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...

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...

Aberrations and the distortions due to imaging optics can be compensated in quantitative phase microscopy of thin objects by digital holography using a single hologram. The reconstructed distribution map directly corrected image plane numerical method. To remove this unwanted aberration, special case objects, authors perform two-dimensional fit with Zernike polynomials unwrapped phase. Subtraction fitted polynomial from original gives free aberrations.

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...

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....

The ability to see behind flames is a key challenge for the industrial field and particularly safety field. Development of new technologies detect live people through smoke in fire scenes an extremely desirable goal since it can save human lives. latest technologies, including equipment adopted by departments, use infrared bolometers digital cameras that allow users smoke. However, such detectors are blinded flame-emitted radiation. Here we show completely different approach makes lensless...

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...

Lens effect was obtained in an open microfluidic system by using a thin layer of liquid on polar electric crystal like LiNbO3. An array micro-lenses generated electrowetting pyroelectric periodically poled crystals. Compared to conventional devices, the allowed have electrode-less and circuit-less configuration. interferometric technique used characterize curvature corresponding results are presented discussed. The preliminary concerning imaging capability micro-lens also reported.

Quantitative Phase Microscopy (QPM) by interferometric techniques can require a multiwavelength configuration to remove π 2 ambiguity and improve accuracy.However, severe chromatic aberration affect the resulting phase-contrast map.By means of classical interference microscope it is quite unpractical correct such aberration.We propose demonstrate that Digital Holography (DH) in possible clear out QPM map from simpler more effective way with respect other approaches.The proposed method takes...

A special class of tunable liquid microlenses is presented here. The are generated by an electrowetting effect under electrode-less configuration and they exhibit two different regimes that named here as separated lens regime (SLR) wave-like (WLR). induced the pyroelectricity polar dielectric crystals, was proved in principle a previous work same authors (S. Grilli et al., Opt. Express 16, 8084, 2008). Compared to work, improvements experimental set-up procedure allow reveal which optical...

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,...

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...

The appearance of the first laser approximately 12 years after invention holography by Gabor (1948) revolutionized field optical metrology. In fact, holographic interferometry enabled exploitation on non-mirror surfaces and full-scale objects. holography-based measurement methods has been implemented to several industrial systems or in support R&D with aim improving new products many fields (automotive, aerospace, electronics, etc.). To date, considered an important tool for non-destructive...

Abstract In‐flow phase‐contrast tomography provides a 3D refractive index of label‐free cells in cytometry systems. Its major limitation, as with any quantitative phase imaging approach, is the lack specificity compared to fluorescence microscopy, thus restraining its huge potentialities single‐cell analysis and diagnostics. Remarkable results introducing are obtained through artificial intelligence (AI), but only for adherent cells. However, accessing ground truth obtaining accurate...

In color digital holography the recording and reconstruction of three holograms, by an optical setup operating in multi-wavelength mode, is necessary. Incorrect superposition different images at various wavelengths may occur due to presence chromatic aberrations. We demonstrate that compensation aberrations can be achieved using all information contained holograms. Chromatic aberration extracted from phase maps reconstructed wavelengths, resulting perfect full amplitude reconstructions. The...