WO1997038128A1 - Technique de differenciation immediate de bacteries et appareil correspondant - Google Patents

Technique de differenciation immediate de bacteries et appareil correspondant Download PDF

Info

Publication number
WO1997038128A1
WO1997038128A1 PCT/JP1996/002370 JP9602370W WO9738128A1 WO 1997038128 A1 WO1997038128 A1 WO 1997038128A1 JP 9602370 W JP9602370 W JP 9602370W WO 9738128 A1 WO9738128 A1 WO 9738128A1
Authority
WO
WIPO (PCT)
Prior art keywords
light
fluorescent
bacteria
fungi
cells
Prior art date
Application number
PCT/JP1996/002370
Other languages
English (en)
Japanese (ja)
Inventor
Yoshiyuki Tokuda
Jongchul Park
Original Assignee
Nippon Mizushorigiken Co., Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nippon Mizushorigiken Co., Ltd. filed Critical Nippon Mizushorigiken Co., Ltd.
Priority to AU67547/96A priority Critical patent/AU6754796A/en
Publication of WO1997038128A1 publication Critical patent/WO1997038128A1/fr

Links

Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2304/00—Chemical means of detecting microorganisms
    • C12Q2304/10—DNA staining
    • C12Q2304/13—Propidium

Definitions

  • fungi such as bacteria are adhered or collected, and the fungi are appropriately adhered and collected from a specimen to which the fungi adhere or are mixed, and the bacterial species, the amount of attached fungi, the amount of viable bacteria, and the death Bacteria can be distinguished, and especially for food producers and distributors, it is extremely useful for hygiene management, and the method for immediately distinguishing fungi and its discriminating apparatus are used.
  • Background art fungi such as bacteria are adhered or collected, and the fungi are appropriately adhered and collected from a specimen to which the fungi adhere or are mixed, and the bacterial species, the amount of attached fungi, the amount of viable bacteria, and the death Bacteria can be distinguished, and especially for food producers and distributors, it is extremely useful for hygiene management, and the method for immediately distinguishing fungi and its discriminating apparatus are used.
  • Foods especially fresh foods and processed foods with a high moisture content, have conditions that facilitate the propagation of fungi such as bacteria and bacteria, and these foods contain fungi from the beginning or In the process leading to its distribution and consumption, it adheres or falls into the water, which not only propagates in a short time and becomes remarkably unhealthy, but also results in food poisoning along with deterioration and spoilage of foods.
  • preservatives have been added to the extent permitted to prevent the growth of fungi.
  • the use of preservatives is considered dangerous, and consumers are forced to select non-additive processed foods.There is a growing demand for these products, so they must deal with them without additives.Product liability to strengthen consumer protection In conjunction with the enforcement of the liability law, food producers and distributors are in high demand for sanitary D control at the time of production or distribution.
  • fungi collected from specimens are inoculated in a coloring medium in which a coloring enzyme is mixed with an appropriate medium, and the incubator is kept at a required temperature for about 24 hours to 48 hours. Incubate for a period of time to form colonies, which are then retained by Escherichia coli, etc .; S-galactosidase enzyme
  • a fluorescent dye is mixed with physiological saline to an appropriate degree.
  • the luminescence energy is also weak, so a high-precision high-power microscope must be used, and the device for discrimination must use an extremely expensive and large-sized device.
  • the present invention provides a method of appropriately depositing and collecting bacteria from a specimen to which bacteria such as bacteria and the like are contaminated, and using a simple method and an inexpensive device to immediately kill live bacteria.
  • the dye can be penetrated into fungal cells, and in the viable cell ⁇ , the dye is dispersed and emits fluorescence with the absorption of the excitation light, and in the dead cell ⁇ , the dye is taken into the cell tissue to absorb the excitation light.
  • a fluorescent dye consisting of fluorescein or a derivative thereof, which cannot emit fluorescence, and an aropidum iodide which cannot penetrate living cells but penetrates only dead cells and emits fluorescence by absorbing excitation light.
  • Fluorescent dye is mixed with physiological saline at a concentration of 3 to 1 o1 / m1 and from salts, chitinus or cellulase so as to increase the penetration into the cells and to effectively stain the cells with the fluorescent dye.
  • a fungus collected from a specimen is mixed into a staining solution obtained by mixing 1 to 1 Ojmo IZm1 with a staining accelerator, and the mixture is stained while being heated to a required temperature.
  • Fluorescent dye consisting of fluorescein or its derivative penetrated into fungal cells by irradiating it with excitation light having a central wavelength of 488 nm from the lower side, It emits green fluorescent light with a slightly yellowish color of 520 nm, and the fluorescent dye consisting of propidium iodide gives a strong reddish orange fluorescent light with a central wavelength of 625 nm. It emits light.
  • FIG. 1 is an explanatory diagram of a method for instantaneously identifying fungi according to the present invention.
  • FIG. 2 is an explanatory diagram of an apparatus for instantly identifying fungi according to the present invention.
  • FIG. 3 is a diagram for immediately identifying fungi by computer-graphics processing.
  • FIG. 3 is an explanatory view of the apparatus, and shows the best form for implementing the ⁇ invention.
  • FIG. 1 shows a flow chart of a method for immediately discriminating fungi.
  • Bacterial susceptible bacteria collected by the method described in (1) include, for example, adding a sterilized halo stick to physiological saline and attaching it to the surface of the sample.
  • the method includes contacting a water-soluble medium with the surface of a sample and collecting the sample.
  • the fungus 1 thus collected can penetrate the physiological saline 2 into the cells of live and dead bacteria, and is dispersed in the viable cells, and is taken up and dispersed in the dead cells in the dead cells.
  • a fluorescent dye 3 consisting of fluorophore or its derivative, which emits strong fluorescence with a center wavelength of 520 nm due to absorption of excitation light having a center wavelength of 488 nm; It can not penetrate into cells but can die and can penetrate into cells, and can absorb and excite excitation light with a center wavelength of 488 nm, and emit strong fluorescence with a center wavelength of 625 nm
  • Uruvide Fluorescent dyes 4 consisting of umodide, 3 mo 1 / m 1 each The mixture is mixed within the range of 15 mo 1 ⁇ ⁇ 1, and the physiological saline solution 2 is made flexible so that the fluorescent dyes 3 and 4 can sufficiently penetrate and disperse in the fungal cells.
  • Staining solution 20 is prepared by mixing 3 mo 1 / m 1 to 15 mo 1 Nom 1 of dyeing accelerator 5 consisting of salt, chiness or cellulase.
  • Specific examples of the salts used for the dye accelerator 5 include sodium chloride and magnesium chloride.
  • the staining solution 20 formed as described above is mixed with the fungus 1 collected from the specimen to perform staining. In this case, the staining solution 20 is heated to an appropriate temperature in order to enhance the staining property. It is preferable to use a temperature of 25 ° C. to 35 ° C. in general, and the time required for dyeing at such a temperature is about 5 to 15 minutes.
  • the fungi 1 mixed into the staining solution 20 are not affected by the stain-promoting agent 5, but the softening of the membrane promotes the penetration of the fluorescent dyes 3 and 4 into the cells. In this case, the fluorescent dyes 3 and 4 that have permeated into the cells are washed away again, resulting in indistinguishability due to a decrease in fluorescence emission.
  • Fig. 2 is an explanatory diagram of a discrimination device that discriminates the species, attached amount, viable bacteria, and dead bacteria of fungus 1 collected from a sample using the stained bacterial solution 21 in which the staining is stabilized. Since the cells of the fungus 1 stained with the fluorescent dyes 3 and 4 are microscopic, it is necessary to create fluorescent light as highly visible as possible from the fluorescent dyes 3 and 4 in the cells. By the way, the fluorescent dye 3 consisting of fluorescein or its derivative, which penetrates and disperses into viable fungal cells of fungi 1, absorbs the excitation light having a center wavelength of 488 nm and absorbs the center of the fluorescent dye according to the S1 ⁇ 1x rule.
  • the fluorescent dye 4 composed of propidum iodide, which emits strong fluorescent light with a wavelength of 520 nm and which can penetrate and disperse into dead cells but cannot penetrate into viable cells, has a central wave It emits strong fluorescent light with a center wavelength of 625 nm as it absorbs excitation light having a length of 488 nm.
  • an excitation light source 31 capable of irradiating excitation light having a center wavelength of 488 nm is provided below the casing 30 so that the inside thereof is held by a dark room streak.
  • a magnifying lens 32 of a required magnification, preferably about 100 to 500 times, and a light scattering process is provided at an intermediate position of the irradiation optical axis search.
  • the light-transmitting plate 33 is provided with a light-transmitting plate 33, and the light-scattering of the light-transmitting plate 33 preferably has a light scattering rate of about 5 to 50 times.
  • a specific example is frosted glass.
  • the stained bacterial solution 21 is dropped on the light-transmitting plate 33, and the excitation light source 31 provided therebelow is irradiated with excitation light having a center wavelength of 488 nm to thereby perform the staining.
  • Fluorescein or fluorescein which is permeated and stained into cells of fungus 1 mixed in bacterial liquid 21 and penetrates and disperses from viable bacteria as it absorbs excitation light, From the fluorescent dye 3 composed of the derivative, the fluorescence emission of the bacterial species and the fluorescence emission number related to the number of bacteria having a fluorescence emission color having a center wavelength of 50 nm and having different emission shapes and emission sizes are different.
  • a fluorescent dye composed of aropidum iodide is used to generate a fluorescent color having a central wavelength of 625 nm and a fluorescent color related to a bacterial species having a different luminescent shape and luminescent intensity.
  • the number of luminescence and the number of fluorescent light related to the number of bacteria are expanded by light scattering.
  • the large lens 3 2 It will be recognized and can be determined.
  • FIG. 3 is an explanatory diagram of a discrimination device for processing and displaying and storing the fluorescence emission light ⁇ from the fungus 1 visually discriminated by the magnifying lens 3 2 as an image or data in the instant fungus discrimination device of the present invention.
  • a band-pass filter 40 that transmits fluorescent light with a wavelength of 52 nm and a band-pass filter 41 that transmits fluorescent light with a wavelength of 65 nm can be freely used.
  • the filter is provided with a band-pass switching filter 4 that can be switched between the fluorescent light-emitting lights, and the fluorescent light that has passed through the magnifying lens 32 and passed through the band-pass filter 40 or 41 has a focal position at which the fluorescent light is emitted.
  • a video camera 5 is provided for converting the shape and size of the light beam, the number of fluorescent light emission or the number of fluorescent light emission into an electric signal, and the video camera 5 converts the converted electric signal.
  • Wakashi stored ⁇ required surface image processing Ku is connected to a computer 6 which is programmed to data processing is performed.
  • the fluorescent light having passed through the magnifying lens 32 has an image formed by the fluorescent light having a wavelength of 520 nm and an image formed by the fluorescent light having a wavelength of 625 ⁇ by switching the band-pass switching filter 4.
  • the fluorescent light having passed through the magnifying lens 32 has an image formed by the fluorescent light having a wavelength of 520 nm and an image formed by the fluorescent light having a wavelength of 625 ⁇ by switching the band-pass switching filter 4.
  • an image visually recognized by the magnifying lens 32 can be displayed and stored as a photograph or data.
  • a video camera 5 equipped with a band-pass filter 40 with a wavelength of 52 nm and a band-pass filter 141 with a wavelength of 6 25 ⁇ ⁇ were installed.
  • the same result can be obtained by using two video cameras 5 and processing the electric signal from each video camera 5 with the computer 6.
  • a fluorescent dye that emits a different fluorescent color in live and dead cells can be permeated and dispersed in a short time.
  • Stained bacteria solution that can prevent staining of the permeated and dispersed fluorescent dye is dropped onto a light-transmissive light-transmissive plate, and excitation is performed with a center wavelength of 488 nm from below.
  • fluorescence is emitted with a center wavelength of 250 nm, which is clear and highly visible from live bacteria, and a center wavelength of 625 nm, which is clear and highly visible from dead bacteria.
  • Fluorescence is emitted, and the emitted light ⁇ ⁇ is scattered by the light-transmitting plate and is enlarged in advance, so that even a relatively low-magnification magnifying lens can determine not only viable cells and dead cells, but also the shape and shape of the fluorescent light.
  • Bacterial species can be distinguished by the size, and the number of bacteria can be Since the distinction is clearly visible, hygiene management can be performed continuously without stopping the product even in the production and distribution processes of foods, and in the present invention, the determination results can be recorded and stored in the form of photographs and data. As a result, it is possible to appropriately deal with the occurrence of hygiene issues with consumers.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • General Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Toxicology (AREA)
  • Optics & Photonics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

L'invention porte sur une technique permettant une différenciation immédiate de bactéries consistant à introduire un échantillonnage de bactéries dans une solution de coloration issue du mélange d'une solution saline physiologique, d'un colorant fluorescent tel que de la fluorescéine ou un dérivé de cette substance, d'un autre colorant fluorescent contenant un iodure de propidium, chacun selon la concentration exigée, et d'un accélérateur de coloration selon une concentration donnée, à chauffer le mélange obtenu à la température requise pour colorer des bactéries vivantes et mortes, ces bactéries se colorant du fait de la pénétration et de la dispersion des colorants fluorescents dans leurs cellules et ce, sélectivement. On ajoute alors au mélange résultant un inhibiteur d'écoulement, selon une concentration requise, afin de préparer une solution contenant les bactéries colorées, puis l'on fait tomber goutte-à-goutte cette solution sur une plaque translucide dont on a fait en sorte qu'elle diffuse la lumière. On soumet cette solution aux rayons d'une lumière d'excitation ayant une longueur d'onde centrale de 488 nm et dont la source se trouve en dessous de la plaque. On est ainsi en mesure d'observer, à l'aide d'un microscope, la lumière fluorescente émise du fait de l'absorption de la lumière d'excitation, la configuration et les dimensions de cette émission fluorescente ainsi que le nombre des points fluorescents, ce qui permet de constater les différences existant entre les bactéries vivantes et mortes, de déterminer le type de bactérie et d'établir la numération bactérienne. L'invention porte également sur un appareil permettant d'effectuer une différenciation immédiate de bactéries, lequel appareil comporte une source de lumière d'excitation, d'une longueur d'onde centrale de 488 nm, située dans la partie inférieure d'un boîtier dont l'intérieur est maintenu dans des conditions similaires de celles d'une chambre noire, un microscope se trouvant au-dessus de l'axe optique de la lumière d'excitation et une plaque translucide dont on a fait en sorte qu'elle diffuse la lumière, cette plaque se trouvant dans la partie intermédiaire de l'appareil. Elle porte également sur un autre appareil permettant d'effectuer cette différenciation immédiate, lequel appareil est, en outre, pourvu d'un filtre passe-bande surmontant le microscope et servant à sélectionner des ondes d'une longueur comprise entre 520 et 625 nm ainsi que d'une caméra vidéo située au-dessus du filtre, ce qui permet de convertir les images ayant traversé le microscope en signaux électriques et de synthétiser ces signaux au moyen d'un ordinateur pour traitement d'image.
PCT/JP1996/002370 1996-04-11 1996-08-26 Technique de differenciation immediate de bacteries et appareil correspondant WO1997038128A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU67547/96A AU6754796A (en) 1996-04-11 1996-08-26 Method of immediately discriminating bacteria and apparatus therefor

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP8125218A JP2979383B2 (ja) 1996-04-11 1996-04-11 菌類の即時判別方法
JP8/125218 1996-04-11

Publications (1)

Publication Number Publication Date
WO1997038128A1 true WO1997038128A1 (fr) 1997-10-16

Family

ID=14904787

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP1996/002370 WO1997038128A1 (fr) 1996-04-11 1996-08-26 Technique de differenciation immediate de bacteries et appareil correspondant

Country Status (3)

Country Link
JP (1) JP2979383B2 (fr)
AU (1) AU6754796A (fr)
WO (1) WO1997038128A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6251624B1 (en) 1999-03-12 2001-06-26 Akzo Nobel N.V. Apparatus and method for detecting, quantifying and characterizing microorganisms
US6309835B1 (en) 1999-05-27 2001-10-30 Koninkiijke Philips Electronics N.V. Methods for quantitating the efficacy of oral care products
US6979828B2 (en) * 2001-02-15 2005-12-27 Nippon Mizushori Giken Co. Ltd. Method and apparatus for immediately determining microorganism
GB2510365A (en) * 2013-01-31 2014-08-06 Blood Analysis Ltd Quantification of bacteria
CN108350409A (zh) * 2015-12-28 2018-07-31 日本理化学开发公司 活菌/死菌状态判定装置以及使用该装置的活菌/死菌状态判定方法
CN111006991A (zh) * 2018-10-30 2020-04-14 江南大学 一种确定污水处理好氧反硝化菌最适保存温度的方法

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003102496A (ja) * 2001-09-27 2003-04-08 Matsushita Ecology Systems Co Ltd 微生物検査システム
JP4876251B2 (ja) * 2006-08-29 2012-02-15 国立大学法人山口大学 生菌、死菌及び疑似生菌の存在割合判別方法
CN101225428B (zh) * 2007-01-19 2012-01-25 华南农业大学 一种染色液及其染色方法和应用
CN113533004A (zh) * 2021-07-30 2021-10-22 深圳联合医学科技有限公司 一种多重荧光染色液及其制备方法和使用方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BY THE FOUNDATION JAPAN CHEMICAL SOCIETY, "New Biochemical Experimental Lecture 17 Microbial Experimental Method", (23-03-92); & TOKYO KAGAKU DOJIN CO. LTD., p. 66-69. *
CYTOMETRY, Vol. 15, (1994), M.J. HUMPHREYS et al., "Determination of the Viability of Trichomonas Vaginalis Using Flow Cytometry", p. 343-348. *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6251624B1 (en) 1999-03-12 2001-06-26 Akzo Nobel N.V. Apparatus and method for detecting, quantifying and characterizing microorganisms
US6416969B2 (en) 1999-03-12 2002-07-09 Akzo Nobel N.V. Susceptibility plates for microbial antibiotic susceptibility testing
US6309835B1 (en) 1999-05-27 2001-10-30 Koninkiijke Philips Electronics N.V. Methods for quantitating the efficacy of oral care products
US6979828B2 (en) * 2001-02-15 2005-12-27 Nippon Mizushori Giken Co. Ltd. Method and apparatus for immediately determining microorganism
GB2510365A (en) * 2013-01-31 2014-08-06 Blood Analysis Ltd Quantification of bacteria
WO2014118544A1 (fr) * 2013-01-31 2014-08-07 Blood Analysis Ltd Quantification de bactéries
CN108350409A (zh) * 2015-12-28 2018-07-31 日本理化学开发公司 活菌/死菌状态判定装置以及使用该装置的活菌/死菌状态判定方法
CN108350409B (zh) * 2015-12-28 2022-06-14 日本理化学开发公司 活菌/死菌状态判定装置以及使用该装置的活菌/死菌状态判定方法
CN111006991A (zh) * 2018-10-30 2020-04-14 江南大学 一种确定污水处理好氧反硝化菌最适保存温度的方法
CN111006991B (zh) * 2018-10-30 2021-06-25 江南大学 一种确定污水处理好氧反硝化菌最适保存温度的方法

Also Published As

Publication number Publication date
AU6754796A (en) 1997-10-29
JP2979383B2 (ja) 1999-11-15
JPH09275998A (ja) 1997-10-28

Similar Documents

Publication Publication Date Title
US6914250B2 (en) Fluorometric detection using visible light
Rost Fluorescence microscopy
US9964489B2 (en) System and method for controlling depth of imaging in tissues using fluorescence microscopy under ultraviolet excitation following staining with fluorescing agents
CN109154569A (zh) 用于对未切片组织样本成像的方法和装置
JP6851668B2 (ja) 生死菌の状態判定装置を用いた生死菌状態判定方法
WO1997038128A1 (fr) Technique de differenciation immediate de bacteries et appareil correspondant
RU2374643C2 (ru) Способ оценки свежести рыбного продукта
JP5717375B2 (ja) 真珠品質の非破壊判定方法
US10511820B2 (en) Pseudo HandE image producing method and optical system using same
US5213830A (en) Method of detecting worms in meat
JP3734080B2 (ja) 菌類の即時判別方法及び装置
JPH11178568A (ja) 菌類の即時判別装置
JP2017169455A (ja) 培養併用蛍光インサイチューハイブリダイゼーション法を用いた食品中の細菌からのシグナル検出方法
JP2019028062A (ja) 蛍光に基づく検知を使用する組織切片の調製
JP3055017B2 (ja) 菌類の即時判別装置
KAUPPI et al. Determination of the distribution of lichen substances in the thallus by fluorescence microscopy
US10665000B2 (en) Pseudo H and E image producing method and optical system using same
JPH09318617A (ja) 食品の鮮度判定方法
Van der Ploeg et al. Reflection versus fluorescence: A note on the physical backgrounds of two types of light microscopy
Model et al. Observation of living organisms in environmental samples by transmission-through-dye microscopy
JPH0349560B2 (fr)
JP2000232897A (ja) 菌類の即時判別方法
Phipps et al. [14] Deconvolution fluorescence microscopy for observation and analysis of membrane biofilm architecture
JP2006329775A (ja) 化合物の観察方法及び該方法に用いる着色溶液
US5952192A (en) Method of fluorescent analysis of biological sample utilizing biebrich scarlet

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CN KR US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
122 Ep: pct application non-entry in european phase