186 research outputs found
Exclusion-zone dynamics explored with microfluidics and optical tweezers
The exclusion zone (EZ) is a boundary region devoid of macromolecules and microscopic particles formed spontaneously in the vicinity of hydrophilic surfaces. The exact mechanisms behind this remarkable phenomenon are still not fully understood and are debated. We measured the short- and long-time-scale kinetics of EZ formation around a Nafion gel embedded in specially designed microfluidic devices. The time-dependent kinetics of EZ formation follow a power law with an exponent of 0.6 that is strikingly close to the value of 0.5 expected for a diffusion-driven process. By using optical tweezers we show that exclusion forces, which are estimated to fall in the sub-pN regime, persist within the fully-developed EZ, suggesting that EZ formation is not a quasi-static but rather an irreversible process. Accordingly, the EZ-forming capacity of the Nafion gel could be exhausted with time, on a scale of hours in the presence of 1 mM Na2HPO4. EZ formation may thus be a non-equilibrium thermodynamic cross-effect coupled to a diffusion-driven transport process. Such phenomena might be particularly important in the living cell by providing mechanical cues within the complex cytoplasmic environment. © 2014 by the authors; licensee MDPI, Basel, Switzerland
Exclusion zone phenomena in water -- a critical review of experimental findings and theories
The existence of the exclusion zone (EZ), a layer of water in which plastic
microspheres are repelled from hydrophilic surfaces, has now been independently
demonstrated by several groups. A better understanding of the mechanisms which
generate EZs would help with understanding the possible importance of EZs in
biology and in engineering applications such as filtration and microfluidics.
Here we review the experimental evidence for EZ phenomena in water and the
major theories that have been proposed. We review experimental results from
birefringence, neutron radiography, nuclear magnetic resonance, and other
studies. Pollack and others have theorized that water in the EZ exists has a
different structure than bulk water, and that this accounts for the EZ. We
present several alternative explanations for EZs and argue that Schurr's theory
based on diffusiophoresis presents a compelling alternative explanation for the
core EZ phenomenon. Among other things, Schurr's theory makes predictions about
the growth of the EZ with time which have been confirmed by Florea et al. and
others. We also touch on several possible confounding factors that make
experimentation on EZs difficult, such as charged surface groups, dissolved
solutes, and adsorbed nanobubbles.Comment: 14 pg
Physical properties of red blood cells in aggregation
Red blood cells (RBC) are micron-sized biological objects and the main corpuscular constituent of blood. It flows from larger arteries to very small capillaries. Utilizing a physical approach, this work aims to assess properties that govern blood flows and in particular the disaggregation and aggregation mechanisms of RBC at a single cell level. The interactions of RBCs are thus, investigated experimentally by measuring adhesive forces in the pN range in various model solutions thanks to optical tweezers. While two models for aggregation have been proposed: bridging and depletion, experimental evidence is still lacking to decide which mechanism prevails. The research presented here provides a new insight on the aggregation of RBCs and shows that the two models may not be exclusive. A complete 3-dimensional phase diagram of doublets has been established and confirmed by experiments by varying the adhesive forces and reduced cell volumes. Besides, the effect of aggregation was studied in vitro in a bifurcating microcapillary network and the distribution of aggregates and their stability in such a geometry are reported. Finally, experiments in flow allowed the characterization of the flow field around single RBCs at different velocities. Interesting vortical fluid structures have been also observed thanks to tracer nanoparticles.Rote Blutkörperchen (Erythrozyten) sind biologische Objekte im Mikrometerbereich und der korpuskuläre Hauptbestandteil des Blutes. Es fließt aus größeren Arterien in sehr kleine Kapillaren. Unter Verwendung eines physikalischen Ansatzes zielt diese Arbeit darauf ab, die Eigenschaften zu bewerten, die den Blutfluss und insbesondere die Disaggregations- und Aggregationsmechanismen der RBC auf Einzelzellebene regeln. Die Interaktionen der Erythrozyten werden daher experimentell untersucht, indem Adhäsionskräfte im pN-Bereich in verschiedenen Modelllösungen mit Hilfe einer optischen Pinzette gemessen werden. Während mit Bridging und Depletion zwei Modelle für die Aggregation vorgeschlagen wurden, fehlen noch experimentelle Beweise, um zu entscheiden, welcher Mechanismus vorherrscht. Die hier vorgestellte Forschung liefert neue Erkenntnisse über die Aggregation von RBCs und zeigt, dass die beiden Modelle möglicherweise nicht exklusiv sind. Es wurde ein vollständiges dreidimensionales Phasendiagramm von Dubletten erstellt und experimentell durch Variation der Adhäsionskräfte und reduzierte Zellvolumina bestätigt. Außerdem wurde der Effekt der Aggregation in vitro in einem sich gabelförmigen Mikrokapillarnetz untersucht, und es wird über die Verteilung der Aggregate und ihre Stabilität in einer solchen Geometrie berichtet. Schließlich erlaubten Strömungsexperimente die Charakterisierung des Strömungsfeldes um einzelne RBCs bei unterschiedlichen Geschwindigkeiten. Dank Tracer-Nanopartikeln konnten auch interessante wirbelartige Fluidstrukturen beobachtet werden
Generation of versatile ss-dsDNA hybrid substrates for single-molecule analysis.
Here, we describe a rapid and versatile protocol to generate gapped DNA substrates for single-molecule (SM) analysis using optical tweezers via site-specific Cas9 nicking and force-induced melting. We provide examples of single-stranded (ss) DNA gaps of different length and position. We outline protocols to visualize these substrates by replication protein A-enhanced Green Fluorescent Protein (RPA-eGFP) and SYTOX Orange staining using commercially available optical tweezers (C-TRAP). Finally, we demonstrate the utility of these substrates for SM analysis of bidirectional growth of RAD-51-ssDNA filaments. For complete details on the use and execution of this protocol, please refer to Belan et al. (2021)
Size-dependent particle migration and trapping in 3D microbubble streaming flows
Acoustically actuated sessile bubbles can be used as a tool to manipulate
microparticles, vesicles and cells. In this work, using acoustically actuated
sessile semi-cylindrical microbubbles, we demonstrate experimentally that
finite-sized microparticles undergo size-sensitive migration and trapping
towards specific spatial positions in three dimensions with high
reproducibility. The particle trajectories are successfully reproduced by
passive advection of the particles in a steady three-dimensional streaming flow
field augmented with volume exclusion from the confining boundaries. For
different particle sizes, this volume exclusion mechanism leads to three
regimes of qualitatively different migratory behavior, suggesting applications
for separating, trapping, and sorting of particles in three dimensions.Comment: 12 pages, 7 figure
Advances in Optofluidics
Optofluidics a niche research field that integrates optics with microfluidics. It started with elegant demonstrations of the passive interaction of light and liquid media such as liquid waveguides and liquid tunable lenses. Recently, the optofluidics continues the advance in liquid-based optical devices/systems. In addition, it has expanded rapidly into many other fields that involve lightwave (or photon) and liquid media. This Special Issue invites review articles (only review articles) that update the latest progress of the optofluidics in various aspects, such as new functional devices, new integrated systems, new fabrication techniques, new applications, etc. It covers, but is not limited to, topics such as micro-optics in liquid media, optofluidic sensors, integrated micro-optical systems, displays, optofluidics-on-fibers, optofluidic manipulation, energy and environmental applciations, and so on
Optical Deformation of Microdroplets at Ultralow Interfacial Tension
What is the shape of a droplet? Its interfacial tension dictates that it is very close to a perfect sphere. Herein, the interfacial tension is reduced to ultralow values (0.1 - 100 uN/m) by careful formulation of surfactant additives, such as for mixtures that form microemulsions. The droplet need not be spherical but can accommodate external forces of a similar magnitude. The control and precision of
forces afforded simply by light - in the form of highly focused Nd:YAG laser beams - are exploited in this work to deform hydrocarbon oil-in-water emulsion droplets of
1-10 um diameter. To this end, a novel, integrated platform for microfluidic generation, optical deformation and 3D fluorescent imaging of droplets is presented. Previous attempts to characterise optically-controlled microdroplet shapes have been limited to 2D projections. Here, that ambiguity is resolved using 3D confocal laser scanning- and structured illumination microscopy. 2D and 3D arrays of up to four Gaussian point traps are generated by holograms and acousto-optics. A variety of regular, prolate, oblate and asymmetric shapes are produced and correlated with parameters such as optocapillary number, trap separation and capillary length. Exotic shapes exhibiting zero or negative mean and Gaussian curvatures are presented
alongside their brightfield counterparts. The complex phase behaviour of emulsion droplets and their parent phases is observed to couple strongly to thermal absorption of the beams. The rich interfacial chemistry, its relation to the forces determining droplet shape and the surprising
ability to create nanofluidic networks between droplets are investigated
Capture probabilities in pair-wise collisions of emulsion drops - measurement and application : report submitted in fulfilment of the requirements for the Doctoral Degree (PhD Course) in the Biophysics and Soft Matter Group, School of Fundamental Sciences, Massey University, New Zealand
This project seeks to measure and model particle interactions under different environmental conditions with a view to being able to control these interactions. The interactions of emulsion drops will be investigated using an optical tweezer set-up and the results considered in the context of measurements of the zeta potential of the emulsion.
Specifically, how the zeta potential of emulsion drops changes with the physio-chemical environment (pH and ionic environment) is captured in a concise mathematical model, the effects of depletion interactions are considered, and a novel experimental procedure is developed to allow hundreds of pairwise stickiness measurements to be taken in an automated fashion.
The major research questions are:
1. Is it possible to address the effects of changes in environmental conditions which are not easily quantifiable with a pragmatic capture probability or pairwise "stickiness" measured at a single particle level?
2. Can we link these pairwise measurements to induced changes in the surface properties and understand how they yield the rheological behaviour of the system
Single-molecule techniques in biophysics : a review of the progress in methods and applications
Single-molecule biophysics has transformed our understanding of the
fundamental molecular processes involved in living biological systems, but also
of the fascinating physics of life. Far more exotic than a collection of
exemplars of soft matter behaviour, active biological matter lives far from
thermal equilibrium, and typically covers multiple length scales from the
nanometre level of single molecules up several orders of magnitude to longer
length scales in emergent structures of cells, tissues and organisms.
Biological molecules are often characterized by an underlying instability, in
that multiple metastable free energy states exist which are separated by energy
levels of typically just a few multiples of the thermal energy scale of kBT,
where kB is the Boltzmann constant and T the absolute temperature, implying
complex, dynamic inter-conversion kinetics across this bumpy free energy
landscape in the relatively hot, wet environment of real, living biological
matter. The key utility of single-molecule biophysics lies in its ability to
probe the underlying heterogeneity of free energy states across a population of
molecules, which in general is too challenging for conventional ensemble level
approaches which measure mean average properties. Parallel developments in both
experimental and theoretical techniques have been key to the latest insights
and are enabling the development of highly-multiplexed, correlative techniques
to tackle previously intractable biological problems. Experimentally,
technological developments in the sensitivity and speed of biomolecular
detectors, the stability and efficiency of light sources, probes and
microfluidics, have enabled and driven the study of heterogeneous behaviours
both in vitro and in vivo that were previously undetectable by ensemble
methods..
Mass Transport via Thermoplasmonics
When a metallic nanoparticle is illuminated with light under resonant conditions, the free electron gas oscillates in such a way that substantial amplification of the local electric field amplitude is achieved – this is known as a plasmonic resonance. This resonance enhances both the optical scattering as well as absorption. In many applications, the enhanced scattering can facilitate efficient coupling between the near-field and the far-field, which enables optical interrogation of nanoscale volumes. Simultaneously, however, the enhanced absorption results in localized heating and substantial temperature gradients. The resulting temperature profile can drive other thermal processes, some beneficial others detrimental. Thermoplasmonics is the study of these plasmonically enhanced thermal processes. Elevated temperatures increase the Brownian motion of small particles. Moreover, if large temperature gradients are present, then a process known as thermophoresis is likely to occur. Thermophoresis tends to cause a local depletion of Brownian particles around a hot region. From the context of “conventional” plasmonic applications (like molecular sensing), these thermally driven mass transport mechanisms are adverse side effects since they reduce the interaction rate between the plasmonic system and the analyte. An investigation of thermal effects in plasmonic optical tweezers showed that the increased Brownian motion essentially negated the optical tweezing effect, resulting in an overall insensitivity between the resonance condition of the antenna and the particle confinement when evaluated in terms of the local temperature increase. Additionally, a significant thermophoretic depletion of analytes occurred, extending tens of microns from the plasmonic structure. This depletion acts in opposition to the plasmonically enhanced optical forces, which are restricted to a region of only a few hundred nanometres.However, thermoplasmonic effects can also be used for advantageous means. Once example is by driving thermocapillary flows directed towards the plasmonic system, thereby facilitating the efficient accumulation of analytes. One method of employing this effect is to superheat a plasmonic particle to a high enough temperature such that a bubble is nucleated. Once a bubble is formed, thermocapillary effects at the bubble interface drive fluid motion with a flow profile similar to that of a Stokeslet. This fluid flow can be utilized for analyte accumulation near the plasmonic structure. In addition to the thermocapillary induced flow, it was found that even more intense flow speeds were achieved immediately upon nucleation due to the mechanical action of the bubble. This transient peak in flow speed was approximately an order of magnitude faster than the subsequent persistent (thermocapillary) flow. By designing the plasmonic nanoparticle so that the Laplace pressure restricted the ultimate bubble size, these bubbles could be kept small enough to permit high modulation rates and maximize the relative effect of the peak transient flow
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