Enhanced differentiation of human embryonic stem cells towards definitive endoderm on ultrahigh aspect ratio nanopillars

Differentiation of human embryonic stem cells is widely studied as a potential unlimited source for cell replacement therapy to treat degenerative diseases such as diabetes. The directed differentiation of human embryonic stem cells relies mainly on soluble factors. Although, some studies have highlighted that the properties of the physical environment, such as substrate stiffness, affect cellular behavior. Here, mass-produced, injection molded polycarbonate nanopillars are presented, where the surface mechanical properties, i.e., stiffness, can be controlled by the geometric design of the ultrahigh aspect ratio nanopillars (stiffness can be reduced by 25.0003). It is found that tall nanopillars, yielding softer surfaces, significantly enhance the induction of definitive endoderm cells from pluripotent human embryonic stem cells, resulting in more consistent differentiation of a pure population compared to planar control. By contrast, further differentiation toward the pancreatic ­endoderm is less successful on “soft” pillars when compared to “stiff” pillars or control, indicating differential cues during the different stages of differentiation. To accompany the mechanical properties of the nanopillars, the concept of surface shear modulus is introduced to describe the characteristics of engineered elastic surfaces through micro or nanopatterning. This provides a framework whereby comparisons can be drawn between such materials and bulk elastomeric materials

An ellipsometer with variable angle of incidence for studies in ultrahigh vacuum

The windows for the vacuum chamber are incorporated into the optical bench system by means of flexible bellows which allow measurements to be made over a large range of angle of incidence, one of which is chosen so that maximum sensitivity is obtained. The principal angle of incidence was determined, and straightforward corrections for strain birefringence of vacuum chamber windows were made. Atomically clean surfaces of sodium chloride and lithium fluoride were investigated to verify the performance of the system. Submonolayer and monolayer coverage of water on these surfaces could be detected. On cleavage planes of NaCl, a first monolayer of adsorbed water is complete at about one torr only

Recent Milestones in Unraveling the Full-Field Structure of Dynamic Shear Cracks and Fault Ruptures in Real-Time: From Photoelasticity to Ultrahigh-Speed Digital Image Correlation

The last few decades have seen great achievements in dynamic fracture mechanics. Yet, it was not possible to experimentally quantify the full-field behavior of dynamic fractures, until very recently. Here, we review our recent work on the full-field quantification of the temporal evolution of dynamic shear ruptures. Our newly developed approach based on digital image correlation combined with ultrahigh-speed photography has revolutionized the capabilities of measuring highly transient phenomena and enabled addressing key ques- tions of rupture dynamics. Recent milestones include the visualization of the complete displacement, particle velocity, strain, stress and strain rate fields near growing ruptures, capturing the evolution of dynamic friction during individual rupture growth, and the detailed study of rupture speed limits. For example, dynamic friction has been the big- gest unknown controlling how frictional ruptures develop but it has been impossible, until now, to measure dynamic friction during spontaneous rupture propagation and to understand its dependence on other quantities. Our recent measurements allow, by simul- taneously tracking tractions and sliding speeds on the rupturing interface, to disentangle its complex dependence on the slip, slip velocity, and on their history. In another application, we have uncovered new phenomena that could not be detected with previous methods, such as the formation of pressure shock fronts associated with “supersonic” propagation of shear ruptures in viscoelastic materials where the wave speeds are shown to depend strongly on the strain rate

An Ultrahigh-throughput Microfluidic Platform for Single-cell Genome Sequencing.

Sequencing technologies have undergone a paradigm shift from bulk to single-cell resolution in response to an evolving understanding of the role of cellular heterogeneity in biological systems. However, single-cell sequencing of large populations has been hampered by limitations in processing genomes for sequencing. In this paper, we describe a method for single-cell genome sequencing (SiC-seq) which uses droplet microfluidics to isolate, amplify, and barcode the genomes of single cells. Cell encapsulation in microgels allows the compartmentalized purification and tagmentation of DNA, while a microfluidic merger efficiently pairs each genome with a unique single-cell oligonucleotide barcode, allowing >50,000 single cells to be sequenced per run. The sequencing data is demultiplexed by barcode, generating groups of reads originating from single cells. As a high-throughput and low-bias method of single-cell sequencing, SiC-seq will enable a broader range of genomic studies targeted at diverse cell populations

Axon diameters and myelin content modulate microscopic fractional anisotropy at short diffusion times in fixed rat spinal cord

Mapping tissue microstructure accurately and noninvasively is one of the frontiers of biomedical imaging. Diffusion Magnetic Resonance Imaging (MRI) is at the forefront of such efforts, as it is capable of reporting on microscopic structures orders of magnitude smaller than the voxel size by probing restricted diffusion. Double Diffusion Encoding (DDE) and Double Oscillating Diffusion Encoding (DODE) in particular, are highly promising for their ability to report on microscopic fractional anisotropy ({\mu}FA), a measure of the pore anisotropy in its own eigenframe, irrespective of orientation distribution. However, the underlying correlates of {\mu}FA have insofar not been studied. Here, we extract {\mu}FA from DDE and DODE measurements at ultrahigh magnetic field of 16.4T in the aim to probe fixed rat spinal cord microstructure. We further endeavor to correlate {\mu}FA with Myelin Water Fraction (MWF) derived from multiexponential T2 relaxometry, as well as with literature-based spatially varying axonal diameters. In addition, a simple new method is presented for extracting unbiased {\mu}FA from three measurements at different b-values. Our findings reveal strong anticorrelations between {\mu}FA (derived from DODE) and axon diameter in the distinct spinal cord tracts; a moderate correlation was also observed between {\mu}FA derived from DODE and MWF. These findings suggest that axonal membranes strongly modulate {\mu}FA, which - owing to its robustness towards orientation dispersion effects - reflects axon diameter much better than its typical FA counterpart. The {\mu}FA exhibited modulations when measured via oscillating or blocked gradients, suggesting selective probing of different parallel path lengths and providing insight into how those modulate {\mu}FA metrics. Our findings thus shed light into the underlying microstructural correlates of {\mu}FA and are (...

UHMWPE/SBA-15 nanocomposites synthesized by in situ polymerization

Different nanocomposites have been attained by in situ polymerization based on ultra-high molecular weight polyethylene (UHMWPE) and mesoporous SBA-15, this silica being used for immobilization of the FI catalyst bis [N-(3-tert-butylsalicylidene)-2,3,4,5,6-pentafluoroanilinato] titanium (IV) dichloride and as filler as well. Two distinct approaches have been selected for supporting the FI catalyst on the SBA-15 prior polymerization. A study on polymerization activity of this catalyst has been performed under homogenous conditions and upon heterogenization. A study of the effect of presence of mesoporous particles and of the immobilization method is also carried out. Moreover, the thermal characterization, phase transitions and mechanical response of some pristine UHMWPEs and UHMWPE/SBA-15 materials have been carried out. Relationships with variations on molar mass, impregnation method of catalyst and final SBA-15 content have been established