showing 30 related works from this author
Physical compensation of phase curvature in digital holographic microscopy by use of programmable liquid lens.
2015
Quantitative phase measurements obtained with digital holographic microscopes are strongly dependent on the optical arrangement of the imaging system. The nontelecentric operation provides phase measurements affected by a parabolic phase factor and requires numerical postprocessing, which does not always remove all the perturbation. Accurate phase measurements are achieved by using the imaging system in telecentric mode. Unfortunately, this condition is not accomplished when a commercial microscope is used as the imaging system. In this paper, we present an approach for obtaining accurate phase measurements in nontelecentric imaging systems without the need for numerical postprocessing. The…
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…
Experimental validation of a customized phase mask designed to enable efficient computational optical sectioning microscopy through wavefront encodin…
2017
In this paper, wavefront-encoded (WFE) computational optical sectioning microscopy (COSM) using a fabricated square cubic (SQUBIC) phase mask, designed to render the system less sensitive to depth-induced aberration, is investigated. The WFE-COSM system is characterized by a point spread function (PSF) that does not vary as rapidly with imaging depth compared to the conventional system. Thus, in WFE-COSM, image restoration from large volumes can be achieved using computationally efficient space-invariant (SI) algorithms, thereby avoiding the use of depth-variant algorithms. The fabricated SQUBIC phase mask was first evaluated and found to have a 75% fidelity compared to the theoretical desi…
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
Phase-shifting by means of an electronically tunable lens: quantitative phase imaging of biological specimens with digital holographic microscopy
2016
The use of an electronically tunable lens (ETL) to produce controlled phase shifts in interferometric arrangements is shown. The performance of the ETL as a phase-shifting device is experimentally validated in phase-shifting digital holographic microscopy. Quantitative phase maps of a section of the thorax of a Drosophila melanogaster fly and of human red blood cells have been obtained using our proposal. The experimental results validate the possibility of using the ETL as a reliable phase-shifter device.
Digital holographic microscopy for diabetes screening
2016
A digital holographic microscope operating in telecentric mode could be used to diagnose diabetes and evaluate long-term glycemic control in patients with diabetes.
Digital holographic microscopy as a screening technology for diabetes
2019
Label-free quantitative phase imaging (QPI) is the hallmark of digital holographic microscopy (DHM). One of the most interesting medical applications of QPI-DHM is that it can be used to analyze illnesses in which the refractive index or/and the morphology of cells/tissues are distorted, from the acquisition of a single image. In this contribution, we obtain the phase maps of red blood cells (RBCs) samples of patients suffering from diabetes mellitus type 1 (DM1) by using a DHM. Our experimental results show that the measured phase values are significantly different between control non-diabetic and diabetic patients. The high correlation coefficient between the phase and the glycated hemogl…
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
Optimal design of incoherent tunable-frequency structured illumination microscope scheme
2018
Structured illumination microscopy (SIM) improves resolution and optical sectioning capability compared to conventional widefield techniques. The main idea of this method is the illumination of the sample with a structured pattern of fixed spatial modulation frequency. Previously, a Fresnel biprism has been implemented in a structured illumination (SI) device providing tunable-frequency sinusoidal patterns. However, the use of this SI system introduces a tradeoff between the visibility and field of view of the illumination fringes. In this contribution, we analyze theoretically this tradeoff and propose the optimal design for the Fresnel biprism-based SIM system.
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.
Spatial light modulator phase mask implementation of wavefront encoded 3D computational-optical microscopy
2015
Spatial light modulator (SLM) implementation of wavefront encoding enables various types of engineered pointspread functions (PSFs), including the generalized-cubic and squared-cubic phase mask wavefront encoded (WFE) PSFs, shown to reduce the impact of sample-induced spherical aberration in fluorescence microscopy. This investigation validates dynamic experimental parameter variation of these WFE-PSFs. We find that particular design parameter bounds exist, within which the divergence of computed and experimental WFE-PSFs is of the same order of magnitude as that of computed and experimental conventional PSFs, such that model-based approaches for solving the inverse imaging problem can be a…
Tunable structured illumination system based on a Wollaston prism
2018
Experimental verification of a simple illumination system to generate a 1D structured pattern with tunable modulation frequency is shown based on a Wollaston prism illuminated by the diffracted field of an incoherent linear source.
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.
Shaded-Mask Filtering for Extended Depth-of-Field Microscopy
2013
This paper proposes a new spatial filtering approach for increasing the depth-of-field (DOF) of imaging systems, which is very useful for obtaining sharp images for a wide range of axial positions of the object. Many different techniques have been reported to increase the depth of field. However the main advantage in our method is its simplicity, since we propose the use of purely absorbing beam-shaping elements, which allows a high focal depth with a minimum modification of the optical architecture. In the filter design, we have used the analogy between the axial behavior of a system with spherical aberration and the transverse impulse response of a 1D defocused system. This allowed us the…
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.
Tunable-frequency three-dimensional structured illumination microscopy with reduced data-acquisition
2018
The performance of a tunable three-dimensional (3D) structured illumination microscope (SIM) system and its ability to provide simultaneously super-resolution (SR) and optical-sectioning (OS) capabilities are investigated. Numerical results show that the performance of our 3D-SIM system is comparable with the one provided by a three-wave interference SIM, while requiring 40% fewer images for the reconstruction and providing frequency tunability in a cost-effective implementation. The performance of the system has been validated experimentally with images from test samples, which were also imaged with a commercial SIM based on incoherent-grid projection for comparison. Restored images from d…
Optical transfer function engineering for a tunable 3D structured illumination microscope
2019
Two important features of three-dimensional structured illumination microscopy (3D-SIM) are its optical sectioning (OS) and super-resolution (SR) capabilities. Previous works on 3D-SIM systems show that these features are coupled. We demonstrate that a 3D-SIM system using a Fresnel biprism illuminated by multiple linear incoherent sources provides a structured illumination pattern whose lateral and axial modulation frequencies can be tuned separately. Therefore, the compact support of the synthetic optical transfer function (OTF) can be engineered to achieve the highest OS and SR capabilities for a particular imaging application. Theoretical performance of our engineered system based on the…
Diabetes screening by telecentric digital holographic microscopy
2015
Diabetes is currently the world's fastest growing chronic disease and it is caused by deficient production of insulin by the endocrine pancreas or by abnormal insulin action in peripheral tissues. This results in persistent hyperglycaemia that over time may produce chronic diabetic complications. Determination of glycated haemoglobin level is currently the gold standard method to evaluate and control sustained hyperglycaemia in diabetic people. This measurement is currently made by high-performance liquid chromatography, which is a complex chemical process that requires the extraction of blood from the antecubital vein. To reduce the complexity of that measurement, we propose a fully-optica…
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.
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.
Evaluation of the use of wavefront encoding to reduce depth-induced aberration in structured-illumination microscopy
2018
Three-dimensional imaging is affected by depth-induced spherical aberration (SA) when imaging deep into an optically thick sample. In this work, we evaluate the impact of SA on the performance of incoherent grating-projection structured illumination microscopy (SIM). In particular, we analyze the reduction of the contrast in the structured pattern and compare the reconstructed SIM images for different amounts of SA. In order to mitigate the impact of SA, we implement and evaluate in SIM a wavefront encoded imaging system using a square cubic (SQUBIC) phase mask, an approach shown previously to be successful in conventional microscopy.
Recent Advances in 3D Structured Illumination Microscopy
2018
In structured illumination microscopy (SIM) the sample under investigation is illuminated using a structured illumination (SI) pattern. This SI pattern encodes high spatial frequencies of fine features within the sample, which usually are not transferred by the conventional three-dimensional (3D) optical transfer function (OTF) of the imaging system and fills the missing cone of frequencies in the OTF for better discrimination of the out-of-focus light. Thereby, SIM provides super-resolution (SR) performance beyond the diffraction limit and optical-sectioning (OS) capability with the use of data post-processing approaches. 3D structured patterns that include lateral and axial variations in …
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…
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…