JP2008096156A - Fine particle detector - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a fine particle detector capable of rapidly separating and detecting a plurality kinds of fine particles or the number of live bacteria and dead bacteria of a target microorganism, at high sensitivity. <P>SOLUTION: The fine particle detector is constituted so as to detect fine particles by treating a sample by fluorescent dyeing selectively bonded, corresponding to the kind of fine particles and subjecting the sample to capillary electrophoresis using a migration liquid and equipped with an exciting light emitter 1 for irradiating the fine particles subjected to fluorescent dyeing with an exciting light; a fluorescence detector 12 for detecting the fluorescence emitted by the irradiation with exciting light and a plurality of kinds of filters 11a, 11b and 11c in parallel arranged, in front of the fluorescence detector 12. The fluorescence of the fine particles that pass through the respective filters 11a, 11b and 11c is formed into an image, in parallel with the detection element of the fluorescence detector 12. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a microparticle detection apparatus for detecting various microparticles such as cells and the like present in a specimen sample using microtubule electrophoresis. The present invention relates to a fine particle detection apparatus capable of separating and detecting with sensitivity.

In general, in order to investigate whether or not desired fine particles are present in a sample, and to examine the type and amount of various fine particles such as microorganisms contained in a sample, or the distinction between live and dead bacteria, By inoculating these samples in an appropriate medium and culturing, it is necessary to grow microparticles that are the target microorganisms.
For this reason, conventionally, detection of microorganisms requires a long time of at least several days or more for culturing, and there has been a problem that results cannot be obtained quickly.
In addition, since microorganism culture is basically performed aseptically, there is a problem that special culture techniques and facilities are required.
On the other hand, there is a method for identifying the type of microorganism from genetic information, but there is a problem that the number of microorganisms and the distinction between live and dead bacteria cannot be identified.

For these reasons, there is a demand for a method that can quickly and easily detect the types and properties of harmful microorganisms that can be contained in a specimen sample without the need for culturing microorganisms and advanced techniques and devices.
For the detection of microorganisms classified into many types, conventionally, after culturing in selective separation culture, it is necessary to catch suspicious colonies and determine with confirmation medium, identification kit or antiserum. It took a lot of experience and skill to identify. Moreover, when time for culture | cultivation etc. was match | combined, the determination result required the long term of 3 to 5 days or more.
For this reason, in the environment / food and medical fields, establishment of a quick and safe method for specifically detecting microorganisms, particularly harmful microorganisms, for each species and strain is required.

By the way, in recent years, as a rapid method for separating and detecting microorganisms, an electrophoresis technique has been proposed in which microorganisms that have been pre-fluorescently stained in a microtubule are electrophoresed and irradiated with excitation light to detect the fluorescence generated at that time ( For example, see Patent Document 1).
Further, although there is a paper on the fine tube isoelectric focusing technique, details of specific detection means are not described (for example, see Non-Patent Document 1).
On the other hand, when migrating and separating microorganisms in a microtubule, the interaction between the microbial cells and the inner wall of the tubule or between the cells and the cells is large. It is effective to contain an alginate (see, for example, Patent Document 2).
In this method, alginate as an additive that improves the separation efficiency of microorganisms does not function in the same manner for all microorganisms, and more types of separation efficiency improvers that can handle a wide variety of microorganism species. The introduction is expected.

By the way, in order to quickly take appropriate measures against environmental water, food contamination and infectious diseases, and to quickly identify and deal with the contamination sources and infection sources, it is important There is a need for a rapid and specific detection method for harmful particulates.
In addition, it is important to confirm whether or not all harmful microorganisms have been killed when such microorganisms are subjected to a treatment such as chlorine sterilization.
In addition, when it is difficult to kill all harmful bacteria, it is necessary to know the status of chlorine sterilization and the sterilization efficiency by checking the number of live and dead bacteria after sterilization.
As a method for detecting these multiple types of microorganisms, an example has been proposed in which only light of a desired wavelength is separated and analyzed using a plurality of reflecting mirrors and filters. Since a plurality of reflecting mirrors are used, there is a problem that the amount of light to be detected is attenuated (see Patent Document 3).
JP 2002-345451 A JP 2002-181781 A US Pat. No. 5,982,497 Microbial electrophoresis separation and detection method (Monthly Food Chemical 2005-2)

  In view of the problems of the conventional fine particle detection apparatus, the present invention can rapidly and highly sensitively detect and detect a plurality of types of fine particles, the number of living microorganisms of a target microorganism, the number of dead bacteria, and the like. An object is to provide a detection device.

  In order to achieve the above object, the microparticle detection apparatus of the present invention treats a sample with fluorescent staining that selectively binds depending on the type of microparticle, and performs microtube electrophoresis of the sample using an electrophoresis solution. In a fine particle detection apparatus for detecting fine particles, an excitation light emitter that irradiates fluorescent dyed fine particles with excitation light, a fluorescence detector that detects fluorescence emitted by excitation light irradiation, and a fluorescence detector that is arranged in front of the fluorescence detector. And a plurality of types of filters that transmit only a specific wavelength region, and the fluorescence of the fine particles that have passed through each filter is imaged in parallel on the detection element of the fluorescence detector.

  In this case, it is possible to provide a fine particle detection control device that distinguishes and detects the fluorescence signal obtained from the fluorescence detector for each filter, and an image signal recording device that records the detection signal of the fine particle detection control device.

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

  According to the fine particle detection apparatus of the present invention, an excitation light emitter that irradiates fluorescence-stained fine particles with excitation light, a fluorescence detector that detects fluorescence emitted by the excitation light irradiation, and a parallel arrangement in front of the fluorescence detector And a plurality of types of filters that transmit only a specific wavelength region, and the fluorescence of the fine particles that have passed through each filter is imaged in parallel on the detection element of the fluorescence detector. In the case of emitting fluorescence of different wavelengths, or even when the same type of fine particles emits fluorescence of different wavelengths by distinguishing between live and dead bacteria, the detection wavelength by the multiple filters from the image signal in the fluorescence detector It is possible to detect microparticles according to the region at the same time. This makes it possible to detect various types of microparticles contained in a sample with high resolution without requiring special techniques. In addition to separation with high reproducibility, the target microparticles are fluorescently stained in advance, so that even minute amounts of microparticles can be detected with high sensitivity without being subjected to growth treatment such as culturing. Fine particles can be detected, and furthermore, fine particles mixed in a sample can be detected not only simply, but also can be quantitatively detected separately for each kind of particles and for each characteristic.

  In this case, a plurality of types can be provided by including a fine particle detection control device that distinguishes and detects the fluorescence signal obtained from the fluorescence detector for each filter and an image signal recording device that records the detection signal of the fine particle detection control device. A plurality of types of fluorescence can be simultaneously detected by this filter, and the image processing result or the particle detection image can be simultaneously displayed or stored and recorded in a memory such as a personal computer.

  Further, by using the fine particle detection signal as the luminance of the image, the area of the image, or the number of fine particles, the detailed data of the detection result of the fine particles becomes clear.

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

The microparticle detection apparatus of the present invention separates a plurality of types of microparticles contained in a sample with high resolution and high reproducibility by utilizing microtubule electrophoresis 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, desired fine particles can be detected with high accuracy and even higher resolution.
Furthermore, the fine particles mixed in the sample can be detected not only simply but also quantitatively.
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) An apparatus for detecting microparticles using microtubule electrophoresis, which is a water tank containing an electrophoretic liquid, a microtubule that concentrates and separates microparticles injected into the electrophoretic liquid, or a detection for detecting concentrated microparticles. Means.
(4) An apparatus for detecting microparticles using microtubule electrophoresis, wherein the microparticles are concentrated by a pH 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 fluorescence emitted from the fluorescence-stained microparticles is detected using an optical system. Detect fine particles.
(6) Plural types of filters that transmit only a specific wavelength region are arranged in parallel on the same plane in front of the fluorescence detector, and the fluorescence of the fine particles that have passed through each filter is simultaneously or Images are sequentially formed.
(7) Further, the image signal recording apparatus for recording the image signal detected by the fluorescence in the optical system and the particle detection control apparatus for recording the particle detection signal for detecting the particle from the image signal are provided.
(8) 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.

FIG. 1 shows a basic configuration of a microtubule isoelectric focusing system of the present invention as an embodiment.
In FIG. 1, 1 is an excitation light emitter such as a diode, 2 is a microtube, 3 is its detection window, 4a and 4b are electrodes, 5a and 5b are solutions, 6 is fine particles contained in the solution, and 7 is excitation light. Excitation light emitted from the light emitter 1, 8 is an objective lens, 9 is fluorescence emitted from fine particles, and 10 is a dichroic filter.
Also, 11a and 11b are two types of filters that allow only specific wavelengths to pass, 11c is a non-filter, that is, a filter that passes fluorescence of all wavelengths, 12 is a fluorescence detector such as a CCD camera, and 13 is particulate detection. A control device, 15 is a display device, and 16 is an image signal recording device.

Microparticles such as harmful bacteria that are detection objects, for example, microorganisms harmful to the human body such as Escherichia coli and Legionella, are preliminarily fluorescently stained so as to selectively bind only to the desired microorganisms. It inject | pours into the inside of the microtube 2 from the solution 5a or the solution 5b installed in the both ends.
The solution 5a or the solution 5b is changed according to the detection stage, and a high voltage power source is applied to both ends of the microtube 2 by the electrodes 4a and 4b.
The high voltage is supplied from a power source (not shown), and the magnitude and application time of the voltage are controlled by the particulate detection control device 13 such as a personal computer, for example.

The excitation light 7 emitted from the excitation light emitter 1 is reflected by the dichroic filter 10 and is irradiated from the objective lens 8 to the fluorescently stained fine particles from the detection window 3 of the fine tube.
The fluorescently stained fine particles emit fluorescent light 9, enter the apparatus main body through the objective lens 8, and pass through the dichroic filter 10.
At this time, the emitted fluorescence 9 has various wavelengths based on the characteristics of the fluorescent staining that binds depending on the type of fine particles. For example, when a microorganism that is fluorescently stained with a LIVE / DEAD reagent (manufactured by Invitrogen) is irradiated with blue excitation light, it emits green fluorescence when viable, and red fluorescence when killed. .
The fluorescence 9 passes through the filters 11a and 11b and the non-filter 11c installed immediately above the fluorescence detector 12 composed of a CCD camera or the like, and then forms an image on the fluorescence detector 12 in parallel. The image signal is sent to the particle detection control device 13 for detection processing.
This detected image signal is recorded in the image signal recording device 16.
The fluorescently stained fine particles are concentrated by a method described later and passed through the detection window 3. The particulate detection operation can be performed during the passage.

FIG. 2 is a plan view of the filters 11a and 11b and the non-filter 11c. Each of the filters 11a, 11b, and 11c is formed in a rectangular shape, and is directly above the CCD light receiving element of the fluorescence detector 12. They are arranged in parallel on the same plane in the longitudinal direction.
When the fluorescence 9 is imaged on the fluorescence detector 12, if each filter 11a, 11b, 11c is configured to have a characteristic that allows only the fluorescence having a desired wavelength to pass therethrough, only the fluorescence having the desired characteristic is selectively acquired. it can. Details of the processing content of the acquired image will be described later.

FIG. 3 shows a migration state of microorganisms that are harmful bacteria in the microtubule 2 as an example of fine particles, and the following detection process is performed.
First, a test solution, which is a sample containing microorganisms as detection targets, is prepared as, for example, a solution 5a and sucked from the solution 5b side to be injected as a target test solution 20 into the microtube 2.
Next, an acidic solution such as phosphoric acid is used as the solution 5a, and an alkaline (basic) solution such as caustic soda is used as the solution 5b. The electrodes 4a and 4b are respectively connected to a high voltage power source (not shown), and the fine particle detection control device 13 uses an anode for the electrode 4a (acidic solution 5a side) and a cathode for the electrode 4b (alkaline solution 5b side). Connection is made so as to apply a voltage of.
Here, both ends of the microtube 2 are respectively immersed in the above acidic or alkaline solution (referred to as an electrophoretic solution), and then when a high voltage power supply is activated, a potential gradient is generated in the sample solution 20 in the microtube 2, As a result, the entire liquid in the migration region (in the microtube) in the microtube generates a pH gradient between the cathode direction and the anode direction due to the isoelectric point phenomenon.

At this time, since the fine particles 21 that may exist in the test solution 20 are diffused to different pH regions, they migrate toward a region having no charge, that is, the isoelectric point position 24 or 25. .
That is, the fine particle 21 moves to a region (isoelectric point position 24 or isoelectric point position 25) having the same pH condition as the isoelectric point that it has.
For example, when the pH range of the fine particles having an isoelectric point pH of pH (A) is the isoelectric point position 24, the fine particles are moved closer to the electrode (cathode) 4b side than the isoelectric point position 24 due to the formed pH gradient. When it exists, it moves in the moving direction 22, and when it exists on the electrode (anode) 4 a side from the isoelectric point position 24, it moves in the moving direction 23 and is concentrated at the isoelectric point position 24.
Similarly, assuming that the pH range of the fine particles having an isoelectric point pH of pH (B) is the isoelectric point position 25, the fine particles are moved from the isoelectric point position 25 to the electrode (cathode) 4b side by the formed pH gradient. Is present in the moving direction 22, and when present on the electrode (anode) 4 a side from the isoelectric point position 25, it moves in the moving direction 23 and is concentrated at the isoelectric point position 25.
That is, the fine particles are accumulated at the isoelectric point position 24 or the isoelectric point position 25 in the microtube 2 by the isoelectric point that the fine particles have, thereby forming a concentrated fine particle lump 24a or fine particle lump 25a. This isoelectric point position varies depending on the type of the target fine particles, the status of live bacteria, and the status of dead bacteria. In general, since the surface charge component of the fine particles has a specific isoelectric point, the fine particles are concentrated to the specific electric point positions 24, 25, etc., which are the specific locations by the pH gradient in the microtube.
In this state, the electrophoretic current between the electrodes 4a and 4b is in a steady state and hardly flows.

Next, as shown in FIG. 4, when the solution 5a is an acidic solution as it is and a voltage of an anode is applied to the electrode 4a and a cathode is applied to the electrode 4b using a salt (mobilizer), for example, The fine particle lump 24a or the fine particle lump 25a concentrated at the isoelectric point position 24 or the isoelectric point position 25 in FIG. 3 is directed toward the electrode (cathode) 4b in the form of being flowed by a mobilizer, for example, due to the potential gradient generated again. Move in the moving direction 26.
At this time, the fine particle masses 24 a and 25 a pass through the detection window 3. This fine particle lump is fluorescently stained, and as shown in FIG. 1, when excitation light is irradiated from the objective lens 8, a fluorescent color having a wavelength corresponding to the characteristic of the fluorescent staining is emitted.

The method of injecting the specimen sample into the microtube 2 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 4a and 4b and the high-voltage power source exemplified above, the potential gradient of the strength necessary for the fine particles injected into the microtube to migrate in the electrophoretic solution in the microtube, It is a means for applying with respect to a liquid.
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 selected.

FIG. 5 shows, for example, an enlarged microbe when applied to a microbe as the microparticles 21 to be detected here. The antibody 30 and the antibody 30 that selectively bind to the outer wall 21a are fluorescently labeled. A fluorescent dye 31 and the like are shown.
Although not shown, in addition to the antibody and fluorescent dye 31 that are bound to the detection target fine particle 21, fluorescence is also emitted from the antibody and fluorescent dye that are floating in the solution without being bound to the fine particle. 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. 6 is a flowchart showing the process of detecting the target fine particles described above.
First, a sample solution is injected into a microtube (step 32), and a fine particle is subjected to isoelectric focusing by providing a pH gradient to form a fine particle mass (concentration) (step 33).
Next, the fine particle lump is migrated in a fine tube to search for the fine particle lump (step 34), and the fine particle is detected by a CCD camera or the like through a filter (step 35).
In the above process, the target fine particles are detected.

FIG. 7 is a diagram showing a fine particle image processing display example in the fine particle detection control device 13 such as a personal computer.
Main keys include a start key 40, a stop key 41, an end key 42, and the like.
When the particle detection operation is started by clicking the start key 40, the status display 43 is being detected, the elapsed time is displayed in the comment column 44, and whether or not there is an abnormality (normal here) is displayed in the comment column 45. .
Click the stop key 41 to stop the detection operation in the middle of detection, and click the end key 42 to end the detection operation.

In FIG. 7, an image obtained from the CCD element immediately below the filter 11 a is shown as a region 46 on the left side of the screen, and an image obtained from the CCD element just below the filter 11 b is shown as a center region 47 on the screen. In addition, an image obtained from the CCD element immediately below the non-filter 11c is shown as a region 48 on the right side of the screen.
Here, 51a and 51b indicate the wall of the fine tube, and when the fluorescently stained fine particles are detected, the fine particles 52, 53 and 54 are displayed on the image.
Image tracking is performed from the brightness, shape, area, spatial position, direction of movement, and the like of these detected images, and a unique number is assigned to the detected fine particles, and the number is counted.
In this embodiment, three fine particles 52 are detected in the region 46, two fine particles 53 are detected in the region 47, and two fine particles 54 are detected in the region 48.
That is, the fine particles 52 pass through the first filter 11a, the fine particles 53 pass through the second filter 11b, and the fine particles 54 show the non-filter 11c, that is, the fine particles when the filter is not applied. . These move rightward with time, and disappear or appear based on the combination of the wavelength condition of the fine particles and the filter.

The image brightness, the area, etc. of the fine particles to be detected vary depending on the intensity of the excitation light 7, the fluorescence 9, 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 (▽ is black in the figure. The same applies hereinafter).
Further, the adjustment of the image area to be detected is changed by clicking ▽ of the area key 61.
Such a particle detection technique by image processing is an existing technique and can be easily realized.

Furthermore, in the present embodiment, the total luminance value 62 detected in the region 46, the total area 63 of the detected fine particles, the instantaneous value 64 of the number of detected fine particles, and the total sum 65 of the fine particles detected after the start of the detection operation are shown. The total luminance value 66 detected in the region 47, the total area 67 of the detected fine particles, the instantaneous value 68 of the number of detected fine particles, and the total number 69 of fine particles detected after the start of the detection operation are displayed. Further, although it is possible to display data detected in the area 48, an example in which the data is not displayed is shown here.
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 particle detection control device 13.

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 16 is preferably a DVD deck or the like capable of recording many image signals.
Further, the fine particle detection control device 13 can sufficiently function on a normal personal computer, but since it is difficult to record an image for 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 16, the fine particle detection control device 13 has the image processing results as shown in this embodiment, that is, the image luminance for the target fine particles, the number of fine particles, the size thereof, the electrophoresis parameters, etc. It may be limited to only.

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 or Examples include optical detection methods such as fluorescence detection, luminescence detection, and light scattering detection.
Among them, a fluorescence detection method excellent in detection specificity and sensitivity is preferable, and a laser-induced fluorescence detection method capable of detecting with higher sensitivity is desirable when an inexpensive LED is used as excitation light and the light quantity is insufficient. In the detection based on these fluorescences, particularly in the detection of harmful microorganisms or the like, by appropriately combining reactions such as an immune reaction (antigen-antibody reaction), an intracellular enzyme reaction, and a nucleic acid complementary chain formation reaction according to the target microorganism, The target microorganism can be detected more specifically and with high sensitivity.
Among these methods, the immune reaction is one of the preferred methods because it does not require any special pretreatment on the microorganism sample and is excellent in specificity. Specific examples of such an immune reaction include a method of labeling a target microorganism with fluorescence, an enzyme, or the like 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, the particle detector of the present embodiment includes an excitation light emitter 1 that irradiates fluorescence-stained particles with excitation light, a fluorescence detector 12 that detects fluorescence emitted by excitation light irradiation, and the fluorescence detector 12. A plurality of types of filters 11a, 11b, and 11c that are arranged in parallel in front and transmit only a specific wavelength region are provided, and the fluorescence of the fine particles that have passed through the filters 11a, 11b, and 11c is arranged in parallel with the detection element of the fluorescence detector 12. Therefore, the microparticles such as microorganisms in the specimen sample can be separated, detected and quantified with high accuracy in a short time. That is, it becomes possible to isolate the microorganism species and strains individually. In addition, since it is fluorescently stained so as to selectively bind to the target microorganism, it can be detected easily and accurately in a short time without requiring a special technique and without a culture operation. Accurate microbial testing and monitoring system can be established.
Further, by detecting microorganisms using an image processing technique, the image processing results and microorganism detection images 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 microorganisms, measures against sterilization, and the like.
In addition, since a desired microorganism can be detected with high sensitivity and specificity by using an inexpensive LED or a laser-induced fluorescence detector as a fluorescence detection means, a proliferation process such as culturing a sample is performed. Therefore, a minute amount of microorganisms can be separated and detected with high accuracy.
In addition, since the microparticle detection apparatus of the present embodiment has a quantitative property, it is possible to discriminate and measure (quantitative detection) the number of cells present in the sample, and it is possible to detect 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 detection device of this embodiment is suitable for detection of harmful particulates (qualitative detection, quantitative detection), environmental water and food contaminated with pathogenic microorganisms can be eliminated at an early stage, resulting in the occurrence of disease. It is useful for prevention. 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.

In this case, a fine particle detection control device 13 that distinguishes and detects the fluorescence signal obtained from the fluorescence detector 12 for each filter 11 and an image signal recording device 16 that records the detection signal of the fine particle detection control device 13 are provided. As a result, a plurality of types of fluorescence can be simultaneously detected by the plurality of types of filters 11, and the image processing results or particle detection images can be simultaneously displayed or stored and recorded in a memory such as a personal computer.
Further, by using the fine particle detection signal as the luminance of the image, the area of the image, or the number of fine particles, the detailed data of the detection result of the fine particles becomes clear.

  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 embodiments, and the configuration is appropriately changed without departing from the gist thereof. can do.

  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 fine particles such as harmful microorganisms mixed in a sample sample collected in the above.

It is a schematic block diagram which shows one Example of the microparticle detection apparatus of this invention. It is a top view of a filter. It is explanatory drawing which shows the detection process of the microparticles | fine-particles by microtube electrophoresis. It is explanatory drawing of the next step of the detection process of microparticles | fine-particles by microtube electrophoresis. It is an enlarged view showing details of fine particles. It is a flowchart which shows the detection operation of microparticles | fine-particles. It is a figure which shows the screen which displayed the image processing condition in a microparticle detection control apparatus.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Excitation light emitter 2 Fine tube 3 Detection window 4a Electrode (anode)
4b Electrode (cathode)
5a Solution 5b Solution 6 Fine Particle 7 Excitation Light 8 Objective Lens 9 Fluorescence 10 Dichroic Filter 11a First Filter 11b Second Filter 11c Non-filter 12 Fluorescence Detector 13 Fine Particle Detection Control Device 15 Display Device 16 Image Signal Recording Device 20 Test Solution 21 Fine Particles 24 Isoelectric Point Position 25 Isoelectric Point Position 30 Antibody 31 Fluorescent Dye 40 Start Key 41 Stop Key 42 End Key 43 Status Display 44 Elapsed Time 45 Comment Field 46 First Filter Image Area 47 Second Filter Image Area 48 Non-filter region 51a Fine tube tube wall 51b Fine tube tube wall 52 Fine particle image of first filter region 53 Fine particle image of second filter region 54 Fine particle image of non-filter region 60 Brightness value adjustment key 61 Area adjustment key 62 First Total brightness of filter area 3 Total area of fine particles in the first filter area 64 Instantaneous value of the number of fine particles in the first filter area 65 Total of fine particles in the first filter area 66 Total luminance value of the second filter area 67 Total area of fine particles in the second filter area 68 Instantaneous value of the number of fine particles in the second filter area 69 Sum of fine particles in the second filter area

Claims (3)

  The sample is treated with fluorescent staining that selectively binds depending on the type of fine particles, and the sample is subjected to microtubule electrophoresis using an electrophoresis solution. An excitation light emitter that emits excitation light, a fluorescence detector that detects fluorescence emitted by excitation light irradiation, and a plurality of types of filters that are arranged in parallel in front of the fluorescence detector and transmit only a specific wavelength region. A particulate detection apparatus comprising: a fluorescence of particulates having passed through each filter; and imaged in parallel with a detection element of a fluorescence detector.   6. A fine particle detection control device that distinguishes and detects a fluorescence signal obtained from a fluorescence detector for each filter, and an image signal recording device that records a detection signal of the fine particle detection control device. The fine particle detection apparatus according to 1.   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.

Images