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WO2003033691A1 - Micropuce cellulaire et son utilisation dans un procede d'analyse de cellules vivantes - Google Patents

Micropuce cellulaire et son utilisation dans un procede d'analyse de cellules vivantes Download PDF

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Publication number
WO2003033691A1
WO2003033691A1 PCT/RU2001/000432 RU0100432W WO03033691A1 WO 2003033691 A1 WO2003033691 A1 WO 2003033691A1 RU 0100432 W RU0100432 W RU 0100432W WO 03033691 A1 WO03033691 A1 WO 03033691A1
Authority
WO
WIPO (PCT)
Prior art keywords
gel
microchip
microcircuit
cells
cellular
Prior art date
Application number
PCT/RU2001/000432
Other languages
English (en)
Russian (ru)
Inventor
Andrei Darievich Mirzabekov
Tatyana Vasilievna Nasedkina
Denis Olegovich Fesenko
Original Assignee
Institut Molekulyarnoi Biologii Im. V.A. Engelgardta Rossiiskoi Akademii Nauk
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 Institut Molekulyarnoi Biologii Im. V.A. Engelgardta Rossiiskoi Akademii Nauk filed Critical Institut Molekulyarnoi Biologii Im. V.A. Engelgardta Rossiiskoi Akademii Nauk
Priority to PCT/RU2001/000432 priority Critical patent/WO2003033691A1/fr
Publication of WO2003033691A1 publication Critical patent/WO2003033691A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • 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
    • C12Q1/18—Testing for antimicrobial activity of a material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Definitions

  • the area of technology is related to the field of biological biology, microbiology and biology, and is subject to loss of payment.
  • the method of manufacture of the microchip is written in gel elements of the patient's cells, the immobilized or eukarotic cells are immobilized. It is manufactured by optimizing a gel-containing product that contains immobilized cells. Therefore, gel-forming products must be applied in the form of a microcircuit to be applied prior to polymerization.
  • a number of methods of analysis of hereditary factors, stocks of growth and development, and other properties of industrial and eukaryotic methods are known. Particularly, there is a group of methods for monitoring cell viability under conditions of exposure to different factors.
  • the objective of this invention is to work out a method that would have been easier to: 1. to speed up the analysis of the costs, the cost of the development and the development of development
  • One of the aspects of the invention is a cell phone, which is a simple product that contains gel elements, which are not used in any way.
  • the main gel elements are taken out of the group consisting of acrylamide, alginate, caragenene, collagen, fibrin, agarose, agarose, gelatin, vaginalis,
  • the gel cell element has a size suitable for the supply of healthy substances and substances, the interaction between food products and the consumer.
  • the service provides a free plate
  • the batteries that are used in the cells of the microcircuit may be the owners of both the e-cigarettes, the cigarettes and the cigarettes.
  • the method provides for the use of a portable microchip, which is deployed in this invention.
  • s ⁇ s ⁇ be ma ⁇ e ⁇ ial ⁇ dl ⁇ zh ⁇ i ⁇ le ⁇ chn ⁇ g ⁇ mi ⁇ chi ⁇ a vybi ⁇ ayu ⁇ of g ⁇ u ⁇ y s ⁇ s ⁇ yaschey of s ⁇ e ⁇ la ⁇ azlichny ⁇ s ⁇ v, me ⁇ all ⁇ v, ⁇ e ⁇ amiches ⁇ i ⁇ ma ⁇ e ⁇ ial ⁇ v, ⁇ lime ⁇ ny ⁇ n ⁇ si ⁇ eley ( ⁇ lime ⁇ ilme ⁇ a ⁇ ila ⁇ , ⁇ lis ⁇ i ⁇ l, ⁇ li ⁇ ilen and ⁇ .d.) lyuby ⁇ ⁇ m ⁇ zitsi ⁇ nny ⁇ ma ⁇ e ⁇ ial ⁇ v and ⁇ .d.
  • the method is designed to ensure that • the cages are removed from the group of ecuards or from the group of carriages, due to immobilization not obligatory in a free state.
  • the invention uses a method for manufacturing microchips that are immersed in a gel that is used to process the food. Therefore, gel-containing products containing a suspension are applied to the suspension in the form of a micro-droplet, after which they are applied.
  • Figure 1 is a schematic representation of the incubation chamber, and the numbers mean: 1 - plugs; 2-star glass; 3 - ⁇ emphasize favor; 4 - mikroochi ⁇ ;
  • Figure 3 shows the result of the incubation of acrylamide mixtures in the environment with antibiotic
  • Figure 4 shows the results of the incubation of the microbial in the environment with the antibiotic, where the numbers 1 and 2 are sensitive strains, and the numbers 3 and 4 are indicated;
  • Figure 5 shows the gel elements of the microbial after incubation in a healthy environment with antibiotics, and ⁇ - a cell with a sensitive culture, and B - a cell with a sensitivity.
  • the manufactured microchip may be used immediately, or it may be unavailable.
  • acrylamide gel for the manufacture of mixtures on the basis of live animals is not acceptable, t. ⁇ . causes the death of 100% cells.
  • the engine is the most suitable for this purpose.
  • the problem of fixation of alginate gel elements in order to improve the service is solved by the following method.
  • the processed ⁇ -dyaine is formed with a thin layer (5-10 ⁇ m) of polylamide gel (similar to [ ⁇ .
  • n ⁇ vmes ⁇ mas ⁇ i is ⁇ lzue ⁇ sya ⁇ z ⁇ achn ⁇ e ⁇ va ⁇ tsev ⁇ e s ⁇ e ⁇ l ⁇ ) for ⁇ y nan ⁇ sya ⁇ sya mi ⁇ a ⁇ li algina ⁇ a with ⁇ le ⁇ ami ⁇ lime ⁇ izuyu ⁇ sya in SaS1 2.
  • gel elements are generally only available to the user.
  • a more technical solution is to apply a microproil to a chemically modified glass. For specialists in the area of chemistry, it may be difficult to find a means of chemical alteration of the glass.
  • the proposed method makes it possible in one stage to apply the microcircuit with a high density of the elements of the matrix, which is not necessary for larger manipulations.
  • the proposed method makes it possible to use 8
  • a reagent which is slightly modified by unsaturated groups; it is a For the manufacture of a portable microchip on a basic bacterium or other, it is predominantly used as a part of a large number of acrylamides.
  • cordless processors are provided on the basis of the US. The writing of these data should not be used to limit the use of this patent; In order to show the possibility of researching the properties of the mains plugs using a microchip, it is possible to carry out various property tests.
  • ⁇ a ⁇ a ⁇ e ⁇ ny e ⁇ s ⁇ nentsialny ⁇ s ⁇ signal ⁇ is ⁇ di ⁇ in ⁇ ezul ⁇ a ⁇ e ⁇ azmn ⁇ zheniya us ⁇ ychivy ⁇ ⁇ an ⁇ ibi ⁇ i ⁇ u ba ⁇ e ⁇ y in yachey ⁇ a ⁇ mi ⁇ chi ⁇ a and na ⁇ leniya them ⁇ lu ⁇ estsen ⁇ n ⁇ g ⁇ ⁇ asi ⁇ elya, s ⁇ de ⁇ zhascheg ⁇ sya in ⁇ i ⁇ a ⁇ eln ⁇ y s ⁇ ede ( ⁇ ivaya 1 ⁇ ig. 2.). For this, in the gel cell, separate fractions are used.
  • Example 2 Gel-forming mixture, at 37 ° ⁇ and containing
  • Bacterial cultures isolated from patients are primarily grown on a solid medium. All the interested parts remove the lugs and insert them into the separate parts with the gel-forming solution, carefully suspend.
  • the automatic microprocessor selects 0.1–0.2 microns from each appliance and, in a separate order, is applied to the appliance, it is free of charge. They are used in parishes ⁇ a (see Example 1).
  • the number of manufactured products is determined by the number of antibiotics, the sensitivity to inadvertent process and the inability to process it.
  • Each microscope uses a droplet (which covers all the elements) of the liquid medium containing a separate concentration of antigens.
  • the cameras are placed in a humid chamber and incubated for 3-4 hours at 37 ° ⁇ .
  • the recording of the results is carried out as follows. Cleaners wash the water, place a microcircuit (400 x ) and evaluate the presence of seeds in each microcircuit cell (Fig. 5 ⁇ ). The absence of cells in the cell (Fig. 5B) indicates that this antibiotic is effective in this culture.
  • Sources and publications listed in the text are an integral part of it, as long as all their contents were included in the text.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Organic Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Cell Biology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Food Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biophysics (AREA)
  • Toxicology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)

Abstract

Cette invention concerne une micropuce pouvant être utilisée dans les domaines de la médecine, la biologie moléculaire, la microbiologie et la biotechnologie notamment pour déterminer l'activité anti-bactérienne d'antibiotiques en pharmacologie expérimentale, ou pour analyser l'environnement. Cette invention concerne également un procédé de fabrication de cette micropuce consistant à immobiliser, dans des éléments sous forme de gel de cette micropuce, des cellules procaryotes et eucaryotes au moyen de la polymérisation d'une solution gélifiante contenant les cellules immobilisées. Des solutions gélifiantes se présentant sous la forme de micro-gouttes sont appliquées sur un substrat avant l'étape de polymérisation.
PCT/RU2001/000432 2001-10-19 2001-10-19 Micropuce cellulaire et son utilisation dans un procede d'analyse de cellules vivantes WO2003033691A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/RU2001/000432 WO2003033691A1 (fr) 2001-10-19 2001-10-19 Micropuce cellulaire et son utilisation dans un procede d'analyse de cellules vivantes

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/RU2001/000432 WO2003033691A1 (fr) 2001-10-19 2001-10-19 Micropuce cellulaire et son utilisation dans un procede d'analyse de cellules vivantes

Publications (1)

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WO2003033691A1 true WO2003033691A1 (fr) 2003-04-24

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011097664A2 (fr) 2010-02-10 2011-08-18 Forschungsholding Tu Graz Gmbh Dispositif de test

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5037740A (en) * 1985-03-13 1991-08-06 Asama Chemical Co., Ltd. Novel immobilized cells and fermentation method utilizing the same
RU2041262C1 (ru) * 1993-08-11 1995-08-09 Институт молекулярной биологии им.В.А.Энгельгардта РАН Способ иммобилизации водорастворимых биоорганических соединений на капиллярно-пористый носитель
SU1264575A1 (ru) * 1984-10-23 1996-02-10 Институт биологической физики АН СССР Способ иммобилизации клеток
RU2086652C1 (ru) * 1993-10-01 1997-08-10 Александр Людвигович Симонян Способ количественного определения l-пролина
RU2157385C1 (ru) * 1999-07-19 2000-10-10 Институт молекулярной биологии им. В.А. Энгельгардта РАН Способ изготовления микрочипов на основе олигонуклеотидов
WO2000065098A2 (fr) * 1999-04-27 2000-11-02 University Of Chicago Extension nucleotidique sur un micro-arrangement d'amorces immobilisees au moyen d'un gel

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SU1264575A1 (ru) * 1984-10-23 1996-02-10 Институт биологической физики АН СССР Способ иммобилизации клеток
US5037740A (en) * 1985-03-13 1991-08-06 Asama Chemical Co., Ltd. Novel immobilized cells and fermentation method utilizing the same
RU2041262C1 (ru) * 1993-08-11 1995-08-09 Институт молекулярной биологии им.В.А.Энгельгардта РАН Способ иммобилизации водорастворимых биоорганических соединений на капиллярно-пористый носитель
RU2086652C1 (ru) * 1993-10-01 1997-08-10 Александр Людвигович Симонян Способ количественного определения l-пролина
WO2000065098A2 (fr) * 1999-04-27 2000-11-02 University Of Chicago Extension nucleotidique sur un micro-arrangement d'amorces immobilisees au moyen d'un gel
RU2157385C1 (ru) * 1999-07-19 2000-10-10 Институт молекулярной биологии им. В.А. Энгельгардта РАН Способ изготовления микрочипов на основе олигонуклеотидов

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
EGOROV N.S.: "Mikroby antogonisty I biologicheskie metody opredeleniya antibioticheskoi aktivnosti", VYSCHAYA SHKOLA, M., 1965, pages 83 *
ZVYAGINTSEVA D.G. PROF.: "Moskovskogo universiteta", MOSKOVSKOGO UNIVERSITETA, 1980, pages 27 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2011097664A2 (fr) 2010-02-10 2011-08-18 Forschungsholding Tu Graz Gmbh Dispositif de test
AT509355B1 (de) * 2010-02-10 2012-04-15 Univ Graz Tech Testanordnung
US8785142B2 (en) 2010-02-10 2014-07-22 Eva Sigl Test arrangement

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