Resolution gain in space-time digital holography by self-assembling of the object frequencies.

Space-time digital holography (STDH) exploits the object motion to record the hologram in a hybrid ST domain. This representation adds new capabilities to conventional DH, e.g., unlimited field of view and variable phase shifting. This is the best candidate for imaging biological samples flowing in microfluidic channels. Here, we show that STDH is able to improve the spatial resolution as well. Different from other super-resolution approaches, stitching between holograms or their spectra is no longer required. Moreover, we introduce a new oblique STDH modality to record and process hybrid ST representations. This allows improving resolution in 2D with one single object scan, paving the way to the use of STDH for super-resolution imaging onboard microfluidic devices.

Digital holography as 3D tracking tool for assessing acoustophoretic particle manipulation.

The integration of digital holography (DH) imaging and the acoustic manipulation of micro-particles in a microfluidic environment is investigated. The ability of DH to provide efficient 3D tracking of particles inside a microfluidic channel is exploited to measure the position of multiple objects moving under the effect of stationary ultrasound pressure fields. The axial displacement provides a direct verification of the numerically computed positions of the standing wave's node, while the particles' transversal movement highlights the presence of nodes in the planar direction. Moreover, DH is used to follow the aggregation dynamics of trapped spheres in such nodes by using aggregation rate metrics.

The lived experience of adults with heart failure: a phenomenological study.

BACKGROUND: Although a number of studies have been conducted on patients with Heart Failure (HF), they have not given a rigorous comprehensive description of what it is like to live with HF. The objective of this study was to describe the lived experience of adults with HF. STUDY DESIGN: A hermeneutic phenomenological design was used. METHODS: Cohen's method was used to conduct the study. Thirty HF patients were enrolled between February and July 2014 from an outpatient cardiovascular clinic in Tuscany, Italy. Phenomenological interviews took place at patients' homes, and the investigators analyzed verbatim transcripts. Once data saturation was achieved, to ensure data trustworthiness, participants were asked to confirm all the extracted themes. Atals.ti vers.7 was used for data analysis. RESULTS: The patients were mostly male (67%) with a mean age of 71 (SD 9.15) and an age range of 48-86. Seven themes emerged from the phenomenological analysis: 1) important life changes; 2) social isolation caused by the illness; 3) anger and resignation associated with the disease; 4) relief from spirituality; 5) will to live; 6) uncertainty about the future and 7) the inescapability of disease and death. CONCLUSIONS: The meaning that patients attribute to their lived experience helps to create their needs, which are important to direct care. Family support and religious beliefs are an important source for HF patients to better manage their fears and cope with the future. Findings of this study provide nurses with a comprehensive description of what it is like to live with HF, which can be useful in helping to meet patients' needs more effectively and in tailoring interventions.

Forward electrohydrodynamic inkjet printing of optical microlenses on microfluidic devices.

We report a novel method for direct printing of viscous polymers based on a pyro-electrohydrodynamic repulsion system capable of overcoming limitations on the material type, geometry and thickness of the receiving substrate. In fact, the results demonstrate that high viscosity polymers can be easily manipulated for optical functionalizing of lab-on-a-chip devices through demonstration of direct printing of polymer microlenses onto microfluidic chips and optical fibre terminations. The present system has great potential for applications from biomolecules to nano-electronics. Moreover, in order to prove the effectiveness of the system, the optical performance of such microlenses has been characterized by testing their imaging capabilities when the fibroblast cells were allowed to flow inside the microfluidic channel, showing one of their possible applications on-board a LoC platform.

Self-propelling bacteria mimic coherent light decorrelation.

We show here that live e-coli bacterial culture, thanks to the self-propelling feature, can significantly reduce the coherent noise. In fact, the typical self-propelled drive of such microorganisms provides enough time diversity in speckle patterns. Optical properties of a bacteria suspension have been investigated and analyzed thus showing that it behaves as a quite good optical speckle decorrelation device. Samples with different bacteria densities have been studied. The decorrelation effect has been demonstrated by probing the imaging performance in through transmission in coherent microscope configuration.

Optofluidic holographic microscopy with custom field of view (FoV) using a linear array detector.

Simple and effective imaging strategies are of utmost interest for applications on a lab-on-chip scale. In fact, the majority of diagnostic tools for medical as well as biotechnological studies still employ image-based approaches. Having onboard the chip a compact but powerful imaging apparatus with multiple imaging capabilities, such as 3D dynamic focusing along the optical axis, unlimited field of view (FoV) and double outputs, namely, intensity and quantitative phase-contrast maps of biological objects, is of extreme importance for the next generation of Lab-on-a-Chip (LoC) devices. Here we present a coherent 3D microscopy approach with a holographic modality that is specifically suitable for studying biological samples while they simply flow along microfluidic paths. The LoC device is equipped with a compact linear array detector to capture and generate a new conceptual type of a digital hologram in the space-time domain, named here as Space-Time Digital Hologram (STDH). The reported results show that the method is a promising diagnostic tool for optofluidic investigations of biological specimens.

Surface plasmon resonance imaging by holographic enhanced mapping.

We designed, constructed and tested a holographic surface plasmon resonance (HoloSPR) objective-based microscope for simultaneous amplitude-contrast and phase-contrast surface plasmon resonance imaging (SPRi). SPRi is a widely spread tool for label-free detection of changes in refractive index and concentration, as well as mapping of thin films. Currently, most of the SPR sensors rely on the detection of amplitude or phase changes of light. Despite the high sensitivities achieved so far, each technique alone has a limited detection range with optimal sensitivity. Here we use a high numerical aperture objective that avoids all the limitations due to the use of a prism-based configuration, yielding highly magnified and distortion-free images. Holographic reconstructions of SPR images and real-time kinetic measurements are presented to show the capability of HoloSPR to provide a versatile imaging method for high-throughput SPR detection complementary to conventional SPR techniques.

Spatio-temporal scanning modality for synthesizing interferograms and digital holograms.

We investigate the spatio-temporal scanning of a single-pixel row for building up synthetic interferograms or digital holograms, shifted each other of a desired phase step. This unusual recording modality exploits the object movement to synthesize interferograms with extended Field of View and improved noise contrast. We report the theoretical formulation of the synthetizing recording process and experimental evidence of various cases demonstrating quantitative phase retrieval by adopting this intrinsic phase-shifting procedure. The proposed method could be particularly suited in all cases where the object shift is an intrinsic feature of the investigated system, as e.g. in microfluidics imaging.

Imaging through scattering microfluidic channels by digital holography for information recovery in lab on chip.

We tackle the problem of information recovery and imaging through scattering microfluidic chips by means of digital holography (DH). In many cases the chip can become opalescent due to residual deposits settling down the inner channel faces, biofilm formation, scattering particle uptake by the channel cladding or its damaging by corrosive substances, or even by condensing effect on the exterior channels walls. In these cases white-light imaging is severely degraded and no information is obtainable at all about the flowing samples. Here we investigate the problem of counting and estimating velocity of cells flowing inside a scattering chip. Moreover we propose and test a method based on the recording of multiple digital holograms to retrieve improved phase-contrast images despite the strong scattering effect. This method helps, thanks to DH, to recover information which, otherwise, would be completely lost.

Random resampling masks: a non-Bayesian one-shot strategy for noise reduction in digital holography.

Holographic imaging may become severely degraded by a mixture of speckle and incoherent additive noise. Bayesian approaches reduce the incoherent noise, but prior information is needed on the noise statistics. With no prior knowledge, one-shot reduction of noise is a highly desirable goal, as the recording process is simplified and made faster. Indeed, neither multiple acquisitions nor a complex setup are needed. So far, this result has been achieved at the cost of a deterministic resolution loss. Here we propose a fast non-Bayesian denoising method that avoids this trade-off by means of a numerical synthesis of a moving diffuser. In this way, only one single hologram is required as multiple uncorrelated reconstructions are provided by random complementary resampling masks. Experiments show a significant incoherent noise reduction, close to the theoretical improvement bound, resulting in image-contrast improvement. At the same time, we preserve the resolution of the unprocessed image.

Imaging live humans through smoke and flames using far-infrared digital holography.

The ability to see behind flames is a key challenge for the industrial field and particularly for the safety field. Development of new technologies to detect live people through smoke and flames in fire scenes is an extremely desirable goal since it can save human lives. The latest technologies, including equipment adopted by fire departments, use infrared bolometers for infrared digital cameras that allow users to see through smoke. However, such detectors are blinded by flame-emitted radiation. Here we show a completely different approach that makes use of lensless digital holography technology in the infrared range for successful imaging through smoke and flames. Notably, we demonstrate that digital holography with a cw laser allows the recording of dynamic human-size targets. In this work, easy detection of live, moving people is achieved through both smoke and flames, thus demonstrating the capability of digital holography at 10.6 µm.

Visible reconstruction by a circular holographic display from digital holograms recorded under infrared illumination.

A circular holographic display that consists of phase-only spatial light modulators is used to reconstruct images in visible light from digital holograms recorded under infrared (10.6 µm) illumination. The reconstruction yields a holographic digital video display of a three-dimensional ghostlike image of an object floating in space where observers can move and rotate around it.

Multi-wavelengths digital holography: reconstruction, synthesis and display of holograms using adaptive transformation.

A method based on spatial transformations of multiwavelength digital holograms and the correlation matching of their numerical reconstructions is proposed, with the aim to improve superimposition of different color reconstructed images. This method is based on an adaptive affine transform of the hologram that permits management of the physical parameters of numerical reconstruction. In addition, we present a procedure to synthesize a single digital hologram in which three different colors are multiplexed. The optical reconstruction of the synthetic hologram by a spatial light modulator at one wavelength allows us to display all color features of the object, avoiding loss of details.

An automatic method for assembling a large synthetic aperture digital hologram.

A major issue so far for digital holography is the low spatial resolution generally achieved. The numerical aperture is limited by the area of currently available detectors, such as CCD sensors, which is significantly lower than that of a holographic plate. This is an even more severe constraint when IR sensors such as microbolometers are taken into account. In order to increase the numerical aperture of such systems, we developed an automatic technique which is capable of recording several holograms and of stitching them together, obtaining a digital hologram with a synthetic but larger numerical aperture. In this way we show that more detail can be resolved and a wider parallax angle can be achieved. The method is demonstrated for visible as well IR digital holography, recording and displaying large size objects.

Identification of bovine sperm head for morphometry analysis in quantitative phase-contrast holographic microscopy.

An investigation is reported of the identification and measurement of region of interest (ROI) in quantitative phase-contrast maps of biological cells by digital holographic microscopy. In particular, two different methods have been developed for in vitro bull sperm head morphometry analysis. We show that semen analysis can be accomplished by means of the proposed techniques . Extraction and measurement of various parameters are performed. It is demonstrated that both proposed methods are efficient to skim the data set in a preselective analysis for discarding anomalous data.

Driving and analysis of micro-objects by digital holographic microscope in microfluidics.

We propose an optical configuration in which floating particles in a microfluidic chamber can be characterized by an interference microscopy configuration to obtain quantitative phase-contrast maps. The configuration is simply made by two laser beams from the same laser source. One beam provides the optical forces for driving the particle along appropriate paths, but at same time works as the object illumination beam in the holographic microscope. The second beam plays the role of the reference beam, allowing recording of an interference fringe pattern (i.e., the digital hologram) in an out-of-focus image plane. The system and method are illustrated and experimental results are offered for polymeric particles as well as for in vitro cells with the aim to demonstrate the approach.

Automatic focusing in digital holography and its application to stretched holograms.

The searching and recovering of the correct reconstruction distance in digital holography (DH) can be a cumbersome and subjective procedure. Here we report on an algorithm for automatically estimating the in-focus image and recovering the correct reconstruction distance for speckle holograms. We have tested the approach in determining the reconstruction distances of stretched digital holograms. Stretching a hologram with a variable elongation parameter makes it possible to change the in-focus distance of the reconstructed image. In this way, the proposed algorithm can be verified at different distances by dispensing the recording of different holograms. Experimental results are shown with the aim of demonstrating the usefulness of the proposed method, and a comparative analysis has been performed with respect to other existing algorithms developed for DH.

Digital self-referencing quantitative phase microscopy by wavefront folding in holographic image reconstruction.

A completely numerical method, named digital self-referencing holography, is described to easily accomplish a quantitative phase microscopy for microfluidic devices by a digital holographic microscope. The approach works through an appropriate numerical manipulation of the retrieved complex wavefront. The self-referencing is obtained by folding the retrieved wavefront in the image plane. The folding operation allows us to obtain the correct phase map by subtracting from the complex region of interest a flat area outside the microfluidic channel. To demonstrate the effectiveness of the method, quantitative phase maps of bovine spermatozoa and in vitro cells are retrieved.

Dispensing nano-pico droplets and liquid patterning by pyroelectrodynamic shooting.

Manipulating and dispensing liquids on the micrometre- and nanoscale is important in biotechnology and combinatorial chemistry, and also for patterning inorganic, organic and biological inks. Several methods for dispensing liquids exist, but many require complicated electrodes and high-voltage circuits. Here, we show a simple way to draw attolitre liquid droplets from one or multiple sessile drops or liquid film reservoirs using a pyroelectrohydrodynamic dispenser. Local pyroelectric forces, which are activated by scanning a hot tip or an infrared laser beam over a lithium niobate substrate, draw liquid droplets from the reservoir below the substrate, and deposit them on the underside of the lithium niobate substrate. The shooting direction is altered by moving the hot tip or laser to form various patterns at different angles and locations. Our system does not require electrodes, nozzles or circuits, and is expected to have many applications in biochemical assays and various transport and mixing processes.

Controlling depth of focus in 3D image reconstructions by flexible and adaptive deformation of digital holograms.

We show here that through an adaptive deformation of digital holograms it is possible to manage the depth of focus in 3D imaging reconstruction. Deformation is applied to the original hologram with the aim to put simultaneously in focus, and in one reconstructed image plane, different objects lying at different distances from the hologram plane (i.e., CCD sensor). In the same way, by adapting the deformation it is possible to extend the depth of field having a tilted object entirely in focus. We demonstrate the method in both lensless as well as in microscope configuration.