent reaction channel 109a, ATP is converted into AMP by a luminescent reaction solution such as a luciferin-luciferase solution and emits light.

The detection unit 106 is a part that collects ATP that reacts with the luminescence reaction solution and emits light. Since ATP is collected in the detector 106, the amount of emitted light can be increased. The detection unit 106 is preferably formed thicker or larger than the diameter or size of the fluidic channel 104 so that ATP is collected and emitted light can be sufficiently generated. Referring to FIG. 4 , the detection unit 106 is thicker and larger than the emission channel 109a connected to its left side and the discharge channel 109b connected to its right side.

Since the detector 106 is a part that measures the concentration of the bioaerosol using light emitted from ATP, it should be able to generate a measurable amount of light and transmit the light to the outside. Therefore, the detection unit 106 is preferably formed in a shape larger than the diameter or size of the fluid channel 104, for example, in a round shape. The detection unit 106 shown in FIG. 4 has a substantially round shape when viewed from above. In addition, the detector 106 should be transparent so that the fluorescence sensor 180 can measure the light emitted from the detector 106 .

It is preferable that the fluorescence measurement sensor 180 is positioned above the detector 106 to measure light emitted from the detector 106 . The fluorescence measurement sensor 180 measures a Relative Light Unit (RLU) of light emitted from the detector 106 and displays the value on the display unit 195 in real time.

A photomultiplier tube (PMT) may be used as the fluorescence measurement sensor 180 .

On the other hand, since the light emitted from the detection unit 106 spreads in the same direction, accurate measurement is possible when the fluorescence measurement sensor 180 is positioned on a straight optical path of the light spreading in the same direction. However, in order to increase the amount of light transmitted to the fluorescence measurement sensor 180, a light reflection unit 108 may be provided on the lower surface of the detection unit 106.

The light reflection unit 108 is preferably provided at a position facing the fluorescence measuring sensor 180 based on the detection unit 106 . When the light reflection unit 108 is disposed on the lower surface of the detection unit 106 and the fluorescence measurement sensor 180 is disposed on the upper surface of the detection unit 106, the light emitted upward from the detection unit 106 is the fluorescence measurement sensor 180 Light (L, see FIG. 5) transmitted directly to and emitted downward from the detector 106 may be reflected by the light reflector 108 and transmitted to the fluorescence measurement sensor 180. By disposing the light reflection part 108, the amount of light measured by the fluorescence sensor 180 can be increased.

The light reflection part 108 preferably has a flat mirror or concave mirror shape. When the light reflector 108 is formed of a concave mirror, a greater amount of light may be transmitted to the fluorescence measuring sensor 180 than when the light reflector 108 is formed of a flat mirror. As such, the light reflection unit 108 may improve the sensitivity of the fluorescence measurement sensor 180 by reflecting the emitted light.

The fluid containing ATP emitted by the detector 160 passes through the discharge channel 109b and is then discharged to the outside of the body 101 through the fluid discharge unit 109 .

Since the optofluidic bioluminescence detector 100 according to an embodiment of the present invention can form a fluid channel 104 finely, samples (sample solution, solution solution, luminescence reaction solution, etc.) necessary for bioaerosol detection It is possible to use less and realize miniaturization of the system 10 . For example, as shown in FIG. 4 , it is possible to miniaturize the optofluidic bioluminescence detector 100 such that the distance between adjacent fluid channels in the cell lysis region (CLR) is about 5 mm.

Referring to FIG. 5 , when the collected bioaerosol (BA) is injected into the optofluidic bioluminescence detector 100, it is mixed with a Lysis solution, which is a dissolution solution, inside the microfluidic channel 104, and the bioaerosol (BA) is dissolved. In the process, ATP (adenosine triphosphate) is extracted. The extracted ATP is mixed and reacted with a Luciferin-Luciferase solution, which is a luminescent reaction solution, to emit light (L), and the fluorescence measuring sensor 180 measures the light.

6 shows a second embodiment of an optofluidic bioluminescence detector for use in system 10 according to one embodiment of the present invention.

Referring to FIG. 6 , the optofluidic bioluminescence detector 200 includes a body portion 201; a sample liquid injection unit 202 formed in the body unit 201; a solution injection unit 203 formed on one side of the sample solution injection unit 202; a fluid channel 204 formed in the body 201 to communicate with the sample solution injection unit 202 and the solution injection unit 203; a light emitting reaction solution injection unit 205 formed in the body portion 201 to communicate with the fluid channel 204; a fluid outlet 209 formed in the body 201 to communicate with the fluid channel 204; and a detection unit 206 formed in the body 201 to communicate with the light emitting reaction liquid injection unit 209, the fluid channel 204, and the fluid discharge unit 209.

The sample solution channel 202a, the solution channel 203a, the luminescence reaction solution channel 205a, the luminescence reaction channel 209a, the discharge channel 209b, etc. of the optofluidic bioluminescence detector 200 shown in FIG. It is the same as the optofluidic bioluminescence detector 100 shown in FIG. 4 . However, the shape of the fluid channel 204 and the shape of the detector 206 are different from those of the optofluidic bioluminescence detector 100 of FIG. 4 . Hereinafter, the differences are described in detail.

In the case of the optofluidic bioluminescence detector 200 shown in FIG. 6, in the fluid channel 204, a part where the bioaerosol and the lysate are mixed (CLR), and a part where the luminescence reaction liquid and ATP of the bioaerosol are mixed (BLR) It has a repetitive zig-zag pattern.

As described above, it is preferable to secure a time of about 3 minutes in order for the cell walls of the bioaerosol to be dissolved by the lysis solution. For this, the cell lysis region (CLR) where the bioaerosol is mixed with the lysis solution The fluid channel 204 may be formed such that the flow direction repeatedly changes or is switched in the left and right directions as well as the up and down directions.

Referring to FIG. 6, the fluid channel 204 of the cell lysis region (CLR) is formed so that the flow direction repeatedly changes along the vertical direction. The fluid channel 204 provided in the vertical direction has a flow direction in the left and right directions It is arranged in an up and down direction while changing. Therefore, the cell lysis region (CLR) of the optofluidic bioluminescence detector 200 of FIG. 6 is a longer fluidic channel within the same area as the cell lysis region (CLR) of the optofluidic bioluminescence detector 100 of FIG. (204). In addition, if the length of the fluid channel of the cell lysis region (CLR) is the same, the size or area of the cell lysis region (CLR) can be reduced in the case of FIG. 6 compared to the case of FIG. is advantageous for the miniaturization of

In addition, in the optofluidic bioluminescence detector 200 shown in FIG. 6, in the fluid channel 204, a mixture of bioaerosol and dissolving solution (ie, ATP is extracted) and a luminescent reaction solution are mixed and flowed (BLR) may be formed such that the flow direction of the fluid is repeatedly changed or switched.

As will be described later, the optofluidic bioluminescence detector 200 shown in FIG. 6 has a smaller detector 206 than the detector 100 shown in FIG. 4 . Therefore, the reaction time between ATP and the luminescent reaction solution within the detector 206 is not sufficient compared to the detector 100 of FIG. 4 , and in this case, a sufficient amount of light may not be emitted from the detector 206 . In order to solve this problem, the fluidic channel 204 of the optofluidic bioluminescence detector 200 according to the second embodiment shown in FIG. A light emitting response region (BLR) may be included to secure a reaction time.

As shown in FIG. 6, the luminescent response region (BLR) of the fluidic channel 204 is formed from the point where the other end of the luminescent reaction solution channel 205a and the exit channel of the cell lysis region (CLR) join, Similar to the melting region CLR, the flow direction may be changed in the vertical and horizontal directions. Since the luminescence reaction region (BLR) is formed as a sufficiently long fluid channel, ATP can secure sufficient time to react with the luminescence reaction solution while passing through the luminescence reaction region (BLR) and measured by the fluorescence measuring sensor 180 It can produce sufficient amount of light required for

Meanwhile, in the case of the optofluidic bioluminescence detector 100 according to the first embodiment shown in FIG. 4 , air bubbles or bubbles may be generated inside the detector 106 . Because the detection unit 106 is larger than the fluid channel 104 connected to the detection unit 106, the air introduced into the detection unit 106 through the fluid channel 104 does not escape the detection unit 106 and the inside of the detection unit 106 This is because there is enough space to create bubbles in If air bubbles or bubbles exist inside the detection unit 106, the amount of light emitted from the detection unit 106 may change, and accordingly, a measurement result of the fluorescence measurement sensor 180 may not be accurate.

The optofluidic bioluminescence detector 200 according to the second embodiment shown in FIG. 6 is formed so that air bubbles or bubbles are not generated inside the detector 206 . That is, the detection unit 206 located between the luminescent reaction channel 209a where the ATP of the bioaerosol, the lysate, and the luminescent reaction liquid are mixed and flowing and the fluid discharge unit 209 increases in diameter or size more gently than the fluid channel, or It can be formed in a decreasing form.

Referring to FIG. 6, the detection unit 206 is formed long along the longitudinal direction of the emission channel 209a or the discharge channel 209b, but the diameter or size of the detection unit 206 gradually increases and then gradually decreases. Since the detection unit 206 has an elongated shape along the flow direction of the fluid, the wetting characteristics of the fluid introduced into the detection unit 206 extends throughout the detection unit 206, so that bubbles or bubbles are formed inside the detection unit 206. An environment that is difficult to exist can be created.

Although not shown in FIG. 6 , a temperature control unit (not shown) may be provided on the rear side of the cell lysis region CLR, and a light reflection unit (not shown) may be provided on the rear side of the detection unit 206 . The temperature control unit and the light reflection unit provided in the optofluidic bioluminescence detector (200) shown in FIG. is the same as

Meanwhile, FIG. 7 is a graph showing the total collection efficiency and the size of fine particles of the collection unit 110 provided as a wet cyclone. There was no significant difference in collection efficiency depending on the particle size, and it was confirmed that the total collection efficiency was 99.5%.

8 is a graph showing bioaerosol detection sensitivity according to various bioaerosol concentrations of the bioaerosol real-time detection system 10 according to an embodiment of the present invention. In the air, Gram-positive bioaerosols (S.epidermidis, B.subtilis, M.luteus) and Gram-negative bacteria bioaerosols (E.coli, Enterobacter arrowroot) E. aerogens and K. pneumoniae) were sprayed and collected by the collecting unit 110 of the wet cyclone, and then the evaluation was conducted. As a result, it was confirmed that the RLU value of the bioaerosol real-time detection system 10 tended to increase as the bioaerosol concentration increased. This is a result showing that the higher the bioaerosol concentration, the more ATP is extracted and the bioluminescence increases.

9 is a graph showing the real-time monitoring sensitivity of the bioaerosol real-time detection system 10 for bioaerosol concentration changes in a laboratory. This is the result of measuring the real-time monitoring sensitivity of the detection system 10 while stopping the spraying of the bioaerosol when the concentrations of the bioaerosol reached 130/cm ³ and 200/cm ³ , respectively. As an example of the bioaerosol, Staphylococcus epidermidis was used. As a result, it was confirmed that the bioaerosol real-time detection system 10 quickly and accurately tracks the concentration change of the bioaerosol in the air.

As described above, in one embodiment of the present invention, specific details such as specific components and limited embodiments and drawings have been described, but this is only provided to help a more general understanding of the present invention, and the present invention is based on the above embodiments. It is not limited, and those skilled in the art can make various modifications and variations from these descriptions. Therefore, the spirit of the present invention should not be limited to the described embodiments and should not be determined, and all things equivalent or equivalent to the claims as well as the following claims belong to the scope of the present invention.

10: bioaerosol real-time detection system
100,200: optofluidic bioreaction detector
101,201: body part 102,202: sample liquid injection part
103,203: solution injection unit 104,204: fluid channel
105,205: light emitting reaction liquid injection unit 106,206: detection unit
107: temperature control unit 108: light reflection unit
109,209: fluid discharge unit 110: collection unit
180: fluorescence measurement sensor 190: control unit

Claims (12)

body part;
a sample liquid injection unit formed in the body;
a solution injection unit formed on one side of the sample solution injection unit;
a fluid channel formed in the body so as to communicate with the sample solution injection unit and the solution injection unit;
a light-emitting reaction solution injection unit formed in the body portion to communicate with the fluid channel;
a fluid outlet formed in the body to communicate with the fluid channel; and
A detection unit formed in the body so as to communicate with the light emitting reaction solution injection unit, the fluid channel, and the fluid discharge unit;
The bioaerosol included in the sample solution injected from the sample solution injection unit and the lysis solution injected from the lysis solution injection unit are positioned between the sample solution injection unit, the lysis solution injection unit, and the luminescence reaction solution injection unit. While flowing through the fluid channel, the light emitting reaction liquid injected from the light emitting reaction liquid injection unit and the detector are introduced in a state of being mixed and reacting with each other,
The detection unit located between the fluid discharge unit and the fluid channel in which the bioaerosol, the dissolution solution, and the luminescent reaction solution are mixed and flows has a diameter or size gradually increased from the fluid channel so that air bubbles or bubbles are not generated therein. The optofluidic bioluminescence detector, characterized in that it gradually decreases toward the fluid outlet and is formed in an elongated shape along the flow direction of the fluid flowing inside the fluid channel.
According to claim 1,
A portion positioned between the sample solution injection unit, the solution injection unit, and the luminescent reaction solution injection unit among the fluid channels is formed such that the flow direction of the fluid flowing inside the fluid channel is repeatedly changed or switched. An optofluidic bioluminescence detector made with
According to claim 2,
The optofluidic bioluminescence detector, characterized in that the portion of the fluid channel located between the sample solution injection unit, the lysis solution injection unit, and the luminescent reaction solution injection unit is formed to be the longest.
According to claim 2,
A portion of the fluid channel positioned between the sample solution injection unit, the solution injection unit, and the luminescent reaction solution injection unit is formed to repeatedly change in the flow direction along the vertical and horizontal directions. Fluid bioluminescence detector.
According to claim 4,
The optofluidic bioluminescence detector, characterized in that the detection unit is formed thicker or larger than the diameter or size of the fluid channel.
According to claim 4,
Of the fluid channel, in the part where the mixed solution of the bioaerosol and the dissolution solution and the light emitting reaction solution are mixed and flowed, the flow direction of the fluid is in the vertical direction and the left and right directions. An optofluidic bioluminescence detector characterized in that it is formed to be repeatedly changed or switched.
delete According to claim 2,
The optofluidic bioluminescence detector, characterized in that it comprises a temperature control unit for controlling the temperature of a portion of the fluid channel positioned between the sample solution injection unit, the solution injection unit, and the luminescent reaction solution injection unit.
According to claim 2,
Optofluidic bioluminescence detector, characterized in that the light reflection unit is provided on the lower surface of the detection unit.
an inlet through which air containing bioaerosol is introduced;
a collection unit communicating with the suction unit to collect the bioaerosol;
a water storage unit supplying water to the collecting unit;
The optofluidic bioluminescence detector according to any one of claims 1 to 6, 8 and 9, in which the bioaerosol collected by the collecting unit is injected together with water;
a lysis solution storage unit supplying lysis solution to the optofluidic bioluminescence detector;
a luminescence reaction solution storage unit supplying a luminescence reaction solution to the optofluidic bioluminescence detector; and
a fluorescence measuring sensor for detecting light generated from the detection unit of the optofluidic bioluminescence detector;
Bioaerosol real-time detection system comprising a.
According to claim 10,
The bioaerosol real-time detection system, characterized in that the collecting unit is provided as a wet cyclone.
According to claim 11,
The bioaerosol real-time detection system, characterized in that the temperature control element for controlling the refrigerated storage temperature of the lysate and the luminescent reaction liquid is formed in the lysate storage unit and the luminescent reaction liquid storage unit, respectively.

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