Digital holographic microscopy for live cell applications and technical inspection (original) (raw)
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Digital Holographic Microscopy
Optik & Photonik, 2007
Digital holographic microscopy provides quantitative phase contrast imaging that is suitable for high resolving investigations on reflective surfaces as well as for marker-free analysis of living cells. Results from engineered surfaces and living cells demonstrate applications of digital holographic microscopy for technical inspection and life cell imaging.
Parameter-Optimized Digital Holographic Microscope for High-Resolution Living-Cell Analysis
Applied Optics, 2004
A parameter-optimized off-axis setup for digital holographic microscopy is presented for simultaneous, high-resolution, full-field quantitative amplitude and quantitative phase-contrast microscopy and the detection of changes in optical path length in transparent objects, such as undyed living cells. Numerical reconstruction with the described nondiffractive reconstruction method, which suppresses the zero order and the twin image, requires a mathematical model of the phase-difference distribution between the object wave and the reference wave in the hologram plane. Therefore an automated algorithm is explained that determines the parameters of the mathematical model by carrying out the discrete Fresnel transform. Furthermore the relationship between the axial position of the object and the reconstruction distance, which is required for optimization of the lateral resolution of the holographic images, is derived. The lateral and the axial resolutions of the system are discussed and quantified by application to technical objects and to living cells.
Application of Digital Holographic Microscopy for Imaging Biological Samples
DOAJ (DOAJ: Directory of Open Access Journals), 2013
Red blood cells are able to undergo shape change from the "normal" discocyte to either echinocytes or stomatocytes depending on a large variety of membrane and cytoplasmic parameters. Such shape changes can be relatively fast (within seconds) during the sedimentation of the cells in suspension or after the cells are getting in contact with artificial surfaces. High resolution digital holographic microscopy has been applied to study these processes. This method represents a new setup allowing a contact-less and marker-free quantitative phase-contrast imaging of living cells under conventional laboratory conditions. With the applied technique we were able to detect and analyse fast shape changes of red blood cells.
Digital holographic microscopy–innovative and non-destructive analysis of living cells
2010
Abstracts of proposals for the book titled: Microscopy: Science, Technology, Applications and Education Digital holography is a novel technique that has been developed recently to study living cells. The technique is an innovative, non-destructive method with possibilities to study living cells over time. We are investigating cell number, growth, viability and death of adherent cells using digital holography, which is a novel, label-free, imaging technique for biological applications. We have recently demonstrated that digital holography is highly comparable to the conventional manual cell counting method using a hemocytometer . Digital holography is a method that gives us information about the refractive index of cells, which can change under different circumstances. The technique is cheap, fast and simple to use. The unique measurable parameters are the cell number, cell area, thickness and volume, which can be transformed to proliferation, migration, viability and cell death. The digital holographic images produced can provide both quantitative and qualitative phase information from a single hologram. Future applications can include real-time cell monitoring of various parameters of cells of different diseases in response to clinically relevant compounds.
Photonics Letters of Poland
The statistical analysis and comparison of biophysical parameters of living and fixed, mouse embryonic fibroblasts cells are presented. The parameters are calculated based on phase measurements performed by means of a digital, holographic microscope. The phases are retrieved from off-axis, image plane holograms, followed by custom image segmentation and statistical analysis of cells’ surface, phase volume and dry mass. The results indicated statistically significant differences between fixed and living cell parameters, which is an important message for setting methodology for further diagnosis based on quantitative phase (label-free) analysis.Full Text: PDF References:K. Alm, et al. "Cells and Holograms – Holograms and Digital Holographic Microscopy as a Tool to Study the Morphology of Living Cells", InTech, 2013. [CrossRef]Y. Rivenson, Y. Wu, A. Ozcan, Light: "Deep learning in holography and coherent imaging", Science & Applications, 8, Art. No. 85 (2019) [Cross...
Optics Letters, 2005
We have developed a digital holographic microscope (DHM), in a transmission mode, especially dedicated to the quantitative visualization of phase objects such as living cells. The method is based on an original numerical algorithm presented in detail elsewhere [Cuche et al., Appl. Opt. 38, 6994 (1999)]. DHM images of living cells in culture are shown for what is to our knowledge the first time. They represent the distribution of the optical path length over the cell, which has been measured with subwavelength accuracy. These DHM images are compared with those obtained by use of the widely used phase contrast and Nomarski differential interference contrast techniques.
Digital Holographic Microscopy for Quantitative Live Cell Imaging and Cytometry
Methods and Disease Diagnoses, 2011
Digital holographic microscopy allows determination of dynamic changes in the optical thickness profile of a transparent object with subwavelength accuracy. Here, we report a quantitative phase laser microsurgery system for evaluation of cellular/ sub-cellular dynamic changes during laser micro-dissection. The proposed method takes advantage of the precise optical manipulation by the laser microbeam and quantitative phase imaging by digital holographic microscopy with high spatial and temporal resolution. This system will permit quantitative evaluation of the damage and/or the repair of the cell or cell organelles in real time.
Digital Holographic Microscopy (DHM)
Imaging & Microscopy, 2006
Digital Holographic Microscopes (DHM) enables strictly noninvasive visualisation of unstained transparent and partially reflective specimens, in real time, by providing simultaneously amplitude and phase changes of a light wave transmitted or reflected. They are used for characterisation of samples at the nanometer scale, for quality control on production line, and for dynamical analysis of biological specimen and micro systems. more than 15 reconstructions per second for 512 x 512 pixels holograms with a standard personal computer.
Biophotonics and New Therapy Frontiers, 2006
We have developed a digital holographic microscope (DHM), in a transmission mode, adapted to the quantitative study of cellular dynamics. Living cells are optically probed by measuring the phase shift they produce on the transmitted wave front. The high temporal stability of the phase signal, equivalent to λ/1800, and the low acquisition time (down to 20 µs) enables to monitor cellular dynamics processes. An experimental procedure allowing to calculate both the intracellular refractive index and the cellular thickness (morphometry) from the measured phase shift is presented.