JP2008000038A - Microbe detection filter device - Google Patents

Description

  The present invention relates to a microorganism used in a microbe counting apparatus capable of coloring microorganisms and measuring microorganisms at a time from a sample having a plurality of kinds of microorganisms, that is, microorganisms such as bacteria and fungi attached or mixed therein. The present invention relates to a filter device for detection.

FIG. 4 shows an example of a microorganism number counting apparatus that can color microorganisms and measure microorganisms at a time from a sample in which a plurality of types of microorganisms, that is, microorganisms such as bacteria and fungi may be attached or mixed. 1 is a configuration diagram of a microorganism count device disclosed in Patent Document 1 (Republished Patent (A1) WO2003 / 0667230).
The microorganism count device shown in FIG. 4 includes a light source 10 that emits excitation light, an episcopic microscope 6, 4, 3, 5, 11, 12 for observing the fluorescence state of the specimen 1 by the excitation light, a CCD, (Imaging device) 7, and the fluorescence state by the excitation light is imaged by the CCD 7, and the bacteria are counted by image processing in the calculation unit 8. Reference numeral 9 denotes a stage moving mechanism.

As a preparatory work prior to starting measurement with such a microbe counting apparatus, it is necessary to first collect microorganisms on a solid surface such as a membrane filter, and then contact the microorganisms with a chromogenic substance to stain them. For this purpose, pretreatment work is performed using a filter or a pipette.
FIG. 5 is an overall configuration diagram of a filtration device disclosed in Patent Document 2 (Japanese Patent Laid-Open No. 5-52850) as an example of a filtration device for extracting microorganisms used in this pretreatment, and FIG. 6 shows the filtration device. It is sectional drawing which shows the detail of the filtration part 2 in an apparatus.
5 and 6, the membrane filter 4 is connected between the funnel 1 for pouring liquid and the branch flask 3 by a stationary scissors 8. A support screen 5 made of, for example, SUS304 is fixed to the branch flask 3, and the membrane filter 4 is filtered. The support screen 5 has a structure having a large number of small holes in a disk so that vacuum filtration is possible.

In such a filtration device, a liquid (specimen) containing microorganisms poured into the funnel 1 is sucked from the branch flask 3 with a reduced pressure source such as a vacuum pump, and the microorganisms are collected on the membrane filter 4. Further, a coloring material is poured into the funnel 1 and is dyed by contacting with the microorganism.
Next, the membrane filter 4 that has collected the microorganisms is picked from the filter with tweezers, placed on a preparation, for example, and set in the microorganism count device shown in FIG.
The preparation comprising the membrane filter 4 in which the stained microorganisms are collected is then brightened by the CCD 7 in the microorganism counting device with automatic focusing, so-called autofocus as shown in FIG. The number of microorganisms can be counted by performing image processing on the calculation unit 8.
Republished patent (A1) WO2003 / 067230 JP-A-5-52850

In order to collect microorganisms on a membrane filter and perform image analysis with a microorganism counter using the means of Patent Document 1 and Patent Document 2 described above, there are the following problems.
(1) When the membrane filter 4 collecting microorganisms is picked up from the filter by tweezers or the like, a problem that the membrane filter 4 is broken is likely to occur.
This is because it is difficult to peel off the membrane filter 4 because the membrane filter 4 comes into close contact with the branch flask 3 by vacuum filtration.
Normally, the above problem can be solved by providing a valve on the suction side of the branch flask 3 and opening the inside of the flask 3 to atmospheric pressure. However, the membrane filter 4 and the support screen 5 The above problem cannot be fundamentally solved because the surface tension of the liquid remaining between them sticks.
(2) Further, since the membrane filter 4 that collects such microorganisms is intended to be used in a microorganism counting device, in the microorganism counting device equipped with autofocus, if the membrane filter 4 has slack or undulation, It is difficult to auto-focus and causes a counting error in the microorganism counter.
(3) Furthermore, since it is necessary to manually place the membrane filter 4 on a preparation from a filter after collecting microorganisms, the work requires time and skill.

  The present invention has been made in view of such circumstances, and its purpose is to capture microorganisms and detect microorganisms without causing damage to the membrane filter and without using special tools such as tweezers. To provide a filter device for detecting microorganisms that can be attached to a microorganism counting apparatus with a very simple work procedure and can be measured quickly, and can improve the counting accuracy of microorganisms in the microorganism counting apparatus. is there.

  In order to solve the above-described problems of the prior art, the present invention is a filter device for detecting microorganisms used in a microorganism counting device for analyzing images of microorganisms, and includes a membrane filter for collecting microorganisms. A filter device for detecting microorganisms, comprising: a support screen having a small hole for filtering a specimen; and a ring-shaped member that holds an outer peripheral portion of the membrane filter, and the membrane filter is disposed on the support screen In addition, the outer peripheral portion of the support screen is supported by the ring-shaped member.

In the present invention, it is preferable that the support screen is specifically configured as follows.
(1) The support screen is a metal plate having chemical resistance, and includes a plurality of small holes for filtering the specimen.
(2) The support screen is a glass plate and has a plurality of small holes for filtering the specimen.
(3) The support screen is configured by meshing a fine wire material such as a metal wire.
(4) A non-reflective coating is applied to the surface of the support screen.
(5) An uneven surface is formed on the surface of the support screen.

In the present invention, the support screen is preferably attached as follows.
(1) The outer periphery of the support screen is fixed to the ring-shaped member to which the membrane filter is fixed.
(2) The support screen is fixed by sandwiching an outer peripheral portion between the membrane filter fixed to the ring-shaped member and the ring-shaped member.

  As described above, in the filter device for detecting microorganisms according to the present invention, the membrane filter and the support screen are fixed to the ring-shaped member at the outer peripheral portion thereof, or the support screen is interposed between the membrane filter and the ring-shaped member. Since the sample is filtered through a membrane filter and a support screen, the unitized microorganism detection filter device can be counted without using tweezers. It can be set in the device. Therefore, by using tweezers as in the prior art, it is possible to prevent problems such as breakage of the membrane filter and to stably set the microbe detection filter device to the microbe counting device.

Further, in the present invention, the membrane filter is arranged on the support screen, and the outer peripheral part of both is supported by one ring-shaped member, so that the membrane filter is slackened and undulated even after collecting the microorganisms. The membrane filter can always be kept flat.
As a result, when the microbe detection filter device is attached to a microbe counting device equipped with autofocus, autofocus is easily applied at the time of microbe counting, which is accompanied by the occurrence of slack and undulation in the membrane filter as in the prior art. It is possible to prevent a decrease in the accuracy of the microorganism counting in the microorganism counting apparatus. Therefore, the blur or the focus shift of the fluorescent image due to the autofocus in the microorganism counter is eliminated, and the microorganism can be observed or counted with high accuracy.

In the present invention, the support screen is a metal plate having chemical resistance and has a plurality of small holes for filtering the specimen, so that the support screen can be supported by press working or drilling. The screen can be easily manufactured and the durability is increased. Moreover, the following effects can be further achieved by configuring as follows.
If the support screen is a glass plate and includes a plurality of small holes for filtering the specimen, it is possible to suppress background noise.
In addition, if the support screen is configured by forming a fine wire material such as a metal wire into a mesh shape, the fluorescence generated by the microorganisms is reflected by the support screen and the influence of background noise is removed. Is possible.

Furthermore, if the surface of the support screen is provided with a non-reflective coating, the influence of reflected light on the surface of the support screen can be suppressed.
If an uneven surface is formed on the surface of the support screen, the influence of reflected light on the surface can be suppressed by irregularly reflecting the uneven surface.

On the other hand, in the present invention, as an attachment mode of the support screen, if the outer periphery of the support screen is bonded to a ring-shaped member to which the membrane filter is fixed, and the support screen is fixed, The support screen can be firmly held between the membrane filter and the ring-shaped member, and the microorganism detecting filter device can be securely attached to the microorganism counting device.
Further, as an attachment mode of the support screen, the support screen is fixed by sandwiching the outer peripheral portion of the support screen between the membrane filter fixed to the ring-shaped member and the ring-shaped member. If so, the filter device for detecting microorganisms can be configured by simply sandwiching the support screen between the membrane filter and the ring-shaped member. As a result, the man-hour for bonding and fixing the support screen itself to the fixing ring can be omitted, and the completed microorganism detection filter device can be obtained with a small man-hour.

  Hereinafter, embodiments of the filter for detecting microorganisms according to the present invention will be described in detail.

[First Embodiment]
1A and 1B show a microorganism detection filter according to a first embodiment of the present invention, in which FIG. 1A is a sectional view and FIG. 1B is a perspective view. FIG. 2 is an explanatory view showing the attachment procedure of the microorganism detection filter in the first embodiment.
Prior to starting measurement with a microbe counting device that can color microorganisms from a sample that may have multiple types of microorganisms such as bacteria and fungi attached or mixed, and measure the microorganisms at once, Microorganisms are collected on a solid surface such as a membrane filter, and a pretreatment is performed in which the microorganisms are contacted with a chromogenic substance and stained.
As a filtration apparatus for extracting microorganisms used in this pretreatment, a filter apparatus for detecting microorganisms that collects microorganisms on a solid surface such as a membrane filter is used.
The present invention relates to a pretreatment microbe detection filter device.

1 and 2 showing the details of the microorganism detection filter according to the first embodiment of the present invention, 1 is a funnel, 3 is a flask, and the funnel 1 is a liquid (specimen) containing microorganisms toward the flask 3. Is used to inject.
Reference numeral 10 denotes a microorganism detection filter device. As shown in FIG. 2, the specimen in the funnel 1 is injected into the flask 3 through the microorganism detection filter device 10.
The microorganism detection filter device 10 shown in FIGS. 1 and 2 includes a disk-shaped membrane filter 4 that filters a specimen and collects microorganisms, and a fixing ring 6. The fixing ring 6 includes: It is formed in an annular shape so that vacuum filtration is possible. The membrane filter 4 has a large number of small holes 4a so that the specimen can be filtered, and the outer peripheral portion thereof is fixed to the upper surface of the fixing ring 6 by adhesion or the like.

A disk-like support screen 5 made of a metal plate having chemical resistance, such as SUS304 material, which is used in vacuum filtration or the like, is disposed in the vicinity of the membrane filter 4 immediately below the membrane filter 4. ing. That is, the membrane filter 4 is placed on the support screen 5. The support screen 5 has a large number of small holes (not shown) like the membrane filter 4, and the outer peripheral portion thereof is fixed to the fixing ring 6 by bonding or the like.
In addition to the metal plate, the support screen 5 may be formed by forming a large number of small holes in a fluorescent observation slide glass. Thus, if the support screen 5 is made of a slide glass, background noise can be suppressed.
As shown in FIG. 2, the microorganism detection filter device 10 configured as described above is set on the flask 3, thereby allowing the specimen to be injected from the funnel 1.

  In such a microorganism detection filter device 10, a microorganism-containing sample poured into the funnel 1 is passed through the membrane filter 4 of the microorganism detection filter device 10, whereby microorganisms in the sample are captured on the membrane filter 4. Be collected. Then, the sample after filtration in the flask 3 is aspirated by a reduced pressure source such as a vacuum pump. Thereafter, if a coloring material is poured into the funnel 1 and the coloring material is brought into contact with the microorganism, the microorganism is stained.

  The microorganism detection filter device 10 according to the first embodiment of the present invention is unitized by attaching and fixing the outer peripheral portion of the membrane filter 4 and the support screen 5 to the fixing ring 6. As a result, after the sample is filtered through the membrane filter 4 and the support screen 5, the microorganism detecting filter device 10 unitized as described above can be set in the microorganism counting device without using tweezers. It becomes. Therefore, according to the microorganism detection filter device 10 of the present embodiment, it is possible to prevent the occurrence of problems such as the above-described prior art such as the membrane filter 4 being damaged by using tweezers.

A support screen 5 is disposed immediately below the membrane filter 4 in the vicinity of the membrane filter 4, and the membrane filter 4 is placed on the support screen 5. Since the membrane filter 4 is bonded and fixed to one fixing ring 6, the membrane filter 4 does not sag or swell even after microorganisms are collected, and the membrane filter 4 can be kept flat at all times.
Therefore, when the microbe detection filter device 10 is attached to a microbe counting device equipped with autofocus, it becomes easy to perform autofocus during microbe count, and slack and undulation occur in the membrane filter 4 as in the prior art. Therefore, it is possible to prevent a decrease in the accuracy of the microorganism counting in the microorganism counting apparatus. As a result, the fluorescence image is not blurred or out of focus due to autofocusing in the microorganism counter, so that microorganisms can be observed or counted with high accuracy.
Note that when observing an image by fluorescence development with a microbe counting apparatus, the excitation light and the fluorescence have different wavelengths, and thus the reflection by the support screen 5 does not affect the observation result.

On the other hand, the support screen 5 can be configured as follows.
(1) The support screen 5 is configured by meshing a fine wire material such as a metal wire. As a result, the fluorescence generated by the microorganisms is reflected by the support screen 5, so that it becomes possible to remove the influence of background noise.
(2) The support screen 5 is formed by applying a non-reflective coating to the surface of a metal plate. Thereby, the influence of the reflected light on the surface of the support screen 5 can be suppressed.
(3) The surface of the support screen 5 is an uneven surface on which unevenness is formed. Thereby, the influence of the reflected light on the surface can be suppressed by providing irregularities and irregular reflection.

[Second Embodiment]
FIG. 3 is a cross-sectional view of a microorganism detection filter device according to a second embodiment of the present invention.
In the microorganism detection filter device 10 according to the second embodiment, the support screen 5 disposed in the vicinity of a portion immediately below the membrane filter 4 fixed to the upper surface of the fixing ring 6 by adhesion or the like is provided on the fixing ring 6. It is mounted by sandwiching it between the membrane filter 4 fixed to the plate and the step portion 6a of the fixing ring 6. Other configurations are the same as those in the first embodiment.
In this case, the man-hour for bonding and fixing the support screen 5 itself to the fixing ring 6 as in the first embodiment can be omitted.

  While the embodiments of the present invention have been described above, the present invention is not limited to the above-described embodiments, and various modifications and changes can be made based on the technical idea of the present invention.

The filter for microorganisms detection concerning a 1st embodiment of the present invention is shown, (A) is a sectional view and (B) is a perspective view. It is explanatory drawing which shows the attachment point of the filter for microorganisms detection in the said 1st Embodiment. It is a figure corresponding to Drawing 1 showing the filter for microorganisms detection concerning a 2nd embodiment of the present invention. It is a block diagram which shows an example of the microorganisms count device which concerns on a prior art. It is a whole block diagram of the microorganisms filtration apparatus concerning a prior art. It is sectional drawing which shows the detail of the filtration part of the filtration apparatus which concerns on a prior art.

Explanation of symbols

1 funnel 3 flask 4 membrane filter 5 support screen 6 fixing ring

Claims (8)

  A microbe detection filter device used in a microbe counting device that performs image analysis of microbes, including a membrane filter for collecting microbes, and having a small hole for filtering a specimen A support screen and a ring-shaped member for holding the outer peripheral portion of the membrane filter, the membrane filter is disposed on the support screen, and the outer peripheral portion of the support screen is disposed on the ring-shaped member. A filter device for detecting microorganisms, characterized by being supported by.   2. The filter device for detecting microorganisms according to claim 1, wherein the support screen is a metal plate having chemical resistance and has a plurality of small holes for filtering a specimen.   2. The filter device for detecting microorganisms according to claim 1, wherein the support screen is a glass plate material and has a plurality of small holes for filtering a specimen.   2. The filter device for detecting microorganisms according to claim 1, wherein the support screen is formed by meshing a fine wire material such as a metal wire.   4. The filter device for detecting microorganisms according to claim 1, wherein a surface of the support screen is provided with an anti-reflective coating.   The microbe detection filter device according to any one of claims 1 to 3, wherein an uneven surface is formed on a surface of the support screen.   The filter device for microorganism detection according to any one of claims 1 to 6, wherein an outer peripheral portion of the support screen is fixed to the ring-shaped member to which the membrane filter is fixed.   7. The support screen is fixed by sandwiching an outer peripheral portion between the membrane filter fixed to the ring-shaped member and the ring-shaped member. A filter device for detecting microorganisms.

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