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WO2000037920A1 - Procede et dispositif pour la spectroscopie electro-optique de particules individuelles - Google Patents

Procede et dispositif pour la spectroscopie electro-optique de particules individuelles Download PDF

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Publication number
WO2000037920A1
WO2000037920A1 PCT/EP1999/010278 EP9910278W WO0037920A1 WO 2000037920 A1 WO2000037920 A1 WO 2000037920A1 EP 9910278 W EP9910278 W EP 9910278W WO 0037920 A1 WO0037920 A1 WO 0037920A1
Authority
WO
WIPO (PCT)
Prior art keywords
particle
rotation
optical
spectra
particles
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.)
Ceased
Application number
PCT/EP1999/010278
Other languages
German (de)
English (en)
Inventor
Günter FUHR
Torsten Müller
Thomas Schnelle
Gabriele Gradl
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Evotec Biosystems GmbH
Original Assignee
Evotec Biosystems GmbH
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 Evotec Biosystems GmbH filed Critical Evotec Biosystems GmbH
Publication of WO2000037920A1 publication Critical patent/WO2000037920A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C5/00—Separating dispersed particles from liquids by electrostatic effect
    • B03C5/02—Separators
    • B03C5/022—Non-uniform field separators
    • B03C5/028—Non-uniform field separators using travelling electric fields, i.e. travelling wave dielectrophoresis [TWD]
    • 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
    • G01N15/1023—Microstructural devices for non-optical measurement
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography

Definitions

  • the invention relates to methods for dielectric single particle spectroscopy in microsystems and devices for their implementation.
  • the particle misaligned or the measuring time is shortened uncontrollably. Both stand in the way of an automatic measurement.
  • Cell movement can also be determined automatically using dynamic light scattering methods [GIMSA, J., PRUGER, B., EPPMANN, P. and DONATH, E., Electrorotation of particles measured by dynamic light scattering - a new dielect ⁇ c spectroscopy technique, m " Colloids and Surfaces A ", Vol. 98, 243-249, 1995].
  • this method cannot be used on individual objects, but provides average values for all particles that are in the laser beam. As a rule, this is a few hundred or more.
  • the exact positioning at one point in the electrical rotation field was achieved by using 3-dimensional electrode arrangements, so-called field cages, and the alternating application of a centering field and a rotation field.
  • optical field traps also called “optical tweezers”, “laser tweezers” or “optical traps”
  • laser tweezers optical traps
  • optical traps which have been used for about two decades in the fields of biotechnology, medicine and molecular biology and in other technical fields for positioning and manipulating micrometer-sized and submicron-sized particles are used [G. Weber et al. in "Int. Rev. Cytol.” Vol. 131, 1992, p. 1; S.M. Block m “Nonmvasive Techniques m Cell Biology", Wiley-Liss., New York 1990, p. 375].
  • the development of the laser tweezers goes back mainly to A. Ashkm [A. Ashkm m "Phys. Rev. Lett.”, Vol.
  • the principle of particle capture by optically induced forces is based on the fact that, in addition to the light pressure, which always pushes a particle away from the light source, gradient forces occur which lead to a particle getting a focus or being held stable with it or with it is moved.
  • the prerequisite is that the absorption and reflection of the particle is low, while the difference in the refractive index from the surrounding solution should be as large as possible.
  • the invention has for its object to provide new methods for dielectric single particle spectroscopy in microsystems and devices for their implementation, with which the above. Problems can be solved and, in particular, the particles in a rotating field, regardless of whether attractive or repulsive dielectrophoretic forces occur, are kept floating in a solution at any point with an accuracy below the particle radius in the rotating field, without reducing the speed of rotation.
  • a suspended particle which can be artificial or also biological in nature, is caught in a strongly focused laser beam, as is known from optical tweezers.
  • the capture point of the laser with the particles in it is now guided between microelectrodes, which are usually planarly applied to a smooth substrate, until the Particles are located in the area of the electric field spreading out in the solution, provided the electrodes are exposed to high-frequency, phase-shifted alternating voltage signals in a suitable manner.
  • the particle is in a free suspension state. It is held by the interaction of electrical and optical forces.
  • the laser focus is expediently positioned on a line which is perpendicular to the point which denotes the field minimum between the electrodes. Even if forces are developed by the rotating electric field that want to pull the particle to the electrodes, they act at this location in all electrode directions pretty evenly, so that, according to the invention, only very small forces are required to keep the particle stable in spite of the field-induced attractive forces Hold laser focus. On the other hand, the intensity of the laser must be chosen so high that the particle is raised. If the particle is repelled by the electrodes via the electrically induced polarization forces, the forces of the optical field can be selected even less, since the particle itself centers on the designated line of symmetry. Here, however, it is raised and pushed out of the electrode area.
  • This force must be compensated for by the choice of the intensity of the laser beam.
  • This particle which is caught very optically by m free solution, experiences a torque through the rotating electrical field and can be shifted depending on the frequency m in the manner known per se m slow rotation.
  • FIG. 1 an overview representation for holding a particle according to the invention in a quadrupole arrangement for dielectric spectroscopy with laser tweezers;
  • Figure 2 is a graph showing the dependence of the torque or the pushing or pulling forces on the frequency f of
  • FIG. 3 an overview representation for the combination according to the invention of a microsystem for dielectric spectroscopy with a microscope arrangement
  • Figure 4 a schematic illustration of a microsystem for Screenmg tasks.
  • FIG. 1 shows a perspective view of the arrangement. Details of the microsystem that are known per se are not shown.
  • a particle 11, suspended in an ambient solution 12, is located in the radiation field of a strongly focused laser beam 13 and is caught in the focus 14.
  • Four planar electrodes 16a to 16d, which are usually planar on a substrate 15, are phase-shifted signals by 90 degrees (phase angle 0 °, 90 °, 180 °, 270 °) of the same frequency (amplitudes, for example, about 1 to 20 V) controlled so that a rotating field m of the xy plane is created.
  • the captured particle rotates much more slowly than the field rotates compared to the suspension liquid 12.
  • the speed of rotation of the object as a function of frequency is determined by measuring or observing the particle and provides the desired rotation spectra.
  • 3 or more electrodes can also be used in one plane, of greater thickness as well as in a multilevel arrangement.
  • the microsystem shown in FIG. 1 can advantageously be equipped with resonance devices for forming a resonant increase or damping of the field strength of the alternating electrical fields at predetermined frequencies, as described in PCT / EP96 / 05244.
  • the content of patent application PCT / EP96 / 05244 is hereby incorporated in its entirety by express reference to the content of the present description. This applies in particular to all measures for generating resonance Nanzeschemieux m particle suspensions in microelectrode arrangements.
  • FIG. 2 shows a rotation spectrum (curve 21, describing the spectrum of a living cell) and the associated dielectrophoretic force (curve 22). It can be seen that the cell was pulled to the electrodes without the optical capture field in the frequency range ( ⁇ ) between 20 Hz and 1 GHz.
  • the curves shown are a measurement on a 20 ⁇ m cell in an aqueous solution with a conductivity of ImS / m, as is typical for algae. As a result, it has hitherto not been possible to measure in this frequency range or only with reduced accuracy, as was explained above.
  • the force force represented by curve 22 is compensated for with the laser tweezers. Accordingly, the laser tweezers are used with such operating parameters that a sufficiently large trapping force is exerted on the particle.
  • FIG. 3 shows a device that is transparent at least on one side, with further details with which the rotation measurements can be carried out.
  • Planar electrodes 32a to 32d are processed on a substrate 31 (for example glass) using the means of semiconductor technology and AC signals are applied via the feed lines 33a to 33d for generating the rotating field.
  • a channel is formed through the side walls 34a, 34b and the cover plate 35, which is less than 250 ⁇ m thick, through which the particle suspension can be wound (36, arrow direction).
  • the channel ceiling is made of glass, so that a lens 37 with a high numerical aperture, for example also as an olim ersionsobj ektiv (01 38) can generate a highly focused laser focus in the channel interior, with which the particle 39 is captured.
  • FIG. 4 shows a microsystem which is designed for the construction of a test system (assay system) for the high-throughput screenmg.
  • the microsystem 40 has a channel structure 41a, 41b.
  • a suspension with particles that are to be tested flows through the first channel 41a.
  • the particles include, for example, biological cells or modified synthetic particles or combinations of biological cells and synthetic particles.
  • the second channel 41b through which a solution or suspension of a test substance flows, flows into the first channel 41a.
  • the test substance preferably comprises ligands, for example antibodies.
  • a first electrode system 42a is attached to the first channel 41a, which is constructed, for example, like the microelectrode system according to FIG. 1.
  • a second electrode system 42b is arranged downstream of the mouth point.
  • the electrode systems 42a, 42b are designed for rotational measurements on the particles 43 before or after the interaction with the test substance.
  • Each electrode arrangement is equipped with a laser device and a microscope arrangement for generating the optical traps (see FIG. 3).
  • a test procedure could be implemented as follows, for example.
  • the inflowing particles 43 in the first electrode system 42a are subjected to a first rotation measurement (with simultaneous holding with an optical trap). After mixing with the test substance there are interactions (eg particles 43a), the influence of which on the particle properties is detected by the second rotation measurement in the electrode system 42b.
  • the changed properties can be absolutely by characteristic spectra or relatively by comparing the spectra of the first and second rotation measurements can be determined.
  • the particles are released from the second electrode system, possibly subjected to further rotation measurements in further electrode arrangements (not shown) and, depending on the measurement result, subjected to a sorting process.
  • Biomolecules and associated application examples can be: antigen antibodies, in studies for the development of immunoreagents and immunoassays, epitope mapping and screen mg from phage libraries; Ligands and their cell membrane receptors; Cell adhesion molecules and their ligands, for example in studies of the affinity between cadherms and integers and their receptors located in the cell membrane; Membrane molecules, such as lipids or glycoprotems, when studying the interaction of these molecules with other soluble or cell membrane-containing biomolecules; extracellular matrix molecules and their soluble or cell membrane-bound ligands; intracellular messenger substances, in the investigation of signal transmission within the cell through the interaction of molecules in a signal transmission cascade; soluble proteins and peptides, in the monitoring of the production of proteins and peptides in
  • test system according to the invention can also be constructed from a multiplicity of microsystems according to FIG. 4, which interact in series or in parallel.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Dispersion Chemistry (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Electrochemistry (AREA)
  • Molecular Biology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)
  • Microscoopes, Condenser (AREA)

Abstract

Selon l'invention, pour effectuer une spectroscopie diélectrique sur au moins une particule (11) en suspension dans un microsystème, on expose ladite particule (11) à des champs électriques haute fréquence tournants, dans un dispositif à électrodes (16), et on la maintient dans le foyer (14) d'un piège optique.
PCT/EP1999/010278 1998-12-22 1999-12-21 Procede et dispositif pour la spectroscopie electro-optique de particules individuelles Ceased WO2000037920A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19859460A DE19859460C2 (de) 1998-12-22 1998-12-22 Verfahren und Vorrichtung zur elektro-optischen Einzelpartikelspektroskopie
DE19859460.7 1998-12-22

Publications (1)

Publication Number Publication Date
WO2000037920A1 true WO2000037920A1 (fr) 2000-06-29

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PCT/EP1999/010278 Ceased WO2000037920A1 (fr) 1998-12-22 1999-12-21 Procede et dispositif pour la spectroscopie electro-optique de particules individuelles

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DE (1) DE19859460C2 (fr)
WO (1) WO2000037920A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2370520A (en) * 2000-12-21 2002-07-03 Univ St Andrews Optical rotation of microscopic particles
EP1413911A1 (fr) * 2002-10-25 2004-04-28 Evotec Technologies GmbH Procédé et dispositif d'imagerie tridimensionelle d'objets microscopiques suspendus permettant une microscopie haute résolution
WO2007088517A3 (fr) * 2006-02-01 2007-11-15 Ecole Polytech Appareil de manipulation, modification et caractérisation de particules dans un micro canal
US7366377B2 (en) 2003-12-04 2008-04-29 Commissariat A L'energie Atomique Particle concentration method
US7402795B2 (en) 2003-12-04 2008-07-22 Commissariat A L'energie Atomique Particle sorting method
US7511263B2 (en) 2003-12-04 2009-03-31 Commissariat A L'energie Atomique Object separation device using optical method
DE102014005219A1 (de) * 2014-03-28 2015-10-01 Josip Mihaljevic Verfahren und System zum Bilden einer optischen Falle
WO2019227563A1 (fr) * 2018-05-29 2019-12-05 清华大学 Dispositif microfluidique représentant des paramètres multiples pour cellule unique

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DD256192A1 (de) * 1985-09-30 1988-04-27 Univ Berlin Humboldt Verfahren und vorrichtung zur messung des rotationsspektrums dielektrischer objekte
WO1996041154A1 (fr) * 1995-06-07 1996-12-19 UNITED STATES GOVERNMENT, as represented by THE SECRETARY OF COMMERCE NATIONAL INSTITUTE OF STANDARDS AND TECHNOLOGY Piege optique de detection et de quantification de quantites subzeptomolaires d'analytes

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3325843A1 (de) * 1983-07-18 1985-02-07 Kernforschungsanlage Jülich GmbH, 5170 Jülich Verfahren und vorrichtung zur unterscheidung von in einem medium befindlichen teilchen oder partikeln
DE19653659C1 (de) * 1996-12-20 1998-05-20 Guenter Prof Dr Fuhr Elektrodenanordnung für Feldkäfige

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DD256192A1 (de) * 1985-09-30 1988-04-27 Univ Berlin Humboldt Verfahren und vorrichtung zur messung des rotationsspektrums dielektrischer objekte
WO1996041154A1 (fr) * 1995-06-07 1996-12-19 UNITED STATES GOVERNMENT, as represented by THE SECRETARY OF COMMERCE NATIONAL INSTITUTE OF STANDARDS AND TECHNOLOGY Piege optique de detection et de quantification de quantites subzeptomolaires d'analytes

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
GASPERIS DE G ET AL: "AUTOMATED ELECTROROTATION: DIELECTRIC CHARACTERIZATION OF LIVING CELLS BY REAL-TIME MOTION ESTIMATION", MEASUREMENT SCIENCE AND TECHNOLOGY,GB,IOP PUBLISHING, BRISTOL, vol. 9, no. 3, 1 March 1998 (1998-03-01), pages 518 - 529, XP000777488, ISSN: 0957-0233 *

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2370520A (en) * 2000-12-21 2002-07-03 Univ St Andrews Optical rotation of microscopic particles
GB2370520B (en) * 2000-12-21 2003-08-06 Univ St Andrews Optical rotation of microscopic particles
EP1413911A1 (fr) * 2002-10-25 2004-04-28 Evotec Technologies GmbH Procédé et dispositif d'imagerie tridimensionelle d'objets microscopiques suspendus permettant une microscopie haute résolution
WO2004038484A3 (fr) * 2002-10-25 2004-05-27 Evotec Technologies Gmbh Procede et dispositif d'imagerie tridimensionnelle pour micro-objets suspendus engendrant une microscopie a haute resolution
US7738695B2 (en) 2002-10-25 2010-06-15 Institut Pasteur Method and device for 3 dimensional imaging of suspended micro-objects providing high-resolution microscopy
US7366377B2 (en) 2003-12-04 2008-04-29 Commissariat A L'energie Atomique Particle concentration method
US7402795B2 (en) 2003-12-04 2008-07-22 Commissariat A L'energie Atomique Particle sorting method
US7511263B2 (en) 2003-12-04 2009-03-31 Commissariat A L'energie Atomique Object separation device using optical method
WO2007088517A3 (fr) * 2006-02-01 2007-11-15 Ecole Polytech Appareil de manipulation, modification et caractérisation de particules dans un micro canal
US9995668B2 (en) 2006-02-01 2018-06-12 Ecole polytechnique fédérale de Lausanne (EPFL) Apparatus for manipulating, modifying and characterizing particles in a micro channel
DE102014005219A1 (de) * 2014-03-28 2015-10-01 Josip Mihaljevic Verfahren und System zum Bilden einer optischen Falle
WO2019227563A1 (fr) * 2018-05-29 2019-12-05 清华大学 Dispositif microfluidique représentant des paramètres multiples pour cellule unique

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Publication number Publication date
DE19859460A1 (de) 2000-07-27
DE19859460C2 (de) 2001-02-22

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