Twostep phase shifting differentialrecording digital. Applications of holographic microscopy in life sciences intechopen. Optical scanning holography with matlab r by tingchung. We present a technique of threedimensional 3d differential imaging by the way of incoherent digital holography. Inline holography with electrons was revived in the late 1980s by fink,14 stocker et al. Digital reconstruction is accomplished using fourier transforms. The program starts by switching on the vcsel, which is moved to the first. These techniques represent a fascinating alternative to standard interferometry and digital holography. Digital holography comes in different flavors, such as offaxis fresnel, fourier, image plane, inline, gabor and phaseshifting digital holography, 2 depending on the optical setup. The method, developed at kyoto institute of technology in japan, could.
Compact inline lensfree digital holographic microscope. Realtime 3d imaging with differential digital holography microscopy. Differential digital holography is an evolution of. Twostep phase shifting differentialrecording digital holographic. It use fourier transform to process the origin interference imageby fourier lens, and captured by ccd, and reappear by inverse fourier transform.
Here, we propose a digital holographic autofocusing method that computes the focused distance using the first longitudinal difference of the magnitude of the. Osa applications of digital holography in biomedical microscopy. Hodhmit01 is an inverted type digital holography microscope which can be used for the quantitative phase imaging for live cells. While phasesensitive imaging methodologies such as darkfield, phase contrast and differential interference contrast are known for several decades, they cannot provide quantitative phase information. Retinal blood flow in the optic disc revealed by digital holography duration. Figure 1 illustrates the schematic diagram of tpddh in microscopy. In differential interference contrast dic microscopy, the interference of two sheared. May 16, 2017 twostep phase shifting differential recording digital holographic microscopy. F152f157 march 2017 with 192 reads how we measure reads. Cellbased assay, differential interference contrast, digital holography, ion channel, highcontent screening, imagebased screen, phase contrast, phenotypic drug discovery. Digital holography dh is a potentially disruptive new technology for many areas of imaging science, especially in microscopy and metrology. One or more connected components are identified in each of the plurality of dhm images and one or more training white blood cell images are generated. Twostep phase shifting differential recording digital holographic microscopy.
They enable wave field sensing without the requirement of a particular reference wave, thus allowing the use of low brilliance light sources and even liquidcrystal displays lcd for interferometric applications. In particular the strength of dhm principle allows unique software compensations. However, a picture is the twodimensional projection. V alidated with almost every type of transparent cell culture vessel, ovizios devices can track cell density, cell morphology as well as cell viability with the highest reproducibility at. A million frames per second has been achieved in 3d digital holography of moving objects. Inline digital holographic microscopy dhm is a powerful imaging technique that can be realized around a. Among them, digital holographic microscopy dhm is a wellknown. The first mentioned type of digital holography finds applications in digital holographic microscopy, particle analysis, and interferometric form and deformation measurement, while the second type constitutes the basis for holographic 3d tv. The t 2 signal measured with the long acquisition time the fourth trace is considerably greater than the t 2 signal measured with the short acquisition time the fifth trace, and the separation between the two the sixth trace indicates distinct characteristics of a gas layer. Digital holography dh is a promising technique for modern threedimensional 3d imaging.
Dec 28, 2009 this paper presents a complete procedure of the differential digital holography process. This paper presents a complete procedure of the differential digital holography process. Digital holography is an emerging field of new paradigm in general imaging applications. Osa digital holography and threedimensional imaging 2016. Digital holographic microscopes dhm enables strictly noninvasive. In comparison to the software unwrapping, dualwavelength optical. Digital holographic microscopy is a new imaging technology applied to optical microscopy. Optical aberrations, digital image focusing, and numerical compensation for sample tilt and environmental disturbances can be. Computer simulation of differential digital holography. Digital holography digital hologram recording, numerical.
A possibility of applying differential digital holography in. This technique is called interferometric microscopy. To verify the effectiveness of the proposed approach, a series of simulation experiments have been conducted using matlab at first. Pdf color reconstructions of real digital holograms in holographic display with white light led illumination tomasz kozacki, maksymilian chlipala, weronika zaperty, and piotr makowski.
Dh offers a number of significant advantages such as the ability to acquire holograms rapidly, availability of complete amplitude and phase information of the optical field, and versatility of the interferometric and image processing techniques. Threedimensional microscopy by laser scanning and multi. Official web site of the ozcan research group, the research group of prof. Based on selfinterference of two beamsplit copies of the objects.
In the path of object wave, a test specimen, such as a microlens array, is placed in the front focal plane of microscope objective mo. He wrote the program that was used to reconstruct all of the holograms in this project. Goethe, faust an old dream of mankind and a sign of culture is the conservation of moments by taking an image of the world around. A method for analyzing digital holographic microscopy dhm data for hematology applications includes receiving a plurality of dhm images acquired using a digital holographic microscopy system. Fullcolor, threedimensional images of objects under incoherent illumination are obtained by a digital holography technique. Holography is an advanced form of photography that allows an image to be recorded in three dimensions.
Ovizios microscopes offer an automated, labelfree tool designed for viable cell counting. Imaging of various cell types, including skov3 ovarian cancer cells, fibroblast cells, testate amoeba, diatom skeletons, and red blood cells. Optical scanning holography with matlab consists of tutorials with numerous matlab examples throughout the text, research material, as well as new ideas and insights that are useful for engineering or physics students, scientists, and engineers working in the fields of fourier optics, optical scanning imaging and holography. Transparent objects, like living biological cells, are traditionally viewed in a phasecontrast microscope or in a differential interference. A possibility of applying differential digital holography. The main contribution of this paper is proposed method called differential digital holography. Due to hologram characteristic to project every part of image in every part of hologram, even small defect is amplified. Osa fast autofocusing in digital holography using the. Principles and techniques of digital holographic microscopy. The first part deals with optical foundations and the theory of holography.
The difference between holography and photography is best understood by considering what a black and white photograph actually is. This digital approach to holography allows the application of computer based procedures at a level unreached in videomicroscopy. Overview holography digital holographic transformation differential digital holography information coding with holography holography holography was invented in 1947 by hungarian physicist dennis gabor 19001979, discovery was an unexpected result of research into improving electron microscopes at the british thomsonhouston company in. Improved phase reconstruction using linear programming for.
University of south florida scholar commons graduate theses and dissertations graduate school 9122008 threedimensional microscopy by laser scanning. Schnars and juptner applied their method to interferome. F152f157 march 2017 with 188 reads how we measure reads. Dyi digital holography stanislovas zacharovas1a, andrej nikolskija, jevgenij kuchin a ageola digital uab, naugarduko str. Part of theamerican studies commons this dissertation is brought to you for free and open access by the graduate school at scholar commons. Digital holographic microscopy dhm is a labelfree imaging technique allowing. Digital recording and numerical reconstruction of holograms. Results of experiment show possibility for small ceramic tiles pattern defect detection.
The method is based on digital hologram construction and reconstruction. A sample with height map of 2500x 2y 2 nm is tested as shown in fig. Us10176363b2 analyzing digital holographic microscopy data. Digital holography as well as transmitted led bright field observation. Also, the explanation of software solution is included. Introduction phenotypic screens 1 3 rely on imagebased cellular assays for analyzing cell morphological variations or alterations.
This book presents a selfcontained treatment of the principles and major applications of digital hologram recording and numerical reconstruction digital holography. Image processing in differential digital holography ddh iajit. Within the scope of this article digital sampling and numerical hologram reconstruction is called digital holography. This program is used to reappear digital holography. V alidated with almost every type of transparent cell culture vessel, ovizios devices can. Aydogan ozcan from ucla, working on nanotechnology, biotechnology, nanophotonics and biophotonics. Typical methods of automatic estimation of focusing in digital holography calculate every single reconstructed frame to get a critical function and then ascertain the focal plane by finding the extreme value of that function. Threedimensional microscopy by laser scanning and multiwavelength digital holography alexander khmaladze university of south florida follow this and additional works at. Based on patented differential digital holography technology, advanced algorithms extract meaningful quantitative data about complex cellular dynamics. Holograms and reconstructs are calculated by applying fourier transformation on bitmap images. The software can analyze more images and more details of each imageresulting in more and better information on a continuous basis. Coherent holography records the complex amplitude of a 3d object holographically, giving speckle noise upon reconstruction and presenting a serious drawback inherent in coherent optical systems.
In digital holography, a scattered light beam reflected by an illuminated object interferes with a reference beam on a ccd camera allowing for a 3d numerical reconstruction of the object. Digital holography comes in different flavors, such as offaxis fresnel, fourier, image plane, inline, gabor and phaseshifting digital holography, depending on the optical setup. Principles and techniques of digital holographic microscopy myung k. Digital holographic microscopy dhm is digital holography applied to microscopy. Pdf fast autofocusing in digital holography using the. Digital holographic microscopy is an emerging modality that offers capability of quantitative phase imaging qpi of transparent unstained cells in their most natural state. The ozcan research group refereed journal publications. Pdf selfinterference incoherent digital holography for. Phase contrast microscopy 1 and differential interference contrast dic. Digital holography and image processing methods for.
The digital analysis of a set of holograms recorded from different directions or with different direction of the reference wave allows the numerical emulation of an objective with large numerical aperture, leading to corresponding enhancement of the resolution. The method, developed at kyoto institute of technology in japan, could be used to analyse highspeed. We demonstrate that the method is suitable for fluorescence microscopy without. Digital holography matlab version about this program. It is a transmission type inverted digital holography microscope with balanced afocal configuration that uses infinity corrected plan achromatic objectives in both reference and object beams. Realtime digital holography of the retina by eigendecomposition. Fast autofocusing in digital holography using the magnitude differential article pdf available in applied optics 56. The spacebandwidthproblem occuring in this context is addressed and first partial solutions are presented. Digital holography is an emerging field of new paradigm in general imaging appli. Kim university of south florida, department of physics, 4202 e.
Nov 06, 2014 a million frames per second has been achieved in 3d digital holography of moving objects. A new approach is based on differential digital holography. Digital holography using a laser pointer and consumer digital. Sag ich zum augenblicke verweile doch, du bist so schon j. Ovizios differential digital holographic microscopy, patented by the ulb, is an evolution of this base technology bringing increased stability, flexibility and an important reduction of size of the instruments while using lower cost, noninvasive led light sources. Digital holograms of a target acquired with a sweptsource laser. 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.
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