The laser induced plasma spectroscopy was applied on particles attached on substrate represented by a silica wafer covered with a thin oil film. The substrate itself weakly interacts with a ns Nd:YAG ...laser (1064nm) while presence of particles strongly enhances the plasma emission, here detected by a compact spectrometer array. Variations of the sample mass from one laser spot to another exceed one order of magnitude, as estimated by on-line photography and the initial image calibration for different sample loadings. Consequently, the spectral lines from particles show extreme intensity fluctuations from one sampling point to another, between the detection threshold and the detector's saturation in some cases. In such conditions the common calibration approach based on the averaged spectra, also when considering ratios of the element lines i.e. concentrations, produces errors too large for measuring the sample compositions. On the other hand, intensities of an analytical and the reference line from single shot spectra are linearly correlated. The corresponding slope depends on the concentration ratio and it is weakly sensitive to fluctuations of the plasma temperature inside the data set. A use of the slopes for constructing the calibration graphs significantly reduces the error bars but it does not eliminate the point scattering caused by the matrix effect, which is also responsible for large differences in the average plasma temperatures among the samples. Well aligned calibration points were obtained after identifying the couples of transitions less sensitive to variations of the plasma temperature, and this was achieved by simple theoretical simulations. Such selection of the analytical lines minimizes the matrix effect, and together with the chosen calibration approach, allows to measure the relative element concentrations even in highly unstable laser induced plasmas.
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•Here used sample preparation traps the particles and enhances laser-particle interaction.•Particle weights on the substrate and inside the laser spot are estimated by on-line photography.•Calibration based on growth among two lines is weakly sensitive to temperature fluctuations.•Different plasma temperatures among the samples produce scattering in the calibration graphs.•Matrix effect is reduced by a suitable choice of the line pairs less affected by the plasma temperature.
Cell-cell interactions are an observable manifestation of underlying complex biological processes occurring in response to diversified biochemical stimuli. Recent experiments with microfluidic ...devices and live cell imaging show that it is possible to characterize cell kinematics via computerized algorithms and unravel the effects of targeted therapies. We study the influence of spatial and temporal resolutions of time-lapse videos on motility and interaction descriptors with computational models that mimic the interaction dynamics among cells. We show that the experimental set-up of time-lapse microscopy has a direct impact on the cell tracking algorithm and on the derived numerical descriptors. We also show that, when comparing kinematic descriptors in two diverse experimental conditions, too low resolutions may alter the descriptors' discriminative power, and so the statistical significance of the difference between the two compared distributions. The conclusions derived from the computational models were experimentally confirmed by a series of video-microscopy acquisitions of co-cultures of unlabelled human cancer and immune cells embedded in 3D collagen gels within microfluidic devices. We argue that the experimental protocol of acquisition should be adapted to the specific kind of analysis involved and to the chosen descriptors in order to derive reliable conclusions and avoid biasing the interpretation of results.
We propose and analyze a class of vectorial crystallization problems, with applications to crystallization of anisotropic molecules and collective behavior such as birds flocking and fish schooling. ...We focus on two-dimensional systems of “oriented” particles: Admissible configurations are represented by vectorial empirical measures with density in
S
1
. We endow such configurations with a graph structure, where the bonds represent the “convenient” interactions between particles, and the proposed variational principle consists in maximizing their number. The class of bonds is determined by hard sphere type pairwise potentials, depending both on the distance between the particles and on the angles between the segment joining two particles and their orientations, through threshold criteria. Different ground states emerge by tuning the angular dependence in the potential, mimicking ducklings swimming in a row formation and predicting as well, for some specific values of the angular parameter, the so-called
diamond formation
in fish schooling.
Organs On a Chip (OOCs) represent a sophisticated approach for exploring biological mechanisms and developing therapeutic agents. In conjunction with high-quality time-lapse microscopy (TLM), OOCs ...allow for the visualization of reconstituted complex biological processes, such as multi-cell-type migration and cell–cell interactions. In this context, increasing the frame rate is desirable to reconstruct accurately cell-interaction dynamics. However, a trade-off between high resolution and carried information content is required to reduce the overall data volume. Moreover, high frame rates increase photobleaching and phototoxicity. As a possible solution for these problems, we report a new hybrid-imaging paradigm based on the integration of OOC/TLMs with a Multi-scale Generative Adversarial Network (GAN) predicting interleaved video frames with the aim to provide high-throughput videos. We tested the performance of the predictive capability of GAN on synthetic videos, as well as on real OOC experiments dealing with tumor–immune cell interactions. The proposed approach offers the possibility to acquire a reduced number of high-quality TLM images without any major loss of information on the phenomena under investigation.
The incremented uptake provided by time-lapse microscopy in Organ-on-a-Chip (OoC) devices allowed increased attention to the dynamics of the co-cultured systems. However, the amount of information ...stored in long-time experiments may constitute a serious bottleneck of the experimental pipeline. Forward long-term prediction of cell trajectories may reduce the spatial-temporal burden of video sequences storage. Cell trajectory prediction becomes crucial especially to increase the trustworthiness in software tools designed to conduct a massive analysis of cell behavior under chemical stimuli. To address this task, we transpose here the exploitation of the presence of "social forces" from the human to the cellular level for motion prediction at microscale by adapting the potential of Social Generative Adversarial Network predictors to cell motility. To demonstrate the effectiveness of the approach, we consider here two case studies: one related to PC-3 prostate cancer cells cultured in 2D Petri dishes under control and treated conditions and one related to an OoC experiment of tumor-immune interaction in fibrosarcoma cells. The goodness of the proposed strategy has been verified by successfully comparing the distributions of common descriptors (kinematic descriptors and mean interaction time for the two scenarios respectively) from the trajectories obtained by video analysis and the predicted counterparts.
By mimicking naturally occurring superhydrophobic surfaces, scientists can now realize artificial surfaces on which droplets of a few microliters of water are forced to assume an almost spherical ...shape and an extremely high contact angle. In recent decades, these surfaces have attracted much attention due to their technological applications for anti-wetting and self-cleaning materials. Very recently, researchers have shifted their interest to investigate whether superhydrophobic surfaces can be exploited to study biological systems. This research effort has stimulated the design and realization of new devices that allow us to actively organize, visualize and manipulate matter at both the microscale and nanoscale levels. Such precise control opens up wide applications in biomedicine, as it allows us to directly manipulate objects at the typical length scale of cells and macromolecules. This progress report focuses on recent biological and medical applications of superhydrophobicity. Particular regard is paid to those applications that involve the detection, manipulation and study of extremely small quantities of molecules, and to those that allow high throughput cell and biomaterial screening.
In this paper we present a simple and robust method to realize highly ordered arrays of stretched and suspended DNA molecules over the millimeter length scale. To this end we used an ad hoc designed ...superhydrophobic surface made of high aspect-ratio silicon pillars, where we deposited a droplet containing genomic DNA. A precise positioning of DNA strands was achieved by shaping the silicon pillars so that sharpened features resembling tips were included. Such features allowed us to accurately control the droplet de-wetting dynamics, pinning DNA strands in a well-defined position above pillars. The proposed technique has the potential to positively impact on the development of novel DNA chips for genetic analysis.
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► A superhydrophobic structure has been realized for wet sample handling in X-ray measurements. ► First measuring tests has been carried on for X-ray fluorescence, phase contrast ...imaging, and X-ray diffraction. ► Test measurements on ferritin protein sample has been done.
In this work we propose a wet sample handling technique which enables the simultaneous collection of X-ray fluorescence (XRF) spectra and X-ray phase contrast imaging (XPCI) using a few microliters drop confined on a superhydrophobic surface. To this purpose, we fabricated and tested a superhydrophobic patterned surface entailing an hydrophilic region which leads to pinning of the drop, enabling X-ray beam effortless alignment and measuring in liquid phase. Our technique allows to acquire capillary-free XRF spectra, resulting in a significant fluorescence detection gain, which is particularly interesting for light elements. Simultaneous XPCI provides sample geometry assuring a fine control of the experimental conditions and allowing real time monitoring of the drop during measurements. As an additional advantage – after solvent evaporation – the solute is deposited on a precise spot, greatly increasing its concentration allowing further measurements, such as X-ray microanalysis and X-ray diffraction.
These results could have potential applications in the study of blood proteins, such as ferritin and low density lipoprotein, which are usually available in very limited quantity.
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