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1.
Opt Express ; 31(17): 28382-28399, 2023 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-37710893

RESUMO

Optical diffraction tomography (ODT) solves an inverse scattering problem to obtain label-free, 3D refractive index (RI) estimation of biological specimens. This work demonstrates 3D RI retrieval methods suitable for partially-coherent ODT systems supported by intensity-only measurements consisting of axial and angular illumination scanning. This framework allows for access to 3D quantitative RI contrast using a simplified non-interferometric technique. We consider a traditional iterative tomographic solver based on a multiple in-plane representation of the optical scattering process and gradient descent optimization adapted for focus-scanning systems, as well as an approach that relies solely on 3D convolutional neural networks (CNNs) to invert the scattering process. The approaches are validated using simulations of the 3D scattering potential for weak phase 3D biological samples.

2.
Appl Opt ; 61(5): B132-B146, 2022 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-35201134

RESUMO

Multi-wavelength digital holographic microscopy (MWDHM) provides indirect measurements of the refractive index for non-dispersive samples. Successive-shot MWDHM is not appropriate for dynamic samples and single-shot MWDHM significantly increases the complexity of the optical setup due to the need for multiple lasers or a wavelength tunable source. Here we consider deep learning convolutional neural networks for computational phase synthesis to obtain high-speed simultaneous phase estimates on different wavelengths and thus single-shot estimates of the integral refractive index without increased experimental complexity. This novel, to the best of our knowledge, computational concept is validated using cell phantoms consisting of internal refractive index variations representing cytoplasm and membrane-bound organelles, respectively, and a simulation of a realistic holographic recording process. Specifically, in this work we employed data-driven computational techniques to perform accurate dual-wavelength hologram synthesis (hologram-to-hologram prediction), dual-wavelength phase synthesis (unwrapped phase-to-phase prediction), direct phase-to-index prediction using a single wavelength, hologram-to-phase prediction, and 2D phase unwrapping with sharp discontinuities (wrapped-to-unwrapped phase prediction).


Assuntos
Holografia , Simulação por Computador , Holografia/métodos , Lasers , Redes Neurais de Computação , Refratometria/métodos
3.
Appl Opt ; 60(4): A21-A37, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33690351

RESUMO

In recent years, research efforts in the field of digital holography have expanded significantly, due to the ability to obtain high-resolution intensity and phase images. The information contained in these images have become of great interest to the machine learning community, with applications spanning a wide portfolio of research areas, including bioengineering. In this work, we seek to demonstrate a high-fidelity simulation of holographic recording. By accurately and numerically simulating the propagation of a coherent light source through a series of optical elements and the object itself, we accurately predict the optical interference of the object and reference wave at the recording plane, including diffraction effects, aberrations, and speckle. We show that the optical transformation that predicts the complex field at the recording plane can be generalized for arbitrary holographic recording configurations using a matrix method. In addition, we provide a detailed description of digital phase reconstruction and aberration compensation for a variety of off-axis holographic configurations. Reconstruction errors are presented for the various holographic recording geometries and complex field objects. While the primary objective of this work is not to evaluate phase reconstruction approaches, the reconstruction of simulated holograms provides validation of the generalized simulation method. The long-term goal of this work is that the generalized holographic simulation motivates the use of phase reconstruction of the simulated holograms to populate databases for training machine-learning algorithms aimed at classifying relevant objects recorded through a variety of holographic setups.

4.
Methods Mol Biol ; 2644: 247-266, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37142927

RESUMO

Digital holographic microscopy is an imaging technique particularly well suited to the study of living cells in culture, as no labeling is required and computed phase maps produce high contrast, quantitative pixel information. A full experiment involves instrument calibration, cell culture quality checks, selection and setup of imaging chambers, a sampling plan, image acquisition, phase and amplitude map reconstruction, and parameter map post-processing to extract information about cell morphology and/or motility. Each step is described below, focusing on results from imaging four human cell lines. Several post-processing approaches are detailed, with an aim of tracking individual cells and dynamics of cell populations.


Assuntos
Holografia , Microscopia , Humanos , Microscopia/métodos , Linhagem Celular , Interpretação de Imagem Assistida por Computador/métodos , Técnicas de Cultura de Células
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