High-throughput single-cell analysis is a challenging task. Label-free tomographic phase microscopy an excellent candidate to perform this However, in-line tomography very difficult implement in practice because it requires complex set-up for rotating the sample and examining cell along several directions. We demonstrate that by exploiting random rolling of cells while they are flowing microfluidic channel, possible obtain phase-contrast tomography, if smart strategies wavefront adopted. In...

Particle tracking is a fundamental technique for investigating variety of biophysical processes, from intracellular dynamics to the characterization cell motility and migration. However, observing three-dimensional (3D) trajectories particles in general challenging task classical microscopy owing limited imaging depth field commercial optical microscopes, which represents serious drawback analysis time-lapse image data. Therefore, numerous automated particle-tracking approaches have been...

Digital holography (DH) has emerged as one of the most effective coherent imaging technologies. The technological developments digital sensors and optical elements have made DH primary approach in several research fields, from quantitative phase to metrology 3D display technologies, name a few. Like many other techniques, must cope with issue speckle artifacts, due nature required light sources. Despite complexity recently proposed de-speckling methods, not yet attained level effectiveness....

Abstract Accurate depiction of waves in temporal and spatial is essential to the investigation interactions between physical objects waves. Digital holography (DH) can perform quantitative analysis wave–matter interactions. Full detector-bandwidth reconstruction be realized based on in-line DH. But overlapping twin images strongly prevents analysis. For off-axis DH, object wave detector bandwidth need satisfy certain conditions accurately. Here, we present a reliable approach involving...

Abstract Quantitative phase imaging has gained popularity in bioimaging because it can avoid the need for cell staining, which, some cases, is difficult or impossible. However, as a result, quantitative does not provide labelling of various specific intracellular structures. Here we show novel computational segmentation method based on statistical inference that makes possible techniques to identify nucleus. We demonstrate approach with refractive index tomograms stain-free cells...

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

The perspective of using live cells as lenses could open new revolutionary and intriguing scenarios in the future biophotonics biomedical sciences for endoscopic vision, local laser treatments via optical fibres diagnostics. Here we show that a suspended red blood cell (RBC) behaves an adaptive liquid-lens at microscale, thus demonstrating its imaging capability tunable focal length. In fact, thanks to intrinsic elastic properties, RBC can swell up from disk volume 90 fl sphere reaching 150...

Sperm morphology is regarded as a significant prognostic factor for fertilization, abnormal sperm structure one of the most common factors in male infertility. Furthermore, obtaining accurate morphological information an important issue with strong implications zoo-technical industries, example to perform sorting species X from Y. A challenging step forward would be availability fast, high-throughput and label-free system measurement physical parameters visualization 3D shape such biological...

One of the main drawbacks Digital Holography (DH) is coherent nature light source, which severely corrupts quality holographic reconstructions. Although numerous techniques to reduce noise in DH have provided good results, suppression remains a challenging task. We propose novel framework that combines concepts encoding multiple uncorrelated digital holograms, block grouping and collaborative filtering achieve quasi noise-free The optimized joint action these different image-denoising...

Microplastics (MPs) are a major environmental concern due to their possible impact on water pollution, wildlife, and the food chain. Reliable, rapid, high‐throughput screening of MPs from other components sample after sieving and/or digestion is still highly desirable goal avoid cumbersome visual analysis by expert users under optical microscope. Here, new approach presented that combines 3D coherent imaging with machine learning (ML) achieve accurate automatic detection in filtered samples...

Several automatic approaches have been proposed in the past to compute refocus distance digital holography (DH). However most of them are based on a maximization or minimization suitable amplitude image contrast measure, regarded as function reconstruction parameter. Here we show that, by using sparsity measure coefficient refocusing criterion holographic reconstruction, it is possible recover focus plane and, at same time, establish degree holograms, when samples diffraction Fresnel...

Abstract Three dimensional (3D) morphometric analysis of flowing and not‐adherent cells is an important aspect for diagnostic purposes. However, diagnostics tools need to be quantitative, label‐free and, as much possible, accurate. Recently, a simple holographic approach, based on shape from silhouette algorithm, has been demonstrated accurate calculation biovolume displaying their 3D shapes. Such approach adopted in combination with optical tweezers successfully applied convex shape....

We report a reliable full-angle tomographic phase microscopy (FA-TPM) method for flowing quasi-spherical cells along microfluidic channels. This lies in completely passive optical system, i.e. mechanical scanning or multi-direction probing of the sample is avoided. It exploits engineered rolling while they are channel. Here we demonstrate significant progress with respect to state art in-flow TPM by showing general extension having almost spherical shapes suspension. In fact, adopted...

Abstract Circulating tumor cells (CTCs) are rare released from primary, metastatic, or recurrent tumors in the peripheral blood of cancer patients. CTCs isolation and their molecular characterization represent a new marker screening, diagnostic tool called “liquid biopsy” (LB). Compared to traditional tissue biopsy that is invasive does not reveal heterogeneity, LB noninvasive reflects “real‐time” dynamism drug sensitivity. In frame LB, paradigm based on single‐cell label‐free analysis...

Tomographic flow cytometry by digital holography is an emerging imaging modality capable of collecting multiple views moving and rotating cells with the aim recovering their refractive index distribution in 3D. Although this allows us to access high-resolution high-throughput, huge amount time-lapse holographic images be processed (hundreds holograms per cell) constitutes actual bottleneck. This prevents system from being suitable for lab-on-a-chip platforms real-world applications, where...

Abstract Digital holography is a technique that provides non-invasive, label-free, quantitative, and high-resolution imaging employable in biological science of matter fields, but not only. In the last decade, digital (DH) has undergone very significant signs progress made it one most powerful metrology tools. However, important issues to be afforded solved for obtaining quantitative phase information about analyzed specimen related aberrations. Sources aberrations can diverse, several...

Abstract Image-based identification of circulating tumor cells in microfluidic cytometry condition is one the most challenging perspectives Liquid Biopsy scenario. Here we show a machine learning-powered tomographic phase imaging flow system capable to provide high-throughput 3D phase-contrast tomograms each single cell. In fact, that discrimination against white blood potentially achievable with aid artificial intelligence label-free flow-cyto-tomography method. We propose hierarchical...

A 3D scene is synthesized combining multiple optically recorded digital holograms of different objects. The novel idea consists compositing moving objects in a dynamic using process that analogous to stop-motion video. However this case the movie has exciting attribute it can be displayed and observed 3D. We show scenes projected as an alternative complicated heavy computations needed generate realistic-looking computer generated holograms. key tool for creating action based on new concept...

Holographic imaging may become severely degraded by a mixture of speckle and incoherent additive noise. Bayesian approaches reduce the noise, but prior information is needed on noise statistics. With no knowledge, one-shot reduction highly desirable goal, as recording process simplified made faster. Indeed, neither multiple acquisitions nor complex setup are needed. So far, this result has been achieved at cost deterministic resolution loss. Here we propose fast non-Bayesian denoising method...

Today, fast and accurate diagnosis through portable cheap devices is in high demand for the general healthcare. Lab-on-chips (LoCs) have undergone a great growth this direction, supported by optical imaging techniques more refined. Here we present recent progresses developing tools based on coherent microscopy that can be very useful when applied into biomicrofluidics. In some cases, tweezers (OT) technique combined with digital holography (DH), thus offering possibility to manipulate,...

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

In digital holography (DH) a mixture of speckle and incoherent additive noise, which appears in numerical as well optical reconstruction, typically degrades the information object wavefront. Several methods have been proposed order to suppress noise contributions during recording or even reconstruction steps. Many them are based on combination multiple holographic reconstructions achieving remarkable improvement, but only i.e. visualization pc monitor. So far, it has not shown direct...

The 3D tracking of micro-objects, based on digital holography, is proposed through the analysis complex wavefront light scattered by micro-samples. Exploiting advantages off-axis full-field holographic interferometry, multiple objects achieved a direct at focal plane overcoming limitation conventional back interferometry in which only one object time can be tracked. Furthermore, method and demonstrated here step forward with respect to other tools. approach tested two experiments, first...

Water plastic pollution is a serious problem affecting sealife, marine habitats, and the food chain. Artificial intelligence-enabled coherent imaging has recently shown exciting advances in field of environmental monitoring, portable holographic microscopes are good candidates to map microparticles content waters. The "holographic fingerprint" due light diffraction rich information, fully encoded into complex wavefront scattered by sample. Hence, proper analysis wavefronts reconstructed from...