Surgical Excision of Keloids Followed by In-office Superficial Radiation Therapy: Prospective Study Examining Clinical Outcomes.

BACKGROUND: Keloids are benign proliferative scars that often occur among individuals of color, and are thought to be the result of excessive collagen deposition that occurs after injury to the skin. The treatment of these scars is difficult with often poor outcomes. This study aimed to evaluate the effectiveness of surgical excision followed by in-office superficial radiation therapy (SRT) as a method to improve keloid remission. METHODS: Participants for this study were recruited from June 2016 through February 2017 with 48 subjects enrolled and completed this study. All keloids were surgically resected and participants received 3 consecutive days of a customized dose of SRT, with a maximum cumulative dosage of 18 Gy. Patients were followed over the course of 12 months to monitor outcomes. RESULTS: In this cohort, we found 39 (81%) to have achieved successful remission with 9 (19%) being classified as refractory. There were no adverse effects or medical complications reported as a part of this study. CONCLUSION: Study outcomes support the clinical benefits of surgical excision followed by SRT as a practical and efficient treatment for keloids.

Biophysical investigation of living monocytes in flow by collaborative coherent imaging techniques.

We implemented a completely label-free biophysical (morphometric and optical) property characterization of living monocytes in flow, using measurements obtained from two coherent imaging techniques: a pure light scattering approach to obtain an optical signature (OS) of cells, and a digital holography (DH) approach to achieve optical cell reconstructions in flow. A precise 3D cell alignment platform, taking advantage of viscoelastic fluid properties and microfluidic channel geometry, was used to investigate the OS of cells to achieve their refractive index, ratio of the nucleus over cytoplasm, and overall cell dimension. Further quantitative phase-contrast reconstructions by DH were employed to calculate surface area, dry mass, and biovolume of monocytes by using the OS outcomes as input parameters. The results show significantly different biophysical cell properties, confirming the possibility to differentiate monocytes from other cell classes in flow, thus avoiding chemical cell staining or labeling, which are nowadays used.

Label-free analysis of mononuclear human blood cells in microfluidic flow by coherent imaging tools.

The investigation of the physical properties of peripheral blood mononuclear cells (PBMC) is of great relevance, as they play a key role in regulating human body health. Here we report the possibility to characterize human PBMC in their physiological conditions in a microfluidic-based measurement system. A viscoelastic polymer solution is adopted for 3D alignment of individual cells inflow. An optical signature (OS) acquisition of each flowing cell is performed using a wide angle light scattering apparatus. Besides, a quantitative phase imaging (QPI) holographic system is employed with the aim (i) to check the position in flow of individual cells using a holographic 3D cell tracking method; and (ii) to estimate their 3D morphometric features, such as their refractive index (RI). Results obtained by combining OS and QPI have been compared with literature values, showing good agreement. The results confirm the possibility to obtain sub-micrometric details of physical cell properties in microfluidic flow, avoiding chemical staining or fluorescent labelling.

Quasi noise-free digital holography.

One of the main drawbacks of Digital Holography (DH) is the coherent nature of the light source, which severely corrupts the quality of holographic reconstructions. Although numerous techniques to reduce noise in DH have provided good results, holographic noise suppression remains a challenging task. We propose a novel framework that combines the concepts of encoding multiple uncorrelated digital holograms, block grouping and collaborative filtering to achieve quasi noise-free DH reconstructions. The optimized joint action of these different image-denoising methods permits the removal of up to 98% of the noise while preserving the image contrast. The resulting quality of the hologram reconstructions is comparable to the quality achievable with non-coherent techniques and far beyond the current state of art in DH. Experimental validation is provided for both single-wavelength and multi-wavelength DH, and a comparison with the most used holographic denoising methods is performed.

Holographic tracking of living cells by three-dimensional reconstructed complex wavefronts alignment.

We propose here a new three-dimensional (3D) holographic tracking method capable to track, simultaneously and in a single step, all the spatial coordinates of micro-objects. The approach is based on the enhanced correlation coefficient (ECC) maximization method but applied, for the first time to the best of our knowledge, directly on the holographic reconstructed complex wave fields. The key novelty of the proposed strategy is its ability to calculate simultaneously the 3D coordinates of cells, without decoupling the contribution of amplitude and phase. The proposed strategy is tested on living cells (i.e., NIH 3T3 mouse fibroblast) flowing into a microfluidic channel and compared with classical holographic tracking approach. Theoretical description and experimental validation of the proposed strategy are reported.

Quantitative phase maps denoising of long holographic sequences by using SPADEDH algorithm.

We propose a denoising method for digital holography mod 2π wrapped phase map by using an adaptation of the SPArsity DEnoising of Digital Holograms (SPADEDH) algorithm. SPADEDH is a l(1) minimization algorithm able to suppress the noise components on digital holograms without any prior knowledge or estimation about the statistics of noise. We test our algorithm with either general numerical simulated wrapped phase, quantifying the performance with different efficiency parameters and comparing it with two popular denoising strategies, i.e., median and Gaussian filters, and specific experimental tests, by focusing our attention on long-sequence wrapped quantitative phase maps (QPMs) of in vitro cells, which aim to have uncorrupted QPMs. In addition, we prove that the proposed algorithm can be used as a helper for the typical local phase unwrapping algorithms.

Enhancing depth of focus in tilted microfluidics channels by digital holography.

In this Letter we propose a method to enhance the limited depth of field (DOF) in optical imaging systems, through digital holography. The proposed approach is based on the introduction of a cubic phase plate into the diffraction integral, analogous to what occurs in white-light imaging systems. By this approach we show that it is possible to improve the DOF and to recover the extended focus image of a tilted object in a single reconstruction step. Moreover, we demonstrate the possibility of obtaining well-focused biological cells flowing into a tilted microfluidic channel.

On the holographic 3D tracking of in vitro cells characterized by a highly-morphological change.

Digital Holography (DH) in microscopic configuration is a powerful tool for the imaging of micro-objects contained into a three dimensional (3D) volume, by a single-shot image acquisition. Many studies report on the ability of DH to track particle, microorganism and cells in 3D. However, very few investigations are performed with objects that change severely their morphology during the observation period. Here we study DH as a tool for 3D tracking an osteosarcoma cell line for which extensive changes in cell morphology are associated to cell motion. Due to the great unpredictable morphological change, retrieving cell's position in 3D can become a complicated issue. We investigate and discuss in this paper how the tridimensional position can be affected by the continuous change of the cells. Moreover we propose and test some strategies to afford the problems and compare it with others approaches. Finally, results on the 3D tracking and comments are reported and illustrated.

Terahertz tuning of whispering gallery modes in a PDMS stand-alone, stretchable microsphere.

We report on tuning the optical whispering gallery modes (WGMs) in a poly dimethyl siloxane-based (PDMS) microsphere resonator by more than 1 THz. The PDMS microsphere system consists of a solid spherical resonator directly formed with double stems on either side. The stems act like tie-rods for simple mechanical stretching of the microresonator, resulting in tuning of the WGMs by one free spectral range. Further investigations demonstrate that the WGM shift has a higher sensitivity (0.13 nm/µN) to an applied force when the resonator is in its maximally stretched state compared to its relaxed state.

Clear coherent imaging in turbid microfluidics by multiple holographic acquisitions.

Recently it has been demonstrated that digital holography is a powerful means allowing imaging of both amplitude and phase objects in turbid flowing media. However, in quasi-static turbid microfluidics, multiple scattering contributions through the colloids superimpose coherently to the recording device, resulting in speckle noise and hindering a clear vision of the objects. In this Letter we exploit the Brownian motion of the colloidal particles to get multiple uncorrelated holograms, and we combine them to reduce the speckle contrast. In this way we get a multi-look gain without losing image resolution.

Microscopy imaging and quantitative phase contrast mapping in turbid microfluidic channels by digital holography.

We show that sharp imaging and quantitative phase-contrast microcopy is possible in microfluidics in flowing turbid media by digital holography. In fact, in flowing liquids with suspended colloidal particles, clear vision is hindered and cannot be recovered by any other microscopic imaging technique. On the contrary, using digital holography, clear imaging is possible thanks to the Doppler frequency shift experienced by the photons scattered by the flowing colloidal particles, which do not contribute to the interference process, i.e. the recorded hologram. The method is illustrated and imaging results are demonstrated for pure phase objects, i.e. biological cells in microfluidic channels.

3D lithography by rapid curing of the liquid instabilities at nanoscale.

In liquids realm, surface tension and capillarity are the key forces driving the formation of the shapes pervading the nature. The steady dew drops appearing on plant leaves and spider webs result from the minimization of the overall surface energy [Zheng Y, et al. (2010) Nature 463:640-643]. Thanks to the surface tension, the interfaces of such spontaneous structures exhibit extremely good spherical shape and consequently worthy optical quality. Also nanofluidic instabilities generate a variety of fascinating liquid silhouettes, but they are however intrinsically short-lived. Here we show that such unsteady liquid structures, shaped in polymeric liquids by an electrohydrodynamic pressure, can be rapidly cured by appropriate thermal treatments. The fabrication of many solid microstructures exploitable in photonics is demonstrated, thus leading to a new concept in 3D lithography. The applicability of specific structures as optical tweezers and as novel remotely excitable quantum dots-embedded microresonators is presented.

Dynamic DIC by digital holography microscopy for enhancing phase-contrast visualization.

Differential image contrast (DIC), through the numerical managing and manipulation of complex wavefronts obtained by digital holography (DH), is investigated. We name the approach Dynamical Differential Holographic Image Contrast (DDHIC). DDHIC dispenses from special optics and/or complex setup configurations with moveable components, as usually occurs in classical DIC, that is not well-suited for investigating objects experiencing dynamic evolution during the measurement. In fact, the technique presented here, is useful for floating samples since it allows, from a single recording, to set a posteriori the best conditions for DIC imaging in conjunction with the numerical focusing feature of DH. By DDHIC, the movies can be easily built-up to offering dynamic representation of phase-contrast along all directions, thus improving the visualization. Furthermore, the dynamic representation is useful for making the proper choice of other key parameters of DIC such as the amount of shear and the bias, with the aim to optimize the visualized phase-contrast imaging as favorite representation for bio-scientists. Investigation is performed on various biological samples.

Twin-beams digital holography for 3D tracking and quantitative phase-contrast microscopy in microfluidics.

We report on a compact twin-beam interferometer that can be adopted as a flexible diagnostic tool in microfluidic platforms with twofold functionality. The novel configuration allows 3D tracking of micro-particles and, at same time, can simultaneously furnish Quantitative Phase-contrast maps of tracked micro-objects by interference microscopy, without changing the configuration. Experimental demonstration is given on for in vitro cells in a microfluidic environment.

Compression of digital holograms via adaptive-sparse representation.

Efficient storage and transmission of digital holograms (DHs) requires the development of appropriate compression techniques for such a special class of images. In this Letter, we investigate a method to compress DHs using a sparse matrix representation. Using digital holography to numerically manage complex wave fields, we are able to apply an adaptive mask, based on a threshold filter, to the object wave field. From there, we store the result of this filtering by sparse representation. In this Letter, we demonstrate that using sparse representation allows for a high compression factor with minimal loss in the quality of the reconstructed image. This technique is efficient for storage and transmission of DHs.

Synthesis and display of dynamic holographic 3D scenes with real-world objects.

A 3D scene is synthesized combining multiple optically recorded digital holograms of different objects. The novel idea consists of compositing moving 3D objects in a dynamic 3D scene using a process that is analogous to stop-motion video. However in this case the movie has the exciting attribute that it can be displayed and observed in 3D. We show that 3D dynamic scenes can be projected as an alternative to complicated and heavy computations needed to generate realistic-looking computer generated holograms. The key tool for creating the dynamic action is based on a new concept that consists of a spatial, adaptive transformation of digital holograms of real-world objects allowing full control in the manipulation of the object's position and size in a 3D volume with very high depth-of-focus. A pilot experiment to evaluate how viewers perceive depth in a conventional single-view display of the dynamic 3D scene has been performed.

Light induced patterning of poly(dimethylsiloxane) microstructures.

A new method for direct patterning of Poly(dimethylsiloxane) (PDMS) microstructures is developed by taking advantage of photorefractive effect in a functionalized substrate. Here we show that when a x-cut Iron doped Lithium Niobate (LN) crystal is exposed to appropriate structured laser light, a charge density pattern builds-up in the crystal and a space charge field arise that is able to induce self-patterning of the PDMS liquid film deposited on its surface via the dielectrophoretic effects. Proper heating treatment allows to achieve polymeric linking process creating a solid and stable PDMS microstructures. The self-patterned structures replicate the illuminating light pattern. We show that 1D and 2D patterning of PDMS gratings can be achieved. This new soft-lithographic approach can pave the way for realizing PDMS micro-structures with high degree of flexibility that avoids the need of moulds fabrication.

Optical reconstruction of digital holograms recorded at 10.6 microm: route for 3D imaging at long infrared wavelengths.

We demonstrate the optical reconstruction in the visible range (0.532 microm) of digital holograms recorded at long IR wavelengths (10.6 microm) by means of a spatial light modulator. By using an integrated recording-reconstruction system, it is, in fact, feasible to achieve direct imaging of holograms acquired outside the visible range, i.e., in the IR spectrum. By choosing a Fourier recording configuration, the reconstructed image, obtained at about a 20 times shorter wavelength than the acquisition image, exhibits minor aberrations, which do not significantly affect the optical reconstruction. The high NA achievable at a long IR wavelength allows us to image large objects at reasonable distances.

Exploiting nanotechnologies and TRPV1 channels to investigate the putative anandamide membrane transporter.

BACKGROUND: Considerable efforts have been made to characterize the pathways regulating the extracellular levels of the endocannabinoid anandamide. However, none of such pathways has been so argued as the existence of a carrier-mediated transport of anandamide across the membrane. Apart from the lack of molecular evidence for such a carrier, the main reasons of this controversy lie in the methodologies currently used to study anandamide cellular uptake. Furthermore, the main evidence in favor of the existence of an "anandamide transporter" relies on synthetic inhibitors of this process, the selectivity of which has been questioned. METHODOLOGY/PRINCIPAL FINDINGS: We used the cytosolic binding site for anandamide on TRPV1 channels as a biosensor to detect anandamide entry into cells, and exploited nanotechnologies to study anandamide membrane transport into intact TRPV1-overexpressing HEK-293 cells. Both fluorescence and digital holographic (DH) quantitative phase microscopy were used to study TRPV1 activation. Poly-epsilon-caprolactone nanoparticles (PCL-NPs) were used to incorporate anandamide, which could thus enter the cell and activate TRPV1 channels bypassing any possible specific protein(s) involved in the uptake process. We reasoned that in the absence of such protein(s), pharmacological tools previously shown to inhibit the "anandamide transporter" would affect in the same way the uptake of anandamide and PCL-NP-anandamide, and hence the activation of TRPV1. However, when masked into PCL-NPs, anandamide cellular uptake became much less sensitive to these agents, although it maintained the same pharmacokinetics and pharmacodynamics as that of "free" anandamide. CONCLUSIONS: We found here that several agents previously reported to inhibit anandamide cellular uptake lose their efficacy when anandamide is prevented from interacting directly with plasma membrane proteins, thus arguing in favor of the specificity of such agents for the putative "anandamide transporter", and of the existence of such mechanism.

Spatially resolved refractive index profiles of electrically switchable computer-generated holographic gratings.

We describe a spatially resolved interferometric technique combined with a phase reconstruction method that provides a quantitative two-dimensional profile of the refractive index and spatial distribution of the optical contrast between the on-off states of electrically switchable diffraction gratings as a function of the external electric field. The studied structures are holographic gratings optically written into polymer/liquid crystal composites through single-beam spatial light modulation by means of computer-generated holograms. The electro-optical response of the gratings is also discussed. The diffraction efficiency results to be dependent on the incident light polarization suggesting the possibility to develop polarization dependent switching devices.