0000000000124185
AUTHOR
Emilio Sánchez-ortiga
showing 42 related works from this author
Axial resonance of periodic patterns by using a Fresnel biprism.
2013
This paper proposes a method for the generation of high-contrast localized sinusoidal fringes with spatially noncoherent illumination and relatively high light throughput. The method, somehow similar to the classical Lau effect, is based on the use of a Fresnel biprism. It has some advantages over previous methods for the noncoherent production of interference fringes. One is the flexibility of the method, which allows the control of the fringe period by means of a simple axial shift of the biprism. Second is the rapid axial fall-off in visibility around the high-contrast fringe planes. And third is the possibility of creating fringes with increasing or with constant period as the light bea…
Enhancing spatial resolution in digital holographic microscopy by biprism structured illumination.
2014
A novel and efficient architecture of a structured-illumination digital holographic microscope (DHM) is presented. As the DHM operates at the diffraction limit, its spatial resolution on label-free imaging of transparent samples is improved by illuminating the sample with a structured illumination produced by a Fresnel's biprism. The theoretical analysis of the method forecasts a twofold improvement of the spatial resolution. The proposed method requires only two images to improve the spatial resolution, which eases the process of unmixing the high-resolution components by means of an unknown phase-shift procedure. Numerical modeling and experimental results validate the theoretical finding…
Mapping electron-beam-injected trapped charge with scattering scanning near-field optical microscopy.
2016
Scattering scanning near-field optical microscopy (s-SNOM) has been demonstrated as a valuable tool for mapping the optical and optoelectronic properties of materials with nanoscale resolution. Here we report experimental evidence that trapped electric charges injected by an electron beam at the surface of dielectric samples affect the sample-dipole interaction, which has direct impact on the s-SNOM image content. Nanoscale mapping of the surface trapped charge holds significant potential for the precise tailoring of the electrostatic properties of dielectric and semiconductive samples, such as hydroxyapatite, which has particular importance with respect to biomedical applications. The meth…
THREE-DIMENSIONAL INTEGRAL MICROSCOPY WITH ENHANCED RESOLUTION AND DEPTH OF FIELD
2016
In this contribution we explain two new techniques developed by our group, which permit to increase the two-dimensional spatial resolution of the computed depth images in integral microscopy.
Free-depths reconstruction with synthetic impulse response in integral imaging
2015
Integral Imaging provides spatial and angular information of three-dimensional (3D) objects, which can be used both for 3D display and for computational post-processing purposes. In order to recover the depth information from an integral image, several algorithms have been developed. In this paper, we propose a new free depth synthesis and reconstruction method based on the two-dimensional (2D) deconvolution between the integral image and a simplified version of the periodic impulse response function (IRF) of the system. The period of the IRF depends directly on the axial position within the object space. Then, we can retrieve the depth information by performing the deconvolution with compu…
Time-multiplexing Integral Microscopy
2014
Conventional microscopes do not capture the 3D information of thick specimens. In order to avoid this limitation Integral Microscopy was proposed. An integral microscope is implemented by inserting a microlens array between the microscope objective and the camera sensor. Although this device captures multiperspective information of the specimen, the small number of microlenses limits the lateral resolution. In this paper we propose to improve the resolution by time multiplexing. Specifically, we propose an electrically addressable device that permits to obtain three sheared versions of the microscopic plenoptic map. Digital processing algorithm applied to the maps provides images with resol…
Fourier-domain lightfield microscopy: a new paradigm in 3D microscopy
2020
Recently, integral (also known as lightfield or plenoptic) imaging concept has been applied successfully to microscopy. The main advantage of lightfield microscopy when compared with conventional 3D imaging techniques is that it offers the possibility of capturing the 3D information of the sample after a single shot. However, integral microscopy is now facing many challenges, like improving the resolution and depth of field of the reconstructed specimens or the development and optimization of specially-adapted reconstruction algorithms. This contribution is devoted to review a new paradigm in lightfield microscopy, namely, the Fourier-domain integral microscope (FiMic), that improves the ca…
Improvement of two-dimensional structured illumination microscopy with an incoherent illumination pattern of tunable frequency.
2018
In two-dimensional structured illumination microscopy (2D-SIM), high-resolution images with optimal optical sectioning (OS) cannot be obtained simultaneously. This tradeoff can be overcome by using a tunable-frequency 2D-SIM system and a proper reconstruction method. The goal of this work is twofold. First, we present a computational approach to reconstruct optical-sectioned images with super-resolution enhancement (OS-SR) by using a tunable SIM system. Second, we propose an incoherent tunable-frequency 2D-SIM system based on a Fresnel biprism implementation. Integration of the proposed computational method with this tunable structured illumination (SI) system results in a new 2D-SIM system…
Subtractive imaging in confocal scanning microscopy using a CCD camera as a detector
2012
[EN] We report a scheme for the detector system of confocal microscopes in which the pinhole and a large-area detector are substituted by a CCD camera. The numerical integration of the intensities acquired by the active pixels emulates the signal passing through the pinhole. We demonstrate the imaging capability and the optical sectioning of the system. Subtractive-imaging confocal microscopy can be implemented in a simple manner, providing superresolution and improving optical sectioning. (C) 2012 Optical Society of America
Stable and simple quantitative phase-contrast imaging by Fresnel biprism
2018
Digital holographic (DH) microscopy has grown into a powerful nondestructive technique for the real-time study of living cells including dynamic membrane changes and cell fluctuations in nanometer and sub-nanometer scales. The conventional DH microscopy configurations require a separately generated coherent reference wave that results in a low phase stability and a necessity to precisely adjust the intensity ratio between two overlapping beams. In this work, we present a compact, simple, and very stable common-path DH microscope, employing a self-referencing configuration. The microscope is implemented by a diode laser as the source and a Fresnel biprism for splitting and recombining the be…
Resolution enhancement in integral microscopy by physical interpolation
2015
Integral-imaging technology has demonstrated its capability for computing depth images from the microimages recorded after a single shot. This capability has been shown in macroscopic imaging and also in microscopy. Despite the possibility of refocusing different planes from one snap-shot is crucial for the study of some biological processes, the main drawback in integral imaging is the substantial reduction of the spatial resolution. In this contribution we report a technique, which permits to increase the two-dimensional spatial resolution of the computed depth images in integral microscopy by a factor of √2. This is made by a double-shot approach, carried out by means of a rotating glass…
Non-Homogeneity of Lateral Resolution in Integral Imaging
2013
We evaluate the lateral resolution in reconstructed integral images. Our analysis takes into account both the diffraction effects in the image capture stage and the lack of homogeneity and isotropy in the reconstruction stage. We have used Monte Carlo simulation in order to assign a value for the resolution limit to any reconstruction plane. We have modelled the resolution behavior. Although in general the resolution limit increases proportionally to the distance to the lens array, there are some periodically distributed singularity planes. The phenomenon is supported by experiments.
Digital holographic microscopy with pure-optical spherical phase compensation
2011
Telecentric architecture is proposed for circumventing, by the pure-optical method, the residual parabolic phase distortion inherent to standard configuration of digital holographic microscopy. This optical circumvention produces several important advantages. One is that there is no need for computer compensation of the parabolic phase during the phase map recovering procedure. The other is that in off-axis configuration, the spatial frequency useful domain is enlarged. The validity of the method is demonstrated by performing quantitative measurement of depth differences with high axial resolution. (C) 2011 Optical Society of America
Multispectral integral imaging acquisition and processing using a monochrome camera and a liquid crystal tunable filter
2012
This paper presents an acquisition system and a procedure to capture 3D scenes in different spectral bands. The acquisition system is formed by a monochrome camera, and a Liquid Crystal Tunable Filter (LCTF) that allows to acquire images at different spectral bands in the [480, 680]nm wavelength interval. The Synthetic Aperture Integral Imaging acquisition technique is used to obtain the elemental images for each wavelength. These elemental images are used to computationally obtain the reconstruction planes of the 3D scene at different depth planes. The 3D profile of the acquired scene is also obtained using a minimization of the variance of the contribution of the elemental images at each …
Investigating the performance of reconstruction methods used in structured illumination microscopy as a function of the illumination pattern's modula…
2016
Surpassing the resolution of optical microscopy defined by the Abbe diffraction limit, while simultaneously achieving optical sectioning, is a challenging problem particularly for live cell imaging of thick samples. Among a few developing techniques, structured illumination microscopy (SIM) addresses this challenge by imposing higher frequency information into the observable frequency band confined by the optical transfer function (OTF) of a conventional microscope either doubling the spatial resolution or filling the missing cone based on the spatial frequency of the pattern when the patterned illumination is two-dimensional. Standard reconstruction methods for SIM decompose the low and hi…
Integral imaging with Fourier-plane recording
2017
Integral Imaging is well known for its capability of recording both the spatial and the angular information of threedimensional (3D) scenes. Based on such an idea, the plenoptic concept has been developed in the past two decades, and therefore a new camera has been designed with the capacity of capturing the spatial-angular information with a single sensor and after a single shot. However, the classical plenoptic design presents two drawbacks, one is the oblique recording made by external microlenses. Other is loss of information due to diffraction effects. In this contribution report a change in the paradigm and propose the combination of telecentric architecture and Fourier-plane recordin…
Aberration compensation for objective phase curvature in phase holographic microscopy: comment
2014
In a recent Letter by Seo et al. [Opt. Lett. 37, 4976 (2012)], the numerical correction of the quadratic phase distortion introduced by the microscope objective in digital holographic microscopy (DHM) has been presented. In this comment, we would like to draw to the attention of the authors and the readers in general that this approach could not be the optimal solution for maintaining the accuracy of the quantitative phase via DHM. We recall that the use of telecentric imaging systems in DHM simplifies the numerical processing of the phase images and produces more accurate measurements.
Fast and robust phase-shift estimation in two-dimensional structured illumination microscopy.
2019
A method of determining unknown phase-shifts between elementary images in two-dimensional Structured Illumination Microscopy (2D-SIM) is presented. The proposed method is based on the comparison of the peak intensity of spectral components. These components correspond to the inherent structured illumination spectral content and the residual compo- nent that appears from wrongly estimated phase-shifts. The estimation of the phase-shifts is carried out by finding the absolute maximum of a function defined as the normalized peak intensity difference in the Fourier domain. This task is performed by an optimization method providing a fast estimation of the phase-shift. The algorithm stability an…
Resolution limit in opto-digital systems revisited
2023
The resolution limit achievable with an optical system is a fundamental piece of information when characterizing its performance, mainly in case of microscopy imaging. Usually this information is given in the form of a distance, often expressed in microns, or in the form of a cutoff spatial frequency, often expressed in line pairs per mm. In modern imaging systems, where the final image is collected by pixelated digital cameras, the resolution limit is determined by the performance of both, the optical systems and the digital sensor. Usually, one of these factors is considered to be prevalent over the other for estimating the spatial resolution, leading to the global performance of the imag…
Lightfield microscopy, an emerging tool for real-time 3D imaging
2020
Integral, or lightfield, microscopy offers the possibility of capturing and processing in real time multiple views of 3D fluorescent samples captured with a single shot. In this contribution we review the recent advances in lightfield microscopy and enunciate the forthcoming challenges.
Machine Learning-Based View Synthesis in Fourier Lightfield Microscopy
2022
Current interest in Fourier lightfield microscopy is increasing, due to its ability to acquire 3D images of thick dynamic samples. This technique is based on simultaneously capturing, in a single shot, and with a monocular setup, a number of orthographic perspective views of 3D microscopic samples. An essential feature of Fourier lightfield microscopy is that the number of acquired views is low, due to the trade-off relationship existing between the number of views and their corresponding lateral resolution. Therefore, it is important to have a tool for the generation of a high number of synthesized view images, without compromising their lateral resolution. In this context we investigate h…
Recent Advances in Digital Holographic Microscopy
2018
In digital holographic microscopy (DHM) a hologram is captured in the image space provided by a microscope. The transfer of the phase and amplitude structure in the original sample to the hologram is in fact strongly affected by the use of the imaging microscope. A big research effort has been devoted to correct these distortions both by numerical and optical compensation. In this contribution, we present several proposals to improve the performance of classical DHM architectures by an a priori approach to compensate physically these perturbations. Experimental results are also presented to validate the proposed techniques.
New Analytical Tools for Evaluation of Spherical Aberration in Optical Microscopy
2010
The required tightly focused spots in three-dimensional (3D) scanning optical techniques are usually achieved by high-NA immersion lenses. The refractive index mismatch between the sample and the immersion medium introduces an important amount of spherical aberration when imaging deep inside the specimen, spreading out the focusing response. Since this aberration depends on the focalization depth, it is not possible to simultaneously achieve a global compensation for the whole scanned sample. In this way, the design of pupil elements that increase the tolerance to this aberration is of great interest. We present a new formalism for the evaluation and the design of filters that decrease the …
View images with unprecedented resolution in integral microscopy
2018
Integral microscopy is a novel technique that allows the simultaneous capture of multiple perspective images of microscopic samples. This feature is achieved at the cost of a significant reduction of the spatial resolution. In fact, it is assumed that in the best cases the resolution is reduced by a factor that is not smaller than ten, what poses a hard drawback to the utility of the technique. However, to the best of our knowledge, this resolution limitation has never been researched rigorously. For this reason, the aim of this paper is to explore the real limitations in resolution of integral microscopy and to obtain optically, without the need of any image-processing algorithm, perspecti…
Off-axis digital holographic microscopy: practical design parameters for operating at diffraction limit.
2014
The utilization of microscope objectives (MOs) in digital holographic microscopy (DHM) has associated effects that are not present in conventional optical microscopy. The remaining phase curvature, which can ruin the quantitative phase imaging, is the most evident and analyzed. As phase imaging is considered, this interest has made possible the development of different methods of overcoming its undesired consequences. Additionally to the effects in phase imaging, there exist a set of less obvious conditions that have to be accounted for as MOs are utilized in DHM to achieve diffraction-limit operation. These conditions have to be considered even in the case in which only amplitude or intens…
Optical-sectioning microscopy by patterned illumination
2010
We propose a very simple method for the flexible production of 1D structured illumination for high resolution 3D microscopy. Specifically, we propose the insertion of a Fresnel biprism after a monochromatic point source for producing a pair of twin, fully coherent, virtual point sources. The resulting interference fringes are projected into the 3D sample and, by simply varying the distance between the biprism and the point source, one can tune the period of the fringes, while keeping their contrast, in a very versatile and efficient way.
Three-dimensional real-time darkfield imaging through Fourier lightfield microscopy
2020
We report a protocol that takes advantage of the Fourier lightfield microscopy concept for providing 3D darkfield images of volumetric samples in a single-shot. This microscope takes advantage of the Fourier lightfield configuration, in which a lens array is placed at the Fourier plane of the microscope objective, providing a direct multiplexing of the spatio-angular information of the sample. Using the proper illumination beam, the system collects the light scattered by the sample while the background light is blocked out. This produces a set of orthographic perspective images with shifted spatial-frequency components that can be recombined to produce a 3D darkfield image. Applying the ade…
Depth rendering of large incoherent scenes from integral images
2016
Integral imaging is a technique that provides the spatial and angular information of three-dimensional (3D) scenes through a single shot. Taking advantage of this capability, different applications have been developed. Some of these applications are the 3D display and digital post-processing, in particular depth-reconstruction from integral images.
Reduction of spherical-aberration impact in microscopy by wavefront coding
2009
In modern high-NA optical scanning instruments, like scanning microscopes, the refractive-index mismatch between the sample and the immersion medium introduces a significant amount of spherical aberration when imaging deep inside the specimen, spreading out the impulse response. Since such aberration depends on the focalization depth, it is not possible to achieve a static global compensation for the whole 3D sample in scanning microscopy. Therefore a depth-variant impulse response is generated. Consequently, the design of pupil elements that increase the tolerance to this aberration is of great interest. In this paper we report a hybrid technique that provides a focal spot that remains alm…
Free segmentation in rendered 3D images through synthetic impulse response in integral imaging
2016
Integral Imaging is a technique that has the capability of providing not only the spatial, but also the angular information of three-dimensional (3D) scenes. Some important applications are the 3D display and digital post-processing as for example, depth-reconstruction from integral images. In this contribution we propose a new reconstruction method that takes into account the integral image and a simplified version of the impulse response function (IRF) of the integral imaging (InI) system to perform a two-dimensional (2D) deconvolution. The IRF of an InI system has a periodic structure that depends directly on the axial position of the object. Considering different periods of the IRFs we …
Fast Axial Scanning in 3D Imaging
2016
We establish the conditions for obtaining fast axial 3D microscopy with constant magnification, invariant resolution, and symmetrical displacement range. We demonstrate that liquid lenses permit the axial scanning free of mechanical movements and vibrations. This allows speeding-up the process and avoiding distortionsin the image of dynamic specimens. Article not available.
Shift-variant digital holographic microscopy: inaccuracies in quantitative phase imaging
2013
Inaccuracies introduced in quantitative phase digital holographic microscopy by the use of nontelecentric imaging systems are analyzed. Computer modeling of the experimental result shows that even negligible errors in the radius and center of curvature of the numerical compensation needed to get rid of the remaining quadratic phase factor introduce errors in the phase measurements; these errors depend on the position of the object in the field-of-view. However, when a telecentric imaging system is utilized for the recording of the holograms, the numerical modeling and experimental results show the shift-invariant behavior of the quantitative-phase digital holographic microscope.
Scanning microscopy with spatial sampling of the detector plane
2013
We present the implementation of a confocal scanning microscope in which the signal detection is performed through a matrix sensor, specifically, a CCD camera. This kind of detection has several advantages over the conventional detection in confocal microscopes. One of those advantages is the possibility to recover information of the sample that vanishes when the confocal image is directly acquired by the integration of light into a signal. We demonstrate the applicability of the system which allows implementing super-resolution techniques in a very easy manner.
Optical sectioning with a Wiener-like filter in Fourier integral imaging microscopy
2018
Non-scanning, single-shot, 3D integral microscopy with optical sectioning is presented. The method is based on the combination of Fourier-mode integral microscopy with a 3D deconvolution technique. Specifically, the refocused volume provided by a regular back-projection algorithm is 3D deconvolved with a synthetic 3D impulse response function that takes into account the number and positions of the elemental images. The use of this hybrid technique provides a stack of true-color depth-refocused images with significant gain of optical sectioning. The stack can be used, among other applications, to inspect inside the thick microscope specimen, to calculate collections of perspective views wit…
Confocal scanning microscope using a CCD camera as a pinhole-detector system
2011
We report a scheme for a detector system of confocal microscopes. In our scheme the pinhole and the large area detector are subtituted by a CCD camera. The numerical integration of the intensities acquired by the active pixels emulates the signal acquired by the detector. To demonstrate the utility of the system we efficiently performed an experiment of substractive-image confocal microscopy
Optically-undistorted digital holographic microscopy for quantitative phase-contrast imaging
2011
We propose a telecentric architecture for circumventing, by a pure-optical method, the residual phase distortion inherent to standard configuration of digital holographic microscopy (DHM). With this proposal there is no need for computer compensation of the parabolic phase during the phase map recovering procedure. Futhermore, in off-axis configuration, the spatial frequency useful domain is enlarged. The validity of the method is demonstrated by performing quantitative measurements of depth differences
Fast Axial-Scanning Widefield Microscopy With Constant Magnification and Resolution
2015
In this paper, we propose the use of electrically-addressable lens devices for performing fast non-mechanical axial scanning when imaging three-dimensional samples. This non-mechanical method can be implemented in any commercial microscope. The approach is based on the insertion of the tunable lens at the aperture stop of the microscope objective. By tuning the voltage, a stack of depth images of 3D specimens can be captured in real time and with constant magnification and resolution. The main advantage of our technique is the possibility of performing fast axial scanning free of mechanical vibrations.
FIMic: design for ultimate 3D-integral microscopy of in-vivo biological samples
2018
In this work, Fourier integral microscope (FIMic), an ultimate design of 3D-integral microscopy, is presented. By placing a multiplexing microlens array at the aperture stop of the microscope objective of the host microscope, FIMic shows extended depth of field and enhanced lateral resolution in comparison with regular integral microscopy. As FIMic directly produces a set of orthographic views of the 3D-micrometer-sized sample, it is suitable for real-time imaging. Following regular integral-imaging reconstruction algorithms, a 2.75-fold enhanced depth of field and [Formula: see text]-time better spatial resolution in comparison with conventional integral microscopy is reported. Our claims …
3D Integral Microscopy based in far-field detection
2018
Lately, Integral-Imaging systems have shown very promising capabilities of capturing the 3D structure of micro- scopic and macroscopic scenes. The aim of this work is to provide an optimal design for 3D-integral microscopy with extended depth of field and enhanced lateral resolution. By placing an array of microlenses at the aperture stop of the objective, this setup provides a set of orthographic views of the 3D sample. Adopting well known integral imaging reconstruction algorithms it can be shown that the depth of field as well as spatial resolution are improved with respect to conventional integral microscopy imaging. Our claims are supported on theoretical basis and experimental images …
Accurate single-shot quantitative phase imaging of biological specimens with telecentric digital holographic microscopy
2014
The advantages of using a telecentric imaging system in digital holographic microscopy (DHM) to study biological specimens are highlighted. To this end, the performances of nontelecentric DHM and telecentric DHM are evaluated from the quantitative phase imaging (QPI) point of view. The evaluated stability of the microscope allows single-shot QPI in DHM by using telecentric imaging systems. Quantitative phase maps of a section of the head of the drosophila melanogaster fly and of red blood cells are obtained via single-shot DHM with no numerical postprocessing. With these maps we show that the use of telecentric DHM provides larger field of view for a given magnification and permits more acc…
Fast and robust wave optics-based reconstruction protocol for Fourier lightfield microscopy
2023
Fourier lightfield microscopy (FLMic) is a powerful technique to record 3D images of thick dynamic samples. Belonging FLMic to the general class of computational imaging techniques, its efficiency is determined by several factors, like the optical system, the calibration process, the reconstruction algorithm, or the computation architecture. In the case of FLMic the calibration and the reconstruction algorithm should be fully adapted to the singular features of the technique. To this end, and concerning the reconstruction, we discard the use of experimental PSFs, and propose the use of a synthetic one, which is calculated on the basis of paraxial optics and taking into account the equal inf…
Study of spatial lateral resolution in off-axis digital holographic microscopy
2015
The lateral resolution in digital holographic microscopy (DHM) has been widely studied in terms of both recording and reconstruction parameters. Although it is understood that once the digital hologram is recorded the physical resolution is fixed according to the diffraction theory and the pixel density, still some researches link the resolution of the reconstructed wavefield with the recording distance as well as with the zero-padding technique. Aiming to help avoiding these misconceptions, in this paper we analyze the lateral resolution of DHM through the variation of those two parameters. To support our outcomes, we have designed numerical simulations and experimental verifications. Both…