DE19906352A1 - Apparatus to identify and count biological microparticles - Google Patents
Apparatus to identify and count biological microparticlesInfo
- Publication number
- DE19906352A1 DE19906352A1 DE19906352A DE19906352A DE19906352A1 DE 19906352 A1 DE19906352 A1 DE 19906352A1 DE 19906352 A DE19906352 A DE 19906352A DE 19906352 A DE19906352 A DE 19906352A DE 19906352 A1 DE19906352 A1 DE 19906352A1
- Authority
- DE
- Germany
- Prior art keywords
- particles
- capillary
- ferromagnetic
- metal coil
- antibodies
- Prior art date
- Legal status (The legal status 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 status listed.)
- Withdrawn
Links
- 239000011859 microparticle Substances 0.000 title claims abstract 6
- 239000002245 particle Substances 0.000 claims abstract 16
- 230000005294 ferromagnetic effect Effects 0.000 claims abstract 9
- 239000002184 metal Substances 0.000 claims abstract 6
- 239000007788 liquid Substances 0.000 claims abstract 3
- 241000700605 Viruses Species 0.000 claims abstract 2
- 230000010355 oscillation Effects 0.000 claims abstract 2
- 229920003023 plastic Polymers 0.000 claims abstract 2
- 239000004033 plastic Substances 0.000 claims abstract 2
- 238000004062 sedimentation Methods 0.000 claims abstract 2
- 238000000034 method Methods 0.000 claims 2
- 101710132601 Capsid protein Proteins 0.000 claims 1
- 101710094648 Coat protein Proteins 0.000 claims 1
- 102100021181 Golgi phosphoprotein 3 Human genes 0.000 claims 1
- 101710125418 Major capsid protein Proteins 0.000 claims 1
- 101710141454 Nucleoprotein Proteins 0.000 claims 1
- 101710083689 Probable capsid protein Proteins 0.000 claims 1
- 238000001514 detection method Methods 0.000 claims 1
- 230000006698 induction Effects 0.000 abstract 1
- 230000001939 inductive effect Effects 0.000 abstract 1
- 102000004169 proteins and genes Human genes 0.000 abstract 1
- 108090000623 proteins and genes Proteins 0.000 abstract 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/04—Investigating sedimentation of particle suspensions
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/1031—Investigating individual particles by measuring electrical or magnetic effects
-
- 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
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/06—Investigating concentration of particle suspensions
- G01N15/0656—Investigating concentration of particle suspensions using electric, e.g. electrostatic methods or magnetic methods
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
- G01N2015/019—Biological contaminants; Fouling
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/0098—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Dispersion Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analyzing Materials By The Use Of Magnetic Means (AREA)
Abstract
Description
Die Zählung von Bakterien, Blutzellen oder Zellbestandteilen in wäßrigen Lösungen erfolgt bisher mittels Durchflußzytometer oder Coultercounter. Hier werden die entsprechenden Partikel gefärbt und anhand von optischen Signalen identifiziert oder durch kapazitive Messungen gezählt.Bacteria, blood cells or cell components in aqueous solutions have so far been counted using a flow cytometer or Coulter counter. Here the corresponding particles are colored and identified on the basis of optical signals or counted by capacitive measurements.
Um den apparativen Aufwand bei der optischen Messung zu umgehen und eine höhere Spezifität gegenüber der kapazitiven Messung zu erreichen, wird für den Nachweis des einzelnen Partikels ein anderes Meßprinzip eingesetzt: Die Messung der Induktivitätsänderung einer Mikrospule aus Metall. Da biologische Partikel aber eine Permeabilitätskonstante µ von annähernd 1 haben, müssen diese zum Nachweis und zur Zählung mittels Spule zuvor mit induktivitätsändernden Substanzen markiert werden. Diese Markierung geschieht durch die immunologische, phagologische oder molekularbiologische Ankopplung von ferromagnetischen Partikeln, welche monovalent entweder mit Antikörpern, mit Virus-Andockmolekülen oder mit Gensonden an Spacermolekülen verbunden sind.In order to avoid the expenditure on equipment for optical measurement and a higher specificity compared to the capacitive measurement, is used for the detection of the individual particle Another measuring principle used: The measurement of the change in inductance of a metal micro coil. Since biological particles have a permeability constant µ of approximately 1, they have to previously marked with inductivity-changing substances for detection and counting by means of a coil become. This marking happens through the immunological, phagological or molecular biological coupling of ferromagnetic particles which are monovalent with either Antibodies are connected with virus docking molecules or with gene probes on spacer molecules.
Die Kopplung der ferromagnetischen Marker erfolgt in einer Vorrichtung, welche gleichzeitig eine Anreicherung der zu zählenden Partikel ermöglicht: Die Marker werden in einer Teflonkapillare mittels einem Elektromagnet als Sorptions-Schicht festgehalten, bis die gesamte Probe in die Kapillare gepumpt wurde und gleichzeitig die überschüssige Probe aus der Kapillare herausgelaufen ist. Hierauf wird der Magnet ausgeschaltet, damit die Marker frei diffundieren und die Oberfläche der biologischen Partikel sättigen können. Darauf wird der Kapillaren-Inhalt mit einer piezoelektrischen Pumpe durch eine Metallspule gepumpt, die als Spirale auf eine Leiterplatte geätzt wurde und mit Kondensator und Widerstand als Schwingkreis geschaltet ist. Der Schwingkreis wird mit einer Frequenz angeregt die derjenigen Eigenschwingfrequenz entspricht, welche entsteht, wenn sich ein durchschnittlich markierter biologischer Mikropartikel in der Spule befindet. Dadurch entsteht im Schwingkreis immer dann eine Resonanzschwingung, wenn ein entsprechender Mikropartikel durch die Spule tritt.The coupling of the ferromagnetic markers takes place in a device which is also a Enrichment of the particles to be counted enables: The markers are placed in a Teflon capillary an electromagnet as a sorption layer until the entire sample is held in the capillary was pumped and at the same time the excess sample ran out of the capillary. The magnet is then switched off so that the markers diffuse freely and the surface of the can saturate biological particles. The capillary content is then covered with a piezoelectric Pump pumped through a metal coil, which was etched as a spiral on a printed circuit board and with Capacitor and resistor is connected as a resonant circuit. The resonant circuit is with a Frequency excited which corresponds to the natural vibration frequency which arises when a average labeled biological microparticles in the coil. This creates in The resonant circuit always resonates when a corresponding microparticle passes through the coil kicks.
Ein Beispiel für die Anwendung dieses Verfahrens ist der Nachweis von Kolibakterien in Wasserproben (siehe schematische Zeichnung, Vermerk 1). Hierzu werden monovalente primäre E.-coli-spezifische Antikörper mit an magnetische Beads gekoppelten sekundären Antikörpern konjugiert. Die Suspension dieser Konjugate (2) wird in die Teflon-Kapillare (3) gepumpt und mittels Elektromagnet (4) dort fixiert. Beim Durchströmen der Kapillare mit der zu untersuchenden Wasserprobe werden Kolibakterien über die primären Antikörper an den Konjugaten festgehalten. Nach dem Abschalten des Magnetes kann die Suspension von magnetisch markierten Kolibakterien durch die Meßspule (5) gepumpt werden. Die Anzahl der Resonanz-Ereignisse im angeschlossenen Schwingkreises entspricht der Anzahl der Kolibakterien in der ursprünglichen Wasserprobe. Durch den Einsatz dieses Gerätes und der entsprechenden Konjugate ist es möglich, ohne den aufwendigen Einsatz der Durchflußzytometrie Bakterien automatisch zu zählen. Des weiteren ist es möglich, mit dieser Meßmethode eine Miniaturisierung des Nachweisgerätes zu erreichen.An example of the application of this method is the detection of coli bacteria in water samples (see schematic drawing, note 1 ). For this purpose, monovalent primary E. coli-specific antibodies are conjugated with secondary antibodies coupled to magnetic beads. The suspension of these conjugates ( 2 ) is pumped into the Teflon capillary ( 3 ) and fixed there by means of an electromagnet ( 4 ). When the water sample to be examined flows through the capillary, coli bacteria are retained on the conjugates via the primary antibodies. After the magnet has been switched off, the suspension of magnetically marked coli bacteria can be pumped through the measuring coil ( 5 ). The number of resonance events in the connected resonant circuit corresponds to the number of coli bacteria in the original water sample. By using this device and the corresponding conjugates, it is possible to automatically count bacteria without the complex use of flow cytometry. It is also possible to use this measuring method to miniaturize the detection device.
Claims (5)
Priority Applications (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19906352A DE19906352A1 (en) | 1999-02-17 | 1999-02-17 | Apparatus to identify and count biological microparticles |
DE19939208A DE19939208C2 (en) | 1999-02-17 | 1999-08-18 | Process for displaying biologically activated inductivity-changing particles for their detection and counting and device therefor |
DE19946656A DE19946656A1 (en) | 1999-02-17 | 1999-09-29 | Production of biologically activated magnetic particles, useful for detection and quantification of e.g. bacteria, by attachment of specific antibody |
PCT/EP2000/001214 WO2000049407A2 (en) | 1999-02-17 | 2000-02-15 | Method for representing biologically activated inductance-altering particles and device for carrying out the method |
CA002370745A CA2370745A1 (en) | 1999-02-17 | 2000-02-15 | Method for representing biologically activated inductance-altering particles and device for carrying out the method |
EP00920436A EP1198712A2 (en) | 1999-02-17 | 2000-02-15 | Method for representing biologically activated inductance-altering particles and device for carrying out the method |
AU41020/00A AU4102000A (en) | 1999-02-17 | 2000-02-15 | Method for representing biologically activated inductance-altering particles anddevice for carrying out the method |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19906352A DE19906352A1 (en) | 1999-02-17 | 1999-02-17 | Apparatus to identify and count biological microparticles |
Publications (1)
Publication Number | Publication Date |
---|---|
DE19906352A1 true DE19906352A1 (en) | 1999-07-22 |
Family
ID=7897605
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DE19906352A Withdrawn DE19906352A1 (en) | 1999-02-17 | 1999-02-17 | Apparatus to identify and count biological microparticles |
DE19939208A Expired - Fee Related DE19939208C2 (en) | 1999-02-17 | 1999-08-18 | Process for displaying biologically activated inductivity-changing particles for their detection and counting and device therefor |
DE19946656A Withdrawn DE19946656A1 (en) | 1999-02-17 | 1999-09-29 | Production of biologically activated magnetic particles, useful for detection and quantification of e.g. bacteria, by attachment of specific antibody |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DE19939208A Expired - Fee Related DE19939208C2 (en) | 1999-02-17 | 1999-08-18 | Process for displaying biologically activated inductivity-changing particles for their detection and counting and device therefor |
DE19946656A Withdrawn DE19946656A1 (en) | 1999-02-17 | 1999-09-29 | Production of biologically activated magnetic particles, useful for detection and quantification of e.g. bacteria, by attachment of specific antibody |
Country Status (1)
Country | Link |
---|---|
DE (3) | DE19906352A1 (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2000049407A2 (en) * | 1999-02-17 | 2000-08-24 | Kilian Hennes | Method for representing biologically activated inductance-altering particles and device for carrying out the method |
DE10129815A1 (en) * | 2001-06-24 | 2003-01-09 | Profos Ag | Process for the purification of bacterial cells and cell components |
CN108169480A (en) * | 2018-02-07 | 2018-06-15 | 上海澜澈生物科技有限公司 | A kind of detection method of molecular amounts of biomarker and system, chip |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2002029430A1 (en) * | 2000-10-04 | 2002-04-11 | Evotec Technologies Gmbh | Method and device for examining biological and/or chemical samples by means of giant magneto-impedance (gmi) |
DE10111520B4 (en) * | 2001-03-09 | 2004-01-15 | Chemagen Biopolymer-Technologie Aktiengesellschaft | Process for the purification of biomolecules using magnetic particles |
AT501194A1 (en) * | 2004-12-30 | 2006-07-15 | Thomas Dr Schlederer | METHOD FOR ISOLATING CELLS AND VIRUSES |
US20110263044A1 (en) * | 2008-07-31 | 2011-10-27 | Eads Deutschland Gmbh | Device and method for the automatic detection of biological particles |
DE102013109467A1 (en) * | 2013-08-30 | 2015-03-05 | MRB Forschungszentrum für Magnet - Resonanz - Bayern e.V. | Method and device for analyzing a sample volume comprising magnetic particles |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SU907484A1 (en) * | 1980-06-30 | 1982-02-23 | Рижский Краснознаменный Институт Инженеров Гражданской Авиации Им.Ленинского Комсомола | Device for checking ferromagnetic particle content in liquid |
US4913883A (en) * | 1987-07-20 | 1990-04-03 | Hitachi, Ltd. | Particle agglutination immunoassay apparatus |
DE68919565T2 (en) * | 1988-07-20 | 1995-06-29 | Olympus Optical Co | Immunoassay method using magnetic marker particles. |
US5385707A (en) * | 1988-12-28 | 1995-01-31 | Stefan Miltenyi | Metal matrices for use in high gradient magnetic separation of biological materials and method for coating the same |
WO1996005326A1 (en) * | 1994-08-17 | 1996-02-22 | Fox John S | Method and apparatus for magnetically detecting proteins and nucleic acids |
FR2732116B1 (en) * | 1995-03-21 | 1997-05-09 | Bio Merieux | METHOD AND DEVICE FOR THE QUALITATIVE AND / OR QUANTITATIVE DETERMINATION OF AN ANALYTE, IN PARTICULAR OF A BACTERIA, IN A SAMPLE, BY MAGNETIC WAY |
-
1999
- 1999-02-17 DE DE19906352A patent/DE19906352A1/en not_active Withdrawn
- 1999-08-18 DE DE19939208A patent/DE19939208C2/en not_active Expired - Fee Related
- 1999-09-29 DE DE19946656A patent/DE19946656A1/en not_active Withdrawn
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2000049407A2 (en) * | 1999-02-17 | 2000-08-24 | Kilian Hennes | Method for representing biologically activated inductance-altering particles and device for carrying out the method |
WO2000049407A3 (en) * | 1999-02-17 | 2002-02-07 | Kilian Hennes | Method for representing biologically activated inductance-altering particles and device for carrying out the method |
DE10129815A1 (en) * | 2001-06-24 | 2003-01-09 | Profos Ag | Process for the purification of bacterial cells and cell components |
US7579142B2 (en) | 2001-06-24 | 2009-08-25 | Profos Ag | Methods for purification of bacterial cells and components |
US8932580B2 (en) | 2001-06-24 | 2015-01-13 | Hyglos Invest Gmbh | Methods for purification of bacterial cells and cell components |
CN108169480A (en) * | 2018-02-07 | 2018-06-15 | 上海澜澈生物科技有限公司 | A kind of detection method of molecular amounts of biomarker and system, chip |
CN108169480B (en) * | 2018-02-07 | 2024-03-08 | 上海澜澈生物科技有限公司 | Method, system and chip for detecting molecular number of biomarker |
Also Published As
Publication number | Publication date |
---|---|
DE19946656A1 (en) | 2000-08-24 |
DE19939208A1 (en) | 2000-09-07 |
DE19939208C2 (en) | 2001-05-17 |
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Legal Events
Date | Code | Title | Description |
---|---|---|---|
OAV | Publication of unexamined application with consent of applicant | ||
OP8 | Request for examination as to paragraph 44 patent law | ||
OR8 | Request for search as to paragraph 43 lit. 1 sentence 1 patent law | ||
8122 | Nonbinding interest in granting licences declared | ||
8105 | Search report available | ||
8143 | Lapsed due to claiming internal priority |