1,251 research outputs found
Enhancing single-molecule photostability by optical feedback from quantum-jump detection
We report an optical technique that yields an enhancement of single-molecule
photostability, by greatly suppressing photobleaching pathways which involve
photoexcitation from the triplet state. This is accomplished by dynamically
switching off the excitation laser when a quantum-jump of the molecule to the
triplet state is optically detected. This procedure leads to a lengthened
single-molecule observation time and an increased total number of detected
photons. The resulting improvement in photostability unambiguously confirms the
importance of photoexcitation from the triplet state in photobleaching
dynamics, and may allow the investigation of new phenomena at the
single-molecule level
An evaporation-based model of thermal neutron induced ternary fission of plutonium
Ternary fission probabilities for thermal neutron induced fission of
plutonium are analyzed within the framework of an evaporation-based model where
the complexity of time-varying potentials, associated with the neck collapse,
are included in a simplistic fashion. If the nuclear temperature at scission
and the fission-neck-collapse time are assumed to be ~1.2 MeV and ~10^-22 s,
respectively, then calculated relative probabilities of ternary-fission
light-charged-particle emission follow the trends seen in the experimental
data. The ability of this model to reproduce ternary fission probabilities
spanning seven orders of magnitude for a wide range of light-particle charges
and masses implies that ternary fission is caused by the coupling of an
evaporation-like process with the rapid re-arrangement of the nuclear fluid
following scission.Comment: 25 pages, 12 figures, accepted for publication in IJMP
Tissue-specific expression of high-voltage-activated dihydropyridine-sensitive L-type calcium channels
The cloning of the cDNA for the α1 subunit of L-type calcium channels revealed that at least two genes (CaCh1 and CaCh2) exist which give rise to several splice variants. The expression of mRNA for these α1 subunits and the skeletal muscle α2/δ, β and γ subunits was studied in rabbit tissues and BC3H1 cells. Nucleic-acid-hybridization studies showed that the mRNA of all subunits are expressed in skeletal muscle, brain, heart and aorta. However, the α1-, β- and γ-specific transcripts had different sizes in these tissues. Smooth muscle and heart contain different splice variants of the CaCh2 gene. The α1, β and γ mRNA are expressed together in differentiated but not in proliferating BC3H1 cells. A probe specific for the skeletal muscle α2/δ subunit did not hybridize to poly(A)-rich RNA from BC3H1 cells. These results suggest that different splice variants of the genes for the α1, β and γ subunits exist in tissues containing L-type calcium channels, and that their expression is regulated in a coordinate manner
Fluorescence Photooxidation with Eosin - a Method for High-Resolution Immunolocalization and in-Situ Hybridization Detection for Light and Electron-Microscopy
A simple method is described for high-resolution light and electron microscopic immunolocalization of proteins in cells and tissues by immunofluorescence and subsequent photooxidation of diaminobenzidine tetrahydrochloride into an insoluble osmiophilic polymer. By using eosin as the fluorescent marker, a substantial improvement in sensitivity is achieved in the photooxidation process over other conventional fluorescent compounds. The technique allows for precise correlative immunolocalization studies on the same sample using fluorescence, transmitted light and electron microscopy. Furthermore, because eosin is smaller in size than other conventional markers, this method results in improved penetration of labeling reagents compared to gold or enzyme based procedures. The improved penetration allows for three-dimensional immunolocalization using high voltage electron microscopy. Fluorescence photooxidation can also be used for high resolution light and electron microscopic localization of specific nucleic acid sequences by in situ hybridization utilizing biotinylated probes followed by an eosin-streptavidin conjugate
Changes in intracellular ion activities induced by adrenaline in human and rat skeletal muscle
To study the stimulating effect of adrenaline (ADR) on active Na+/K+ transport we used double-barrelled ion-sensitive micro-electrodes to measure the activities of extracellular K+ (aKe) and intracellular Na+ (aNai) in isolated preparations of rat soleus muscle, normal human intercostal muscle and one case of hyperkalemic periodic paralysis (h.p.p.). In these preparations bath-application of ADR (10−6 M) resulted in a membrane hyperpolarization and transient decreasesaKe andaNai which could be blocked by ouabain (3×10−4 M). In the h.p.p. muslce a continuous rise ofaNai induced by elevation ofaKe to 5.2 mM could be stopped by ADR. In addition, the intracellular K+ activity (aKi), the free intracellular Ca2+ concentration (pCai) and intracellular pH (pHi) were monitored in rat soleus muscle. During ADRaKi increased, pHi remained constant and intracellular Ca2+ apparently decreased. In conclusion, our data show that ADR primarily stimulates the Na+/K+ pump in mammalian skeletal muscle. This stimulating action is not impaired in the h.p.p. muscle
Perfect Fluid Theory and its Extensions
We review the canonical theory for perfect fluids, in Eulerian and Lagrangian
formulations. The theory is related to a description of extended structures in
higher dimensions. Internal symmetry and supersymmetry degrees of freedom are
incorporated. Additional miscellaneous subjects that are covered include
physical topics concerning quantization, as well as mathematical issues of
volume preserving diffeomorphisms and representations of Chern-Simons terms (=
vortex or magnetic helicity).Comment: 3 figure
Fluorescent Protein-Based Methods for On-Plate Screening of Gene Insertion
Unlike the commonly used method of blue-white screening for gene insertion, a fluorescent protein-based screening method offers a gain-of-function screening process without using any co-factors and a gene fusion product with a fluorescent protein reporter that is further useful in cell imaging studies. However, complications related to protein-folding efficiencies of the gene insert in fusion with fluorescent protein reporters prevent effective on-plate bacterial colony selection leading to its limited use.Here, we present three methods to tackle this problem. Our first method promotes the folding of the gene insert by using an N-terminal protein such as calmodulin that is well folded and expressed. Under this method, fluorescence was increased more than 30x over control allowing for enhanced screening. Our second method creates a fluorescent protein that is N-terminal to the gene upon insertion, thereby reducing the dependency of the fluorescent protein reporter on the folding of the gene insert. Our third method eliminates any dependence of the fluorescent protein reporter on the folding of the gene insert by using a stop and start sequence for protein translation.The three methods together will expand the usefulness of fluorescence on-plate screening and offer a powerful alternative to blue-white screening
Advanced optical imaging in living embryos
Developmental biology investigations have evolved from static studies of embryo anatomy and into dynamic studies of the genetic and cellular mechanisms responsible for shaping the embryo anatomy. With the advancement of fluorescent protein fusions, the ability to visualize and comprehend how thousands to millions of cells interact with one another to form tissues and organs in three dimensions (xyz) over time (t) is just beginning to be realized and exploited. In this review, we explore recent advances utilizing confocal and multi-photon time-lapse microscopy to capture gene expression, cell behavior, and embryo development. From choosing the appropriate fluorophore, to labeling strategy, to experimental set-up, and data pipeline handling, this review covers the various aspects related to acquiring and analyzing multi-dimensional data sets. These innovative techniques in multi-dimensional imaging and analysis can be applied across a number of fields in time and space including protein dynamics to cell biology to morphogenesis
Determining the neurotransmitter concentration profile at active synapses
Establishing the temporal and concentration profiles of neurotransmitters during synaptic release is an essential step towards understanding the basic properties of inter-neuronal communication in the central nervous system. A variety of ingenious attempts has been made to gain insights into this process, but the general inaccessibility of central synapses, intrinsic limitations of the techniques used, and natural variety of different synaptic environments have hindered a comprehensive description of this fundamental phenomenon. Here, we describe a number of experimental and theoretical findings that has been instrumental for advancing our knowledge of various features of neurotransmitter release, as well as newly developed tools that could overcome some limits of traditional pharmacological approaches and bring new impetus to the description of the complex mechanisms of synaptic transmission
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