Showing papers by "Pietro Ferraro published in 2015"

Recent advances in holographic 3D particle tracking

Abstract: Particle tracking is a fundamental technique for investigating a variety of biophysical processes, from intracellular dynamics to the characterization of cell motility and migration. However, observing three-dimensional (3D) trajectories of particles is in general a challenging task in classical microscopy owing to the limited imaging depth of field of commercial optical microscopes, which represents a serious drawback for the analysis of time-lapse microscopy image data. Therefore, numerous automated particle-tracking approaches have been developed by many research groups around the world. Recently, digital holography (DH) in microscopy has rapidly gained credit as one of the elective techniques for these applications, mainly due to the uniqueness of the DH to provide a posteriori quantitative multiple refocusing capability and phase-contrast imaging. Starting from this paradigm, a huge amount of 3D holographic tracking approaches have been conceived and investigated for applications in various branches of science, including optofluids, microfluidics, biomedical microscopy, cell mechano-trasduction, and cell migration. Since a wider community of readers could be interested in such a review, i.e., not only scientists working in the fields of optics and photonics but also users of particle-tracking tools, it should be very beneficial to provide a complete review of state-of-the-art holographic 3D particle-tracking methods and their applications in bio-microfluidics.

Red blood cell as an adaptive optofluidic microlens.

Abstract: The perspective of using live cells as lenses could open new revolutionary and intriguing scenarios in the future of biophotonics and biomedical sciences for endoscopic vision, local laser treatments via optical fibres and diagnostics. Here we show that a suspended red blood cell (RBC) behaves as an adaptive liquid-lens at microscale, thus demonstrating its imaging capability and tunable focal length. In fact, thanks to the intrinsic elastic properties, the RBC can swell up from disk volume of 90 fl up to a sphere reaching 150 fl, varying focal length from negative to positive values. These live optofluidic lenses can be fully controlled by triggering the liquid buffer's chemistry. Real-time accurate measurement of tunable focus capability of RBCs is reported through dynamic wavefront characterization, showing agreement with numerical modelling. Moreover, in analogy to adaptive optics testing, blood diagnosis is demonstrated by screening abnormal cells through focal-spot analysis applied to an RBC ensemble as a microlens array.

Diagnostic Tools for Lab-on-Chip Applications Based on Coherent Imaging Microscopy

Abstract: Today, fast and accurate diagnosis through portable and cheap devices is in high demand for the general healthcare. Lab-on-chips (LoCs) have undergone a great growth in this direction, supported by optical imaging techniques more and more refined. Here we present recent progresses in developing imaging tools based on coherent imaging microscopy that can be very useful when applied into biomicrofluidics. In some cases, the optical tweezers (OT) technique is combined with digital holography (DH), thus offering the possibility to manipulate, analyze, and measure fundamental parameters of different kinds of cells. This approach can open the route for rapid and high-throughput analysis in label-free microfluidic devices and for prognostic based on cell examination, thus allowing advancements in biomedical science.

Reversible Holographic Patterns on Azopolymers for Guiding Cell Adhesion and Orientation.

Abstract: Topography of material surfaces is known to influence cell behavior at different levels: from adhesion up to differentiation. Different micro- and nanopatterning techniques have been employed to create patterned surfaces to investigate various aspects of cell behavior, most notably cellular mechanotransduction. Nevertheless, conventional techniques, once implemented on a specific substrate, fail in allowing dynamic changes of the topographic features. Here we investigated the response of NIH-3T3 cells to reversible topographic signals encoded on light-responsive azopolymer films. Switchable patterns were fabricated by means of a well-established holographic setup. Surface relief gratings were realized with Lloyd’s mirror system and erased with circularly polarized or incoherent light. Cell cytoskeleton organization and focal adhesion assembly proved to be very sensitive to the underlying topographic signal. Thereafter, pattern reversibility was tested in air and wet environment by using temperature or lig...

Effects of Lithium Niobate Polarization on Cell Adhesion and Morphology.

Abstract: Understanding how the interfacial effects influence cell adhesion and morphology is of fundamental interest for controlling function, growth, and movement of cells in vitro and in vivo. In particular, the influence of surface charges is well-known but still controversial, especially when new functional materials and methods are introduced. Here, the influence of the spontaneous polarization of ferroelectric lithium niobate (LN) on the adhesion properties of fibroblast cells is investigated. The spontaneous polarization of LN has one of the largest known magnitudes at room temperature (∼78 μC/cm2), and its orientation can be patterned easily by an external voltage, this motivating highly the investigation of its interaction with cells. Immunofluorescence and migration assays show strong evidence that the surface polarity regulates the adhesion functions, with enhanced spreading of the cytoskeleton on the negative face. The results suggest the potential of LN as a platform for investigating the role of char...

Holographic imaging of unlabelled sperm cells for semen analysis: a review.

Abstract: Quantitative phase maps of living swimming spermatozoa. Male reproductive health in both humans and animals is an important research field in biological study. In order to characterize the morphology, the motility and the concentration of the sperm cells, which are the most important parameters to feature them, digital holography demonstrated to be an attractive technique. Indeed, it is a label-free, non-invasive and high-resolution method that enables the characterization of live specimen. The review is intended both for summarizing the state-of-art on the semen analysis and recent achievement obtained by means of digital holography and for exploring new possible applications of digital holography in this field.

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

Abstract: 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.

Optical signature of erythrocytes by light scattering in microfluidic flows

Abstract: A camera-based light scattering approach coupled with a viscoelasticity-induced cell migration technique has been used to characterize the morphological properties of erythrocytes in microfluidic flows. We have obtained the light scattering profiles (LSPs) of individual living cells in microfluidic flows over a wide angular range and matched them with scattering simulations to characterize their morphological properties. The viscoelasticity-induced 3D cell alignment in microfluidic flows has been investigated by bright-field and holographic microscopy tracking, where the latter technique has been used to obtain precise cell alignment profiles in-flow. Such information allows variable cell probability control in microfluidic flows at very low viscoelastic polymer concentrations, obtaining cell measurements that are almost physiological. Our results confirm the possibility of precise, label-free analysis of individual living erythrocytes in microfluidic flows.

Self-propelling bacteria mimic coherent light decorrelation.

Abstract: 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.

Impact damage investigation on composite laminates: comparison among different NDT methods and numerical simulation

Abstract: The aim of this paper is to investigate the ability of different NDT techniques to detect and evaluate barely visible and non-visible impact damage on composite laminates. Firstly, a conventional ultrasound technique was adopted to investigate the delamination in carbon fibre laminates after low velocity impact s. Then the results were compared with a thermographic and holographic analysis, as well as a theoretical simulation of the expected delamination. The results were compared and discussed. Overall a good agreement was found between the data obtained by the different techniques. Furthermore, the true values of the damage parameters were confirmed by DT performed on the samples.

Surface Plasmon Resonance Imaging by Holographic Enhanced Mapping

Abstract: 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.

Improving holographic reconstruction by automatic Butterworth filtering for microelectromechanical systems characterization.

Abstract: Digital holographic microscopy is an important interferometric tool in optical metrology allowing the investigation of engineered surfaces with microscale lateral resolution and nanoscale axial precision. In particular, microelectromechanical systems (MEMS) surface analysis, conducted by holographic characterization, requires high accuracy for functional testing. The main issues related to MEMS inspection are the superficial roughness and the complex geometry resulting from the several fabrication steps. Here, an automatic procedure, particularly suited in the case of high-roughness surfaces, is presented to selectively filter the spectrum, providing very low-noise reconstructed images. The numerical procedure is based on Butterworth filtering, and the obtained results demonstrate a significant increase in the images’ quality and in the accuracy of the measurements, making our technique highly applicable for quantitative phase imaging in MEMS analysis. Furthermore, our method is fully tunable to the spectrum under investigation and automatic. This makes it highly suitable for real-time applications. Several experimental tests show the suitability of the proposed approach.

Investigation on cone jetting regimes of liquid droplets subjected to pyroelectric fields induced by laser blasts

Abstract: Electrical conductivity and viscosity play a major role in the tip jetting behaviour of liquids subjected to electrohydrodynamic (EHD) forces, thus influencing significantly the printing performance. Recently, we developed a nozzle- and electrode-free pyro-EHD system as a versatile alternative to conventional EHD configurations and we demonstrated different applications, including inkjet printing and three-dimensional lithography. However, only dielectric fluids have been used in all of those applications. Here, we present an experimental characterization of the pyro-EHD jetting regimes, induced by laser blasts, of sessile drops in case of dielectric and conductive liquids in order to extend the applicability of the system to a wider variety of fields including biochemistry and biotechnology where conductive aqueous solutions are typically used.

Nanocomposite polymer carbon-black coating for triggering pyro-electrohydrodynamic inkjet printing

Abstract: The pyro-electrohydrodynamic (EHD) manipulation of liquids has been discovered and demonstrated recently as a high resolution printing technique avoiding the use of nozzles and external electrodes. The activation of the pyro-electric effect is usually achieved on ferroelectric crystals by an external heating source or by an infrared laser. Here, we show an original modality for triggering the pyro-EHD process through a light-absorbing polymer nanocomposite thin layer deposited on the ferroelectric substrate, thus overcoming some limitations of the previous configuration. Significant simplification and compactness of the set-up is achieved thanks to the nanocomposite coating, since a commercial low-cost white-light halogen lamp can be adopted to trigger the pyro-jetting process from a liquid reservoir. Remarkably, high resolution is achieved in dispensing very high viscous liquids. Practical demonstrations in polymer optical microlenses direct printing using polydimethylsiloxane and poly(methyl methacrylate) are finally reported to validate the approach in handling high-viscous polymers for practical applications.

Electrohydrodynamic Assembly of Multiscale PDMS Microlens Arrays

Abstract: In this paper, we introduce an easy multiscale approach for the fabrication of polymer microlens arrays through a self-assembling process driven by the electrohydrodynamic (EHD) pressure. This method represents a simple alternative to the conventional soft lithography techniques. A thin layer of liquid polymer is deposited on a microengineered ferroelectric crystal and can be self-assembled and cross-linked in a single-step process as a consequence of the pyroelectric effect activated by simply heating the substrate. Although the EHD instability induced by the pyroelectric effect was discovered in principle few years ago, here we demonstrate a systematic investigation for fabrication of microlens arrays in a multiscale range (i.e., between 25 to 200 μm diameter) with high degree of uniformity. By controlling the polymer instability driven by EHD, we report on two different microoptical shapes can be obtained spontaneously, i.e., spherical or toroidal. Here, we show how the geometrical properties and the focal length of the lens array are modulated by controlling two appropriate parameters. Such microlenses can be useful also as polymer patterned arrayed microstructures for optical data interconnections, OLEDs efficient light extraction, concentrating light in energy solar cells, imaging and 3-D display solutions, and other photonics applications.

Numerical tools for the characterization of microelectromechanical systems by digital holographic microscopy

Abstract: Digital holography (DH) in microscopy became an important interferometric tool in optical metrology when camera sensors reached a higher pixel number with smaller size and high-speed computers became able to process the acquired images. This allowed the investigation of engineered surfaces on microscale, such as microelectromechanical systems (MEMS). In DH, numerical tools perform the reconstruction of the wave field. This offers the possibility of retrieving not only the intensity of the acquired wavefield, but also the phase distribution. This review describes the principles of DH and shows the most important numerical tools discovered and applied to date in the field of MEMS. Both the static and the dynamic regimes can be analyzed by means of DH. Whereas the first one is mostly related to the characterization after the fabrication process, the second one is a useful tool to characterize the actuation of the MEMS.

Portable IR Laser System for Real-Time Display of Alive People in Fire Scenes

Abstract: An innovative approach is shown that can allow search-and-rescue personnel to find survivors trapped in fire scenarios through active laser scanning. This new but simple imaging technique permits to see live people through a flame wall by portable system suitable for on-field applications. Combining IR laser raster scanning with a bandpass frequency filter and an IR camera, we show that a direct useful signature of a scattered human target can be detected while discarding the flame emission. Several experiments are reported also by using a cheap, light-weight and low-resolution camera to demonstrate that the proposed approach allows real-time operation by portable light-weight device, thus being useful in the hands of firefighters.

Automatic Frames Extraction and Visualization From Noisy Fringe Sequences for Data Recovering in a Portable Digital Speckle Pattern Interferometer for NDI

Abstract: We report on a method for recovering data from a simple portable Digital Speckle Pattern Interferometer, we used for monitoring structural behavior of a painting on wood, hanging on a wall, outside of laboratory conditions, without anti-vibration devices. In such a situation, fringes produced by the object displacements were affected by unpredictable distortions caused by environment vibrations. However, an sufficient number of suitable, i.e., undistorted or barely distorted, fringe patterns usable for processing was found. We performed multiple acquisitions at a frame rate as high as possible. The main task was picking out usable interferograms from large amount of frames. We developed a PC-based method based on jointly analysis of spectral content and fringe image sharpness as selection rules. The selected frames were utilized for off-line processing by using an approach based on Hilbert Transform and Phase Unwrapping via MAx-flow (PUMA) algorithm. We obtained a collection of displacement-maps, that allowed for evaluating the whole structure deformations, caused by environmental thermo-hygrometric fluctuations.

Volume holographic gratings: fabrication and characterization

Abstract: Volume Holographic Gratings is designed and fabricated to obtain a simple, lightweight and cheap light deflector. The entire process, starting from the chemical preparation of the photosensitive material, to the recording of Volume Holographic Gratings and their appropriate characterization, is reported. The recording material was a new photopolymer sensitive to light at 532nm. Results showed that the recorded Volume Holographic Grating presents a very high value of the diffraction efficiency, up to 94%. In addition, a flexible material is used to write Volume Holographic Gratings.

Coding Color Three-Dimensional Scenes and Joining Different Objects by Adaptive Transformations in Digital Holography

Abstract: We show that a three-dimensional (3D) scene can be coded by joining different objects by combining multiple optically recorded color digital holograms. Recently, an adaptive scheme based on affine transformations, able to correct defocus aberrations of digital holograms, was demonstrated, that we name here Adaptive Transformation in Digital Holography (ATDH). We propose an effective framework to create dynamic color holographic 3D scene by using a generalization of such scheme aided by a speckle reduction method, called Multilevel Bi-dimensional Empirical Mode Decomposition (MBEMD), used for the first time in color holography. We also demonstrate its feasibility to the synthesis of multiple Color Computer Generated Holograms (CCGHs).

Investigation on Axicon Transformation in Digital Holography for Extending the Depth of Focus in Bio-Microfluidics Applications

Abstract: Limited depth of focus (DOF) is one main shortage in traditional optical microscopy systems that severely affect simultaneous visualization of objects at different depths in the same field of view. In this paper, we propose a novel method to enhance the DOF in digital holography that consists of applying a numerical axicon transformation to the hologram during the reconstruction process. The idea behind this is to exploit the well-known ability of an axicon lens to create long and narrow focal lines along the optical axis. By this approach, we demonstrate that it is possible to obtain an extended focused image in which objects located at different depths are simultaneously visualized in good focus. First, the proposed method is tested in a case study of three different wires, positioned on different planes and recorded in lensless configuration. A comparison with a common DOF extension approach based on cubic phase function is performed. Finally, experiments of motile cells, flowing in a microfluidic channel and at different depths, are investigated for demonstrating the effectiveness of the proposed approach in bio-microfluidics.

Monitoring cell morphology during necrosis and apoptosis by quantitative phase imaging

Abstract: Cellular morphology changes and volume alterations play significant roles in many biological processes and they are mirrors of cell functions. In this paper, we propose the Digital Holographic microscope (DH) as a non-invasive imaging technique for a rapid and accurate extraction of morphological information related to cell death. In particular, we investigate the morphological variations that occur during necrosis and apoptosis. The study of necrosis is extremely important because it is often associated with unwarranted loss of cells in human pathologies such as ischemia, trauma, and some forms of neurodegeneration; therefore, a better elucidation in terms of cell morphological changes could pave the way for new treatments. Also, apoptosis is extremely important because it’s involved in cancer, both in its formation and in medical treatments. Because the inability to initiate apoptosis enhances tumour formation, current cancer treatments target this pathway. Within this framework, we have developed a transmission off-axis DH apparatus integrated with a micro incubator for investigation of living cells in a temperature and CO2 controlled environment. We employ DH to analyse the necrosis cell death induced by laser light (wavelength 473 nm, light power 4 mW). We have chosen as cellular model NIH 3T3 mouse embryonic fibroblasts because their adhesive features such as morphological changes, and the time needed to adhere and spread have been well characterized in the literature. We have monitored cell volume changes and morphological alterations in real time in order to study the necrosis process accurately and quantitatively. Cell volume changes were evaluated from the measured phase changes of light transmitted through cells. Our digital holographic experiments showed that after exposure of cells to laser light for 90-120 min., they swell and then take on a balloon-like shape until the plasma membrane ruptures and finally the cell volume decreases. Furthermore, we present a preliminary study on the variation of morphological parameters in case of cell apoptosis induced by exposure to 10 μM cadmium chloride. We employ the same cell line, monitoring the process for 18 hours. In the vast group of environmental pollutants, the toxic heavy metal cadmium is considered a likely candidate as a causative agent of several types of cancers. Widely distributed and used in industry, and with a broad range of target organs and a long half-life (10-30 years) in the human body, this element has been long known for its multiple adverse effects on human health, through occupational or environmental exposure. In apoptosis, we measure cell volume decrease and cell shrinking. Both data of apoptosis and necrosis were analysed by means of a Sigmoidal Statistical Distribution function, which allows several quantitative data to be established, such as swelling and cell death time, flux of intracellular material from inside to outside the cell, initial and final volume versus time. In addition, we can quantitatively study the cytoplasmatic granularity that occurs during necrosis. As a future application, DH could be employed as a non-invasive and label-free method to distinguish between apoptosis and necrosis in terms of morphological parameters.

Sparsity promoting automatic focusing in digital holography

Abstract: Sparsity properties of digital holograms have been investigated for application in compressive holography, permitting the discovery of the sparsest reconstruction plane in which the recovery of digital holograms is suitable. Recent approaches for denoising and phase retrieval are also proposed exploiting the sparsity properties of digital holograms. Thus it can be shown a strong correlation between holograms sparsity and focal plane detection, making a sparsity measure coefficient as a candidate to be used for focus plane calculation. Here we implement different sparsity metrics, that are able to measure a degree of sparsity of reconstructed digital hologram and we investigate their relation with the automatic focusing criterions, highlighting the possibility to use a sparsity measure as refocusing metric as well as the contrary, i.e. using image contrast coefficients as sparsity measures. Our analysis will be reported for digital holograms recorded in both lensless and microscope configuration and for both amplitude and pure-phase objects.

Non-destructive testing of an original XVI century painting on wood by ESPI system

Abstract: Electronic Speckle Pattern Interferometry (ESPI), a non-contact and non-destructive optical techniques, was employed for assessing the conservation state of a XVI Century painting on wood (72x88x1,9 cm). By a long term analysis, the whole structure alterations, induced by the room temperature and relative humidity variations, were evaluated. Measurement of the whole painting structural bends was achieved. Local flaws and hidden detachments of pictorial layers from the support, which cannot be recognized by traditional art-restorer methods, were also revealed. This work was prevalently aimed at achieving a simple approach, in the laboratory practice, to get an intuitively user-friendly method for art conservators, not accustomed to high-tech or math based methods. The results demonstrate that ESPI can largely improve the traditional art conservation survey techniques.

Cells characterization in microfluidic flows by small angle light scattering and 3D holographic technique

Abstract: The Light Scattering Profile (LSP) of an individual cell provides a fast and accurate characterization of its morphological properties. By combining a camera-based small angle light scattering apparatus with a microfluidic-induced particle migration technique, it is possible to characterize cells in microfluidic flows. The scattering profile of an individual cell can be fully characterized by our optimized optical light collection system. Viscoelastic-induced particle migration by polyethylene oxide implemented in a low-cost microfluidic device composed of an alignment section and a measuring section opens the possibility of precise, label-free, individual cell analysis. We have studied living cells in microfluidic flows by our light scattering apparatus and by a Digital Holographic Microscope (DHM) system. Our DHM measurements provided an accurate 3D position tracking even in multiple cell conditions.

Extended field of view space-time digital holograms for lab-on-a-chip microfluidic imaging

Abstract: We propose a new recording modality, named here Space-Time Scanning Interferometry (STSI), which allows to capture interferograms using a linear array detector instead of a common 2D sensor. Object scanning is exploited to perform a different mapping of the holograms in the space-time domain. Three sensor rows are sufficient to yield the whole complex object field from a time sequence of interferograms. This approach is particularly useful in microfluidic microscopy, where the sample motion is intrinsically provided. We then introduce the Space-Time Digital Hologram (STDH), still possessing all the capabilities of a common DH, namely quantitative phase-contrast mapping of the samples and flexible refocusing starting from blind out-of focus recordings, but obtainable using a compact single line detector easily embeddable onboard a Lab-on-a-Chip platform. Above all, the proposed optofluidic approach is able to provide STDHs with unlimited FoV along the flow direction, independently of the set magnification factor and without the need for further processing such as hologram stitching. Hence, thanks to the possibility to refocus multiple flowing objects displaced in a liquid volume, STDH assures drastically enhanced throughput, quantitative and label-free, on-chip microscopy.

Recovering data from noisy fringe patterns from a portable digital speckle pattern interferometer for in-situ inspection of painting hanging on the wall

Abstract: We report on a method for recovering data from a simple portable Digital Speckle Pattern Interferometer intended for utilization outside of laboratory conditions, without anti-vibration devices. We used the system for monitoring the structural behavior of a painting on wood, hanging on a wall. In such a situation, fringes, produced by the object displacements, were affected by random distortions caused by environment noise. However a satisfactory number of undistorted, or barely distorted, fringe patterns were found and utilized for processing. We performed fast continuous acquisitions of consecutive interferograms, picking usable fringe patterns out of a large amount of recorded frames. This is the crucial task in the measurement procedure. For this purpose we developed a software routine, based on jointly analysis of both spectral content and fringe image sharpne ss, as selection rule. From the selected frames, by using a simple approach based on Hilbert Transform and Phase Unwrapping, via MAx-flow (PUMA) algorithm, we were able to evaluate the painting whole structure deformations, caused by environmental thermo-hygrometric fluctuations.

Electrohydrodynamic Dispenser for Delivering Multiphase Samples at Nanoscale

Abstract: Manipulation of liquids on micro- and nanoscale is a key issue in many fields of technology and biotechnology. Electric field induced formation of microliter and nanoliter droplets is very useful in lab-on-chip applications and would represent a new and contact-less way for functionalizing smart materials [1, 2, 3]. Ink-jet printing, manipulation of biomolecules, deposition of inorganic, organic and biological inks [4, 5], dispensing of small amounts of material into well-defined areas would be a further possibility for functionalizing sensing area for lab-on-a-fiber devices and related applications.

Full 3D morphology of diatoms flowing in a microfluidic channel by digital holographic microscopy

Abstract: In this paper, we present a new approach for three-dimensional reconstruction and biovolume estimation of some species of diatoms. An optofluidic platform, composed by an optical tweezer and holographic modulus, is employed to retrieve several holograms at different angular positions, which are processed by the shape from silhouette algorithm to estimate the 3D shape of the cells.

Investigation on cytoskeleton dynamics for non-adherent cells under point-like stimuli

Abstract: In the present paper, Holographic Optical Tweezers (HOT) is employed to trap and manage functionalized micrometric latex beads with the aim at probing cellular forces in no-adherent state. For the first time at best of our knowledge, a suspended cell, subjected to mechanical stress, structures its cytoskeleton when anchored to point-like bonds. We exploit the HOT arrangement to induce mechanical deformation in suspended NIH 3T3 fibroblast. Our investigation is devoted to understand the inner cell mechanism when it is mechanically stressed by point-like stimulus without the substrate influence. In our experiment, cell adhesion is prevented and the stimulus is applied through latex beads trapped by HOT and positioned externally to the cell membrane. Our aims are devoted to analyze cell response during the transition from an homogeneous and isotropic structure (as it’s in suspension) to a mechanically stressed state. To analyze the cell material interaction we combine the HOT arrangement with two imaging systems: a Digital Holography (DH) setup in microscope configuration that is an investigation method useful for quantitative, label-free and full-field analysis of low contrast object and a fluorescence modulus. HOT are exploited to induce cellular response to specific stimuli while DH allows to measure such responses in no-invasive way. Finally, fluorescence imaging is added to discriminate the inner cell structures.