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Extensive research has demonstrated the potential of cell viscoelastic properties as intrinsic indicators of cell state, functionality, and disease. For this, several microfluidic techniques have been developed to measure cell viscoelasticity with high-throughput. However, current microchannel designs introduce complex stress distributions on cells, leading to inaccuracies in determining the stress--strain relationship and, consequently, the viscoelastic properties. Here, we introduce a novel approach using hyperbolic microchannels that enable precise measurements under a constant extensional stress and offer a straightforward stress--strain relationship, while operating at a measurement rate of up to 100 cells per second. We quantified the stresses acting in the channels using mechanical calibration particles made from polyacrylamide (PAAm) and found that the measurement buffer, a solution of methyl cellulose and phosphate buffered saline, shows strain-thickening following a power law up to 200 s-1. By measuring oil droplets with varying viscosities, we successfully detected changes in the relaxation times of the droplets and our approach could be used to get the interfacial tension and viscosity of liquid--liquid droplet systems from the same measurement. We further applied this methodology to PAAm microgel beads, demonstrating the accurate recovery of Young's moduli and the near-ideal elastic behavior of the beads. To explore the influence of altered cell viscoelasticity, we treated HL60 human leukemia cells with latrunculin B and nocodazole, resulting in clear changes in cell stiffness while relaxation times were only minimally affected. In conclusion, our approach offers a streamlined and time-efficient solution for assessing the viscoelastic properties of large cell populations and other microscale soft particles. [ABSTRACT FROM AUTHOR]

Sorting cells is an essential primary step in many biological and clinical applications such as high-throughput drug screening, cancer research and cell transplantation. Cell sorting based on their mechanical properties has long been considered as a promising label-free biomarker that could revolutionize the isolation of cells from heterogeneous populations. Recent advances in microfluidic image-based cell analysis combined with subsequent label-free sorting by on-chip actuators demonstrated the possibility of sorting cells based on their physical properties. However, the high purity of sorting is achieved at the expense of a sorting rate that lags behind the analysis throughput. Furthermore, stable and reliable system operation is an important feature in enabling the sorting of small cell fractions from a concentrated heterogeneous population. Here, we present a label-free cell sorting method, based on the use of focused travelling surface acoustic wave (FTSAW) in combination with real-time deformability cytometry (RT-DC). We demonstrate the flexibility and applicability of the method by sorting distinct blood cell types, cell lines and particles based on different physical parameters. Finally, we present a new strategy to sort cells based on their mechanical properties. Our system enables the sorting of up to 400 particles per s. Sorting is therefore possible at high cell concentrations (up to 36 million per ml) while retaining high purity (>92%) for cells with diverse sizes and mechanical properties moving in a highly viscous buffer. Sorting of small cell fraction from a heterogeneous population prepared by processing of small sample volume (10 μl) is also possible and here demonstrated by the 667-fold enrichment of white blood cells (WBCs) from raw diluted whole blood in a continuous 10-hour sorting experiment. The real-time analysis of multiple parameters together with the high sensitivity and high-throughput of our method thus enables new biological and therapeutic applications in the future. [ABSTRACT FROM AUTHOR]

We investigate the rheological properties of a non-Newtonian glass-former liquid within lithographically defined microchannels in the range of temperatures above the vitrification region. The non-Newtonian behavior of the fluid, as evidenced by rotational rheology, is well described by a power law dependence of the viscosity on the shear rate. Taking into account such non-Newtonian character in the equations for the microfluidic motion, we relate the penetration dynamics into capillaries with the liquid rheological properties. The temperature dependence of the viscosity, determined over 1 order of magnitude in the temperature range 286-333 K and for shear rates between 0.07 and 1 [s.sup.-1], can be described by a Vogel--Fulcher--Tamman law, consistent with the fragile nature of the investigated compound. Microfluidics is a promising analytical approach for the investigation of the rheology of non-Newtonian fluids within confined microenvironments.

Intracellular delivery of cargo molecules such as membrane-impermeable proteins or drugs is crucial for cell treatment in biological and medical applications. Recently, microfluidic mechanoporation techniques have enabled transfection of previously inaccessible cells. These techniques create transient pores in the cell membrane by shear-induced or constriction contact-based rapid cell deformation. However, cells deform and recover differently from a given extent of shear stress or compression and it is unclear how the underlying mechanical properties affect the delivery efficiency of molecules into cells. In this study, we identify cell elasticity as a key mechanical determinant of delivery efficiency leading to the development of "progressive mechanoporation" (PM), a novel mechanoporation method that improves delivery efficiency into cells of different elasticity. PM is based on a multistage cell deformation, through a combination of hydrodynamic forces that pre-deform cells followed by their contact-based compression inside a PDMS-based device controlled by a pressure-based microfluidic controller. PM allows processing of small sample volumes (about 20 μL) with high-throughput (>10 000 cells per s), while controlling both operating pressure and flow rate for a reliable and reproducible cell treatment. We find that uptake of molecules of different sizes is correlated with cell elasticity whereby delivery efficiency of small and big molecules is favoured in more compliant and stiffer cells, respectively. A possible explanation for this opposite trend is a different size, number and lifetime of opened pores. Our data demonstrates that PM reliably and reproducibly delivers impermeable cargo of the size of small molecule inhibitors such as 4 kDa FITC-dextran with >90% efficiency into cells of different mechanical properties without affecting their viability and proliferation rates. Importantly, also much larger cargos such as a >190 kDa Cas9 protein–sgRNA complex are efficiently delivered high-lighting the biological, biomedical and clinical applicability of our findings. [ABSTRACT FROM AUTHOR]

We present a study of the rheological phenomenology of a non-Newtonian glass former within hybrid microchannels above the vitrification region. We determined the temperature behavior of the viscosity, which is well fitted by a Vogel-Fulcher-Tamman law for shear rates between 4×10-2 and 9×10-1 s-1. The microflow viscosity was compared with previously reported conductivity data of the investigated molecular system. Our findings provide an insight into the coupling between the structural dynamics in the bulk and that within the microchannels, suggesting lithographically defined microfluidic systems as promising tools for the investigation of the rheological properties of complex liquids. [ABSTRACT FROM AUTHOR]

Cells alter the path of light, a fact that leads to well‐known aberrations in single cell or tissue imaging. Optical diffraction tomography (ODT) measures the biophysical property that causes these aberrations, the refractive index (RI). ODT is complementary to fluorescence imaging and does not require any markers. The present study introduces RI and fluorescence tomography with optofluidic rotation (RAFTOR) of suspended cells, facilitating the segmentation of the 3D‐correlated RI and fluorescence data for a quantitative interpretation of the nuclear RI. The technique is validated with cell phantoms and used to confirm a lower nuclear RI for HL60 cells. Furthermore, the nuclear inversion of adult mouse photoreceptor cells is observed in the RI distribution. The applications shown confirm predictions of previous studies and illustrate the potential of RAFTOR to improve our understanding of cells and tissues. [ABSTRACT FROM AUTHOR]

We introduce real-time deformability cytometry (RT-DC) for continuous cell mechanical characterization of large populations (>100,000 cells) with analysis rates greater than 100 cells/s. RT-DC is sensitive to cytoskeletal alterations and can distinguish cell-cycle phases, track stem cell differentiation into distinct lineages and identify cell populations in whole blood by their mechanical fingerprints. This technique adds a new marker-free dimension to flow cytometry with diverse applications in biology, biotechnology and medicine. [ABSTRACT FROM AUTHOR]

The potential integration of polymer nanofibers in photonic devices and circuits is a major driver for research on their waveguiding and optical gain properties. Emission in the near-infrared is especially important in this framework in view of the realization of nanofiber-based optical amplifiers. Here, the optical gain properties of electrospun fibers embedding near-infrared light-emitting molecules are investigated. Upon pulsed optical pumping, line narrowing typical of amplified spontaneous emission is observed, with gain of 5.5 cm−1 and threshold fluence down to 0.25 mJ cm−2. Importantly, the stimulated emission characteristics are strongly dependent on individual fiber characteristics and on the mutual alignment of nanofibers in arrays, thus being tailorable through the fiber architecture and assembling. These results open interesting perspectives for the exploitation of electrospun fibers as active components in the near-infrared range. [ABSTRACT FROM AUTHOR]

We present a bio-inspired renal microdevice that resembles the in vivo structure of a kidney proximal tubule. For the first time, a population of tubular adult renal stem/progenitor cells (ARPCs) was embedded into a microsystem to create a bioengineered renal tubule. These cells have both multipotent differentiation abilities and an extraordinary capacity for injured renal cell regeneration. Therefore, ARPCs may be considered a promising tool for promoting regenerative processes in the kidney to treat acute and chronic renal injury. Here ARPCs were grown to confluence and exposed to a laminar fluid shear stress into the chip, in order to induce a functional cell polarization. Exposing ARPCs to fluid shear stress in the chip led the aquaporin-2 transporter to localize at their apical region and the Na+K+ATPase pump at their basolateral portion, in contrast to statically cultured ARPCs. A recovery of urea and creatinine of (20±5)% and (13±5)%, respectively, was obtained by the device. The microengineered biochip here-proposed might be an innovative “lab-on-a-chip” platform to investigate in vitro ARPCs behaviour or to test drugs for therapeutic and toxicological responses. [ABSTRACT FROM AUTHOR]

Spatiotemporal image correlation spectroscopy (STICS) is a simple and powerful technique, well established as a tool to probe protein dynamics in cells. Recently, its potential as a tool to map velocity fields in lab-on-a-chip systems was discussed. However, the lack of studies on its performance has prevented its use for microfluidics applications. Here, we systematically and quantitatively explore STICS microvelocimetry in microfluidic devices. We exploit a simple experimental setup, based on a standard bright-field inverted microscope (no fluorescence required) and a high-fps camera, and apply STICS to map liquid flow in polydimethylsiloxane (PDMS) microchannels. Our data demonstrates optimal 2D velocimetry up to 10 mm/s flow and spatial resolution down to 5 μm. [ABSTRACT FROM AUTHOR]

Artificial surfaces that exhibit unidirectional waterspreadingand superhydrophobicity are obtained by Strelitziareginaeleaves. Both green and dried leaves are used,thus exploiting the plant senescence. We demonstrate that the naturaldrying process of the leaves strongly affects the surface morphologyand wettability. Polymeric stamps from the green leaf show an arrangementof periodic microridges/microgrooves that favor anisotropic wetting,with a water contact angle (WCA) variation of about 21% along thetwo principal directions. Instead, the shrinkage of the leaf tissue,as a consequence of the natural dehydration process, induces an enhancementof the superficial corrugation. This results in the establishmentof a superhydrophobic state, which shows a WCA of up to 160°,and water rolling off. S. reginaeleavesare therefore easily accessible stamps suitable for controlling wettabilityand realizing surfaces that exhibit various wetting behaviors. [ABSTRACT FROM AUTHOR]

Flows in microcapillaries and associated imbibition phenomenaplaya major role across a wide spectrum of practical applications, fromoil recovery to inkjet printing and from absorption in porous materialsand water transport in trees to biofluidic phenomena in biomedicaldevices. Early investigations of spontaneous imbibition in capillariesled to the observation of a universal scaling behavior, known as theLucas–Washburn (LW) law. The LW allows abstraction of manyreal-life effects, such as the inertia of the fluid, irregularitiesin the wall geometry, and the finite density of the vacuum phase (gasor vapor) within the channel. Such simplifying assumptions set a constrainton the design of modern microfluidic devices, operating at ever-decreasingspace and time scales, where the aforementioned simplifications gounder serious question. Here, through a combined use of leading-edgeexperimental and simulation techniques, we unravel a novel interplaybetween global shape and nanoscopic roughness. This interplay significantlyaffects the early-stage energy budget, controlling front propagationin corrugated microchannels. We find that such a budget is governedby a two-scale phenomenon: The global geometry sets the conditionsfor small-scale structures to develop and propagate ahead of the mainfront. These small-scale structures probe the fine-scale details ofthe wall geometry (nanocorrugations), and the additional frictionthey experience slows the entire front. We speculate that such a two-scalemechanism may provide a fairly general scenario to account for extradissipative phenomena occurring in capillary flows with nanocorrugatedwalls. [ABSTRACT FROM AUTHOR]

Producing polymeric or hybrid microfluidic devices operating at high temperatures with reduced or no water evaporation is a challenge for many on-chip applications including polymerase chain reaction (PCR). We study sample evaporation in polymeric and hybrid devices, realized by glass microchannels for avoiding water diffusion toward the elastomer used for chip fabrication. The method dramatically reduces water evaporation in PCR devices that are found to exhibit optimal stability and effective operation under oscillating-flow. This approach maintains the flexibility, ease of fabrication, and low cost of disposable chips, and can be extended to other high-temperature microfluidic biochemical reactors. [ABSTRACT FROM AUTHOR]

Nanoparticle image velocimetry (nano-PIV), based on total internal reflection fluorescent microscopy, is very useful to investigate fluid flows within ∼100 nm from a surface; but so far it has only been applied to flow over smooth surfaces. Here we show that it can also be applied to flow over a topologically structured surface, provided that the surface structures can be carefully configured not to disrupt the evanescent-wave illumination. We apply nano-PIV to quantify the flow velocity distribution over a polydimethylsiloxane surface, with a periodic gratinglike structure (with 215 nm height and 2 μm period) fabricated using our customized multilevel lithography method. The measured tracer displacement data are in good agreement with the computed theoretical values. These results demonstrate new possibilities to study the interactions between fluid flow and topologically structured surfaces. [ABSTRACT FROM AUTHOR]

In this letter, we demonstrate an unexpected surface-acoustic-wave (SAW)-driven pumping effect in hydrophobic polydimethilsiloxane (PDMS)-lithium niobate (LiNbO3) microchannels. Atomization within the fluidic channel followed by SAW-assisted coalescence leads to liquid counterflow with respect to the SAW propagation direction. This physical mechanism is contrasted with the acoustic-streaming process driving isolated drop displacement on piezoelectric substrates. This principle is shown not to be readily applicable to the present microchannel case. The proposed device geometry can be exploited to integrate micropumps into complex microfluidic chips, improving the portability of micro-total-analysis systems. [ABSTRACT FROM AUTHOR]

The authors demonstrate the possibility to control the fluid flow inside microfluidic networks by photoresponsive capillaries. The approach relies on the use of photochromic molecules undergoing reversible changes in their polarity when irradiated with light of specific wavelength, thus varying the wettability of cyclic olefin copolymer microchannels. The realized photoresponsive elements exhibit a decrease up to 20° between the water contact angles of the native and the irradiated surfaces, which could be exploited for enhancing the penetration flow rate of fluids inside microfluidic channels up to 25%. The photocontrollable microfluidic circuitry presents on-off valve behavior, allowing or blocking liquid filling processes on the base of optical control, thus allowing one to manipulate liquid flow within microfluidic networks without mechanical actuation parts. [ABSTRACT FROM AUTHOR]

The mechanical properties of cancer cells and their microenvironment contribute to breast cancer progression. While mechanosensing has been extensively studied using 2D substrates, much less is known about it in a physiologically more relevant 3D context. Here it is demonstrated that breast cancer tumor spheroids, growing in 3D polyethylene glycol‐heparin hydrogels, are sensitive to their environment stiffness. During tumor spheroid growth, compressive stresses of up to 2 kPa build up, as quantitated using elastic polymer beads as stress sensors. Atomic force microscopy reveals that tumor spheroid stiffness increases with hydrogel stiffness. Also, constituent cell stiffness increases in a Rho associated kinase (ROCK)‐ and F‐actin‐dependent manner. Increased hydrogel stiffness correlated with attenuated tumor spheroid growth, a higher proportion of cells in G0/G1 phase, and elevated levels of the cyclin‐dependent kinase inhibitor p21. Drug‐mediated ROCK inhibition not only reverses cell stiffening upon culture in stiff hydrogels but also increases tumor spheroid growth. Taken together, a mechanism by which the growth of a tumor spheroid can be regulated via cytoskeleton rearrangements in response to its mechanoenvironment is revealed here. Thus, the findings contribute to a better understanding of how cancer cells react to compressive stress when growing under confinement in stiff environments. [ABSTRACT FROM AUTHOR]

We report on the unique emission features of light-emitting fibers made of a prototype conjugated polymer, namely, poly[(9,9-dioctylfluorenyl-2,7-diyl)-co-(1,4-benzo-{2,1′-3}-thiadiazole)] (F8BT), realized by electrospinning with diameters in the range of 500-1000 nm. The fibers display emission polarized along their axis, evidencing a favoured alignment of the polymer molecules. Emission efficiency and time resolved measurements reveal an enhancement of both the quantum efficiency and the radiative rate (up to 22.5%) of the fibers compared to spin-coated films, shedding more light on their potential as miniaturized photon sources in optoelectronic devices requiring high recombination rates. [ABSTRACT FROM AUTHOR]

Multifunctional capability, flexible design, rugged lightweight construction and self-powered operation are desired attributes for electronics that directly interface with the human body or with advanced robotic systems. For these applications, piezoelectric materials, in forms that offer the ability to bend and stretch, are attractive for pressure/force sensors and mechanical energy harvesters. Here, we introduce a large area, flexible piezoelectric material that consists of sheets of electrospun fibres of the polymer poly[(vinylidenefluoride-co-trifluoroethylene]. The flow and mechanical conditions associated with the spinning process yield free-standing, three-dimensional architectures of aligned arrangements of such fibres, in which the polymer chains adopt strongly preferential orientations. The resulting material offers exceptional piezoelectric characteristics, to enable ultra-high sensitivity for measuring pressure, even at exceptionally small values (0.1?Pa). Quantitative analysis provides detailed insights into the pressure sensing mechanisms, and establishes engineering design rules. Potential applications range from self-powered micro-mechanical elements, to self-balancing robots and sensitive impact detectors. [ABSTRACT FROM AUTHOR]

The throughput of cell mechanical characterization has recently approached that of conventional flow cytometers. However, this very sensitive, label-free approach still lacks the specificity of molecular markers. Here we developed an approach that combines real-time 1D-imaging fluorescence and deformability cytometry in one instrument (RT-FDC), thus opening many new research avenues. We demonstrated its utility by using subcellular fluorescence localization to identify mitotic cells and test for mechanical changes in those cells in an RNA interference screen.

Measuring the average refractive index (RI) of spherical objects, such as suspended cells, in quantitative phase imaging (QPI) requires a decoupling of RI and size from the QPI data. This has been commonly achieved by determining the object's radius with geometrical approaches, neglecting light-scattering. Here, we present a novel QPI fitting algorithm that reliably uncouples the RI using Mie theory and a semi-analytical, corrected Rytov approach. We assess the range of validity of this algorithm in silico and experimentally investigate various objects (oil and protein droplets, microgel beads, cells) and noise conditions. In addition, we provide important practical cues for the analysis of spherical objects in QPI.

Cell stiffness is a sensitive indicator of physiological and pathological changes in cells, with many potential applications in biology and medicine. A new method, real-time deformability cytometry, probes cell stiffness at high throughput by exposing cells to a shear flow in a microfluidic channel, allowing for mechanical phenotyping based on single-cell deformability. However, observed deformations of cells in the channel not only are determined by cell stiffness, but also depend on cell size relative to channel size. Here, we disentangle mutual contributions of cell size and cell stiffness to cell deformation by a theoretical analysis in terms of hydrodynamics and linear elasticity theory. Performing real-time deformability cytometry experiments on both model spheres of known elasticity and biological cells, we demonstrate that our analytical model not only predicts deformed shapes inside the channel but also allows for quantification of cell mechanical parameters. Thereby, fast and quantitative mechanical sampling of large cell populations becomes feasible., (Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.)

Fully controlled liquid injection and flow in hydrophobic polydimethylsiloxane (PDMS) two-dimensional microchannel arrays based on on-chip integrated, low-voltage-driven micropumps are demonstrated. Our architecture exploits the surface-acoustic-wave (SAW) induced counterflow mechanism and the effect of nebulization anisotropies at crossing areas owing to lateral propagating SAWs. We show that by selectively exciting single or multiple SAWs, fluids can be drawn from their reservoirs and moved towards selected positions of a microchannel grid. Splitting of the main liquid flow is also demonstrated by exploiting multiple SAW beams. As a demonstrator, we show simultaneous filling of two orthogonal microchannels. The present results show that SAW micropumps are good candidates for truly integrated on-chip fluidic networks allowing liquid control in arbitrarily shaped two-dimensional microchannel arrays.

This paper presents prototypical microfluidic devices made by hybrid microchannels based on piezoelectric LiNbO(3) and polydimethylsiloxane. This system enables withdrawing micropumping by acoustic radiation in microchannels. The withdrawing configuration, integrated on chip, is here quantitatively investigated for the first time, and found to be related to the formation and coalescence dynamics of droplets within the microchannel, primed by surface acoustic waves. The growth dynamics of droplets is governed by the water diffusion on LiNbO(3), determining the advancement of the fluid front. Observed velocities are up to 2.6 mm s(-1) for 30 dBm signals applied to the interdigital transducer, corresponding to tens of nl s(-1), and the micropumping dynamics is described by a model taking into account an acoustic power exponentially decaying upon travelling along the microchannel. This straighforward and flexible micropumping approach is particularly promising for the withdrawing of liquids in lab-on-chip devices performing cycling transport of fluids and biochemical reactions.

Producing solvent-resistant microfluidic devices is a challenge for analytical chemistry and biochemistry. We demonstrate a simple and low-cost fabrication approach for the realization of solvent-resistant microchannels based on perfluoropolyether elastomers, exhibiting very low aspect ratios (0.01). The strength of the microchannels sealing is evaluated through the maximum internal pressure (1.52 MPa) prior to device failure, due to delamination at the bonded interface. This approach allows the elastic properties of silicone elastomers, suitable for high quality external connections, to be combined with the non-swelling character of perfluoropolyethers.