Stain-free identification of cell nuclei using tomographic phase microscopy in flow cytometry.

Quantitative Phase Imaging (QPI) has gained popularity in bioimaging because it can avoid the need for cell staining, which in some cases is difficult or impossible. However, as a result, QPI does not provide labelling of various specific intracellular structures. Here we show a novel computational segmentation method based on statistical inference that makes it possible for QPI techniques to identify the cell nucleus.

Differential diagnosis of hereditary anemias from a fraction of blood drop by digital holography and hierarchical machine learning.

Anemia affects about the 25% of the global population and can provoke severe diseases, ranging from weakness and dizziness to pregnancy problems, arrhythmias and hearth failures. About 10% of the patients are affected by rare anemias of which 80% are hereditary. Early differential diagnosis of anemia enables prescribing patients a proper treatment and diet, which is effective to mitigate the associated symptoms.

Three-Dimensional Quantitative Intracellular Visualization of Graphene Oxide Nanoparticles by Tomographic Flow Cytometry.

Interaction of nanoparticles (NPs) with cells is of fundamental importance in biology and biomedical sciences. NPs can be taken up by cells, thus interacting with their intracellular elements, modifying the life cycle pathways, and possibly inducing death. Therefore, there is a great interest in understanding and visualizing the process of cellular uptake itself or even secondary effects, for example, toxicity.

Rolling angle recovery of flowing cells in holographic tomography exploiting the phase similarity.

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

Learning Diatoms Classification from a Dry Test Slide by Holographic Microscopy.

Diatoms are among the dominant phytoplankters in marine and freshwater habitats, and important biomarkers of water quality, making their identification and classification one of the current challenges for environmental monitoring. To date, taxonomy of the species populating a water column is still conducted by marine biologists on the basis of their own experience. On the other hand, deep learning is recognized as the elective technique for solving image classification problems.

Gold Nanorods and Nanoprisms Mediate Different Photothermal Cell Death Mechanisms In Vitro and In Vivo.

Photothermal therapy (PTT) is an efficient method of inducing localized hyperthermia and can be achieved using gold nanoparticles as photothermal agents. However, there are many hurdles to get over before this therapy can safely reach the clinics, including nanoparticles' optimal shape and the accurate prediction of cellular responses. Here, we describe the synthesis of gold nanorods and nanoprisms with similar surface plasmon resonances in the near-infrared (NIR) and comparable photothermal conversion efficiencies and characterize the response to NIR irradiation in two biological systems, melanoma cells and the small invertebrate .

Biological Lenses as a Photomask for Writing Laser Spots into Ferroelectric Crystals.

Red blood cells on the surface of a lithium niobate crystal can be used as optical lenses for direct writing of laser-induced refractive index changes. The writing process by such a photomask made of biological lenses is due to the photorefractive effect. Wavefront analysis by a digital holographic microscope is performed for deep and accurate evaluation of local refractive index changes.

In vitro cytotoxicity evaluation of cadmium by label-free holographic microscopy.

Among all environmental pollutants, the toxic heavy metal cadmium is considered as a human carcinogen. Cadmium may induce cell death by apoptosis in various cell types, although the underlying mechanisms are still unclear. In this paper we show how a label-free digital holography (DH)-based technique is able to quantify the evolution of key biophysical parameters of cells during the exposure to cadmium for the first time.

Label-Free Optical Marker for Red-Blood-Cell Phenotyping of Inherited Anemias.

The gold-standard methods for anemia diagnosis are complete blood counts and peripheral-smear observations. However, these do not allow for a complete differential diagnosis as that requires biochemical assays, which are label-dependent techniques. On the other hand, recent studies focus on label-free quantitative phase imaging (QPI) of blood samples to investigate blood diseases by using video-based morphological methods.

Full-angle tomographic phase microscopy of flowing quasi-spherical cells.

We report a reliable full-angle tomographic phase microscopy (FA-TPM) method for flowing quasi-spherical cells along microfluidic channels. This method lies in a completely passive optical system, i.e.

Tomographic flow cytometry by digital holography.

High-throughput single-cell analysis is a challenging task. Label-free tomographic phase microscopy is an excellent candidate to perform this task. However, in-line tomography is very difficult to implement in practice because it requires a complex set-up for rotating the sample and examining the cell along several directions.

Biolens behavior of RBCs under optically-induced mechanical stress.

In this work, the optical behavior of Red Blood Cells (RBCs) under an optically-induced mechanical stress was studied. Exploiting the new findings concerning the optical lens-like behavior of RBCs, the variations of the wavefront refracted by optically-deformed RBCs were further investigated. Experimental analysis have been performed through the combination of digital holography and numerical analysis based on Zernike polynomials, while the biological lens is deformed under the action of multiple dynamic optical tweezers.

Investigation on dynamics of red blood cells through their behavior as biophotonic lenses.

The possibility to adopt biological matter as photonic optical elements can open scenarios in biophotonics research. Recently, it has been demonstrated that a red blood cell (RBC) can be seen as an optofluidic microlens by showing its imaging capability as well as its focal tunability. Moreover, correlation between an RBC’s morphology and its behavior as a refractive optical element has been established and its exploitation for biomedical diagnostic purposes has been foreseen.

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

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.

3D morphometry of red blood cells by digital holography.

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 as possible, accurate. Recently, a simple holographic approach, based on shape from silhouette algorithm, has been demonstrated for accurate calculation of cells biovolume and displaying their 3D shapes.

Imaging adherent cells in the microfluidic channel hidden by flowing RBCs as occluding objects by a holographic method.

Imaging through turbid media is a challenging topic. A liquid is considered turbid when dispersed particles provoke strong light scattering, thus destroying the image formation by any standard optical system. Generally, colloidal solutions belong to the class of turbid media since dispersed particles have dimensions ranging between 0.

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].

Multi-imaging capabilities of a 2D diffraction grating in combination with digital holography.

In this Letter we report on an alternative approach to get multiple images in microscopy, exploiting the capabilities of both a lithium niobate diffraction grating and digital holographic technique. We demonstrate that multi-imaging can be achieved in a lensless configuration by using a hexagonal diffraction grating but overcoming, thanks to digital holography (DH), the many constrains imposed by the grating parameters in multi-imaging with Talbot effect or Talbot array illuminators. In fact, DH permits the numerical reconstruction of the optical field diffracted by the grating, thus obtaining in-focus multiple images in a plane different from the fractional or entire Talbot ones.

Bartonella henselae in Ixodes ricinus ticks (Acari: Ixodida) removed from humans, Belluno province, Italy.

The potential role of ticks as vectors of Bartonella species has recently been suggested. In this study, we investigated the presence of Bartonella species in 271 ticks removed from humans in Belluno Province, Italy. By using primers derived from the 60-kDa heat shock protein gene sequences, Bartonella DNA was amplified and sequenced from four Ixodes ricinus ticks (1.