Tuesday, December 12, 2017

Nature Methods Application Notes e-UPDATE: 12 December 2017

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12 December 2017 
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Hardware triggering: maximizing speed and efficiency for live cell imaging
www.nikoninstruments.com/Products/Inverted-Microscopes/Eclipse-Ti2 >
Live cell imaging experiments now require higher speeds and more data throughput than ever before. Nikon Instruments has robust tools that enable hardware triggering of imaging devices in microscopy via direct signaling between hardware. This minimizes delays, synchronizes devices, and reduces the exposure of specimens to light. This Application Note explains how Nikon's NIS-Elements hardware-triggering workflow operates, and details its benefits for common time-lapse acquisition routines.

The new 2D Superresolution mode for ZEISS Airyscan
www.zeiss.com/airyscan >
Utilizing a pinhole-plane imaging concept, Airyscan allows for simultaneous improvement in resolution and signal-to-noise by capitalizing on an innovative 32-channel GaAsP photomultiplier tube (PMT) array detector. Each detection channel functions as a very small pinhole to increase resolution while the overall detector design delivers better signal-to-noise than traditional GaAsP-based confocal systems. In the past, a stack of at least five z-slices had to be deconvolved to get usable images with an optical section thinner than 1 Airy unit. Now, the new 2D Superresolution mode for Airyscan delivers images with the thinnest optical section (0.2 Airy units) from a single image while maintaining the light collection efficiency of a much larger 1.25-Airy-unit pinhole.

Impact of the Claristep® Filtration System on Recovery and Adsorption of Various Therapeutic Proteins at Low Sample Volumes
www.sartorius.com/claristep >
We tested the novel filter device Claristep® for the preparation of protein samples containing one of four different target molecules, i.e. an anti-malaria vaccine candidate, aviscuminum, interferon alfa-2B or monoclonal antibody (mAb) P2G12. In comparison to the standard protocols for sample preparation prior to analysis by either reversed-phase or SEC HPLC, the Claristep® filter did not exhibit a significant difference in protein concentration if the target molecules were present with at least 1.0 g L-1. A significantly reduced product concentration was observed in the Claristep® filtered samples compared to the standard protocol for mAb P2G12 if the concentrations were lower. The protein adsorption to the filter material was well described by a Langmuir adsorption isotherm. Besides, the Claristep® filters facilitated a rapid sample preparation with minimal volume losses.

Monitoring neurite morphology and synapse formation in primary neurons for neurotoxicity assessments and drug screening
www.thermofisher.com/hcs >
Synapse formation during nervous system development and degeneration in the pathogenesis of human neurological diseases are highly regulated processes. Subtle changes in the environment of the complex neuronal network may cause either breakdown or creation of synaptic connections. Drug discovery screening for neurological diseases and compound neurotoxicity evaluation would benefit from robust, automated, quantitative in vitro assays that monitor neuronal function.

A study on the efficacy of Prime&Bond active, the new active universal adhesive from Dentsply Sirona
www.dentsplysirona.com/en-gb/products/restorative/prime-&-bond-active.html >
Prime&Bond active™ is a one-component universal dental adhesive which brings new advantages to the universal adhesive arena. For effective bonding, conventional Universal Dental adhesives require ideal moisture conditions to form a homogeneous adhesive layer, and in turn a strong bond between tooth and restoration. However, when too much or too little moisture is present this may not happen and bonding can fail. Prime&Bond active contains "Active-Guard" technology, which gives this adhesive smart properties that compensate for variations in moisture levels and ensure complete adhesive coverage of the tooth.

Quantitative live-cell analysis for optimization of culture conditions and evaluation of cell health in human induced pluripotent stem cell-derived neurons
www.essenbioscience.com/en/ >
In this application note, we describe methods and present validation data highlighting optimal culture conditions for evaluation of cell viability and neurite outgrowth in hiPSCderived neurons from Cellular Dynamic International (CDI, iCell Neurons). We also monitor neurite outgrowth and cellular viability in iCell Gluta Neurons from CDI using a quantitative, live-cell imaging and analysis approach with the IncuCyte® S3 over days/weeks in 96-well microplate culture. To exemplify a real-time imaging and analysis approach using hiPSCderived neurons, we assess neuronal excitotoxicity using the IncuCyte® S3 Phase/Fluorescent NeuroTrack applications multiplexed with Annexin V reagents. These assays outline optimal culture conditions for an example iPSC-derived neuronal system and demonstrate the ability of the IncuCyte approach for real-time, long-term quantitative analysis ofiPSC-derived neuronal cell health.

Gibson Assembly® Primer-Bridge End Joining (PBnJ™) Cloning
www.syntheticgenomics.com >
Gibson Assembly® cloning has proven to be useful as a molecular biology technique for the seamless assembly of synthetic and natural genes and large-scale genetic pathways. Notably, Gibson Assembly cloning has enabled the synthesis of the first bacterial genome1, the first synthetic cell2, and the first minimal cell3. Additionally, the Gibson Assembly method has been utilized for genetic recoding (genome-wide codon removal)4, the engineering of Cre recombinase for improved site-specificity5, and has been incorporated into a one-step method for cloning gRNA for the CRISPR-Cas9 system6.


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