Digital holograms measure the interference of a light wave that has been diffracted through a sample and combined with a coherent, non. Digital holography is an emerging field of new paradigm in general imaging. Assisted reproductive technologies seek to improve the success rate of pregnancies. Pdf specklefield digital holographic microscopy semantic. A key advantage of dhm over traditional light microscopy is the simultaneous imaging of a thick sample. Calibrated phaseshifting digital holographic microscope. Lensless digital holographic microscopy and its applications in biomedicine and environmental monitoring yichen wua,b,c, aydogan ozcana,b,c,d. Digital holographic microscopy dhm instruments and koala software system make measurements easy in micro optics.
Digital holographic microscopy dhm is an imaging technique offering both subwavelength resolution and real time observation capabilities. Highresolution quantitative phasecontrast microscopy by. The recording method of digital holographic microscopes is the same as that. Nanometric accuracy can be reached in holographic microscopy with a high numerical aperture na. Direct access to the phase as well as amplitude profiles makes quantitative phase microscopy by digital holography dhqpm particularly powerful and versatile.
Digital holographic microscopy dhm is a technique that uses optical interference patterns to record a threedimensional optical field for imaging, sensing, and microscopy applications. Introduction living cells are largely transparent in ordinary microscopy, and therefore computerized analyses today require the addition of a dye to the cells. In a holographic microscope, the sample is illuminated by laser light, and the. Pdf principles and techniques of digital holographic. Digital holographic microscopy principles, techniques, and. Moliner, 50, 46100 burjassot, spain 2school of engineering, barilan university, ramatgan, 52900 israel corresponding author. The main characteristics of the proposed method, which make it different from the previous works in the field, are the possibility of producing the 3d topography by a single shot over the complete field of view with.
Pdf spacebandwidth conditions for efficient phaseshifting. The quantitative dhm phase contrast image provided is related both to the intracellular refractive index and to cell thickness. The digital holographic microscope has the unique advantage of combining the. Digital holography and digital holographic microscopy, focusing on the reduction of speckle noise and improvement of the capability of imaging resolution. Digital holographic microscopy principles, techniques. Moreover, the possibility of reconstructing automatically, i. Apr 01, 2018 inline digital holographic microscopy is used to obtain a particle size distribution of fly ash after wet electrostatic precipitators. Results from engineered surfaces and living cells demonstrate applications of digital holographic microscopy for technical inspection and life cell imaging. Digital holographic microscopy dhm is a powerful optical technique for the study of microscopic samples by retrieving the amplitude and phase of the wave reflected by the sample or transmitted. Digital holographic microscopy dhm is a potent tool to perform threedimensional imaging and tracking. However, most samples used as test images have been solid or adhered to a surface, making it easy to determine focal planes and correct for chromatic aberration. Moving to the next step of 3d imaging is challenging. It is a quantitative phase microscopy technique capable of being configured for reflection or transmission microscopy, and it has been used for a variety of applications 1.
Within the last decade, quantitative phase imaging qpi has been continuously improved for the purposes of high. It is possible to build a digital holographic microscope by simply recording a digital hologram of the wave field propagated in the imaging process of an object. Kersti alm, phd application specialist at phase holographic imagingspeaker biography. Espci paristech cnrs umr 7587, inserm u 979, upmc, up7. Introduction digital holographic microscopy dhm shows great promise as a tool for investigating soft. Recent progress and perspectives in digital holographic. Morphology scoring is a common approach to evaluate oocyte and embryo viability prior to embryo transfer in utero, but the efficacy of the method is low. Ultrafast transient holographic microscopy nano letters. Nadeau1 abstract detection of extant microbial life on earth and elsewhere in the solar system requires the ability to identify and. Digital holographic microscopy, a method for detection of microorganisms in plume samples from enceladus and other icy worlds manuel bedrossian,1 chris lindensmith,2 and jay l. The first dhm images of living cells were obtained 810 years ago 1, 2.
Digital holographic microscopy characterization of micro optics with digital holographic microscopy dhm instruments and koala software system make measurements easy in micro optics. Singleshot 3d topography of reflective samples with digital. Digital holographic microscopy dhm 2, 3 is an interferometrybased variant of qpi that typically uses a laser as a coherent light source. Nov 01, 2020 lensless digital holographic microscopy and its applications in biomedicine and environmental monitoring methods, 6 2018, pp. Techniques of digital holography are improved in order to obtain highresolution, highfidelity images of quantitative phasecontrast microscopy. Digital holographic microscopy with pureoptical spherical. Dhm provides qpi by detecting specimeninduced optical path length changes against the surrounding environment. Digital holographic microscopy is a fast threedimensional 3d imaging tool with many applications in soft matter physics. Compact lensless digital holography infoscience epfl. Digital holographic microscopy dhm is an emerging highresolution imaging technique that offers realtime imaging and quantitative measurements of physiological parameters without any staining or labeling of cells. Digital holographic microscopy dhm is an emerging technique for 3dimensional imaging and tracking of microorganisms 15 and for in vitro analysis of cellular and subcellular processes such as ion channel function and apoptosis 7, 8. Katz, digital holographic microscope for measuring threedimensional particle distributions and motions, appl. Osa biodynamic digital holographic speckle microscopy for. With calibrated optical imaging systems this feature can be used also to determine the position and axial displacements of a sample.
Pdf digital holographic microscopy dhm is a potent tool to perform threedimensional imaging and tracking. Dimiduk and manoharan 2016 bayesian approach to analyzing hologra. The description and validation of an imagej opensource plugin to numerically simulate and reconstruct digital lensless holographic microscopy dlhm holograms are presented. Introduction combination of digital holography with optical microscopy, or in other words, digital holographic microscopy dhm, avoids the limited resolution imposed by the finite number and size of the pixels in the digital sensor. First, the basic results from the general theory of holography, based on the scalar diffraction theory, are summarized, and a general description of the. By now, digital holography has been confirmed as a useful technique to image and qualitatively analyze blood flows in 2d 27,28. The study of single cells over longer periods of time requires nondisturbing measurement. Here, we address this experimental challenge by combining concepts of ultrafast spectroscopy, widefield nanoscopy, and digital holography. Microscopy is one of the main research and application areas of digital holography. Among them, digital holographic microscopy dhm is a unique method due to its advantages of low cost, ease of use, realtime feedback, labelfree approach and wide field of view 9. Legros, improved threedimensional imaging with a digital holography microscope with a source of partial spatial coherence. Lensless digital holographic microscopy and its applications in biomedicine and environmental monitoring.
Multiwavelength digital holographic microscopy dhm has been used to improve phase reconstructions of digital holograms by reducing 2. In this work, an offaxis digital holographic microscope operating in reflection mode and a telecentric regimen to produce 3d topography of a microscopy sample is shown. This means that samples under investigation are, for example, not influenced by fluorophores or dyes which can change physiological processes or. Quantitative phase digital holographic microscopy qpdhm represents an important and efficient quantitative phase method to explore cell structure and dynamics. Superresolution digital holographic microscopy for three. Basic principles of diffraction and general holography are outlined in sec. Dec 29, 2020 various types of technologies including blood cell counter, phase contrast microscopy and digital holography have been implemented to obtain such information. Lensless digital holographic microscopy and its applications.
A speckle beam of a complex spatial pattern is used for illumination to reduce fixed pattern noise and to improve optical sectioning capability. Digital holographic microscopy, a method for detection of. Pdf principles and techniques of digital holographic microscopy. We report a technique which can overcome these drawbacks, but maintains the advantage of phase microscopy high contrast live cell imaging and 3d imaging for the detail, please see the ref. Unlike other digital holographic microscopes, an onchip microscope does not need to use a small e. Lynceetec is pioneer and leader in digital holographic microscopy. Digital holographic microscopy wiley online library. Specklefield digital holographic microscopy yongkeun park 1,2, wonshik choi 1,3, zahid yaqoob 1, ramachandra dasari 1, kamran badizadegan 1,4, and michael s. Automatic procedure for aberration compensation in digital holographic microscopy and applications to specimen shape compensation, appl.
Wavefront division digital holography aip publishing. Principles and techniques of digital holographic microscopy. Measurements based on digital holographic microscopy 2. Kemper has developed various dhm systems and methods, and he has demonstrated their application capabilities for qpi in multiple application fields. Quantitative phase imaging for labelfree analysis of cancer. Harrison spectroscopy laboratory, mit, 77 massachusetts avenue, cambridge, ma 029, usa. Digital holographic microscopy with dualwavelength phase unwrapping. One embodiment includes obtaining interferometric signals of a specimen, obtaining fluorescence signals emanating from the specimen, recording these signals, and processing these signals so as to reconstruct threedimensional. Pdf digital holography is an emerging field of new paradigm in general imaging applications.
This means that samples under investigation are, for example. The ozcan research group refereed journal publications. Quantitative phasedigital holographic microscopy kaust. Among the different implementations of phase imaging, digital holographic microscopy dhm is a wellknown phase method which provides a quantitative value.
A polydimethylsiloxane microfluidic chip with a cross section of 50. Edge processing by synthetic aperture superresolution in. Principle of digital holographic microscopy digital holography is based on the classic holographic principle, with the difference that the hologram recording is performed by a digital image sensor, e. Javidi spacebandwidth conditions for efficient phaseshifting digital holographic microscopy adrian stern1, and bahram javidi2 1 electro optical unit, ben gurion university of the negev, beersheva 84105, israel 2 department of electrical and computer engineering, university of connecticut, storrs, connecticut 062691157. Review of digital holographic microscopy for three. Autofocusing in digital holographic microscopy springerlink. In a second part, the most relevant qpm applications in the field of cell biology are summarized. We apply biodynamic imaging, based on dynamic light scattering and lowcoherence digital holography, to assess the metabolic activity of oocytes and embryos. Simultaneous threewavelength unwrapping using external digital. Cells and holograms holograms and digital holographic. Onchip differential interference contrast microscopy using lensless digital holography, optics express, vol.
Characterization of micro optics with digital holographic. We introduce an ultrafast holographic transient microscope for widefield transient nanoscale imaging with high frequency alloptical signal demodulation. Instead, the light wave front information originating from the object is digitally recorded as a hologram, from which a computer calculates the object image by using a numerical reconstruction algorithm. Digital holographic microscopy distinguishes itself from other microscopy methods by not recording the projected image of the object. In order to assess the ability of dhm to distinguish distinct phenotypes we measured the zfactor 18 obtained on two representative cell lines, hela round shape and high contrast or h9c2 elongated and. Review of quantitative phasedigital holographic microscopy. Inline holography with spherical waves,asoriginallyproposedbygabor,isthesimplest realization of the holographic method, working without lenses. A method and device for obtaining a sample with threedimensional microscopy, in particular a thick biological sample and the fluorescence field emitted by the sample. Digital holography is an emerging field of new paradigm in general imaging applications. Compressed sensing for digital holographic microscopy marcio m. The utilization of microscope objectives mos in digital holographic microscopy dhm has associated effects that are not present in conventional optical.
How digital holographic microscopy can be used to explore. Color imaging in lensless digital holographic microscopy. Where traditional light microscopy records the intensity image of the object, dhm records the interference of a light beam pass ing through an object the object beam with the light of a reference beam dubois et al. Digital holographic microscopy 61 b16f10 mouse skin electrochemotherapy x perfusion slide digital holographic microscopy 62 m229p, m229r5, m238p, m238r1, m249p, m249r4 human melanoma vemurafenib x x 24well glassbottom plate highspeed live cell, interferometry 63 rm82, cadoes1, vh64, staet1 human ewing sarcoma birc5 knockdown x x. Digital holography is based on the classic holographic principle, with the difference that the hologram recording is performed by a digital image sensor, e.
Frontiers multiwavelength digital holographic imaging. Kim, is intended to provide a brief but consistent introduction to the principles of digital holography as well as to give an organized overview of the large number of techniques and applications being developed. Poon, conventional optical microscopy of colloidal suspensions, adv. Measurement of cell volume using inline digital holography. Osa biodynamic digital holographic speckle microscopy. The principle of recording holograms in holographic microscopy is shown in figure 2. A digital holographic microscope comprises both an optical setup devoted to hologram formation and software specially developed to numerically process the. Digital holographic microscopy dhm captures a hologram with an electronic device such as a cmos or ccd. Digital holography and its multidimensional imaging. Digital holography microscopy in 3d biologic samples analysis.
Compressed sensing for digital holographic microscopy. Inline digital holographic microscopy using a consumer scanner. Quantitative phase imaging for labelfree analysis of. Phase uncertainty in digital holographic microscopy. Lensless digital holographic microscopy and its applications in. Digital holographic microscopy dhm represents one promising approach that has been demonstrated in aqueous environments on earth lakes and oceans jericho et al. Digital holographic microscopy with dualwavelength phase. Digital holographic microscopy 66 hela human cervix x slide multimodal holographic microscopy. Recent studies have shown that electromagnetic scattering solutions can be fit to digital holograms to obtain the 3d positions of isolated colloidal spheres with nanometer precision and millisecond temporal resolution. Sample tube lens hologram beam splitter mirror a new. A particular emphasis is placed on the original biological information. Digital holographic microscopy dhm is a labelfree imaging technique allowing visualization of transparent cells with classical imaging cell culture plates. Frontiers multiwavelength digital holographic imaging and. Digital holographic microscopy dhm is a novelsurface topography technique that has recently become commercially available.
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