848 research outputs found
Quantitative Imaging With DNA-PAINT for Applications in Synaptic Neuroscience
Super-resolution (SR) microscopy techniques have been advancing the understanding of neuronal protein networks and interactions. Unraveling the arrangement of proteins with molecular resolution provided novel insights into neuron cytoskeleton structure and actin polymerization dynamics in synaptic spines. Recent improvements in quantitative SR imaging have been applied to synaptic protein clusters and with improved multiplexing technology, the interplay of multiple protein partners in synaptic active zones has been elucidated. While all SR techniques come with benefits and drawbacks, true molecular quantification is a major challenge with the most complex requirements for labeling reagents and careful experimental design. In this perspective, we provide an overview of quantitative SR multiplexing and discuss in greater detail the quantification and multiplexing capabilities of the SR technique DNA-PAINT. Using predictable binding kinetics of short oligonucleotides, DNA-PAINT provides two unique approaches to address multiplexed molecular quantification: qPAINT and Exchange-PAINT. With precise and accurate quantification and spectrally unlimited multiplexing, DNA-PAINT offers an attractive route to unravel complex protein interaction networks in neurons. Finally, while the SR community has been pushing technological advances from an imaging technique perspective, the development of universally available, small, efficient, and quantitative labels remains a major challenge in the field
Calculations of energy levels and lifetimes of low-lying states of barium and radium
We use the configuration interaction method and many-body perturbation theory
to perform accurate calculations of energy levels, transition amplitudes, and
lifetimes of low-lying states of barium and radium. Calculations for radium are
needed for the planning of measurements of parity and time invariance violating
effects which are strongly enhanced in this atom. Calculations for barium are
used to control the accuracy of the calculations.Comment: 8 page
Dual Magnetic Separator for TRIP
The TRIP facility, under construction at KVI, requires the production
and separation of short-lived and rare isotopes. Direct reactions,
fragmentation and fusion-evaporation reactions in normal and inverse kinematics
are foreseen to produce nuclides of interest with a variety of heavy-ion beams
from the superconducting cyclotron AGOR. For this purpose, we have designed,
constructed and commissioned a versatile magnetic separator that allows
efficient injection into an ion catcher, i.e., gas-filled stopper/cooler or
thermal ionizer, from which a low energy radioactive beam will be extracted.
The separator performance was tested with the production and clean separation
of Na ions, where a beam purity of 99.5% could be achieved. For
fusion-evaporation products, some of the features of its operation as a
gas-filled recoil separator were tested.Comment: accepted by Nucl.Instr. Meth., final versio
Radium single-ion optical clock
We explore the potential of the electric quadrupole transitions
7s\,^2S_{1/2} - 6d\,^2D_{3/2}, 6d\,^2D_{5/2} in radium isotopes as
single-ion optical frequency standards. The frequency shifts of the clock
transitions due to external fields and the corresponding uncertainties are
calculated. Several competitive Ra candidates with 223 - 229 are
identified. In particular, we show that the transition
7s\,^2S_{1/2}\,(F=2,m_F=0) - 6d\,^2D_{3/2}\,(F=0,m_F=0) at 828 nm in
Ra, with no linear Zeeman and electric quadrupole shifts, stands
out as a relatively simple case, which could be exploited as a compact, robust,
and low-cost atomic clock operating at a fractional frequency uncertainty of
. With more experimental effort, the Ra clocks
could be pushed to a projected performance reaching the level.Comment: 20 pages, 1 figur
Super-resolution imaging and estimation of protein copy numbers at single synapses with DNA-PAINT
In the brain, the strength of each individual synapse is defined by the complement of proteins present or the "local proteome." Activity-dependent changes in synaptic strength are the result of changes in this local proteome and posttranslational protein modifications. Although most synaptic proteins have been identified, we still know little about protein copy numbers in individual synapses and variations between synapses. We use DNA-point accumulation for imaging in nanoscale topography as a single-molecule super-resolution imaging technique to visualize and quantify protein copy numbers in single synapses. The imaging technique provides near-molecular spatial resolution, is unaffected by photobleaching, enables imaging of large field of views, and provides quantitative molecular information. We demonstrate these benefits by accessing copy numbers of surface AMPA-type receptors at single synapses of rat hippocampal neurons along dendritic segments
Development of a thermal ionizer as ion catcher
An effective ion catcher is an important part of a radioactive beam facility
that is based on in-flight production. The catcher stops fast radioactive
products and emits them as singly charged slow ions. Current ion catchers are
based on stopping in He and H gas. However, with increasing intensity of
the secondary beam the amount of ion-electron pairs created eventually prevents
the electromagnetic extraction of the radioactive ions from the gas cell. In
contrast, such limitations are not present in thermal ionizers used with the
ISOL production technique. Therefore, at least for alkaline and alkaline earth
elements, a thermal ionizer should then be preferred. An important use of the
TRIP facility will be for precision measurements using atom traps. Atom
trapping is particularly possible for alkaline and alkaline earth isotopes. The
facility can produce up to 10 s of various Na isotopes with the
in-flight method. Therefore, we have built and tested a thermal ionizer. An
overview of the operation, design, construction, and commissioning of the
thermal ionizer for TRIP will be presented along with first results for
Na and Na.Comment: 10 pages, 4 figures, XVth International Conference on Electromagnetic
Isotope Separators and Techniques Related to their Applications (EMIS 2007
Determination of transition frequencies in a single Ba ion
Transition frequencies between low-lying energy levels in a single trapped
Ba ion have been measured with laser spectroscopy referenced to
an optical frequency comb. By extracting the frequencies of one-photon and
two-photon components of the line shape using an eight-level optical Bloch
model, we achieved 0.1 MHz accuracy for the 5d D - 6p
P and 6s S - 5d D transition
frequencies, and 0.2 MHz for the 6s S - 6p P
transition frequency.Comment: 5 pages, 7 figures, submitted to Phys. Rev.
Review
Innovation in genomics, transcriptomics, and proteomics research has created a plethora of state-of-the-art techniques such as nucleic acid sequencing and mass-spectrometry-based proteomics with paramount impact in the life sciences. While current approaches yield quantitative abundance analysis of biomolecules on an almost routine basis, coupling this high content to spatial information in a single cell and tissue context is challenging. Here, current implementations of spatial omics are discussed and recent developments in the field of DNA-barcoded fluorescence microscopy are reviewed. Light is shed on the potential of DNA-based imaging techniques to provide a comprehensive toolbox for spatial genomics and transcriptomics and discuss current challenges, which need to be overcome on the way to spatial proteomics using high-resolution fluorescence microscopy
- …