JPH1183853A - Measuring method and device for floating fine matter in air - Google Patents

Abstract

PROBLEM TO BE SOLVED: To ensure an effective prophylaxis for pollinosis according to distribution of fine floating matters in the air by catching the matters, and obtaining a matrix of the pollen amount of the catched matters and the amount of diesel fine particles. SOLUTION: Fine matters catched are dividedly taken into a test tube 15, then pollen 16 and diesel fine particles 17 are sunk after they are left as they are quietly. After the lapse of a specified time, antigen-antibody reaction is made therewith by use of antibody specifically reacting with those 16, 17. The fluorescent antibody 18 composed of antibody selectively reacting with the allergen of pollen 16b floating in a test tube 15 and a fluorescent marker provided thereto and the fluorescent antibody 19 composed of the antibody specifically reacting with the particles 17 and a fluorescent marker provided thereto are charged into the tube 15 so that they are reacted with each other at a specified temperature and for a specified time. After the reaction, the antibodies 18, 19 which are not reacted are removed by washing. The marked antibodies are made to stick to carriers 20 in order to improve the sensitivity of detection. A pollen antigen-antibody reactive matter 21 and a diesel fine particle antigen-antibody reactive matter 22 are obtained in this way.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION The present invention relates to a process for producing fine substances (for example, pollen and diesel particulates, NO
_X ) and the like and a method for measuring suspended airborne substances in the atmosphere.

[0002]

BACKGROUND ART Conventionally, fine suspended solids (e.g. pollen, mites, house dust, dust, NO _X, etc.) in the air is given a big big impact in recent years on the environment, generated especially particular season Pollen affects the human body and causes, for example, allergic conjunctivitis and the like.

[0003] In recent years, studies on pollen allergy have been repeated, and many patients complain of allergy to cedar pollen and hinoki pollen among the causative substances of pollinosis. It has been presumed that a proposal has been made for a monoclonal antibody recognizing the antigen of cedar pollen (JP-A-4-33589).
No. 6). In addition, a method of analyzing suspended particulate matter in air, such as pollen, mites, and house dust, by flow cytometry using the monoclonal antibody has been proposed (JP-A-4-335896).

[0004]

However, in the above-mentioned method, however, only pollen in the atmosphere is measured, and other allergic factors are detected. Information related to air pollution needs to be promptly obtained. The present invention has been made in view of such circumstances, fine particulate matter floating in the air that can effectively perform prevention of pollinosis from the distribution of suspended matter in the air (e.g., pollen and diesel particulate, NO _X, etc.) distribution amount It is an object of the present invention to provide a measuring device for measuring the temperature and a measuring method thereof.

[0005]

According to a first aspect of the present invention, there is provided an apparatus for measuring suspended fine particles in the air, comprising: a collecting means for collecting the suspended fine substances in the atmosphere; A fluorescent labeled antibody that specifically reacts with pollen and diesel microparticles in the micro-substance that has undergone the antigen-antibody reaction, and a means for performing the antigen-antibody reaction, and excites the fluorescent substance after the antigen-antibody reaction and receives the fluorescence emitted by the fluorescent substance And detecting means for detecting minute substances in the atmosphere.

[0006] In the above-mentioned device for measuring minute substances, the pollen is cedar pollen and cypress pollen, and the diesel fine particles are carbon fine particles.

[0007] The measurement method of the present invention is to collect airborne fine substances in the atmosphere, obtain a matrix of the amount of pollen and the amount of diesel particulates of the collected fine substances, and measure the distribution in the atmosphere. It is characterized by.

[0008]

Embodiments of the present invention will be described below.

FIG. 1 is a schematic view showing a method for decomposing suspended microparticles according to the present invention. As shown in FIG. 1, the water is sucked from the atmosphere by a pump 12 provided outside by a collection tube 11 and directly introduced into collected water 14 in a collection container 13. The collected microscopic substances are separated into test tubes 15 and allowed to stand for pollen 16
And the diesel particulates 17 are settled. After a predetermined time, as shown in FIG.
An antigen-antibody reaction using an antibody that specifically reacts with 7 is performed. A fluorescently labeled antibody (F ₁ antibody) 18 with a fluorescent-labeled antibody which selectively reacts with allergens pollen 16 suspended in the test tube 17, with a fluorescently labeled antibody that specifically reacts with diesel particulate 17 The fluorescent labeled antibody (F ₂ antibody) 19 is charged into the test tube 15 and reacted at a predetermined temperature (for example, 37 ° C.) for a predetermined time (for example, 30 minutes). After completion of the reaction, washing is performed to remove unreacted fluorescently labeled antibodies.

Here, the diesel particulates 17 have a particle diameter of 0.6 to 1.2 μm, which is smaller than the particle diameter of pollen (40 μm). In this case, the sensitivity for receiving the fluorescence is low. Therefore, in order to improve the detection sensitivity, a plurality of the fluorescently labeled antibodies are attached to the carrier 20. Examples of the carrier 20 include beads such as fluoroapatite, latex, gelatin, erythrocytes, kaolin, bentonite, polyamino acids, and silica gel. In this way, the pollen and the diesel particulates are reacted with the fluorescently labeled antibodies 18 and 19 to obtain the pollen antigen-antibody reactant 2
1. After obtaining the diesel particulate antigen-antibody reactant 22, the amount in the air can be measured by the detection means.

As the fluorescent-labeled antibody 18, for example, a protein (Cry j) which is an antigen of cedar pollen, hinoki pollen, etc.
As a fluorescent substance, monoclonal antibodies that selectively react with 1) or antibodies that selectively react with diesel particulates
Reagent (FITC-KW) to which ITC (fluorescein isothiocyanate), phycoerythrin (PE), etc. are attached
-S10: trade name, manufactured by Kowa Corporation).

Since the measurement detects two types of reactants,
Pollen antibodies were labeled with FITC, and diesel particulate antibodies were labeled with PE. Fig. 5 shows FITC
2 shows an excitation spectrum and a fluorescence spectrum of PE and PE. As shown in FIG. 5, the maximum value of the fluorescence spectrum of FITC was around 525 nm (excitation wavelength: 495 nm), and the maximum value of the fluorescence spectrum of PE was around 576 nm (excitation wavelength: 564, 564).
576 nm).

As another fluorescent substance, for example, TRI-COLOR can be mentioned.

[Outline of Detection of Suspended Microsubstances] Next, FIG.
The outline of the process of detecting the collected minute substance will be described with reference to FIG. As shown in FIG. 4, the measuring means includes a micro flow cell (hereinafter, referred to as “cell”) 31 for continuously passing and measuring the solution containing the microsubstance after the antigen-antibody reaction.
A light source 32 having a wavelength necessary to excite the fluorescent substance in the micro-substance-containing solution in the cell 31, and a light-receiving part for receiving the fluorescence emitted from the fluorescent substance attached to the micro-substance after the antigen-antibody reaction. It comprises an element 33 and a counter 34 for counting the output of the light receiving element.

In FIG. 4, reference numerals 36 and 37 are mirrors,
38 is a pinhole slit, 39, 40 and 41 are condenser lenses, 42 is a bandpass filter, and 43 is a field stop.

The cell 31 has a rectangular section whose cross section orthogonal to the flow direction of the object to be measured of the liquid sending pump 35 of the measuring section is long in the horizontal direction and short in the vertical direction, and is irradiated with the excitation light. Reactant 2 from the front that is long in the horizontal direction
The light emitted from the fluorescent substance attached to the light receiving elements
To receive light.

Since FITC and PC are used as fluorescent labels for detecting pollen and diesel particulates, respectively, half mirrors 44 and 45 are provided in the optical path of the reflected light as shown in FIG. The first light receiving element 33A, the second light receiving element 33B, the first pulse counter 34A, the second pulse counter 34B, and the first bandpass filter 4
2A and a second band-pass filter 42B are provided to detect two wavelengths. FIG. 3 shows an example of the detection data. Since FIG. 3 (a) detecting the amount of antibody F _1, one of the amount of pollen has been measured as a pulse, is it detecting (b) an antibody amount of F _2, diesel particulate Indicates the amount of the plurality adhered.

FIG. 3C shows the results of the measurement as a matrix. In this table, the horizontal axis is the fluorescence intensity of pollen,
The case where the vertical axis is the fluorescence intensity of diesel particulates is shown. In the figure, A indicates that the amount of pollen is large, B is slightly large, C is small, and D is not detected. Therefore, statistical observation information can be provided by comparing the amount of pollen and the amount of diesel particulates in the air in various places at a specific time on a specific day. This makes it possible to predict hay fever from the correlation between the amount of pollen and the amount of diesel particulates at the same time.

[0019]

As described above, according to the present invention, the collecting means for collecting airborne fine substances, and the pollen and diesel particulates among the fine substances collected by the collecting means. Means for performing an antigen-antibody reaction using a fluorescent-labeled antibody capable of performing an antigen-antibody reaction, and detection means for exciting a fluorescent substance after the antigen-antibody reaction and receiving fluorescence emitted from the fluorescent substance to detect minute substances in the atmosphere Therefore, the amount of pollen in the atmosphere and the amount of diesel particulates can be measured at the same time, and the prediction of hay fever due to local factors can be made in advance from these relationships.

By collecting airborne fine substances in the atmosphere and obtaining a matrix of the amount of pollen and the amount of diesel particulates of the collected fine substances, the distribution in the air can be measured. Early prediction of the disease is possible.

[Brief description of the drawings]

FIG. 1 is a configuration diagram of a reaction step of the present invention.

FIG. 2 is a conceptual diagram of the antigen-antibody reaction of the present invention.

FIG. 3 is a diagram illustrating an example of a detection result according to the present invention.

FIG. 4 is a configuration diagram of a detection unit of the present invention.

FIG. 5 shows the fluorescence spectra of FITC and PE.

[Explanation of symbols]

REFERENCE SIGNS LIST 11 collection tube 12 pump 13 collection container 14 collected water 15 test tube 16 pollen 17 diesel particulate 18 fluorescent labeled antibody (F ₁ antibody) 19 fluorescent labeled antibody (F ₂ antibody) 20 carrier 21 pollen antigen antibody reactant 22 diesel Microparticle antigen-antibody reactant 31 Micro flow cell (cell) 32 Light source 33A, 33B Light receiving element 34A, 34B Counter 36, 37 Mirror 38 Pinhole slit 39, 40, 41 Condensing lens 42A, 42B Bandpass filter 43 Field stop

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Rieko Endo 12th Nishikicho, Naka-ku, Yokohama-shi, Kanagawa Prefecture

Claims (3)

[Claims] 1. A collecting means for collecting airborne fine substances, and a fluorescently labeled antibody which specifically reacts with pollen and diesel particulates in the fine substances collected by the collecting means. Means for causing an antigen-antibody reaction, and detection means for exciting the fluorescent substance after the antigen-antibody reaction, receiving fluorescence emitted by the fluorescent substance, and detecting minute substances in the air. For measuring suspended microscopic substances. 2. An apparatus for measuring minute suspended matter in air according to claim 1, wherein said pollen is cedar pollen and cypress pollen, and said diesel fine particles are carbon fine particles. 3. A method of collecting airborne fine substances in the air, obtaining a matrix of the amount of pollen and the amount of diesel particulates of the collected fine substances, and measuring the distribution in the air. A method for measuring suspended particles collected from inside.