Dimensions of Cellulose Nanocrystals from Cotton and Bacterial Cellulose: Comparison of Microscopy and Scattering Techniques.

Different microscopy and scattering methods used in the literature to determine the dimensions of cellulose nanocrystals derived from cotton and bacterial cellulose were compared to investigate potential bias and discrepancies. Atomic force microscopy (AFM), small-angle X-ray scattering (SAXS), depolarized dynamic light scattering (DDLS), and static light scattering (SLS) were compared. The lengths, widths, and heights of the particles and their respective distributions were determined by AFM. In agreement with previous work, the CNCs were found to have a ribbon-like shape, regardless of the source of cellulose or the surface functional groups. Tip broadening and agglomeration of the particles during deposition cause AFM-derived lateral dimensions to be systematically larger those obtained from SAXS measurements. The radius of gyration determined by SLS showed a good correlation with the dimensions obtained by AFM. The hydrodynamic lateral dimensions determined by DDLS were found to have the same magnitude as either the width or height obtained from the other techniques; however, the precision of DDLS was limited due to the mismatch between the cylindrical model and the actual shape of the CNCs, and to constraints in the fitting procedure. Therefore, the combination of AFM and SAXS, or microscopy and small-angle scattering, is recommended for the most accurate determination of CNC dimensions.

Effect of in-plane and out-of-plane bifurcated microfluidic channels on the flow of aggregating red blood cells.

The blood flow through our microvascular system is a renowned difficult process to understand because the complex flow behavior of blood is intertwined with the complex geometry it has to flow through. Conventional 2D microfluidics has provided important insights, but progress is hampered by the limitation of 2-D confinement. Here we use selective laser-induced etching to excavate non-planar 3-D microfluidic channels in glass that consist of two generations of bifurcations, heading towards more physiological geometries. We identify a cross-talk between the first and second bifurcation only when both bifurcations are in the same plane, as observed in 2D microfluidics. Contrarily, the flow in the branch where the second bifurcation is perpendicular to the first is hardly affected by the initial distortion. This difference in flow behavior is only observed when red blood cells are aggregated, due to the presence of dextran, and disappears by increasing the distance between both generations of bifurcations. Thus, 3-D structures scramble in-plane flow distortions, exemplifying the importance of experimenting with truly 3D microfluidic designs in order to understand complex physiological flow behavior.

Competition between deformation and free volume quantified by 3D image analysis of red blood cell.

Cells in living organisms are subjected to mechanical strains caused by external forces like overcrowding, resulting in strong deformations that affect cell function. We study the interplay between deformation and crowding of red blood cells (RBCs) in dispersions of nonabsorbing rod-like viruses. We identify a sequence of configurational transitions of RBC doublets, including configurations that can only be induced by long-ranged attraction: highly fluctuating T-shaped and face-to-face configurations at low, and doublets approaching a complete spherical configuration at high, rod concentrations. Complementary simulations are used to explore different energy contributions to deformation as well as the stability of RBC doublet configurations. Our advanced analysis of 3D reconstructed confocal images of RBC doublets quantifies the depletion interaction and the resulting deformation energy. Thus, we introduce a noninvasive, high-throughput platform that is generally applicable to investigate the mechanical response of biological cells to external forces and characterize their mechanical properties.

Synchronized, Spontaneous, and Oscillatory Detachment of Eukaryotic Cells: A New Tool for Cell Characterization and Identification.

Despite the importance of cell characterization and identification for diagnostic and therapeutic applications, developing fast and label-free methods without (bio)-chemical markers or surface-engineered receptors remains challenging. Here, we exploit the natural cellular response to mild thermal stimuli and propose a label- and receptor-free method for fast and facile cell characterization. Cell suspensions in a dedicated sensor are exposed to a temperature gradient, which stimulates synchronized and spontaneous cell-detachment with sharply defined time-patterns, a phenomenon unknown from literature. These patterns depend on metabolic activity (controlled through temperature, nutrients, and drugs) and provide a library of cell-type-specific indicators, allowing to distinguish several yeast strains as well as cancer cells. Under specific conditions, synchronized glycolytic-type oscillations are observed during detachment of mammalian and yeast-cell ensembles, providing additional cell-specific signatures. These findings suggest potential applications for cell viability analysis and for assessing the collective response of cancer cells to drugs.

Dynamics of liquid-liquid phase separation of wheat gliadins.

During wheat seeds development, storage proteins are synthetized and subsequently form dense protein phases, also called Protein Bodies (PBs). The mechanisms of PBs formation and the supramolecular assembly of storage proteins in PBs remain unclear. In particular, there is an apparent contradiction between the low solubility in water of storage proteins and their high local dynamics in dense PBs. Here, we probe the interplay between short-range attraction and long-range repulsion of a wheat gliadin isolate by investigating the dynamics of liquid-liquid phase separation after temperature quench. We do so using time-resolved small angle light scattering, phase contrast microscopy and rheology. We show that gliadins undergo liquid-liquid phase separation through Nucleation and Growth or Spinodal Decomposition depending on the quench depth. They assemble into dense phases but remain in a liquid-like state over an extended range of temperatures and concentrations. The analysis of phase separation kinetics reveals that the attraction strength of gliadins is in the same order of magnitude as other proteins. We discuss the respective role of competing interactions, protein intrinsic disorder, hydration and polydispersity in promoting local dynamics and providing this liquid-like behavior despite attractive forces.

The Connection between Biaxial Orientation and Shear Thinning for Quasi-Ideal Rods.

The complete orientational ordering tensor of quasi-ideal colloidal rods is obtained as a function of shear rate by performing rheo-SANS (rheology with small angle neutron scattering) measurements on isotropic fd-virus suspensions in the two relevant scattering planes, the flow-gradient (1-2) and the flow-vorticity (1-3) plane. Microscopic ordering can be identified as the origin of the observed shear thinning. A qualitative description of the rheological response by Smoluchowski, as well as Doi⁻Edwards⁻Kuzuu theory is possible, as we obtain a master curve for different concentrations, scaling the shear rate with the apparent collective rotational diffusion coefficient. However, the observation suggests that the interdependence of ordering and shear thinning at small shear rates is stronger than predicted. The extracted zero-shear viscosity matches the concentration dependence of the self-diffusion of rods in semi-dilute solutions, while the director tilts close towards the flow direction already at very low shear rates. In contrast, we observe a smaller dependence on the shear rate in the overall ordering at high shear rates, as well as an ever-increasing biaxiality.

X-ray scattering in the vorticity direction and rheometry from confined fluids.

An X-ray flexure-based microgap rheometer (X-FMR) has been designed for combining rheology and in situ small-angle X-ray scattering from the vorticity plane. The gap distance can be varied continuously from 500 µm down to several µm, which provides the unique possibility to generate a strong confinement for many complex fluids. A singular advantage of this setup is the possibility to directly probe the vorticity direction of the flow field with a microfocus X-ray beam and to probe the structural response of the fluid to combined shear and confinement in the vorticity plane. The sliding-plate setup operates over a wide range of shear rates of γ = 10(-3)-10(3) s(-1) and strains in the range of 10(-4)-10(2). The flexure-based bearing maintains the plate parallelism within 10(-5) rad. The X-FMR requires very small sample volumes on the order of 10 µl. The applicability of the device is demonstrated here with limited examples of a nematic suspension of fd virus (rods), and a crystalline suspension containing sterically stabilized polystyrene-butylacrylate latex particles.