118 research outputs found
High-throughput continuous dielectrophoretic separation of neural stem cells.
We created an integrated microfluidic cell separation system that incorporates hydrophoresis and dielectrophoresis modules to facilitate high-throughput continuous cell separation. The hydrophoresis module consists of a serpentine channel with ridges and trenches to generate a diverging fluid flow that focuses cells into two streams along the channel edges. The dielectrophoresis module is composed of a chevron-shaped electrode array. Separation in the dielectrophoresis module is driven by inherent cell electrophysiological properties and does not require cell-type-specific labels. The chevron shape of the electrode array couples with fluid flow in the channel to enable continuous sorting of cells to increase throughput. We tested the new system with mouse neural stem cells since their electrophysiological properties reflect their differentiation capacity (e.g., whether they will differentiate into astrocytes or neurons). The goal of our experiments was to enrich astrocyte-biased cells. Sorting parameters were optimized for each batch of neural stem cells to ensure effective and consistent separations. The continuous sorting design of the device significantly improved sorting throughput and reproducibility. Sorting yielded two cell fractions, and we found that astrocyte-biased cells were enriched in one fraction and depleted from the other. This is an advantage of the new continuous sorting device over traditional dielectrophoresis-based sorting platforms that target a subset of cells for enrichment but do not provide a corresponding depleted population. The new microfluidic dielectrophoresis cell separation system improves label-free cell sorting by increasing throughput and delivering enriched and depleted cell subpopulations in a single sort
Exact Hawking Radiation of Scalars, Fermions, and Bosons Using the Tunneling Method Without Back-Reaction
Hawking radiation is studied for arbitrary scalars, fermions, and spin-1
bosons, using a tunneling approach, to every order in but ignoring
back-reaction effects. It is shown that the additional quantum terms yield no
new contribution to the Hawking temperature. Indeed, it is found that the limit
of small in the standard quantum WKB approximation is replaced by the
near-horizon limit in the gravitational WKB approach.Comment: 8 pages, no figures. v3: Introduction updated. Version to appear in
PL
Hawking Radiation due to Photon and Gravitino Tunneling
Applying the Hamilton--Jacobi method we investigate the tunneling of photon
across the event horizon of a static spherically symmetric black hole. The
necessity of the gauge condition on the photon field, to derive the
semiclassical Hawking temperature, is explicitly shown. Also, the tunneling of
photon and gravitino beyond this semiclassical approximation are presented
separately. Quantum corrections of the action for both cases are found to be
proportional to the semiclassical contribution. Modifications to the Hawking
temperature and Bekenstein-Hawking area law are thereby obtained. Using this
corrected temperature and Hawking's periodicity argument, the modified metric
for the Schwarzschild black hole is given. This corrected version of the
metric, upto order is equivalent to the metric obtained by including
one loop back reaction effect. Finally, the coefficient of the leading order
correction of entropy is shown to be related to the trace anomaly.Comment: LaTex, 19 pages, no figures, extended version, new references added,
to appear in Annals of Phy
Back reaction, emission spectrum and entropy spectroscopy
Recently, an interesting work, which reformulates the tunneling framework to
directly produce the Hawking emission spectrum and entropy spectroscopy in the
tunneling picture, has been received a broad attention. However, during the
emission process, most related observations have not incorporated the effects
of back reaction on the background spacetime, whose derivations are therefore
not the desiring results for the real physical process. With this point as a
central motivation, in this paper we suitably adapt the \emph{reformulated}
tunneling framework so that it can well accommodate the effects of back
reaction to produce the Hawking emission spectrum and entropy spectroscopy.
Consequently, we interestingly find that, when back reaction is considered, the
Parikh-Wilczek's outstanding observations that, an isolated radiating black
hole has an unitary-evolving emission spectrum that is \emph{not} precisely
thermal, but is related to the change of the Bekenstein-Hawking entropy, can
also be reproduced in the reformulated tunneling framework, meanwhile the
entropy spectrum has the same form as that without inclusion of back reaction,
which demonstrates the entropy quantum is \emph{independent} of the effects of
back reaction. As our final analysis, we concentrate on the issues of the black
hole information, but \emph{unfortunately} find that, even including the
effects of back reaction and higher-order quantum corrections, such tunneling
formalism can still not provide a mechanism for preserving the black hole
information.Comment: 16 pages, no figure, use JHEP3.cls. to be published in JHE
Quantum corrections and black hole spectroscopy
In the work \cite{BRM,RBE}, black hole spectroscopy has been successfully
reproduced in the tunneling picture. As a result, the derived entropy spectrum
of black hole in different gravity (including Einstein's gravity,
Einstein-Gauss-Bonnet gravity and Ho\v{r}ava-Lifshitz gravity) are all evenly
spaced, sharing the same forms as , where physical process is only
confined in the semiclassical framework. However, the real physical picture
should go beyond the semiclassical approximation. In this case, the physical
quantities would undergo higher-order quantum corrections, whose effect on
different gravity shares in different forms. Motivated by these facts, in this
paper we aim to observe how quantum corrections affect black hole spectroscopy
in different gravity. The result shows that, in the presence of higher-order
quantum corrections, black hole spectroscopy in different gravity still shares
the same form as , further confirming the entropy quantum is universal
in the sense that it is not only independent of black hole parameters, but also
independent of higher-order quantum corrections. This is a desiring result for
the forthcoming quantum gravity theory.Comment: 14 pages, no figure, use JHEP3.cls. to be published in JHE
In situ functional dissection of RNA cis-regulatory elements by multiplex CRISPR-Cas9 genome engineering.
RNA regulatory elements (RREs) are an important yet relatively under-explored facet of gene regulation. Deciphering the prevalence and functional impact of this post-transcriptional control layer requires technologies for disrupting RREs without perturbing cellular homeostasis. Here we describe genome-engineering based evaluation of RNA regulatory element activity (GenERA), a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 platform for in situ high-content functional analysis of RREs. We use GenERA to survey the entire regulatory landscape of a 3'UTR, and apply it in a multiplex fashion to analyse combinatorial interactions between sets of miRNA response elements (MREs), providing strong evidence for cooperative activity. We also employ this technology to probe the functionality of an entire MRE network under cellular homeostasis, and show that high-resolution analysis of the GenERA dataset can be used to extract functional features of MREs. This study provides a genome editing-based multiplex strategy for direct functional interrogation of RNA cis-regulatory elements in a native cellular environment
Tunnelling Methods and Hawking's radiation: achievements and prospects
The aim of this work is to review the tunnelling method as an alternative
description of the quantum radiation from black holes and cosmological
horizons. The method is first formulated and discussed for the case of
stationary black holes, then a foundation is provided in terms of analytic
continuation throughout complex space-time. The two principal implementations
of the tunnelling approach, which are the null geodesic method and the
Hamilton-Jacobi method, are shown to be equivalent in the stationary case. The
Hamilton-Jacobi method is then extended to cover spherically symmetric
dynamical black holes, cosmological horizons and naked singularities. Prospects
and achievements are discussed in the conclusions.Comment: Topical Review commissioned and accepted for publication by
"Classical and Quantum Gravity". 101 pages; 6 figure
Nasal DNA methylation at three CpG sites predicts childhood allergic disease
Childhood allergic diseases, including asthma, rhinitis and eczema, are prevalent conditions that share strong genetic and environmental components. Diagnosis relies on clinical history and measurements of allergen-specific IgE. We hypothesize that a multi-omics model could accurately diagnose childhood allergic disease. We show that nasal DNA methylation has the strongest predictive power to diagnose childhood allergy, surpassing blood DNA methylation, genetic risk scores, and environmental factors. DNA methylation at only three nasal CpG sites classifies allergic disease in Dutch children aged 16 years well, with an area under the curve (AUC) of 0.86. This is replicated in Puerto Rican children aged 9-20 years (AUC 0.82). DNA methylation at these CpGs additionally detects allergic multimorbidity and symptomatic IgE sensitization. Using nasal single-cell RNA-sequencing data, these three CpGs associate with influx of T cells and macrophages that contribute to allergic inflammation. Our study suggests the potential of methylation-based allergy diagnosis
Dynamic and Polarized Muscle Cell Behaviors Accompany Tail Morphogenesis in the Ascidian Ciona intestinalis
BACKGROUND: Axial elongation is a key morphogenetic process that serves to shape developing organisms. Tail extension in the ascidian larva represents a striking example of this process, wherein paraxially positioned muscle cells undergo elongation and differentiation independent of the segmentation process that characterizes the formation of paraxial mesoderm in vertebrates. Investigating the cell behaviors underlying the morphogenesis of muscle in ascidians may therefore reveal the evolutionarily conserved mechanisms operating during this process. METHODOLOGY/PRINCIPLE FINDINGS: A live cell imaging approach utilizing subcellularly-localized fluorescent proteins was employed to investigate muscle cell behaviors during tail extension in the ascidian Ciona intestinalis. Changes in the position and morphology of individual muscle cells were analyzed in vivo in wild type embryos undergoing tail extension and in embryos in which muscle development was perturbed. Muscle cells were observed to undergo elongation in the absence of positional reorganization. Furthermore, high-speed high-resolution live imaging revealed that the onset and progression of tail extension were characterized by the presence of dynamic and polarized actin-based protrusive activity at the plasma membrane of individual muscle cells. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that in the Ciona muscle, tissue elongation resulted from gradual and coordinated changes in cell geometry and not from changes in cell topology. Proper formation of muscle cells was found to be necessary not only for muscle tissue elongation, but also more generally for completion of tail extension. Based upon the characterized dynamic changes in cell morphology and plasma membrane protrusive activity, a three-phase model is proposed to describe the cell behavior operating during muscle morphogenesis in the ascidian embryo
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