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...

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 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...

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...

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...

Interaction of nanoparticles (NPs) with cells is fundamental importance in biology and biomedical sciences. NPs can be taken up by cells, thus interacting their intracellular elements, modifying the life cycle pathways, possibly inducing death. Therefore, there a great interest understanding visualizing process cellular uptake itself or even secondary effects, for example, toxicity. Nowadays, no method reported yet which 3D imaging distribution achieved suspended flow-cytometry. Here we show...

Fourier ptychographic microscopy probes label-free samples from multiple angles and achieves super resolution phase-contrast imaging according to a synthetic aperture principle. Thus, it is particularly suitable for high-resolution of tissue slides over wide field view. Recently, in order make the optical setup robust against misalignments-induced artefacts, numerical multi-look has been added conventional phase retrieval process, thus allowing elimination related errors but at cost long...

The most recent discoveries in the biochemical field are highlighting increasingly important role of lipid droplets (LDs) several regulatory mechanisms living cells. LDs dynamic organelles and therefore their complete characterization terms number, size, spatial positioning relative distribution cell volume can shed light on roles played by LDs. Until now, fluorescence microscopy transmission electron assessed as gold standard methods for identifying due to high sensitivity specificity....

Unbiased learning pipeline for label-free single-cell classification.

Abstract Graphene is considered a possible drug deliver in nanomedicine for its mechanical, physical and chemical characteristics. Thus, studying graphene biocompatibility pivotal to contribute the modern nano-therapy science. The coexistence between cells should be analysed using non-invasive technologies thus quantitative phase imaging (QPI) modalities are suitable investigate morphometric evolution of under nanomaterial exposure. Here, we show how multimodal QPI approach can furnish...

Abstract Accumulation of bioavailable heavy metals in aquatic environment poses a serious threat to marine communities and human health due possible trophic transfers through the food chain toxic, non-degradable, exogenous pollutants. Copper (Cu) is one most spread water, can severely affect primary producers at high doses. Here we show novel imaging test assay dose-dependent effects Cu on live microalgae identifying stress conditions when they are still capable sustaining positive growth....

Holographic tomography allows the 3D mapping of refractive index biological samples thanks to reconstruction methods based on knowledge illumination directions or rotation angles imaged sample. Recently, phase contrast tomographic flow cytometry by digital holography has been demonstrated reconstruct three-dimensional distribution single cells while they are flowing along microfluidic channels. In this system, direction is fixed sample's not deterministically known a priori but induced...

The label-free single cell analysis by machine and Deep Learning, in combination with digital holography transmission microscope configuration, is becoming a powerful framework exploited for phenotyping biological samples. Usually, quantitative phase images of cells are retrieved from the reconstructed complex diffraction patterns used as inputs deep neural network. However, retrieval process can be very time consuming prone to errors. Here we address classification using learning strategies...

Abstract Lysosomes are the terminal end of catabolic pathways in cell, as well signaling centers performing important functions such recycling macromolecules, organelles, and nutrient adaptation. The importance lysosomes human health is supported by fact that deficiency most lysosomal genes causes monogenic diseases called a group Lysosomal Storage Diseases (LSDs). A common phenotypic hallmark LSDs expansion compartment can be detected using conventional imaging methods based on...

Abstract Single-cell phase-contrast tomography promises to become decisive for studying 3D intracellular structures in biology. It involves probing cells with light at wide angles, which unfortunately requires complex systems. Here we show an intriguing concept based on inherent natural process plants biology, i.e., dehydration, allowing us easily obtain 3D-tomography of onion-epidermal cells’ nuclei. In fact, the loss water reduces turgor pressure and recognize it induces significant...

Tomographic phase microscopy (TPM) in flow cytometry is one of the most promising computational imaging techniques for quantitative 3-dimensional (3D) analysis unstained single cells. Continuous cells’ flow, combined with stain-free mode, can assure high-throughput collection and informative 3D data. TPM promises to allow rapid screening by a nondestructive technique statistically relevant The current leading-edge research aimed at developing systems has already demonstrated possibility...

To efficiently tackle certain tumor types, finding new biomarkers for rapid and complete phenotyping of cancer cells is highly demanded. This especially the case most common pediatric solid sympathetic nervous system, namely, neuroblastoma (NB). Liquid biopsy in principle a very promising tool this purpose, but usually enrichment isolation circulating such patients remain difficult due to unavailability universal NB cell-specific surface markers. Here, we show that screening through...

Despite remarkable progresses in quantitative phase imaging (QPI) microscopes, their wide acceptance is limited due to the lack of specificity compared with well-established fluorescence microscopy. In fact, absence fluorescent tag prevents identify subcellular structures single cells, making challenging interpretation label-free 2D and 3D phase-contrast data. Great effort has been made by many groups worldwide address overcome such limitation. Different computational methods have proposed...

Lysosomal storage diseases (LSDs) are genetic disorders caused by enzyme deficiencies that lead to lysosomal dysfunction and progressive cell damage. Accurate visualization quantification of lysosomes essential for understanding disease progression developing effective therapies. Here, the first time, we successfully identified characterized using an innovative Holo-Tomographic Flow Cytometry (HTFC) technique, which allows label-free, high-content, high-throughput 3D imaging compartments in...

In recent years, intracellular LDs have been discovered to play an important role in several pathologies. Therefore, detection of would provide in-demand diagnostic tool if coupled with flow-cytometry give significant statistical analysis and especially the diagnosis is made full non-invasive mode. Here we combine experimental results in-flow tomographic phase microscopy a suited numerical simulation demonstrate that can be easily detected through label-free approach based on direct 2D...