WO2014192787A1

WO2014192787A1 - Detection device

Info

Publication number: WO2014192787A1
Application number: PCT/JP2014/064065
Authority: WO
Inventors: 克佳高橋; 大樹奥野; 藤岡　一志; 伸佳石野
Original assignee: シャープ株式会社
Priority date: 2013-05-31
Filing date: 2014-05-28
Publication date: 2014-12-04

Abstract

This detection device (100) comprises: a collection substrate (10) for collecting particles in a fluid on the surface thereof; a heating unit (20) for heating the collection substrate; a detection unit (30) for detecting particles derived from an organism among the particles collected on the collection substrate; and a control device (50). The collection substrate is formed from a material that has a heat resistance of at least 270°C. The control device controls the heating unit so that the collection substrate is heated at a temperature lower than 270°C. The detection unit comprises a light source (31) for irradiating the surface of the collection substrate with excitation light and a light receiving element (32) for receiving fluorescence. The detection unit detects particles derived from an organism on the basis of the fluorescence from the surface of the collection substrate after heating.

Description

Detection device

The present invention relates to a detection device, and more particularly to a detection device for collecting and detecting biological particles.

Conventionally, as a method for counting microorganisms in a sample such as air, a method is generally used in which microorganisms are collected on a medium in a petri dish and then cultured, and colonies formed by the microorganisms are manually counted.

For example, Japanese Patent Application Laid-Open No. 2009-55790 (hereinafter referred to as Patent Document 1) discloses a pseudo medium composed of silicone rubber for collecting aerosols which are microorganisms in the air. Patent Document 1 discloses that microorganisms collected on a pseudo medium are collected by a collecting liquid and then counted by a measuring instrument such as flow cytometry.

JP 2009-55790 A

However, the method of culturing in a medium and counting colonies has a problem that it takes a long time to obtain a detection result, that is, the detection result cannot be obtained in a short time. In addition to the configuration for detection, there is a problem that it is necessary to maintain an environment and a facility for culturing microorganisms and an environment for maintaining the medium in an appropriate state.

Also, there is a problem that complicated manual work from collection to detection is necessary and automation is difficult. For example, the technique of Patent Document 1 requires a series of complicated operations of pre-fluorescence staining of microorganisms before measurement by flow cytometry, and measurement after collection and staining. Further, the technique disclosed in Patent Document 1 has a problem that the running cost for the detection operation is high because the detection operation requires consumables such as a collected liquid and a fluorescent reagent. Furthermore, in the technique of the above-mentioned patent document 1, since the microorganisms collected by the collected liquid are diluted, the problem that the detection may be difficult, and the used collected liquid cannot be returned to the air. There is also a problem that it must be discarded.

The present invention has been made in view of such problems, and an object of the present invention is to provide a detection device that can easily and accurately detect biologically-derived particles in a specimen.

In order to achieve the above object, according to one aspect of the present invention, the detection apparatus includes a collection substrate for collecting particles in the fluid on the surface, a heating mechanism for heating the collection substrate, A detection mechanism for detecting biologically-derived particles among the particles collected on the collecting substrate. The collection substrate is formed of a material having heat resistance of at least 270 ° C., and the heating mechanism includes a control mechanism for controlling the collection substrate to be heated at a temperature lower than 270 ° C. The detection mechanism includes a light source for irradiating excitation light onto the surface of the collection substrate and a light receiving element for receiving fluorescence, and is based on fluorescence from the surface of the collection substrate after being heated by the heating mechanism. To detect biological particles.

Preferably, the light reflectance of the collection substrate is 60% or more.
Preferably, the detection device further includes a collection mechanism for collecting particles with the collection substrate by introducing a fluid toward the collection substrate.

More preferably, the detection device further includes a movement mechanism for moving the collection substrate between a position where the collection mechanism is provided and a position where the detection mechanism is provided.

Preferably, the collection mechanism includes a nozzle substrate having one or more holes drilled toward the collection substrate, and sucks fluid toward the collection substrate through the holes drilled in the nozzle substrate.

According to the present invention, it is possible to easily and accurately detect biologically derived particles in a specimen using the detection device.

It is the schematic showing the structure of the detection apparatus concerning embodiment. It is the schematic showing the other example of the structure of the detection apparatus. It is the schematic which shows an example of a structure of the collection board | substrate contained in a detection apparatus. It is the schematic showing the 1st example of the collection part contained in a detection apparatus. It is the schematic of the specific example of the nozzle (hole) shape of the nozzle substrate contained in a collection part. It is the schematic showing the 2nd example of the collection part. It is the schematic showing the other example of the structure of the detection part contained in a detection apparatus. It is a block diagram which shows the specific example of a structure of the control apparatus contained in a detection apparatus. It is a flowchart showing the flow of control in a control apparatus. It is a block diagram which shows the specific example of a function structure of a control apparatus. Heat resistance and microorganism capture for PDMS (polydimethylsiloxane) as an example of thermosetting resin, PMMA (polymethyl methacrylate) as an example of thermoplastic resin, glass, gold as an example of metal, and agar medium It is a figure showing the evaluation result of collectivity. It is a figure showing the bright-field image (A) and fluorescence image (B) of mold | fungi when heating temperature is 25 degreeC. It is a figure showing the bright-field image (A) and fluorescence image (B) of mold | fungi when heating temperature is 50 degreeC. It is a figure showing the bright-field image (A) and fluorescence image (B) of mold | fungi when heating temperature is 100 degreeC. It is a figure showing the bright-field image (A) and fluorescence image (B) of mold | fungi when heating temperature is 130 degreeC. It is a figure showing the bright-field image (A) and fluorescence image (B) of mold | fungi when heating temperature is 150 degreeC. It is a figure showing the bright-field image (A) and fluorescence image (B) of mold | fungi when heating temperature is 200 degreeC. It is a figure showing the bright-field image (A) and fluorescence image (B) of mold | fungi when heating temperature is 250 degreeC. It is a figure showing the bright-field image (A) and fluorescence image (B) of mold | fungi when heating temperature is 270 degreeC. It is a figure showing the bright-field image (A) and fluorescence image (B) of mold | fungi when heating temperature is 280 degreeC. It is a figure showing the bright-field image (A) and fluorescence image (B) of mold | fungi when heating temperature is 300 degreeC. It is the figure showing the relationship between the heating temperature and the fluorescence intensity after a heating obtained by experiment. It is a figure showing the fluorescence image of colon_bacillus | E._coli before (A) and after (B) heating at 200 degreeC. It is a figure showing the fluorescence image of the fluorescent dust before (A) and after (B) heating at 200 degreeC.

Hereinafter, embodiments of the present invention will be described with reference to the drawings. In the following description, the same parts and components are denoted by the same reference numerals. Their names and functions are also the same. Therefore, these descriptions will not be repeated.

<Device configuration>
(overall structure)
FIG. 1 is a schematic diagram illustrating a configuration of a detection device 100 according to the present embodiment. Referring to FIG. 1, a detection device 100 includes a collection substrate 10 for collecting particles in the air, which is a fluid, on a surface, and a heating unit 20 that is a heating mechanism for heating the collection substrate 10. The detection unit 30 that is a detection mechanism for detecting biologically-derived particles among the particles collected on the collection substrate 10 and a control device 50 for controlling heating in the heating unit 20 are included. The detection unit 30 includes a light source 31 for irradiating the surface of the collection substrate 10 with excitation light and a light receiving element 32 for receiving fluorescence. Preferably, the detection apparatus 100 further includes a collection unit 40 that is a collection mechanism for collecting particles with the collection substrate 10 by introducing air toward the collection substrate 10.

FIG. 2 is a schematic diagram illustrating another example of the configuration of the detection apparatus 100. With reference to FIG. 2, the collection part 40, the heating part 20, and the detection part 30 may be installed in the position which left | separated. By doing in this way, the damage to the detection part 30 by the heating can be suppressed.

In this case, the detection apparatus 100 moves the moving unit 60 that is a moving mechanism for moving the collection substrate 10 between the position where the collection unit 40 and the heating unit 20 are provided and the position where the detection unit 30 is provided. In addition. The control device 50 is also electrically connected to the moving unit 60 and controls the movement of the collection substrate 10 by the moving unit 60. As the moving unit 60, for example, an electric stage (linear movement, rotation) or the like is preferably used. When the detection device 100 has such a configuration, the degree of freedom of arrangement of the optical system (detection unit 30) can be improved. As a result, fluorescence from biologically derived particles can be detected with higher accuracy, and detection accuracy can be improved.

Moreover, it is good also as a position which left | separated the collection part 40 and the heating part 20 by the said structure. In this case, the moving unit 60 may be a moving mechanism for moving between the position where the collecting unit 40 is provided, the position where the heating unit 20 is provided, and the position where the detecting unit 30 is provided. By doing in this way, the damage by the heating to the collection part 40 can be suppressed.

(Collection substrate 10)
The collection substrate 10 is formed of a heat resistant material. FIG. 3 is a schematic diagram illustrating an example of the configuration of the collection substrate 10. That is, as shown in FIG. 3, the collection substrate 10 may have a heat resistant substrate 11 having heat resistance formed on the support substrate 12. The characteristics of the collection substrate 10 will be described later.

As will be described later, in the detection apparatus 100, particles in the air are collected on one surface of the collection substrate 10, and fluorescence from the particles is received by the light receiving element 32 disposed on the one surface side. Therefore, if the collection substrate 10 transmits light, about 50% of the fluorescence from the particles is transmitted to the surface opposite to the one surface, and the light receiving element 32 receives light. Not. That is, in this case, the light receiving element 32 receives only about 50% of the fluorescence from the particles.

Therefore, preferably, the surface of the collection substrate 10 has light reflectivity. The structure of the collection substrate 10 whose surface is reflective is the case where the surface of the collection substrate 10 (the surface on the one surface side) itself is made of a material having light reflectivity, as shown in FIG. In this case, the heat resistant substrate 11 transmits light, but the surface on the heat resistant substrate 11 side of the support substrate 12 has light reflectivity. In the latter case, for example, silicon or metal is preferably used as the material of the support substrate 12. In this case, the heat-resistant substrate 11 is formed on the support substrate 12 by separately forming and bonding the support substrate 12 and the heat-resistant substrate 11, and forming the heat-resistant substrate 11 on the support substrate 12 as a thin film (spin And the like, and a method of forming the support substrate 12 in a thin film (such as spin coating) on the bottom surface of the heat-resistant substrate 11 are preferably used.

The reflectance Y (%) of the surface of the collection substrate 10 is preferably a reflectance at which the light receiving element 32 can receive fluorescence of 80% or more of the total light emission amount. At this time, since 50% (upper) + (50% (lower) × Y / 100) ≧ 80%, Y = 60, that is, the reflectance Y (%) of the surface of the collection substrate 10 is 60% or more. .

If the surface of the collection substrate 10 has light reflectivity, the excitation light emitted from the light source 31 may be reflected by the surface of the collection substrate 10 and enter the light receiving element 32 as stray light. In this case, as will be described later, it can be prevented by arranging an optical filter to cut the excitation light or arranging the light source 31 and the light receiving element 32 on an oblique axis.

(Heating unit 20)
As the heating unit 20, for example, a ceramic heater, a far infrared heater, a far infrared lamp, or the like is preferably used. The heating unit 20 heats the collection substrate 10 according to a control signal from the control device 50.

(Collector 40)
FIG. 4 is a schematic diagram illustrating a first example of the collection unit 40. The configuration shown in FIG. 4 is a configuration for collecting particles in the air by the collection substrate 10 using the inertial collision method. Referring to FIG. 4, the collection unit 40 includes a nozzle substrate 41 that is arranged in parallel or substantially in parallel with the collection substrate 10, a pressure control device 42 that is a mechanism for controlling the pressure in the detection device 100, and including.

For example, a fan, a pump, a pressure controller, or the like is preferably used as the pressure control device 42, and the outside air is taken into the detection device by exhausting the air inside the detection device 100, and the outside air is directly taken into the detection device 100. There is something to introduce. In the former case, the pressure control device 42 is arranged on the suction side with the collection substrate 10 sandwiched between the nozzle substrate 41 (see FIG. 4). In the latter case, the pressure control device 42 is disposed on the exhaust side with the nozzle substrate 41 sandwiched between the pressure control device 42 and the collection substrate 10.

The pressure control device 42 operates according to a control signal from the control device 50. In the former case, when the pressure control device 42 is operated, an air flow as indicated by the dotted arrow in FIG. 4 is generated in the device of the detection device 100. That is, outside air is introduced to the collection substrate 10 through the nozzles of the nozzle substrate 41, and is exhausted outside the apparatus through the collection substrate 10. The particles in the outside air collide with the surface of the collection substrate 10 due to the inertial force caused by the air flow and are collected.

By using the pressure control device 42, more specimens (air) can be guided onto the collection substrate 10 as compared with the case where the pressure control device 42 is naturally dropped and collected. For example, when a suction fan of 100 L / min is used, 1000 L (1 m ³ ) of specimen (air) can be introduced as a measurement target with 10 minutes of suction, so that the amount of biological particles can be accurately calculated. Can do.

The material of the nozzle substrate 41 is not limited to a specific material. For example, stainless steel, resin, glass, and other metals are preferably used. Preferably, the diameter of the nozzle (hole) is about 0.01 mm to 10 mm, and the nozzle length (that is, the thickness of the nozzle substrate 41) is about 0.1 mm to 50 mm. The distance from the nozzle substrate 41 to the collection substrate 10 is preferably about 0.01 mm to 10 mm. The nozzle shape is not limited. For example, the nozzle substrate 41 may be provided with circular nozzles (holes) as shown in FIG. 5A, or rectangular or slits as shown in FIG. 5B. A shaped nozzle (hole) may be provided. In addition, a taper shape etc. may be sufficient. Also, the number is not limited and may be about 1 to 1000.

FIG. 6 is a schematic diagram illustrating a second example of the collection unit 40. The configuration shown in FIG. 6 is a configuration for collecting particles in the air by the collection substrate 10 using electrostatic induction. With reference to FIG. 6, the collection unit 40 includes a discharge electrode 43 instead of the nozzle substrate 41 of FIG. 4. The discharge electrode 43 is electrically connected to the negative electrode of the high voltage power source. The positive electrode of the high voltage power supply is grounded. Thereby, particles in the air in the detection device 100 are negatively charged in the vicinity of the discharge electrode 43. The collection substrate 10 is grounded. As a result, the negatively charged particles in the air move toward the collection substrate 10 by electrostatic force, and are adsorbed and collected on the surface thereof. The relationship between the negative electrode and the positive electrode may be reversed.

(Detector 30)
The detection unit 30 includes a light source 31 and a light receiving element 32. As the light source 31, for example, a semiconductor laser, an LED (Light Emitting Diode), a lamp, or the like is preferably used. The wavelength of the excitation light may be in the ultraviolet or visible region as long as it excites biological fine particles and emits fluorescence. Preferably, it is 300 nm to 450 nm, which is contained in the microorganism and excites the fluorescent tryptophan, NADH, riboflavin and the like efficiently. As the light receiving element 32, for example, a photodiode, an image sensor, an area sensor, or the like is preferably used. The light receiving element 32 outputs a signal corresponding to the received light intensity to the control device 50.

When the collection part 40 is the structure which collects the particle | grains in the air with the collection board | substrate 10 using the inertial collision method represented by FIG. 4, as mentioned above, the collection board | substrate 10 from the nozzle board | substrate 41 is used. The distance between the light source 31 and the light receiving element 32 is preferably as shown in FIG. 1 in order to avoid the influence of the nozzle substrate 41 such as scattering. An area away from directly above the collection substrate 10 is arranged at an angle with respect to the normal direction of the collection substrate 10, that is, on the oblique axis.

As shown in FIG. 2, when the detection device 100 is configured to be installed at a position where the collection unit 40 and the detection unit 30 are separated from each other, preferably, as shown in FIG. 2, The light source 31 and the light receiving element 32 are arranged so that the excitation light does not enter the light receiving element 32 directly, that is, the oblique axis, that is, the optical axes do not coincide with each other.

More preferably, as shown in FIG. 7A, the detection unit 30 uses a light source to collect the excitation light from the light source 31 in a region where particles on the collection substrate 10 are collected. A lens 33 disposed in the vicinity of 31, a fluorescence detection lens 34 disposed in the vicinity of the light receiving element 32 in order to collect fluorescence from biological particles on the light receiving element 32, and the excitation light is cut off. And a fluorescence detection filter 35 disposed in the vicinity of the light receiving element 32 for allowing the fluorescence to pass therethrough. The same applies to the case where the detection device 100 illustrated in FIG. 2 is configured to be installed at a position where the collection unit 40 and the detection unit 30 are separated from each other.

Further, as described with reference to FIG. 7B, when the detection device 100 is configured to be installed at a position where the collection unit 40 and the detection unit 30 are separated from each other, the detection unit 30 is further dichroic. The light source 31 and the light receiving element 32 may be arranged coaxially by using an optical element 36 for selecting a wavelength, such as a mirror.

(Control device 50)
The control device 50 is preferably a general computer (for example, a personal computer).

The control device 50 is electrically connected to the heating unit 20 and controls heating in the heating unit 20. The control device 50 is further electrically connected to the heating unit 20 and the detection unit 30, and controls heating by the heating unit 20 and light emission / light reception by the detection unit 30. Further, the control device 50 detects biologically-derived particles from the surface of the collection substrate 10 based on the fluorescence from the surface of the collection substrate 10 after being heated by the heating unit 20 using the signal from the light receiving element 32. To do.

FIG. 8 is a block diagram showing a specific example of the configuration of the control device 50, and shows the configuration of a general computer. That is, referring to FIG. 8, control device 50 includes a CPU (Central Processing Unit) 50a which is an arithmetic device for controlling the entire device, and a ROM which is a memory for storing programs executed by CPU 50a. (Read Only Memory) 51, RAM (Random Access Memory) 52, which is a memory used as a work area when the CPU 50a executes a program, and HDD (Hard Disk Drive) 53, a memory for storing detection results and the like. And a communication interface (I / F) 54 for transmitting a control signal and receiving an input of a detection signal by communicating with the heating unit 20 and the like. In addition, the control device 50 may include an input device for receiving a user operation, a display device for displaying a detection result, a transmission device for transmitting the detection result to another device, and the like.

<Detection principle>
The fluorescence intensity from biologically derived particles has the property of increasing with heating. On the other hand, the fluorescence intensity from particles that are not derived from organisms such as fluorescent dust does not increase by heating. This is presumably because a brown substance (melanoidin) having fluorescence is formed by the Maillard reaction generated by heating the sugar and amino acids contained in the biological particles. In addition, the detection principle which isolate | separates and detects from the non-biological origin particle | grains using the increase in the fluorescence intensity from a biological origin particle | grain by heating is also disclosed by international publication WO2011 / 104770 by this applicant. Yes. Therefore, using this characteristic, the detection device 100 separates and detects biologically derived particles from dust that emits fluorescence among the collected particles using at least the fluorescence intensity after heating. Preferably, the detection device 100 detects biological particles based on changes in fluorescence intensity before and after heating.

FIG. 9 is a flowchart showing a control flow in the control device 50. Referring to FIG. 9, when starting the detection operation, control device 50 first executes a collection operation (step S1). In the collecting operation in step S1, when the collecting unit 40 has the configuration shown in FIG. 4, the pressure control device 42 is operated at a specified control amount, and the outside air is taken into the detection device 100 and the nozzle substrate. In this operation, the nozzle 41 is passed through and introduced to the collection substrate 10. When the collection unit 40 has the configuration shown in FIG. 6, the collection operation in step S <b> 1 takes the outside air into the detection device 100 by operating the pressure control device 42 with a specified control amount, and discharge electrodes. This is an operation in which the particles in the vicinity are negatively charged by being applied to 43 and adsorbed to the collection substrate 10 by electrostatic force.

The control device 50 preferably performs the measurement operation after performing the collection operation of step S1 for a predetermined period (step S2). The measurement operation in step S <b> 2 is an operation of irradiating excitation light from the light source 31 for a predetermined time and receiving an input of a signal corresponding to the fluorescence intensity received from the light receiving element 32. When the detection device 100 is configured to be installed at a position where the collection unit 40, the heating unit 20, and the detection unit 30 are separated as shown in FIG. An operation for moving the collection substrate 10 set at the position of the unit 40 and the heating unit 20 to the position of the detection unit 30 is included. That is, the measurement in step S <b> 2 is performed after the collection substrate 10 has moved to the detection unit 30.

When performing the measurement operation in step S2, the control device 50 calculates the fluorescence intensity F1 using the signal input from the light receiving element 32 (step S3). In step S3, for example, the control device 50 stores a relational expression between the signal intensity and the fluorescence intensity in advance, and obtains the fluorescence intensity F1 before heating by substituting the signal intensity from the light receiving element 32 into the expression. be able to.

The control apparatus 50 performs the operation | movement which heats the collection board | substrate 10 (step S4). The heating operation in step S4 is an operation for causing the heating unit 20 to generate heat with a predetermined amount of heat. After the heating operation in step S4, the control device repeats the same measurement operation and calculation operation as in steps S2 and S3 to obtain the fluorescence intensity F2 after heating (steps S5 and S6).

The control device 50 calculates the amount of biological particles using at least the fluorescence intensity F2 after heating (step S7). Preferably, the control device 50 executes the measurement operation and the calculation operation in steps S2 and S3 to obtain the fluorescence intensity F1 before heating, and calculates the amount of biological particles using the fluorescence intensity F1 and F2 before and after heating. (Step S7).

In step S7, for example, the control device 50 stores in advance a relational expression between the fluorescence intensity F2 after heating and the amount of biological particles (concentration, etc.), and the fluorescence intensity F1 obtained from the relational expression is stored. By substituting, the amount of biological particles can be obtained. Further, for example, the control device 50 stores a relational expression between the difference ΔF between the fluorescence intensities F1 and F2 before and after heating and the amount of biological particles, and substitutes the calculated difference ΔF into the relational expression. The amount of biological particles can be obtained.

<Functional configuration>
FIG. 10 is a block diagram illustrating a specific example of a functional configuration of the control device 50 for performing the above operation. Each function of FIG. 10 is mainly realized by the CPU 50a by the CPU 50a of the control device 50 reading the program stored in the ROM 51 onto the RAM 52 and executing it. However, at least a part of the functions may be realized by a hardware configuration of the control device 50 shown in FIG. 8 or a hardware configuration such as an electric circuit (not shown).

Referring to FIG. 10, the CPU 50 a outputs a control signal to the collection unit 40 via the communication I / F 54 to control the collection operation and the light source 31. A light emission control unit 502 for controlling the irradiation of excitation light from the light source 31 by outputting a control signal, and a heating control unit for controlling the heating operation by outputting a control signal to the heating unit 20 503 and the movement control unit 504 for controlling the movement of the collection substrate 10 by outputting a control signal to the movement unit 60 when the movement unit 60 is connected, and the communication I / F 54 A signal input unit 505 for receiving an input of a signal corresponding to the fluorescence intensity from the light receiving element 32, a fluorescence intensity calculating unit 506 for calculating the fluorescence intensity using a signal from the light receiving element 32, and the above relational expression in advance Remember Fluorescence intensity after heating or heating by using the difference of the fluorescence intensity before and after, and a particle amount calculation unit 507 for calculating the amount of particles from organisms.

<Selection of collection substrate material>
Since the above-described operation is performed by the detection apparatus 100 to detect biologically-derived particles, the collection substrate 10 has heat resistance (high melting point, non-water content), high collection property (stickiness) against microorganisms, flatness. Performance (improvement of collection efficiency, focus position at detection, no scattering due to unevenness), workability, low cost, and low autofluorescence are required. When the collection substrate 10 has the configuration shown in FIG. 3, that is, when the heat-resistant substrate 11 is formed on the support substrate 12, the heat-resistant substrate 11 further has optical transparency ( Transparency) is also required.

When the collection substrate 10 has the configuration shown in FIG. 3, the support substrate 12 has heat resistance, mechanical strength, adhesion to the collection substrate, workability, formability of the collection substrate, Flatness, low cost, low autofluorescence, and high reflectivity are required. Therefore, silicon, glass, and metal (gold, silver, platinum, etc.) are preferably used for the support substrate 12. In particular, silicon is preferably used.

The inventor uses PDMS (polydimethylsiloxane) as an example of a thermosetting resin, PMMA (polymethyl methacrylate) as an example of a thermoplastic resin, glass, gold as an example of a metal, and Each agar medium was evaluated for heat resistance and ability to collect microorganisms. In this evaluation, in order to examine the collection efficiency on the surface of the collection substrate 10, the collection efficiency when the inertial collision method shown in FIG. 4 is used is particularly mentioned. This is because when the electrostatic induction shown in FIG. 6 is used, the collection efficiency is 100% if there is conductivity capable of applying a voltage. FIG. 11 is a diagram showing characteristics of each material. Referring to FIG. 11, among the selected materials, it was found that the material satisfying the heat resistance and the ability to collect microorganisms was PDMS as an example of a thermosetting resin.

Further, the inventor formed PDMS on a silicon substrate by spin coating in order to specify the heat resistance of the collection substrate 10 (particularly the heat-resistant substrate 11), and then thermally cured to obtain the collection substrate 10 as shown in FIG. Using the detection apparatus 100 configured as described above, an experiment for detecting biological particles was performed. In this experiment, fungi are used as an example of biologically derived particles. The experiment is carried out in the following steps;
Step 1: Air containing mold bacteria is sucked into the detection device 100 through the nozzle substrate 41 and collected on the collection substrate 10.
Step 2: heating the collection substrate 10 to a predetermined temperature by the heating unit 20;
Step 3: The light receiving element 32 acquires a bright field image and a fluorescent image of mold.

FIG. 12 to FIG. 21 are bright-field images and fluorescent images obtained by experiments, with heating temperatures of 25 ° C. (room temperature without heating), 50 ° C., 100 ° C., 130 ° C., 150 ° C., and 200 ° C., respectively. , 250 ° C., 270 ° C., 280 ° C., and 300 ° C., showing a bright-field image (A) and a fluorescent image (B). FIG. 22 is a diagram showing the relationship between the heating temperature and the fluorescence intensity after heating, obtained by experiments.

From these experimental results, it was found that the fluorescence intensity from mold fungi greatly increased by heating at 150 ° C. to 270 ° C. FIG. 23 is a fluorescence image of E. coli as another example of biological particles before (A) and after (B) heating at 200 ° C. For example, as shown in FIG. 23, it can be seen that the same increase in fluorescence intensity is also observed in particles derived from other organisms other than fungi. For this reason, it was found that the fluorescence intensity from biologically-derived particles is greatly increased by heating at 150 to 270 ° C. FIG. 24 shows fluorescence images of non-living particles (fluorescent dust) before and after heating at 200 ° C. As is clear from comparison of this image with FIGS. 12 to 21 and FIG. 23, it can be seen that the non-living particles do not increase in fluorescence intensity even in the above temperature range. Therefore, it was found that the biological particles can be separated from the non-biological particles and detected by heating in the above temperature range, and the biological particles can be detected with high accuracy.

Therefore, from this experiment, it is assumed that the collection substrate 10 (particularly the heat-resistant substrate 11) is formed of a material having heat resistance of at least 270 ° C. And the control apparatus 50 controls a heating operation so that the collection board | substrate 10 may be heated at temperature lower than the said heat-resistant temperature.

<Effect of Embodiment>
By adopting such a configuration for the detection apparatus 100, it is possible to easily and accurately detect biologically derived particles in the specimen.

In particular, by forming the collection substrate 10 with a material having heat resistance of at least 270 ° C., it is possible to heat the collection substrate 10 at a temperature not exceeding the heat resistance temperature after collecting particles in the specimen. Become. Experiments by the inventors have shown that heating from 150 ° C. to 270 ° C. greatly increases the fluorescence intensity from biologically derived particles. Therefore, by heating the collection substrate 10 to this temperature range, even if the collected particles contain non-living particles that emit fluorescence such as chemical dust, By comparing the fluorescence intensity with the threshold, or by using the difference in fluorescence intensity before and after heating, it is possible to separate biologically-derived particles from such non-biologically-derived particles and accurately detect them. Become.

Furthermore, at that time, by setting the reflectance of light on the surface of the collection substrate 10 to be a high reflectance such as 60% or more, for example, most of the emitted fluorescence (for example, 80% or more) is highly efficient. 32 is received. Therefore, it becomes possible to detect biologically derived particles with higher accuracy.

The embodiment disclosed this time should be considered as illustrative in all points and not restrictive. The scope of the present invention is defined by the terms of the claims, rather than the description above, and is intended to include any modifications within the scope and meaning equivalent to the terms of the claims.

10 collection substrate, 11 heat resistant substrate, 12 support substrate, 20 heating unit, 30 detection unit, 31 light source, 32 light receiving element, 33, 34 lens, 35 filter, 36 optical element, 40 collection unit, 41 nozzle substrate, 42 Pressure control device, 43 discharge electrode, 50 control device, 50a CPU, 51 ROM, 52 RAM, 53 HDD, 54 communication I / F, 60 moving unit, 100 detection device, 501 collection control unit, 502 light emission control unit, 503 Heating control unit, 504 movement control unit, 505 signal input unit, 506 fluorescence intensity calculation unit, 507 particle amount calculation unit.

Claims

A collection substrate for collecting particles in the fluid on the surface;
A heating mechanism for heating the collection substrate;
A detection mechanism for detecting biologically-derived particles among the particles collected on the collection substrate;
The collection substrate is formed of a material having heat resistance of at least 270 ° C.,
The heating mechanism includes a control mechanism for controlling the collection substrate to be heated at a temperature lower than 270 ° C.,
The detection mechanism includes a light source for irradiating excitation light to the surface of the collection substrate and a light receiving element for receiving fluorescence, and from the surface of the collection substrate heated by the heating mechanism. A detection device that detects the particles derived from the organism based on the fluorescence of.
The detection apparatus according to claim 1, wherein the collection substrate has a light reflectance of 60% or more.
The detection device according to claim 1, further comprising a collection mechanism for collecting the particles by the collection substrate by introducing the fluid toward the collection substrate.
The detection apparatus according to claim 3, further comprising a movement mechanism for moving the collection substrate between a position where the collection mechanism is provided and a position where the detection mechanism is provided.
The collection mechanism includes a nozzle substrate having one or more holes drilled toward the collection substrate, and sucks the fluid toward the collection substrate through the holes drilled in the nozzle substrate. The detection device according to claim 3.