JP2008096155A - Fine particle detector - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a fine particle detector for rapidly separating and detecting various kinds of fine particles, at high sensitivity. <P>SOLUTION: The fine particle detector is constituted so as to detect fine particles, by subjecting a sample containing fine particles to fine tube electrophoresis using a migration liquid and equipped with an exciting light emitter 2 for irradiating fine particles subjected to fluorescent dyeing so as to be selectively bonded and a fluorescence detector 5 for detecting the fluorescence emitted from the fine particles, subjected to fluorescence dyeing using an optical system. At least one of either the optical system which contains the exciting light emitter 2 and the fluorescence detector 5, or a fine tube 6 is made movable in a scanning direction and the fine particles concentrated in the fine tube 6 is detected from the image signal of the fluorescence detector 5 for scanning the fine particles, while making them move relatively. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a microparticle detection apparatus for detecting various microparticles such as cells existing in a specimen sample by using microtubule electrophoresis, and particularly, for rapidly and highly sensitively detecting and detecting microorganisms such as Legionella spp. Harmful to the human body. The present invention relates to a fine particle detection apparatus capable of performing the above.

  In recent years, as a separation / detection method for examining the presence or absence of various microparticles such as cells in a sample, and for quickly examining the type and amount of microorganisms contained in a sample, fluorescent staining has been performed in advance in a microtubule. There has been proposed an electrophoresis technique in which microorganisms are electrophoresed and irradiated with excitation light and detected from fluorescence generated at that time (see, for example, Patent Document 1).

  Further, although there is a paper on a technique for concentrating fine particle cells such as microorganisms to the pH region of the isoelectric point of the microbe by means of microtube isoelectric focusing technology, details of the specific detection means are not described (for example, Non-Patent Document 1).

  On the other hand, when the microorganisms are electrophoretically separated in the microtubules, the interaction between the microbial cells and the inner walls of the microtubules or the cells and the cells is large. It is known that it is effective to contain an alginate (see, for example, Patent Document 2).

  In this method, the alginate used as an additive to improve the separation efficiency of microorganisms does not function in the same way for all microorganisms, and more types of separation efficiency can be improved to cope with a wide variety of microorganism species. The introduction of agents is expected.

In order to promptly take appropriate measures against environmental water, food contamination, and infectious diseases, and to quickly identify and deal with the contamination source and infection source, it is necessary to quickly and accurately detect the source of contamination and the source of infection. Specific detection methods are required.
JP 2002-345451 A JP 2002-181781 A Microbial electrophoresis separation and detection method (Monthly Food Chemical 2005-2)

  Accordingly, an object of the present invention is to provide a particle detection apparatus that can rapidly and highly sensitively detect and detect various particles such as microorganisms.

  In order to achieve the above-described object, the particle detector of the present invention is a fluorescent device that selectively binds in a particle detector that detects particles by subjecting a sample containing particles to microtubule electrophoresis using an electrophoresis solution. An excitation light emitter for exciting and irradiating stained fine particles, and a fluorescence detector for detecting fluorescence emitted from the fluorescently stained fine particles using an optical system, and an excitation light emitter and a fluorescence detector are provided. It is characterized in that at least one of the optical system and the microtubule included can be moved in the scanning direction, and the fine particles concentrated in the microtube are detected from the detection signal of the fluorescence detector that scans while relatively moving. And

  In this case, an image signal recording device that records a signal detected by the fluorescence detector as an image signal, and a particle detection control device that records a particle detection signal obtained by detecting particles from the image signal can be provided.

  Further, the particle detection signal can be the luminance of the image, the area of the image, or the number of particles.

  In addition to providing a display device, it is possible to provide an applied voltage control function in microtube electrophoresis, a function of monitoring current flowing through the microtube, or a function of recording a particulate detection signal.

  Further, it is possible to incorporate a waveform control device that can independently vary the magnitude when displaying a recording waveform such as an applied voltage signal, a micro-tube current signal, or a particulate detection signal.

  Moreover, the microparticles | fine-particles which are detection objects can be used as microorganisms harmful | toxic to human bodies, such as microorganisms, for example, Legionella genus bacteria.

  According to the microparticle detection apparatus of the present invention, a microparticle is electrophoresed on a sample containing microparticles by using an electrophoretic solution to excite the fluorescently stained microparticles so as to selectively bind in the microparticle detection apparatus that detects microparticles. An optical system including an excitation light emitter and a fluorescence detector, and an excitation light emitter that irradiates and a fluorescence detector that detects fluorescence emitted from the fluorescently stained fine particles using an optical system. Since at least one of the tubes can be moved in the scanning direction and the fine particles concentrated in the microtube are detected from the detection signal of the fluorescence detector that scans while moving relatively, special technology is required. In addition, the fine particles contained in the sample are separated with high resolving power and high reproducibility, and the target fine particles are fluorescently stained in advance so that even a very small amount of fine particles can be obtained. It can detect with high sensitivity without breeding treatment such as nourishment, and can detect desired fine particles with higher resolution, and use a CCD image sensor or the like as a detector for fine particles mixed in the sample. Thus, not only signal detection but also quantitative detection can be performed from the obtained image signal.

  In addition, an image signal recording device that records an image signal detected by a fluorescence detector and a particle detection control device that records a particle detection signal in which particles are detected from the image signal are provided. Images can be stored and recorded in a memory such as a personal computer.

  Further, it is possible to confirm that the image processing is operating normally by setting the particle detection signal to the brightness of the image, the area of the image, or the number of particles.

  In addition to providing a display device and a function of controlling applied voltage in microtube electrophoresis, a function of monitoring a current flowing through a microtube, or a function of recording a particle detection signal, automatic control of particle detection can be performed in an integrated manner.

  In addition, it has a built-in waveform control device that can independently vary the magnitude when displaying a recording waveform such as an applied voltage signal, a micro-tube current signal, or a particulate detection signal, making it easy to see waveform recordings that become longer over time. Can be displayed on the screen.

  In addition, microparticles that are detection targets are microorganisms, for example, microorganisms that are harmful to the human body such as Legionella, so microorganisms that are harmful to the human body such as Legionella that are likely to live in hot springs or pools Can be detected.

  Hereinafter, embodiments of the particulate detection device of the present invention will be described with reference to the drawings.

The fine particle detection apparatus of the present invention separates various fine particles contained in a sample with high resolution and high reproducibility by utilizing a microtubule electrophoresis method using an electrophoresis solution.
In addition, in this microtubule electrophoresis, by using a fluorescence detector as a detection means, the target microparticles are preliminarily fluorescently stained, so that even a minute amount of microparticles can be highly sensitive without being subjected to growth treatment or the like. Therefore, the desired fine particles can be detected with high accuracy with an even higher resolution.
Furthermore, the fine particles mixed in the sample can be detected not only simply but also quantitatively by performing image processing.
The detection in the present invention includes the concentration of desired fine particles contained in a sample, separation from others, qualitative detection, quantitative detection, screening, life / death discrimination, and the like.

Specifically, for example, it is a fine particle detection device having the following part or all as constituent elements.
(1) A sample containing fine particles is subjected to microtubule electrophoresis using an electrophoresis solution.
(2) A sample containing fine particles is fluorescently stained so as to selectively bind to the fine particles, and this is subjected to microtubule electrophoresis to detect fluorescence of the fluorescently stained fine particles.
(3) A device for detecting microparticles using microtubule electrophoresis, a water tank containing an electrophoretic solution, a microtubule for concentrating and separating microparticles injected into the electrophoretic solution, and a detection for detecting the concentrated microparticles Means.
(4) An apparatus for detecting microparticles using microtubule electrophoresis, wherein the microparticles are concentrated by a voltage gradient provided in the microtube.
(5) A fluorescence detector is provided as a detection means. The fluorescence-stained microparticles are irradiated with excitation light, and the fluorescence emitted from the fluorescence-stained microparticles is detected using an optical system. Detect fine particles.
(6) Further, an image signal recording device that records a signal detected by fluorescence in the optical system as an image signal, and a particle detection control device that records a particle detection signal that detects particles from the image signal are provided.
(7) Further, the fine particle detection signal is the luminance of the image, the area of the image having a signal exceeding the specified luminance, the number of fine particles detected by characterizing the image spatially and temporally, and the like.
(8) The fine particle detection control device has a function of controlling applied voltage in micro tube electrophoresis, a function of monitoring a current flowing through a fine tube, a function of recording a fine particle detection signal, and the like, and recording waveforms such as a current in the fine tube and a fine particle detection signal. It incorporates a waveform control device that allows each display size to be varied independently.

FIG. 1 shows an example of the basic configuration of the microtubule electrophoresis system of the present invention.
In FIG. 1, reference numeral 1 denotes a particle detector main body, which is composed of an excitation light emitter 2 such as a diode and an objective lens 4, for example, a fluorescence detector 5 such as a CCD camera or a photoelectric tube.
Fine particles such as harmful bacteria that are detection objects, for example, microorganisms harmful to the human body such as Escherichia coli are fluorescently stained in advance so as to selectively bind only to the microorganisms, for example, inside the microtubule 6 such as a capillary. The solution 9a or 9b installed at both ends is injected.
The solution 9a or the solution 9b is changed according to the detection stage, and a high voltage power source is applied to both ends of the microtube 6 by the electrodes 7a and 7b. The high voltage is supplied from the high voltage power source 7, and the magnitude of the voltage, the application time, and the like are controlled by the particulate detection control device 8 such as a personal computer, for example.

Excitation light 2 a emitted from the excitation light emitter 2 is irradiated to the fluorescently stained fine particles from the detection window 6 a of the microtube 6.
The fluorescently stained fine particles emit fluorescence 2b by excitation light, enter the apparatus main body through the objective lens 4, and are imaged and detected by a fluorescence detector 5 constituted by, for example, a CCD camera or the like.
The detected image signal detected by the fluorescence detector 5 is recorded in the image signal recording device 5a and connected to the particulate detection control device 8. The fine particle detection control device 8 detects fine particles from this image signal, and records and displays the result.

The fluorescently stained fine particles are concentrated in the detection window 6a by a known electrophoresis technique (Non-Patent Document 1) such as isoelectric focusing.
In order to detect the concentrated fine particles, the micro tube 6 fixed to the arm 11 is formed in a substantially circular shape, and the motor 10 is connected to the center of the micro tube 6 formed in a substantially circular shape, that is, the center 12 of the arm 11. To do.
The arm 11 is rotated by the motor 10, the fine tube 6, the detection window 6a, the solutions 9a, 9b, etc. are integrally rotated about 12, and the distance between the objective lens 4 and the detection window 6a is kept constant, and the detection window 6a Scan while relatively moving from end to end. In this scanning process, a fine particle detection operation is performed.
In this embodiment, the micro tube 6 is rotated by the motor 10. However, the micro tube 6 is fixed, and the motor is attached to the entire optical system including the excitation light emitter 2 and the particle detector 1. You may rotate by the locus | trajectory of the circle used as the center.
That is, the entire optical system including the excitation light emitter 2 and the objective lens 4 and at least one of the detection window 6a can be moved in the scanning direction, and scanning is performed from end to end of the detection window 6a in exactly the same manner. In this scanning process, a fine particle detection operation can be performed.

FIG. 2 shows the state of migration of harmful bacteria in the microtubule 6, and the following detection process is performed. A test solution, which is a sample including the fine particles 21 as a detection target, is prepared as, for example, a solution 9a, and is sucked from the solution 9b side to be injected into the microtube 6 as the target test solution 20.
Next, acidic and alkaline solutions are used as the solutions 9a and 9b, the electrodes 7a and 7b are connected to the high-voltage power source 7, and the fine particle detection control device 8 is connected so as to apply a high voltage to the electrodes 7a and 7b. To do. Next, when the high-voltage power supply 7 is operated, a potential gradient is generated in the sample solution 20 in the microtube 6, and as a result, the entire liquid in the migration region in the microtube has a pH between the cathode direction and the anode direction due to the isoelectric point phenomenon. Generate a gradient.

By this known microtubule isoelectric focusing technique, the fine particles that may be present in the sample solution 20 can be concentrated in a region (isoelectric point position) having the same pH condition as the isoelectric point of the fine particle cells (the isoelectric point position). For example, refer nonpatent literature 1 etc.).
In general, since the surface charge component of the fine particles has a specific isoelectric point, the fine particles are concentrated at the isoelectric point position, which is a specific location, by the pH gradient in the microtube. The isoelectric point position varies depending on the type of target fine particles.
In this state, the electrophoretic current between the electrodes 7a and 7b is in a steady state and hardly flows.

In FIG. 2, the entire optical system including the excitation light emitter and the objective lens 4 is rotated in the direction of 26 along the detection window 6a of the microtube 6 without changing the relative distance between the detection window 6a and the objective lens 4. Although schematically shown as a figure, the microtube 6 may be rotated by the motor 10 as shown in FIG.
At this time, the presence of the concentrated fine particles 27 is detected by the fluorescence detector 5 through the objective lens 4.

  The method of injecting the specimen sample into the microtube 6 is not particularly limited, and any of the conventionally used gravity method, pressurization method, and decompression method can be used. Although the injection amount is not particularly limited, it depends on the size of the microtubule in order to fill the sample in the entire microtubule to be used, and it is 0.1 to 100 μL, preferably 0.5 to 100 μL, more preferably 0.5 to 10 μL. Can be illustrated.

In addition, the pair of electrodes 7a and 7b and the high-voltage power source 7 exemplified above have a potential gradient in the intensity required for the fine particles injected into the microtube to migrate in the electrophoresis solution in the microtube. It is means for applying to the electrophoresis solution.
Any means (other means for applying a potential gradient) can be used without being limited to these (power supply and a pair of electrodes) as long as the object can be achieved.

The voltage applied between the electrodes is generally about 1 kV / m to about 500 kV / m, preferably about 2 kV / m to about 100 kV / m, more preferably about 5 kV / m to about 20 kV / m, relative to the length of the microtube. Can be mentioned.
Examples of the fine tube include a hollow tube having an inner diameter of 1 to 150 μm and a length of 0.1 to 100 cm, preferably an inner diameter of 20 to 100 μm and a length of 1 to 50 cm. The material is not particularly limited, and glass (fused silica) or the like can be arbitrarily exemplified.

FIG. 3 shows, for example, an enlarged microbe when applied to a microbe as the microparticles 21 to be detected here. The antibody 30 selectively binding to the outer wall 21a and the antibody 30 are fluorescently labeled. A fluorescent dye 31 and the like are shown.
Although not shown, fluorescence 2b is also emitted from antibodies and fluorescent dyes floating in the solution without being bound to the fine particles in addition to the antibodies and fluorescent dyes 31 bound to the fine particles 21 to be detected. However, since the concentration position is different from the detection target fine particles, it is possible to separate them by observing the passage of time.

FIG. 4 is a flowchart showing the process of detecting the target fine particles described above.
First, a specimen solution is injected 32 into a microtube, and a fine particle is subjected to isoelectric focusing by providing a potential gradient to form a fine particle mass (concentration) 33. Next, the particle size search 34 is performed by relatively moving the detection window 6a, and the particle detection 35 is performed by the CCD camera or the like through the objective lens. The target fine particles are detected in the above process.

FIG. 5 is a diagram showing a fine particle image processing display example in the fine particle detection control device 8 such as a personal computer.
Main keys include 40 start, 41 stop, 42 waveforms, 43 images, 44 end, and the like.
When the particle detection operation is started by clicking the start key 40, the status display 45 is being detected, the elapsed time is displayed at 46, and the presence or absence of abnormality (normal here) is displayed in the comment field 47.
Click the stop key 41 to stop the detection operation in the middle of detection, and click the end key 44 to end the detection operation.

FIG. 5 is a diagram when the image key 43 is clicked, and when the waveform key 42 is clicked, it will be described later.
When the image key 43 is clicked, the output image of the CCD camera is displayed in the image area 50. Here, 51 indicates the tube wall of the microtubule, and the image between them indicates the electrophoretic solution portion image 52. When fine particles labeled with a fluorescent dye are detected, for example, 53 is displayed.
A unique number is assigned to the detected fine particle 53 from the brightness, shape, area, spatial position, moving direction, and the like of these detected images. In this embodiment, a case where five fine particles are detected is shown, and an example with numbers 21 to 25 is shown.
In the electrophoretic solution portion image 52, the image brightness and area of the detected fine particles vary depending on the intensity of the excitation light 2a and fluorescence 2b and the type of the target fine particles.
Therefore, the level adjustment of the image luminance value detected by the image processing is changed by clicking on the luminance value key 60 (black painting in the figure. The same applies hereinafter). Further, the adjustment of the image area to be detected is changed by clicking the ▽ of the area key 61. Such a particle detection technique based on image processing is an existing technique and can be easily realized.

Further, in this embodiment, a total value 62 of detected luminance, a total area 63 of detected fine particles, an instantaneous value 64 of the number of detected fine particles, and a total sum 65 of fine particles detected after the start of the detection operation are displayed.
Thereby, it can be confirmed that the image processing is operating normally. The obtained data is stored in a waveform control device such as a memory built in the particulate detection control device 8.

FIG. 6 is a diagram showing a display example of signal waveforms related to image processing of fine particles by clicking the waveform key 42 in the fine particle detection control device 8.
When the detection operation is started by the start key 40, a high voltage is applied to the apparatus based on the high voltage application condition at that time. At this time, the horizontal axis can be set to 1 scale, for example, by selecting ▽ at time 70 to select 1 minute, 2 minutes, or the like. At this time, the high voltage monitoring value actually applied can be selected as 1 kV, 2 kV, etc., by clicking on ▽ at 71, and the value of one scale on the vertical axis can be defined.
The monitoring value of the electrophoretic current at this time can be defined as one scale value on the vertical axis by clicking ▽ at 72 and selecting, for example, 500 μA, 1 mA or the like. The image brightness value at this time can be selected as, for example, 10, 15, 20, 30, etc. by clicking ▽ at 73 to define the value of one scale on the vertical axis.

Thus, the parameter unit to be displayed is different for each parameter, and the size of the parameter varies depending on the purpose. Here, in order to clearly indicate one scale on the vertical axis, three types of values, 71a, 72a, and 73a, are displayed side by side.
Here, 71a indicates a scale for high voltage, 72a indicates a scale for electrophoretic current, and 73a indicates a scale for image luminance values. Reference numeral 71b denotes a high voltage waveform, 72b denotes a migration current waveform, and 73b denotes an image luminance waveform over time.

If the elapsed time exceeds 10 minutes, the first panel on the horizontal axis is set to 1 minute in the present embodiment, so that everything cannot be displayed and scrolls automatically.
At this time, a desired time waveform can be displayed by dragging the scroll bar 80 or clicking the symbols 81 and 82.
Reference numeral 83 denotes a vertical scroll bar, which can be dragged to change the vertical display position, that is, the waveform display position. Alternatively, the same function can be used by clicking the symbols 84 and 85.

FIG. 7 shows that in FIG. 6, the value of one scale of the electrophoretic current is changed from 500 μA to 200 μA by 72, that is, the waveform is enlarged, and the first panel of image luminance is changed from 30 to 15 by 73, that is, the waveform is enlarged. A waveform when the scroll bar 83 on the vertical axis is dragged upward is shown.
The waveform 72d of the electrophoretic current is enlarged and displayed vertically, and the waveform 73d of the image luminance is enlarged and displayed vertically. Further, since the entire waveform is dragged upward by the scroll bar 83, the value of the vertical axis changes and changes as 71c, 72c, 73c. However, only the changed electrophoretic current 72c and the image brightness 73c are changed in one scale value.
The electrophoretic current waveform 72b is enlarged to 72d, the image luminance 73b is enlarged to 73d, and the high voltage waveform 71b is scrolled as it is to 71d. In the case of the present embodiment, it can be seen that the image brightness of the fine particles shows the maximum peak at an elapsed time of about 8 minutes.

In this way, the value of one scale on the vertical axis can be changed independently for each parameter, and at the same time, by scrolling the display position in the vertical direction, the electrophoretic parameter of interest is expanded, or It is possible to reduce and display the image freely.
In addition, since a plurality of waveforms such as high voltage, electrophoretic current, and image luminance are displayed at a time, the waveforms, scale values, characters, and the like may be displayed in different colors in order to avoid confusion for the observer.

Next, FIG. 8 shows another embodiment of the particle detection apparatus of the present invention.
In this microparticle detection apparatus, the micropipe 13 is formed in a substantially linear shape, and by rotating the motor 10, the micropipe 13, its detection window 13a, and the solutions 9a and 9b are integrated to form a straight line by a combination of the pinion 14 and the rack 15. The target fine particles are to be detected while the fluorescence detector 5 is relatively scanned while moving in the direction and keeping the relative distance between the objective lens 4 and the detection window 13a constant.
That is, the rotational movement of the circular microtubule 6 in FIG. 1 is changed to the linear movement of the straight tubular microtube 13 in FIG. 8, and other functions and effects are the same in the following. The details are omitted.

FIG. 9 is a schematic diagram showing the relationship between the rack 15 attached to the fine tube 13 and the pinion 14. When the motor 10 is rotated, the pinion 14 is rotated, and the rack 15 meshing with the pin 15, that is, the fine tube 13 is linear. Moving.
8 and 9 show an embodiment in which the microtube 13 is moved by the motor 10, but the microparticle detection apparatus 1 as a whole including the excitation light emitter 2 and the objective lens 4 is illustrated with the microtube 13 fixed. It may be moved in the scanning direction by a method that does not.
That is, the fluorescence detector is obtained by scanning while moving the objective lens relatively while keeping the relative distance between the entire optical system including the excitation light emitter 2 and the objective lens 4 and the detection window 13a of the microtube 13 constant. 5 can detect the target fine particles.
In addition, since the function and effect are the same, the detailed description is abbreviate | omitted.

  Usually, the time required for detecting microparticles such as microorganisms is several minutes to several tens of minutes, so that the image signal recording device 5a is preferably a DVD deck or the like capable of recording many image signals. Further, the fine particle detection control device 8 can sufficiently function on a normal personal computer, but since it is difficult to record an image during the entire detection period, only the images before and after detecting the target fine particles are recorded, or the image is recorded. Dedicated to the signal recording device 5a, the fine particle detection control device 8 has only the image processing results as shown in the present embodiment, that is, the image luminance for the target fine particles, the number of fine particles, the size thereof, the electrophoresis parameters, etc. You may limit to.

In addition, instead of the method of concentrating the fine particles by providing a pH gradient in the microtube, a potential gradient is generated in the microtube in the migration region, and this potential gradient generates an electroosmotic flow that is the flow of the entire liquid in the microtube. Usually, it flows from the anode to the cathode.
The fine particles injected into the microtube move in the direction of the electrode having a polarity opposite to the surface charge of the fine particles in response to the generated potential gradient.
As a result, the migration direction (velocity from the cathode to the anode) and velocity of the fine particles in the microtube are determined by the electroosmotic flow velocity and the mobility of the fine particles, and are concentrated near the opposite end of the microtube (near the anode). The method can be changed as appropriate, such as a method for detecting fine particles by the method.

In addition, as a means for detecting fine particles concentrated and separated in a microtube, generally, any of spectroscopic, electrochemical, and weight detection methods can be used, but preferably UV-visible detection, fluorescence Examples thereof include optical detection methods such as detection, luminescence detection, and light scattering detection.
Among them, a fluorescence detection method excellent in detection specificity and sensitivity is preferable, and a laser excitation detection method capable of detecting with higher sensitivity is desirable when the amount of light in the LED is insufficient. These fluorescence-based detections can be performed by combining reactions such as immune reactions (antigen-antibody reactions), intracellular enzyme reactions, and nucleic acid complementary chain formation reactions as appropriate according to the target microparticles. Can be detected.
Among these methods, the immune reaction is one of the preferred methods because it does not require special pretreatment of the microparticle sample and is excellent in specificity. Specific examples of such an immune reaction include a method of labeling target microparticles with fluorescence or an enzyme using a fluorescent dye-labeled antibody (also simply referred to as a fluorescent antibody), an enzyme-labeled antibody (also simply referred to as an enzyme antibody), or the like. be able to.

Thus, according to the microparticle detection apparatus of the present embodiment, the microparticles in the specimen sample can be separated, detected, and quantified with high accuracy in a short time. That is, it becomes possible to isolate | separate the microbe microbe species and strain individually. In addition, since fluorescent staining is performed so as to selectively bind to the target microparticles, it can be detected easily and accurately in a short time without any special technique and without a culture operation. Accurate particle inspection and monitoring system can be established.
Further, by detecting fine particles using an image processing technique, the image processing result and the fine particle detection image can be stored and recorded in a memory such as a personal computer. Therefore, by recalling the stored data, it is possible to reconfirm the detection result and report the detailed result to the sample solution supplier. In addition, it is possible to contribute to quick measures against fine particles, sterilization measures, and the like.
In addition, by using an inexpensive LED or laser emitter as an excitation light source in the excitation light emitter 2, the desired fine particles can be detected with high sensitivity and specificity. A small amount of fine particles can be separated and detected accurately without any treatment.
In addition, since the fine particle detection apparatus of this embodiment has a quantitative property, it is possible to discriminate and measure (quantitative detection) the number of microorganisms such as microorganisms present in the sample, and to detect the microorganisms in the sample. In addition, it is possible to identify bacterial species by using specific detection reagents (for example, antibodies) specific to various microorganisms.
As described above, since the particulate detector of this embodiment is suitable for the detection of particulates such as harmful microorganisms (qualitative detection, quantitative detection), environmental water and food contaminated with pathogenic microbes can be eliminated at an early stage. Useful in preventing the occurrence of illness. In addition, since early diagnosis of patients with pathogenic microorganisms becomes possible, it helps prevent the spread of damage caused by harmful microorganisms and helps early treatment. Furthermore, the quality control period of the food can be shortened, and the shelf life period can be extended due to the strictness of hygiene management, which has the effect of greatly contributing to the economics of food distribution.

As mentioned above, although the particulate detection device of the present invention has been described based on the embodiments thereof, the present invention is not limited to the configurations described in the above embodiments, and other embodiments are within the scope not departing from the gist thereof. Can be mentioned.
For example, although the image luminance value is shown as one of the waveforms in FIGS. 6 and 7, the present invention is not limited to this, and the (image) area of the detected fine particles, the instantaneous value of the number of fine particles, the detected fine particles Needless to say, the total sum can be freely selected and changed.

  The fine particle detection device of the present invention has the characteristic of separating and detecting various fine particles quickly and with high sensitivity. For example, environmental water such as bath water, pools, cooling water for cooling towers, food and medical fields. For example, it can be suitably used for the purpose of accurately detecting harmful microorganisms, such as Legionella spp.

It is a schematic block diagram which shows one Example of the microparticle detection apparatus of this invention. It is explanatory drawing which shows the detection process of the microparticles | fine-particles by microtube electrophoresis. It is an enlarged view showing details of fine particles. It is an operation | movement flowchart which detects fine particles. It is the screen figure which displayed the image processing condition in a particulate detection control apparatus. It is the screen figure which displayed the waveform condition in a particulate detection control device. It is the screen figure which displayed other waveform situations in a particulate detection control device. It is a schematic block diagram which shows the other Example of the microparticle detection apparatus of this invention. It is a top view which shows the rack and pinion of the Example.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Fine particle detector main body 2 Excitation light emitter 2a Excitation light 2b Fluorescence 4 Objective lens 5 Fluorescence detector 5a Image signal recording device 6 Fine tube 6a Detection window 7a Electrode 7b Electrode 7 High voltage power supply 8 Fine particle detection control device 9a Solution 9b Solution 10 Motor 11 Arm 12 Center 13 Microtube 14 Pinion 15 Rack 20 Sample solution 21 Particle 21a Outer wall of particle 27 Concentrated particle 30 Antibody 31 Fluorescent dye 32 Injection of sample solution 33 Electrophoresis (concentration)
34 Electrophoresis (search for fine particle mass)
35 Particle Detection 40 Start Key 41 Stop Key 42 Waveform Key 43 Image Key 44 End Key 45 Status Display 46 Elapsed Time 47 Comment Field 50 Image Area 51 Microtubule Wall 52 Electrophoresis Solution Part Image 53 Particle Detection Image 60 Brightness Adjustment Key 61 Area adjustment key 62 Total luminance value 63 Total area of fine particles 64 Instantaneous value of the number of fine particles 65 Total sum of fine particles 70 hours 71 High voltage monitoring value 71a High voltage scale 71b High voltage waveform 71c High voltage grid 71d High voltage waveform 72 Electrophoretic current 72a Electrophoretic current scale 72b Electrophoretic current waveform 72c Electrophoretic current scale 72d Electrophoretic current waveform 73 Image luminance value 73a Image luminance value scale 73b Image luminance value waveform 73c Image luminance value scale 73d Image luminance value waveform 80 Scroll bar 81 Symbol 82 Symbol 83 Vertical axis scroll bar 84 No. 85 symbol

Claims (7)

  In a microparticle detection device that detects microparticles by performing microtubule electrophoresis on a sample containing microparticles using an electrophoresis solution, an excitation light emitter that excites and irradiates the microparticles that are fluorescently stained so as to selectively bind, and fluorescent staining And a fluorescence detector that detects fluorescence emitted from the fine particles using an optical system, and at least one of the optical system including the excitation light emitter and the fluorescence detector or a microtube can be moved in the scanning direction. Thus, the fine particle detection apparatus is characterized in that the fine particles concentrated in the fine tube are detected from a fluorescence detector that scans while relatively moving.   2. The image signal recording apparatus for recording a signal detected by a fluorescence detector as an image signal, and a particle detection control apparatus for recording a particle detection signal for detecting particles from the image signal. Fine particle detector.   3. The particle detection apparatus according to claim 1, wherein the particle detection signal is an image brightness, an image area, or the number of particles.   4. The fine particle detection apparatus according to claim 1, further comprising a display device and a function of controlling an applied voltage in microtube electrophoresis, a function of monitoring a current flowing through the microtube, or a function of recording a fine particle detection signal.   A built-in waveform control device that independently varies the magnitude when displaying a recording waveform such as an applied voltage signal, a micro-tube current signal, or a particulate detection signal, or the like, or 4. The fine particle detection apparatus according to 4.   6. The particle detecting apparatus according to claim 1, wherein the particle is a microorganism.   The microparticle detection apparatus according to claim 6, wherein the microbe is a microbe harmful to the human body such as Legionella spp.

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