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WO2001023965A1 - Microscope holographique interferentiel numerique et procedes - Google Patents

Microscope holographique interferentiel numerique et procedes Download PDF

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Publication number
WO2001023965A1
WO2001023965A1 PCT/US2000/026462 US0026462W WO0123965A1 WO 2001023965 A1 WO2001023965 A1 WO 2001023965A1 US 0026462 W US0026462 W US 0026462W WO 0123965 A1 WO0123965 A1 WO 0123965A1
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WO
WIPO (PCT)
Prior art keywords
image
wavelength
radiation
holographic
images
Prior art date
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Ceased
Application number
PCT/US2000/026462
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English (en)
Inventor
Myung K. Kim
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.)
University of South Florida
University of South Florida St Petersburg
Original Assignee
University of South Florida
University of South Florida St Petersburg
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 University of South Florida, University of South Florida St Petersburg filed Critical University of South Florida
Priority to AU77187/00A priority Critical patent/AU7718700A/en
Priority to US10/089,266 priority patent/US7127109B1/en
Publication of WO2001023965A1 publication Critical patent/WO2001023965A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03H—HOLOGRAPHIC PROCESSES OR APPARATUS
    • G03H1/00—Holographic processes or apparatus using light, infrared or ultraviolet waves for obtaining holograms or for obtaining an image from them; Details peculiar thereto
    • G03H1/04—Processes or apparatus for producing holograms
    • G03H1/08—Synthesising holograms, i.e. holograms synthesized from objects or objects from holograms
    • G03H1/0866—Digital holographic imaging, i.e. synthesizing holobjects from holograms
    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03H—HOLOGRAPHIC PROCESSES OR APPARATUS
    • G03H1/00—Holographic processes or apparatus using light, infrared or ultraviolet waves for obtaining holograms or for obtaining an image from them; Details peculiar thereto
    • G03H1/04—Processes or apparatus for producing holograms
    • G03H1/0443—Digital holography, i.e. recording holograms with digital recording means
    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03H—HOLOGRAPHIC PROCESSES OR APPARATUS
    • G03H1/00—Holographic processes or apparatus using light, infrared or ultraviolet waves for obtaining holograms or for obtaining an image from them; Details peculiar thereto
    • G03H1/0005—Adaptation of holography to specific applications
    • G03H2001/005—Adaptation of holography to specific applications in microscopy, e.g. digital holographic microscope [DHM]
    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03H—HOLOGRAPHIC PROCESSES OR APPARATUS
    • G03H1/00—Holographic processes or apparatus using light, infrared or ultraviolet waves for obtaining holograms or for obtaining an image from them; Details peculiar thereto
    • G03H1/26—Processes or apparatus specially adapted to produce multiple sub- holograms or to obtain images from them, e.g. multicolour technique
    • G03H1/2645—Multiplexing processes, e.g. aperture, shift, or wavefront multiplexing
    • G03H2001/266—Wavelength multiplexing
    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03H—HOLOGRAPHIC PROCESSES OR APPARATUS
    • G03H2210/00—Object characteristics
    • G03H2210/30—3D object
    • G03H2210/33—3D/2D, i.e. the object is formed of stratified 2D planes, e.g. tomographic data
    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03H—HOLOGRAPHIC PROCESSES OR APPARATUS
    • G03H2222/00—Light sources or light beam properties
    • G03H2222/10—Spectral composition
    • G03H2222/13—Multi-wavelengths wave with discontinuous wavelength ranges
    • G—PHYSICS
    • G03—PHOTOGRAPHY; CINEMATOGRAPHY; ANALOGOUS TECHNIQUES USING WAVES OTHER THAN OPTICAL WAVES; ELECTROGRAPHY; HOLOGRAPHY
    • G03H—HOLOGRAPHIC PROCESSES OR APPARATUS
    • G03H2223/00—Optical components
    • G03H2223/14—Diffuser, e.g. lens array, random phase mask

Definitions

  • the present invention relates to microscopic imaging, and, more particularly, to
  • holographic microscopy for reconstruction of three-dimensional objects and optical tomographic imaging for selectively imaging cross sections of an object.
  • Imaging of microscopic objects is an essential art, not only in biology and medicine,
  • the lateral resolution can be a fraction of a
  • aperture registers a light signal originating only from the object point.
  • the interference pattern are recorded in a hologram by the interference of an object wave that is to be imaged with a reference wave of simple structure such as a plane or spherical wave.
  • the interference pattern is recorded in a hologram by the interference of an object wave that is to be imaged with a reference wave of simple structure such as a plane or spherical wave.
  • the holographic image retains the phase as well as the amplitude information, a variety of interference experiments can be performed, and this is the basis of many
  • interference pattern of a real object and reference is recorded using an electronic or digital
  • the holographic image is recreated on the computer by numerical calculation.
  • Digital holography alleviates the need for wet chemical processing of a photographic
  • phase information of the light wave is available directly from the numerical reconstruction and greatly simplifies interferometric
  • Holography can be applied to microscopy in two alternative ways. In one, a hologram
  • Holographic microscopy has been particularly useful in particle analysis, where
  • a holographic micrograph freezes the three-dimensional field, and a particle count can
  • the axial magnification goes as the square of the lateral magnification, so that the two
  • image is numerically stored, it can be manipulated by image processing techniques for
  • subwavelength resolution of features and particle analysis and feature recognition can be automated with greater efficiency.
  • laser confocal microscopy (Sheppard and Shotton, 1997) uses aperturing of both the
  • microsca ning using piezo actuators is an
  • the hologram is imaged on a CCD array, replacing photographic plates of conventional holography.
  • the digitally converted hologram is stored in a computer, and
  • the distance information can be
  • Another object is to provide such a method and apparatus wherein the obtained optical
  • sectioned images can be reassembled into a three-dimensional digital model that can be
  • a further obj ect is to provide a method and apparatus for imaging a three-dimensional
  • An additional object is to provide such a method and apparatus for generating a three-
  • the present invention addresses a practical problem in microscopy, where the axial
  • magnification goes as the square of the transverse magnification. Even at moderate
  • focus portions of the object image contribute to blurring and noise of the focal plane image.
  • CSM Confocal scanning microscopy
  • a hologram has depth perception and axial resolution, but determination of axial
  • location in particle analysis depends only on the focusing of the image as the
  • the present invention involves no mechanical motion, and wavelength scanning and
  • the diffracted wave is proportional to E 0 , a replica of the original obj ect wave (or its conjugate E 0 * ).
  • the wave propagates in the general z direction.
  • the factor __(_P) represents the
  • the field at Q is
  • plane distance A can be matched to the axial extent of the object and ⁇ to the desired level of
  • wavelengths N ⁇ is the same as the spectral width of low-coherence or short-pulse lasers in
  • WS-DIH proceeds as follows. A microscopic object is illuminated by a laser, and a laser
  • microscope lens forms a real magnified image of the object. Light from this intermediate
  • the laser wavelength is stepped by ⁇ for the next exposure, and the process is repeated N times, which completes the recording process.
  • the frequency step ⁇ is inversely proportional to the object axial scale A.
  • the set of N digitally stored holograms represents the complete information required for computational reconstruction of the three-dimensional image. For each hologram, after
  • the resultant image is an intensity distribution
  • the result can be displayed as two-dimensional cross sections of the object at an
  • lateral magnification can be processed out by applying corrective scale factors.
  • cross sections can then be reassembled into a three-dimensional computer model with natural aspect ratios.
  • the three-dimensional computer model is then available for application-specific
  • the lateral resolution should be as good as conventional optical microscopy, except
  • the present system is a coherent imaging system, and so one needs to exercise care with speckle noise and other interference effects.
  • the multiple imaging system is a coherent imaging system, and so one needs to exercise care with speckle noise and other interference effects.
  • a confocal system is determined mainly by the focal depth of the illuminated spot
  • the wavelength scanning system of the present invention has no mechanical moving
  • WS-DIH is the same as in SCM: M x * M y * N, where the Ms are the number of pixels
  • the bottleneck may occur at the
  • the system of the present invention is a holographic system, and as such, the complete
  • amplitude and phase information of the light field is available.
  • FIG. 1 is a schematic of an apparatus for multiwavelength digital holography.
  • FIGS.2A-2E are reconstructions of an image of a single object (OBJ1) using a single OBJ1
  • FIG. 2 A reference beam
  • FIG. 2B object beam at the screen
  • FIG. 2C interference between the reference and the obj ect
  • FIG.2D intensity patterns of FIGS .2 A and
  • FIGS. 3A-C are reconstructions of images of two objects (OBJ1 and OBJ2) using a
  • FIG.3A interference pattern between reference and object, minus zero-
  • FIGS. 4A-4E are reconstructed image patterns as functions of image distance.
  • horizontal axis is z in cm
  • vertical axis, in mm is a slice of the reconstructed image
  • FIG.4 A a single wavelength or frequency
  • FIG.4B a single wavelength or frequency
  • FIG. 4D three relative frequencies, 0.0, 1.0, and 2.0 GHz;
  • FIG. 4E eleven relative frequencies, 0.0, 1.0, 2.0,..., 10.0 GHz.
  • FIGS.5A and 5B are reconstructed images with two objects using eleven holograms:
  • FIG. 6 is a schematic of an apparatus for digital interference holography.
  • FIG. 7 A is a direct camera image of a damselfly under laser illumination
  • FIG. 7B is
  • FIG. 7C is an image accumulated
  • FIGS.8A-8C are digitally recorded optical fields: FIG. 8A, a hologram; FIG. 8B, an
  • FIG. 8C a reference, RR*.
  • FIG. 9 is an animation of az-y cross section of the three-dimensional reconstructed
  • FIG. 10A are x-y cross sections of the accumulated array at various axial distances z;
  • FIG. 10B are z-y cross sections of the accumulated array at various x values starting from the
  • FIG. 11 is an animated three-dimensional reconstruction of the insect's illuminated
  • FIG. 1 The apparatus 10 of the present invention is depicted in FIG. 1.
  • beam splitters 16,18 One of these provides the planar reference beam 20, while
  • the object consists of two transparency
  • One target 30 (OBJ1)
  • OBJ2 is a checkerboard pattern with 2.5-mm grid size
  • OBJ2 is a target 32
  • the object 22,24 and reference 20 beams are combined in a Michaelson interferometer
  • the interference pattern on the screen is imaged, for
  • digital camera 36 such as a Kodak DC 120
  • another lens L2 38 for example, by digital camera 36, such as a Kodak DC 120, through another lens L2 38 for
  • the exemplary camera 36 has 960 x 1280 pixels with 10x10 ⁇ m 2 pixel size.
  • the calculations presented here use 256 x 256 pixel images of screen
  • the process is repeated a plurality of times, here up to 11 laser frequencies spaced 1.0 GHz
  • a software package for example, a MatLab program
  • E(x,y;z) exp[ (ik/2z) (x 2 +y 2 ) ] F ⁇ E Q (x 0 ,y 0 ) S (x 0 ,y 0 ; z) ⁇ [ ⁇ ⁇ , ⁇ y ]
  • FIGS.2A-2D illustrate the input images of the reference (__, FIG.2 A), the object (O,
  • FIG. 3 A shows the hologram with both objects OBJ1 and OBJ2 on, after subtraction of reference and object images. The images are reconstructed near the two object distances
  • the axial resolution determined by focal sharpness is at least - 15 cm, as can be seen in FIG. 4A, where the
  • vertical axis is a slice of the reconstructed image along the dotted vertical line of FIG. 3C and
  • the horizontal axis is the image distance z ⁇ from 140 to 190 cm.
  • cm corresponds to the lower left corner of OBJ2's letter "A.”
  • FIG.4E eleven holograms with frequencies 0.0, 1.0, 2.0, ... , 10.0 GHz are combined in FIG.4E, which
  • each of the images contains only one of OBJ1 or OBJ2, and the out-of-focus images are
  • the invention thus demonstrates the use of multiwavelength interference of computer-
  • the apparatus is very simple and amenable to electronic automation without mechanical moving parts. Even
  • FIGS. 4A-4E The main source of imperfection in FIGS. 4A-4E, for example, was the
  • Another embodiment may include,
  • the technique can
  • cross-sectional images can then be recombined with appropriate scaling for the removal of distortion, resulting in a synthesis of three-dimensional models that can be
  • a holographic apparatus 40 (FIG. 6)
  • a laser for example, a ring dye laser 41.
  • the beam 90 is apertured 44 to a desired diameter, here 5 mm, and directed to a first
  • a first portion 91 of the split beam passes through a second neutral density
  • a second portion 93 of the split beam is directed to the object 80, here a damselfly specimen, shown under laser illumination in FIG.
  • the scattered light 94 from the object 80 is combined with the reference beam 92 at
  • a second beam splitter 47 to form an interference beam 95, which then passes through a
  • magnifying lens 48 to image the optical image at the camera' s 49 focal plane 50 onto infinity.
  • the camera 49 for example, a digital camera (such as model DC290, manufactured by Kodak,
  • the object-to-hologram distance 51 here is 195 mm.
  • beam 94 preferably should be apertured so that it only illuminates the area of the object 80 that
  • the digitally recorded images are transferred to a computer 52, where software means
  • a CCD array is used instead of the camera 49, wherein the image
  • the object and reference frames are then numerically subtracted from the hologram
  • the holographic image field is then calculated as above.
  • the numerical reconstruction and digital interference proceeds by starting from a 512
  • This new array then has a field distribution that represents the 3D object structure, as
  • FIG. 7B is an example of a 2D holographic image reconstructed from a single
  • the effect of digital interference is illustrated in FIG. 9.
  • FIGS. 10A and 1 OB show cross-sectional tomographic views of the accumulated field
  • FIG. 10A showing x-y cross sections as the axial distance z is varied from the front
  • FIG. 10B shows
  • holographic field arrays has an additional benefit of averaging out the coherent speckle noise.
  • FIG. 7C is obtained by starting from the accumulated array and summing over the z
  • each object surface element is imaged in focus regardless of the depth of focus of the optical system. This feature is especially beneficial in an embodiment applied to microscopic imaging with a large numerical aperture.

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  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Holo Graphy (AREA)

Abstract

La présente invention concerne un simple appareil holographique numérique et un procédé qui permettent de reconstruire des objets en trois dimensions avec une très faible profondeur focale ou une haute résolution axiale. On génère optiquement un certain nombre d'hologrammes en utilisant différentes longueurs d'onde espacées à intervalles réguliers. Ces hologrammes sont enregistrés, comme sur une caméra numérique, puis ils sont reconstruits numériquement. Les interférences des multiples longueurs d'onde de ces hologrammes donnent des plans de contours de très petite épaisseur et à séparation large. Les objets placés à différentes distances du plan de l'hologramme sont imagés clairement et indépendamment avec suppression complète des images hors foyer. Cette technique n'est disponible que dans le cadre de l'holographie numérique et elle a des applications dans la microscopie holographique.
PCT/US2000/026462 1999-09-27 2000-09-27 Microscope holographique interferentiel numerique et procedes Ceased WO2001023965A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU77187/00A AU7718700A (en) 1999-09-27 2000-09-27 Digital interference holographic microscope and methods
US10/089,266 US7127109B1 (en) 1999-09-27 2000-09-27 Digital interference holographic microscope and methods

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US15625399P 1999-09-27 1999-09-27
US60/156,253 1999-09-27

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102004010091A1 (de) * 2004-02-27 2005-09-15 "Stiftung Caesar" (Center Of Advanced European Studies And Research) Verfahren zur Fokusdetektion
US20100094137A1 (en) * 2007-03-12 2010-04-15 Worcester Polytechnic Institute Methods and systems for observation of tympanic function
WO2011149405A1 (fr) * 2010-05-24 2011-12-01 Phase Holographic Imaging Phi Ab Microscopie holographique numérique d'objets biologiques translucides
DE102016110362A1 (de) * 2016-06-06 2017-12-07 Martin Berz Verfahren zur Bestimmung einer Phase eines Eingangsstrahlenbündels
WO2023015621A1 (fr) * 2021-08-13 2023-02-16 北京航空航天大学 Procédé d'affichage holographique 3d sans diaphonie fondé sur un principe d'imagerie floue de diffraction

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2090838C1 (ru) * 1992-11-11 1997-09-20 Сергей Алексеевич Александров Голографический способ определения рельефа поверхности

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2090838C1 (ru) * 1992-11-11 1997-09-20 Сергей Алексеевич Александров Голографический способ определения рельефа поверхности

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
DATABASE WPI Section EI Week 199820, Derwent World Patents Index; Class S02, AN 1998-229107, XP002156897 *
KIM M K: "WAVELENGTH-SCANNING DIGITAL INTERFERENCE HOLOGRAPHY FOR OPTICAL SECTION IMAGING", OPTICS LETTERS,OPTICAL SOCIETY OF AMERICA, WASHINGTON, US, vol. 24, no. 23, 1 December 1999 (1999-12-01), pages 1693 - 1695, XP000955284, ISSN: 0146-9592 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102004010091A1 (de) * 2004-02-27 2005-09-15 "Stiftung Caesar" (Center Of Advanced European Studies And Research) Verfahren zur Fokusdetektion
US20100094137A1 (en) * 2007-03-12 2010-04-15 Worcester Polytechnic Institute Methods and systems for observation of tympanic function
US9155459B2 (en) * 2007-03-12 2015-10-13 Worcester Polytechnic Institute Methods and systems for observation of tympanic function
WO2011149405A1 (fr) * 2010-05-24 2011-12-01 Phase Holographic Imaging Phi Ab Microscopie holographique numérique d'objets biologiques translucides
DE102016110362A1 (de) * 2016-06-06 2017-12-07 Martin Berz Verfahren zur Bestimmung einer Phase eines Eingangsstrahlenbündels
US10823547B2 (en) 2016-06-06 2020-11-03 Martin Berz Method for determining a phase of an input beam bundle
WO2023015621A1 (fr) * 2021-08-13 2023-02-16 北京航空航天大学 Procédé d'affichage holographique 3d sans diaphonie fondé sur un principe d'imagerie floue de diffraction

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