0000000001301941
AUTHOR
José ÁNgel Picazo-bueno
Effect of counting chamber depth on the accuracy of lensless microscopy for the assessment of boar sperm motility.
Sperm motility is one of the most significant parameters in the prediction of male fertility. Until now, both motility analysis using an optical microscope and computer-aided sperm analysis (CASA-Mot) entailed the use of counting chambers with a depth to 20 µm. Chamber depth significantly affects the intrinsic sperm movement, leading to an artificial motility pattern. For the first time, laser microscopy offers the possibility of avoiding this interference with sperm movement. The aims of the present study were to determine the different motility patterns observed in chambers with depths of 10, 20 and 100 µm using a new holographic approach and to compare the results obtained in the 20-µm c…
Single-Element Reflective Digital Holographic Microscopy
Digital holographic microscopy (DHM) is a well-known microscopy technique using an interferometric architecture for quantitative phase imaging (QPI) and it has been already implemented utilizing a large number of interferometers. Among them, single-element interferometers are of particular interest due to its simplicity, stability, and low cost. Here, we present an extremely simple common-path interferometric layout based on the use of a single one-dimensional diffraction grating for both illuminating the sample in reflection and generating the digital holograms. The technique, named single-element reflective digital holographic microscopy (SER-DHM), enables QPI and topography analysis of r…
Spatially multiplexed interferometric microscopy with partially coherent illumination
We have recently reported on a simple, low cost, and highly stable way to convert a standard microscope into a holographic one [Opt. Express 22, 14929 (2014)]. The method, named spatially multiplexed interferometric microscopy (SMIM), proposes an off-axis holographic architecture implemented onto a regular (nonholographic) microscope with minimum modifications: the use of coherent illumination and a properly placed and selected one-dimensional diffraction grating. In this contribution, we report on the implementation of partially (temporally reduced) coherent illumination in SMIM as a way to improve quantitative phase imaging. The use of low coherence sources forces the application of phase…
Hilbert-Huang single-shot spatially multiplexed interferometric microscopy.
Hilbert-Huang single-shot spatially multiplexed interferometric microscopy (H2S2MIM) is presented as the implementation of a robust, fast, and accurate single-shot phase estimation algorithm with an extremely simple, low-cost, and highly stable way to convert a bright field microscope into a holographic one using partially coherent illumination. Altogether, H2S2MIM adds high-speed (video frame rate) quantitative phase imaging capability to a commercially available nonholographic microscope with improved phase reconstruction (coherence noise reduction). The technique has been validated using a 20×/0.46 NA objective in a regular Olympus BX-60 upright microscope for static, as well as dynamic…
Compact, cost-effective and field-portable microscope prototype based on MISHELF microscopy
AbstractWe report on a reduced cost, portable and compact prototype design of lensless holographic microscope with an illumination/detection scheme based on wavelength multiplexing, working with single hologram acquisition and using a fast convergence algorithm for image processing. All together, MISHELF (initials coming from Multi-Illumination Single-Holographic-Exposure Lensless Fresnel) microscopy allows the recording of three Fresnel domain diffraction patterns in a single camera snap-shot incoming from illuminating the sample with three coherent lights at once. Previous implementations have proposed an illumination/detection procedure based on a tuned (illumination wavelengths centered…
A Novel Marking Reader for Progressive Addition Lenses Based on Gabor Holography
PURPOSE Progressive addition lenses (PALs) are marked with permanent engraved marks (PEMs) at standardized locations. Permanent engraved marks are very useful through the manufacturing and mounting processes, act as locator marks to re-ink the removable marks, and contain useful information about the PAL. However, PEMs are often faint and weak, obscured by scratches, partially occluded, and difficult to recognize on tinted lenses or with antireflection or scratch-resistant coatings. The aim of this article is to present a new generation of portable marking reader based on an extremely simplified concept for visualization and identification of PEMs in PALs. METHODS Permanent engraved marks o…
Simultaneous dual mode imaging platform in lensless holographic microscopy
We present a novel layout capable of dual mode imaging in real time with different magnifications and resolution capabilities in lensless microscopy based on wavelength multiplexing. Experiments using static and dynamic samples are included.
Four channels multi-illumination single-holographic-exposure lensless Fresnel (MISHELF) microscopy
Abstract MISHELF microscopy [Opt. Express 23, 21352 (2015)] has been recently reported as the background technology of a new concept of compact, cost-effective and field-portable lensless microscope [Sci. Rep. 7, 43291 (2017)] based on wavelength multiplexing and a fast and robust algorithm for twin image minimization and noise reduction. In this manuscript, MISHELF microscopy is expanded beyond its actual configuration by considering 4 illumination/detection channels while retaining its working principle concerning single-shot, twin image mitigation and noise averaging. Proof of principle validation of the proposed improvement is conducted through experiments with a resolution test target …
Optical module for single-shot quantitative phase imaging based on the transport of intensity equation with field of view multiplexing
We present a cost-effective, simple, and robust method that enables single-shot quantitative phase imaging (QPI) based on the transport of intensity equation (TIE) using an add-on optical module that can be assembled into the exit port of any regular microscope. The module integrates a beamsplitter (BS) cube (placed in a non-conventional way) for duplicating the output image onto the digital sensor (field of view – FOV – multiplexing), a Stokes lens (SL) for astigmatism compensation (introduced by the BS cube), and an optical quality glass plate over one of the FOV halves for defocusing generation (needed for single-shot TIE algorithm). Altogether, the system provides two laterally separate…
SMIM in reflection imaging mode
We present reflective SMIM (initials incoming from Spatially-Multiplexed Interferometric Microscopy) as an extremely simple and low cost way to convert a standard white-light microscope into a holographic one working under reflection imaging mode.
Single-shot, dual-mode, water-immersion microscopy platform for biological applications
A single-shot water-immersion digital holographic microscope combined with broadband (white light) illumination mode is presented. This double imaging platform allows conventional incoherent visualization with phase holographic imaging of inspected samples. The holographic architecture is implemented at the image space (that is, after passing the microscope lens), thus reducing the sensitivity of the system to vibrations and/or thermal changes in comparison to regular interferometers. Because of the off-axis holographic recording principle, quantitative phase images of live biosamples can be recorded in a single camera snapshot at full-field geometry without any moving parts. And, the use o…
Improved quantitative phase imaging in lensless microscopy by single-shot multi-wavelength illumination using a fast convergence algorithm.
We report on a novel algorithm for high-resolution quantitative phase imaging in a new concept of lensless holographic microscope based on single-shot multi-wavelength illumination. This new microscope layout, reported by Noom et al. along the past year and named by us as MISHELF (initials incoming from Multi-Illumination Single-Holographic-Exposure Lensless Fresnel) microscopy, rises from the simultaneous illumination and recording of multiple diffraction patterns in the Fresnel domain. In combination with a novel and fast iterative phase retrieval algorithm, MISHELF microscopy is capable of high-resolution (micron range) phase-retrieved (twin image elimination) biological imaging of dynam…
Multi-illumination single-holographic-exposure lensless Fresnel (MISHELF) microscopy using 4 channels
MISHELF microscopy is generalized by considering 4 illumination/detection channels while retaining single-shot working principle, twin image mitigation and noise averaging. Proof of principle validation is included considering a resolution test target.
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Comparative between the white light imaging and the retrieved phase distribution from DHM
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2D wrapped phase distribution retrieved from DHM
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3D unwrapped phase distribution retrieved from DHM
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Comparative between the white light imaging and the retrieved phase distribution from DHM
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3D unwrapped phase distribution retrieved from DHM
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2D wrapped phase distribution retrieved from DHM