Probing single cells using flow in microfluidic devices

Enabling fluids to be manipulated on the micron-scale, microfluidic technologies have facilitated major advances in how we study cells. In this review, we highlight key developments in how flow in microfluidic devices is exploited to investigate the behavior of individual cells, from trapping and positioning single cells to probing cell deformability. Exploiting the properties of fluids and flow patterns in microchannels makes it possible to study large populations of single cells at micron-length scales with increased throughput and efficiency

Divalent Cations Crosslink Vimentin Intermediate Filament Tail Domains to Regulate Network Mechanics

Intermediate filament networks in the cytoplasm and nucleus are critical for the mechanical integrity of metazoan cells. However, the mechanism of crosslinking in these networks and the origins of their mechanical properties are not understood. Here, we study the elastic behavior of in vitro networks of the intermediate filament protein vimentin. Rheological experiments reveal that vimentin networks stiffen with increasing concentrations of C

Mechanical Properties of the Cell Nucleus and the Effect of Emerin Deficiency

Nuclear structure and mechanics are gaining recognition as important factors that affect gene expression, development, and differentiation in normal function and disease, yet the physical mechanisms that govern nuclear mechanical stability remain unclear. Here we examined the physical properties of the cell nucleus by imaging fluorescently labeled components of the inner nucleus (chromatin and nucleoli) and the nuclear envelope (lamins and membranes) in nuclei deformed by micropipette aspiration (confocal imaged microdeformation). We investigated nuclei, both isolated and in intact, living cells, and found that nuclear volume significantly decreased by 60–70% during aspiration. While nuclear membranes exhibited blebbing and fluid characteristics during aspiration, the nuclear lamina exhibited behavior of a solid-elastic shell. Under large deformations of GFP-lamin A-labeled nuclei, we observed a decay of fluorescence intensity into the tip of the deformed tongue that we interpreted in terms of nonlinear, two-dimensional elasticity theory. Here we applied this method to study nuclear envelope stability in disease and found that mouse embryo fibroblasts lacking the inner nuclear membrane protein, emerin, had a significantly decreased ratio of the area expansion to shear moduli (K/μ) compared to wild-type cells (2.1 ± 0.2 versus 5.1 ± 1.3). These data suggest that altered nuclear envelope elasticity caused by loss of emerin could contribute to increased nuclear fragility in Emery-Dreifuss muscular dystrophy patients with mutations in the emerin gene. Based on our experimental results and theoretical considerations, we present a model describing how the nucleus is stabilized in the pipette. Such a model is essential for interpreting the results of any micropipette study of the nucleus and porous materials in general

Undersea constellations: The global biology of an endangered marine megavertebrate further informed through citizen science

The whale shark is an ideal flagship species for citizen science projects because of its charismatic nature, its size, and the associated ecotourism ventures focusing on the species at numerous coastal aggregation sites. An online database of whale shark encounters, identifying individuals on the basis of their unique skin patterning, captured almost 30,000 whale shark encounter reports from 1992 to 2014, with more than 6000 individuals identified from 54 countries. During this time, the number of known whale shark aggregation sites (hotspots) increased from 13 to 20. Examination of photo-identification data at a global scale revealed a skewed sex-ratio bias toward males (overall, more than 66%) and high site fidelity among individuals, with limited movements of sharks between neighboring countries but no records confirming large, ocean basin-scale migrations. Citizen science has been vital in amassing large spatial and temporal data sets to elucidate key aspects of whale shark life history and demographics and will continue to provide substantial long-term value

Universal Behavior of Membranes with Sterols

Lanosterol is the biosynthetic precursor of cholesterol and ergosterol, sterols that predominate in the membranes of mammals and lower eukaryotes, respectively. These three sterols are structurally quite similar, yet their relative effects on membranes have been shown to differ. Here we study the effects of cholesterol, lanosterol, and ergosterol on 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine lipid bilayers at room temperature. Micropipette aspiration is used to determine membrane material properties (area compressibility modulus), and information about lipid chain order (first moments) is obtained from deuterium nuclear magnetic resonance. We compare these results, along with data for membrane-bending rigidity, to explore the relationship between membrane hydrophobic thickness and elastic properties. Together, such diverse approaches demonstrate that membrane properties are affected to different degrees by these structurally distinct sterols, yet nonetheless exhibit universal behavior