EP1390719A1 - Appareil et procede d'imagerie - Google Patents

Appareil et procede d'imagerie

Info

Publication number
EP1390719A1
EP1390719A1 EP02718883A EP02718883A EP1390719A1 EP 1390719 A1 EP1390719 A1 EP 1390719A1 EP 02718883 A EP02718883 A EP 02718883A EP 02718883 A EP02718883 A EP 02718883A EP 1390719 A1 EP1390719 A1 EP 1390719A1
Authority
EP
European Patent Office
Prior art keywords
specimen
light source
light
array
polarization
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Withdrawn
Application number
EP02718883A
Other languages
German (de)
English (en)
Other versions
EP1390719A4 (fr
Inventor
Lothar U. Kempen
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.)
Maven Technologies LLC
Original Assignee
Maven Technologies LLC
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
Priority claimed from US09/838,700 external-priority patent/US7023547B2/en
Priority claimed from US10/046,620 external-priority patent/US6833920B2/en
Application filed by Maven Technologies LLC filed Critical Maven Technologies LLC
Publication of EP1390719A1 publication Critical patent/EP1390719A1/fr
Publication of EP1390719A4 publication Critical patent/EP1390719A4/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/55—Specular reflectivity
    • G01N21/552—Attenuated total reflection
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/251—Colorimeters; Construction thereof
    • G01N21/253—Colorimeters; Construction thereof for batch operation, i.e. multisample apparatus

Definitions

  • This invention relates to imaging techniques in conjunction with total internal reflection at the boundary of an optically transparent material and more particularly to the use of such techniques for detecting the presence, composition, quantity, and spatial distribution of substances on optically transparent substrates.
  • Imaging methods are advantageous in contrast to methods performing multiple single-point measurements using a scanning method, because the status of each point of the surface is acquired simultaneously, whereas the scanning process takes a considerable amount of time (for example, some minutes), and creates a time lag between individual point measurements.
  • a time lag between measurements makes it difficult or impossible to acquire the status of the entire surface at any given time.
  • Reported applications of imaging ellipsometry were performed on a silicon surface, with the light employed for the measurement passing through +the surrounding medium, either air or a liquid contained in a cuvette. For applications where the optical properties of the surrounding medium can change during the measurement process, passing light through the medium is disadvantageous because it introduces a disturbance of the measurement.
  • TIR total internal reflection
  • light from a light source member providing an extended, polarized light beam is directed through a transparent substrate and undergoes total internal reflection at the surface of the substrate by a single reflection within the
  • TIR member The reflected light is detected by a polarization-sensitive, two-dimensional array detector.
  • the changes of the local polarization state in the beam's cross-section caused by the total internal reflection are employed to obtain information about the presence and composition in an array of substances on the substrate surface for each point of the surface.
  • Total internal reflection is described in; M. Born, et al., "Principles of Optics", 6 th ed., pp 47-51, Pergamon
  • the light generating element within the light source member is a quasi-monochromatic light source of moderate bandwidth.
  • the light generating element within the light source member is an LED of moderate bandwidth.
  • the light from the light source member is directed through an internal reflection member to reflect off a specimen.
  • the total internal reflection at any point within the cross-section of the light beam causes a phase shift between the light component polarized in the plane of incidence and the component polarized perpendicular to the plane of incidence.
  • the reflected light is detected by a polarization-sensitive, two dimensional array detector and the signal from this detector is then processed in a computer to provide two-dimensional information about substances on the surface of the specimen.
  • Spatially distributed changes in polarization state in the cross-section of the reflected beam are indicative of the substances in the specimen in the location in the specimen array corresponding to a position in the detector.
  • the apparatus and method is especially adapted for imaging material in an aqueous solution. It is furthermore particularly suited for detecting attachment and detachment of analytes to a two-dimensional biomolecular array positioned on the total internal reflection member as part of a biosensor system.
  • a plurality of discrete specimen spots are presented in an array, where the method and apparatus will image the array so as to distinguish each of the discrete specimen spots by an image which represents the change in polarization state within each of the discrete specimen spots. Fluorescence or molecular tagging is not necessary nor practical for use in this invention.
  • Fig. 1 is a block diagram of the invention.
  • Fig. 2 is a block diagram of an embodiment of the invention.
  • Fig. 3 is a block diagram of alternative portions of the invention.
  • Fig. 4 is a block diagram of alternative portions of the invention.
  • Fig. 5 is a block diagram of alternative portions of the invention.
  • the invention comprises a method and apparatus for analyzing a two-dimensional arrangement of chemical substances with an imaging technique.
  • a polarized light source of known polarization state is directed into a total internal reflection member (TIR member) configured for a single reflection at a total internal reflection surface (TIR surface) and then exiting the TIR member.
  • TIR member total internal reflection member
  • TIR surface total internal reflection surface
  • superposition of reflections as encountered at a layered optical structure where the layer thicknesses are smaller than the coherence length of the illuminating light is referred to as a single reflection.
  • the chemical specimen is in place above the TIR surface in the evanescent field of the reflected light beam. After reflection, the beam is passed to a polarization-sensitive two-dimensional detector such as a polarizer and a camera.
  • the beam's content can then be processed to determine the change in polarization state, locally in the two-dimensional cross-section of the beam. This provides a spatially distributed map of change of polarization state in the specimen.
  • a variety of techniques is available to determine the change in polarization such as measuring the deviation from a null condition or by comparing the input polarization state to the output polarization state.
  • the refractive index composition of the materials within the evanescent field determines the change in the polarization state of the beam due to the reflection at the TIR surface.
  • a two-dimensional variation of this composition within the TIR surface is associated with a respective variation of the polarization state spatially distributed across the cross-section of the reflected light beam.
  • the chemical specimen forms a two-dimensional array of molecules (here referred to as receptors) with specific affinities towards respective other molecules (here referred to a ligands).
  • the invention is utilized to indicate the presence or absence of binding between ligands and receptors on the array.
  • Such arrays commonly consist of a plurality of discrete specimen spots.
  • the present method and apparatus will image the array so as to distinguish each of the discrete specimen spots represented by the local change in polarization state in the cross-section of the reflected beam.
  • the invention permits measurement of thickness and/or refractive index composition of the specimen under investigation with a very high resolution, in the sub angstrom range, spatially resolved over an entire area.
  • the invention is particularly useful in applications where the specimen is in an aqueous solution.
  • the present invention is used to determine the presence of biological agents in a solution such as in immunosensor applications by measuring their attachment to antibodies on the TIR surface in the evanescent field.
  • the present invention is used to determine the presence and structure of nucleic acid sequences in a solution by measuring their attachment to other nucleic acid sequences on the TIR surface in the evanescent field. Described in more detail below are different embodiments of the invention.
  • FIG. 1 an apparatus and method is illustrated which implements one embodiment of the invention.
  • the apparatus 10 can be conveniently described as consisting of three general portions.
  • Portion 12 is a polarized light source assembly
  • portion 14 is a total internal reflection assembly
  • portion 16 is a polarization- sensitive two-dimensional array detector assembly.
  • Data from the detector assembly 16 is sent by an electrical signal 24 to processor 18 such as a specially programmed computer and user access system such as a print-out or image display.
  • Data can be presented as an image, a data table, or in other forms.
  • the polarized light source assembly 12 passes polarized light of known polarization state (which may be varied or varying) 20 to the total internal reflection assembly 14 and the reflected light 22 having a changed polarization state passes to the detector assembly 16, where it is recorded spatially over the cross-section of the beam.
  • the recorded data is sent to the processor 18 where the change of polarization state is determined to provide a spatially resolved map of changes in polarization state.
  • each spot will be imaged for its change in polarization state within the spot area.
  • Fig. 2 shows a more detailed preferred embodiment.
  • the polarized light source assembly 12 has a light source 26, a beam forming member 28 (if the nature of the light source is such as to make beam forming useful or necessary) a polarizer 30 and an optical retarder 32.
  • the total internal light reflection assembly 14 has a an optical element 34 which has an optical surface 36. Also shown is a specimen slide 38 on the optical surface 36, and between them an index matching substance 40. Because of the index matching a total internal reflection surface (TIR surface) is defined as the upper surface 39 of the specimen slide 38. A specimen 42 is on the total internal reflection surface 39 of the slide 38.
  • TIR surface total internal reflection surface
  • the optical element 34 is a prism configured along with the index matched slide 38 in relationship to the incoming light beam 20, and the exiting light beam 22 such that the beam reflects only a single time at the TIR surface 39 and then exits the prism. If the specimen is placed directly on the optical surface 36, then the optical surface 36 would be the TIR surface. But this is not the usual application as the specimen (such as a biochip) is usually prepared more conveniently on a specimen slide 38 and placed in the apparatus. In any event, however constructed, there is an optical structure having a TIR surface and the beam reflects only a single time at the TIR surface between entering and leaving the optical structure. In other words, there is a TIR surface in optical contact with the specimen, such that the evanescent field associated with the total internal reflection interacts with the specimen, and there is only a single reflection at that TIR surface.
  • the post reflection detector assembly 16 has a polarizer 44, and a two-dimensional array detector 46, preferably a camera of the CCD type.
  • the processor 18 is a specially programmed computer and output means for processing the imagery into a representation of film thickness variations spatially resolved over the cross-section of the area imaged.
  • the imaging is acquired by detecting changes spatially distributed in the local polarization state in the beam's cross-section caused by the total internal reflection. This provides information about the presence and composition in the array of substances on the substrate surface for each resolvable point on the surface. Different polarization state changes are included in the cross- section of the reflected beam indicative of the substances on the specimen in the location in the specimen array corresponding to a position in the detector.
  • the processor 18 receives the data as an electrical signal 24 and characterizes the change of polarization state spatially over the two-dimensional array.
  • the analysis and processing is done in one embodiment by comparing the known polarization state of the incoming light from the light processing assembly 12 with the changed polarization state of the reflected light 22, spatially resolved two-dimensionally within the beam which provides a map of spatially distributed points or spots in the specimen array.
  • the polarization shift is then analyzed by the processor 18 to provide information of the presence and properties of elements in the chemical specimen.
  • Other known techniques, such as null processing can be used to determine the change in polarization state.
  • the light source member 26 may be an LED, an SLD (Super Luminescent Diode), an incandescent light source, or a laser. If an LED or SLD is used, the set-up shown in Fig. 2 is appropriate, where the beam forming member 28 is a collimator. If an incandescent light source is used, an optical filter is also used.
  • the light source 26 for the apparatus is a quasi-monochromatic light source of moderate bandwidth.
  • the light source 26 is preferably an LED of moderate bandwidth.
  • the bandwidth is a full width half maximum wavelength in the range of about 10nm-50nm, and more preferably a full width half maximum wavelength in the range of about 30nm-50nm.
  • the optical retarder 32 could be placed instead in the exiting beam path 22 before the polarizer 44.
  • a moving diffuser 52 is adapted to produce speckle offsetting fluctuation of the minima and maxima in the speckle pattern caused by the laser.
  • the moving diffuser 52 is attached to a mechanical actuator 54 which is preferably a motor and servo-apparatus for providing the speckle offsetting fluctuations.
  • the beam 20 then proceeds through the beam forming element 28, the polarizer 30 and the optical retarder 32, exiting the light source assembly 20.
  • the polarizer 30 employs a polarizer of selected known polarization state.
  • the polarizer 30 may be of the type having a mechanical actuator driven by a motor control signal so as to enable varying and selecting the polarization state of the light beam 20.
  • the total internal reflection optical element 34 either alone or in combination with an index matched slide may be arranged for use with a specimen in various ways to define a total internal reflection assembly so long as the specimen is in the evanescent field of the reflected beam 20, 22.
  • the specimen 42 could be set directly on the optical surface 36 in which case the optical surface 36 would be the TIR surface but this is inconvenient and repeated use is likely to degrade the optical quality of the optical surface 36, and therefore, consistent with common practice in which a biochip or other chemical assay specimen is provided, a specimen slide 38 or other supporting apparatus is employed. It is common in a biochip to provide an array of discrete specimen spots supported on a structure for obtaining analysis of each spot.
  • the term total internal reflection optical element refers to known optical elements alone or in combination with other elements which provide the phenomenon known as total internal reflection.
  • Fig. 2 shows use of a prism combined with a slide 38, being index matched so that there is a TIR surface 39.
  • Fig. 4 shows an alternative optical arrangement in which a flat optical member 56 having an upper surface 58 is surmounted by a specimen slide 60 and an index matching substance 62 on which is a specimen 64.
  • the TIR surface 66 is the top of the slide 60.
  • the beam 20 enters the assembly, is refracted as it enters, and leaves the optical member 56 after a single reflection at the TIR surface 66 as beam 22.
  • Other mechanisms for providing total internal reflection and an evanescent field can be employed in practicing this invention as long as only a single reflection occurs at the TIR surface upon which the specimen is placed so as to be in the evanescent field associated with the reflection..
  • the post-reflection processing arrangement 16 through which the beam 22 passes can alternatively, consist of a polarizer member 70, a beam forming member 72 and a two-dimensional array detector 74.
  • the method and apparatus can be used in combination with biochips of the type having discrete specimen spots or a micro-titer plate containing an array of discrete spots or locations for analysis, where the detected change in polarization state is spatially related to the discrete locations in the reflected beam. Therefore, as used herein the slide and specimen refers to any type of chemical or biological array which is desired to be examined.

Landscapes

  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Investigating Or Analysing Materials By Optical Means (AREA)

Abstract

La présente invention concerne un appareil (10) et un procédé d'imagerie dans lesquels on utilise le changement de l'état de polarisation d'un faisceau lumineux (20) traversant une structure à réflexion interne totale (RIT) (14) par une réflexion unique au niveau d'une surface RIT (39) dans laquelle un échantillon (42) est placé dans le champ évanescent associé à la réflexion interne totale du faisceau lumineux, l'échantillon (42) étant l'objet d'une étude biologique, chimique ou génétique.
EP02718883A 2001-04-19 2002-01-27 Appareil et procede d'imagerie Withdrawn EP1390719A4 (fr)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US09/838,700 US7023547B2 (en) 2000-07-11 2001-04-19 Apparatus including a biochip for imaging of biological samples and method
US838700 2001-04-19
US10/046,620 US6833920B2 (en) 2000-07-11 2002-01-12 Apparatus and method for imaging
US46620 2002-01-12
PCT/US2002/002662 WO2002086468A1 (fr) 2001-04-19 2002-01-27 Appareil et procede d'imagerie

Publications (2)

Publication Number Publication Date
EP1390719A1 true EP1390719A1 (fr) 2004-02-25
EP1390719A4 EP1390719A4 (fr) 2007-08-29

Family

ID=26724134

Family Applications (1)

Application Number Title Priority Date Filing Date
EP02718883A Withdrawn EP1390719A4 (fr) 2001-04-19 2002-01-27 Appareil et procede d'imagerie

Country Status (5)

Country Link
EP (1) EP1390719A4 (fr)
JP (2) JP4219689B2 (fr)
AU (1) AU2002249994A1 (fr)
IL (1) IL158473A0 (fr)
WO (1) WO2002086468A1 (fr)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7193711B2 (en) * 2000-07-11 2007-03-20 Maven Technologies, Llc Imaging method and apparatus
US20040146917A1 (en) * 2001-08-03 2004-07-29 Nanosphere, Inc. Nanoparticle imaging system and method
CA2455118C (fr) 2001-08-03 2012-01-17 Nanosphere, Inc. Systeme et procede d'imagerie de nanoparticules
US8570525B2 (en) * 2006-06-23 2013-10-29 Optopol Technology S.A. Apparatus for optical frequency domain tomography with adjusting system
WO2009104123A1 (fr) * 2008-02-22 2009-08-27 Koninklijke Philips Electronics N.V. Source de lumière pour biocapteur de spectroscopie infrarouge à transformée de fourier (ftir)
ES2377896B1 (es) * 2009-03-24 2013-02-13 Hanscan Ip B.V. Dispositivo detector de vida.
WO2020014296A1 (fr) 2018-07-12 2020-01-16 Luminex Corporation Systèmes et procédés permettant d'effectuer des processus de préparation et d'analyse d'échantillons variables

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0067921A1 (fr) * 1981-06-22 1982-12-29 Prutec Limited Méthode pour déterminer des substances bioactives
EP0286195A2 (fr) * 1987-04-10 1988-10-12 Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno Méthode et appareil pour détecter des concentrations faibles de composés chimiques ou biologiques dans un milieu d'examen en utilisant la résonance par plasmons de surface
US5313264A (en) * 1988-11-10 1994-05-17 Pharmacia Biosensor Ab Optical biosensor system
US5437840A (en) * 1994-04-15 1995-08-01 Hewlett-Packard Company Apparatus for intracavity sensing of macroscopic properties of chemicals
US5754296A (en) * 1995-03-20 1998-05-19 Kansas State University Research Foundation Ellipsometric microscope

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GB2248497B (en) * 1990-09-26 1994-05-25 Marconi Gec Ltd An optical sensor
GB2254415B (en) * 1991-03-22 1994-10-12 Marconi Gec Ltd An optical sensor
US5485277A (en) * 1994-07-26 1996-01-16 Physical Optics Corporation Surface plasmon resonance sensor and methods for the utilization thereof
EP1650550B1 (fr) * 1996-04-30 2012-10-17 FUJIFILM Corporation Capteur de plasmons de surface

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0067921A1 (fr) * 1981-06-22 1982-12-29 Prutec Limited Méthode pour déterminer des substances bioactives
EP0286195A2 (fr) * 1987-04-10 1988-10-12 Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno Méthode et appareil pour détecter des concentrations faibles de composés chimiques ou biologiques dans un milieu d'examen en utilisant la résonance par plasmons de surface
US5313264A (en) * 1988-11-10 1994-05-17 Pharmacia Biosensor Ab Optical biosensor system
US5437840A (en) * 1994-04-15 1995-08-01 Hewlett-Packard Company Apparatus for intracavity sensing of macroscopic properties of chemicals
US5754296A (en) * 1995-03-20 1998-05-19 Kansas State University Research Foundation Ellipsometric microscope

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DICKE J ET AL: "Ellipsomicroscopy for surface imaging: contrast mechanism, enhancement, and application to CO oxidation on Pt(110)" JOURNAL OF THE OPTICAL SOCIETY OF AMERICA - A, OPTICAL SOCIETY OF AMERICA, WASHINGTON, US, vol. 17, no. 1, January 2000 (2000-01), pages 135-141, XP002182387 ISSN: 1084-7529 *
GANG JIN ET AL: "Imaging ellipsometry for the visualization of bio-molecular layers" ENGINEERING IN MEDICINE AND BIOLOGY SOCIETY, 1998. PROCEEDINGS OF THE 20TH ANNUAL INTERNATIONAL CONFERENCE OF THE IEEE HONG KONG, CHINA 29 OCT.-1 NOV. 1998, PISCATAWAY, NJ, USA,IEEE, US, vol. 2, 29 October 1998 (1998-10-29), pages 581-584, XP010320597 ISBN: 0-7803-5164-9 *
JIN G ET AL: "A BIOSENSOR CONCEPT BASED ON IMAGING ELLIPSOMETRY FOR VISUALIZATION OF BIOMOLECULAR INTERACTIONS" ANALYTICAL BIOCHEMISTRY, ACADEMIC PRESS, SAN DIEGO, CA, US, vol. 232, 1995, pages 69-72, XP001030825 ISSN: 0003-2697 *
See also references of WO02086468A1 *

Also Published As

Publication number Publication date
WO2002086468A8 (fr) 2003-03-20
IL158473A0 (en) 2004-05-12
JP4219689B2 (ja) 2009-02-04
JP2004531719A (ja) 2004-10-14
JP2007178442A (ja) 2007-07-12
EP1390719A4 (fr) 2007-08-29
AU2002249994A1 (en) 2002-11-05
WO2002086468A1 (fr) 2002-10-31

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