Tuesday, August 8, 2017

Nature Methods Application Notes e-UPDATE: 8 August 2017

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8 August 2017 
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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.

A humanized phenotypic screening platform for chronic pain
www.censobio.com >
www.Cellectricon.com >
Censo Biotechnologies and Cellectricon have joined forces to develop a first-in-kind humanized drug discovery platform to target chronic pain. The platform is based on human induced pluripotent stem cell–derived neurons in combination with Cellectricon's Cellaxess® Elektra screening system. The combination of highly functional screening with a human cell type of relevance for chronic pain has the potential to enable the generation of better drug candidates for this condition and other diseases.

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.

Turning neuroscience data into insight in the 21st century
www.metacell.us >
The modern research and industrial approach to neuroscience involves an increasingly vast assortment of data collection methods and data handling approaches. While data collection and accumulation are expanding, approaches to data management and insight gathering are still lacking.

Match & Scratch Barcodes: Tools for the Demultiplexing and Extraction of Target Sequences from PacBio Amplicon Data
www.jax.org >
One of the major challenges for the downstream analysis of amplicon data is to first demultiplex Fastq files based on the ligation of different oligonucleotides combinations. Match & Scratch Barcodes are a set of bioinformatics tools that support the analysis of PacBio sequenced long read amplicon data by detecting forward and reverse end adapter sequences, generic adapters attachedto the region specific oligos, multiple number of region specific oligos of variable length for the extraction of sequences of interest.

INCELLIS: Perfect tool to qualify the cell labelling before using a high end microscope
www.bertin-instruments.com >
Centrosome is an organelle implicated in cell cycle regulation. During cell division, the two centrosomes serve as anchors to guide the chromosomes and divide them into two equitable batches to form the two daughter cells. An aberrant number of centrosomes in a cell is often associated with cancer. Expression of GFP-centrin recombinant protein into in vitro cell culture allows direct visualization of centriole behavior in living cells in different experimental conditions. Nevertheless, as it is a small organelle, imaging of GFP-centrin protein into centrosome requires high sensitivity in fluorescence.

Robotic Microscopy with the Nikon Ti2 for high-content analysis applications
www.nikon.com >
Robotic Microscopy—a combination of high-content screening methods—enables multivariate experimental approaches with large cell populations and member-level sensitivity. Here we explore how the new Nikon Ti2 line of inverted research microscopes is uniquely suited to Robotic Microscopy applications, focusing on work utilizing induced pluripotent stem cells (iPSCs) as disease models in drug screening.


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