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Journal ArticleDOI

Compensation of the inherent wave front curvature in digital holographic coherent microscopy for quantitative phase-contrast imaging

10 Apr 2003-Applied Optics (Optical Society of America)-Vol. 42, Iss: 11, pp 1938-1946
TL;DR: An approach is proposed for removing the wavefront curvature introduced by the microscope imaging objective in digital holography, which otherwise hinders the phase contrast imaging at reconstruction planes and it is shown that a correction effect can be obtained at all reconstruction planes.
Abstract: An approach is proposed for removing the wave front curvature introduced by the microscope imaging objective in digital holography, which otherwise hinders the phase contrast imaging at reconstruction planes. The unwanted curvature is compensated by evaluating a correcting wave front at the hologram plane with no need for knowledge of the optical parameters, focal length of the imaging lens, or distances in the setup. Most importantly it is shown that a correction effect can be obtained at all reconstruction planes. Three different methods have been applied to evaluate the correction wave front and the methods are discussed in detail. The proposed approach is demonstrated by applying digital holography as a method of coherent microscopy for imaging amplitude and phase contrast of microstructures.
Citations
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Journal ArticleDOI
TL;DR: In this article, the least squares surface fitting with points number less than the matrix of the original hologram is performed on the unwrapped phase distribution to remove the unwanted wavefront curvature.

79 citations

Journal ArticleDOI
TL;DR: This study presents a study based on simulations and real measurements describing the shot-noise influence in the quality of the reconstructed phase images, derived from a model for image quality estimation proposed by Wagner and Brown.
Abstract: In digital holographic microscopy, shot noise is an intrinsic part of the recording process with the digital camera. We present a study based on simulations and real measurements describing the shot-noise influence in the quality of the reconstructed phase images. Different configurations of the reference wave and the object wave intensities will be discussed, illustrating the detection limit and the coherent amplification of the object wave. The signal-to-noise ratio (SNR) calculation of the reconstructed phase images based on the decision statistical theory is derived from a model for image quality estimation proposed by Wagner and Brown [Phys. Med. Biol. 30, 489 (1985)]. It will be shown that a phase image with a SNR above 10 can be obtained with a mean intensity lower than 10 photons per pixel and per hologram coming from the observed object. Experimental measurements on a glass-chrome probe will be presented to illustrate the main results of the simulations.

78 citations

Journal ArticleDOI
TL;DR: DHIC allows, from a single recording, to set a posteriori the best conditions for DIC imaging in conjunction with the numerical focusing feature of DH, with the aim to optimize the visualized phase-contrast imaging as favorite representation for bio-scientists.
Abstract: Differential image contrast (DIC), through the numerical managing and manipulation of complex wavefronts obtained by digital holography (DH), is investigated. We name the approach Dynamical Differential Holographic Image Contrast (DDHIC). DDHIC dispenses from special optics and/or complex setup configurations with moveable components, as usually occurs in classical DIC, that is not well-suited for investigating objects experiencing dynamic evolution during the measurement. In fact, the technique presented here, is useful for floating samples since it allows, from a single recording, to set a posteriori the best conditions for DIC imaging in conjunction with the numerical focusing feature of DH. By DDHIC, the movies can be easily built-up to offering dynamic representation of phase-contrast along all directions, thus improving the visualization. Furthermore, the dynamic representation is useful for making the proper choice of other key parameters of DIC such as the amount of shear and the bias, with the aim to optimize the visualized phase-contrast imaging as favorite representation for bio-scientists. Investigation is performed on various biological samples.

75 citations

Journal ArticleDOI
TL;DR: Through frequency analysis and visualization of its 3D optical transfer function, an assessment of the imaging behavior of DHM is given and the results are compared with those from other types of interference microscopy.
Abstract: We present a theoretical formalism for three dimensional (3D) imaging properties of digital holographic microscopy (DHM). Through frequency analysis and visualization of its 3D optical transfer function, an assessment of the imaging behavior of DHM is given. The results are compared with those from other types of interference microscopy. Digital holographic microscopy does not result in true 3D imaging. The main advantage of holographicmicroscopy lies in its quick acquisition of a single 2D image. Full 3D imaging can be obtained with DHM using a broad-band source or tomographic reconstruction.

74 citations

Journal ArticleDOI
TL;DR: This paper investigates the use of a digital holographic microscope, with partial spatial coherent illumination, for the automated detection and tracking of spermatozoa and characterized cell motility on clinical samples of seminal fluid.
Abstract: In this paper we investigate the use of a digital holographic microscope, with partial spatial coherent illumination, for the automated detection and tracking of spermatozoa. This in vitro technique for the analysis of quantitative parameters is useful for assessment of semen quality. In fact, thanks to the capabilities of digital holography, the developed algorithm allows us to resolve in-focus amplitude and phase maps of the cells under study, independently of focal plane of the sample image. We have characterized cell motility on clinical samples of seminal fluid. In particular, anomalous sperm cells were characterized and the quantitative motility parameters were compared to those of normal sperm.

69 citations

References
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Journal ArticleDOI
TL;DR: A new method is proposed in which the distribution of complex amplitude at a plane is measured by phase-shifting interferometry and then Fresnel transformed by a digital computer, which can reconstruct an arbitrary cross section of a three-dimensional object with higher image quality and a wider viewing angle than from conventional digital holography using an off-axis configuration.
Abstract: A new method for three-dimensional image formation is proposed in which the distribution of complex amplitude at a plane is measured by phase-shifting interferometry and then Fresnel transformed by a digital computer. The method can reconstruct an arbitrary cross section of a three-dimensional object with higher image quality and a wider viewing angle than from conventional digital holography using an off-axis configuration. Basic principles and experimental verification are described.

1,813 citations

Journal ArticleDOI
TL;DR: The principle of recording holograms directly on a CCD target is described and a real image of the object is reconstructed from the digitally sampled hologram by means of numerical methods.
Abstract: The principle of recording holograms directly on a CCD target is described. A real image of the object is reconstructed from the digitally sampled hologram by means of numerical methods.

1,444 citations

Journal ArticleDOI
TL;DR: A new application of digital holography for phase-contrast imaging and optical metrology and an application to surface profilometry shows excellent agreement with contact-stylus probe measurements.
Abstract: We present a new application of digital holography for phase-contrast imaging and optical metrology. This holographic imaging technique uses a CCD camera for recording of a digital Fresnel off-axis hologram and a numerical method for hologram reconstruction. The method simultaneously provides an amplitude-contrast image and a quantitative phase-contrast image. An application to surface profilometry is presented and shows excellent agreement with contact-stylus probe measurements.

1,202 citations

Journal ArticleDOI
TL;DR: Off-axis holograms recorded with a magnified image of microscopic objects are numerically reconstructed in amplitude and phase by calculation of scalar diffraction in the Fresnel approximation to show that the transverse resolution is equal to the diffraction limit of the imaging system.
Abstract: We present a digital method for holographic microscopy involving a CCD camera as a recording device. Off-axis holograms recorded with a magnified image of microscopic objects are numerically reconstructed in amplitude and phase by calculation of scalar diffraction in the Fresnel approximation. For phase-contrast imaging the reconstruction method involves the computation of a digital replica of the reference wave. A digital method for the correction of the phase aberrations is presented. We present a detailed description of the reconstruction procedure and show that the transverse resolution is equal to the diffraction limit of the imaging system.

1,174 citations

Journal ArticleDOI
TL;DR: The principles and major applications of digital recording and numerical reconstruction of holograms (digital holography) are described, which are applied to measure shape and surface deformation of opaque bodies and refractive index fields within transparent media.
Abstract: This article describes the principles and major applications of digital recording and numerical reconstruction of holograms (digital holography). Digital holography became feasible since charged coupled devices (CCDs) with suitable numbers and sizes of pixels and computers with sufficient speed became available. The Fresnel or Fourier holograms are recorded directly by the CCD and stored digitally. No film material involving wet-chemical or other processing is necessary. The reconstruction of the wavefield, which is done optically by illumination of a hologram, is performed by numerical methods. The numerical reconstruction process is based on the Fresnel–Kirchhoff integral, which describes the diffraction of the reconstructing wave at the micro-structure of the hologram. In the numerical reconstruction process not only the intensity, but also the phase distribution of the stored wavefield can be computed from the digital hologram. This offers new possibilities for a variety of applications. Digital holography is applied to measure shape and surface deformation of opaque bodies and refractive index fields within transparent media. Further applications are imaging and microscopy, where it is advantageous to refocus the area under investigation by numerical methods.

1,171 citations