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Independent two-color, two-photon uncaging of glutamate and GABA allows autonomous activation and inhibition of neuronal action potentials in brain slices with subcellular resolution.
A chronically implanted biocompatible electrochemical microsensor allows long-term recording of subsecond dopamine dynamics in vivo. The microsensor can reliably detect behaviorally evoked dopamine release from dopamine neurons in the brain over a period of months in rats.
Microscope imaging performance can be seriously degraded by optical inhomogeneities in biological samples. An adaptive optics approach using a spatial light modulator to divide the illumination wavefront into individually controllable subregions recovers near-diffraction–limited two-photon imaging performance in brain tissue.
A mouse strain in which cellular reprogramming factors are expressed from a defined genomic locus is presented. It will enable studies of reprogramming in multiple cell types as well as facilitate comparisons between induced pluripotent stem cells and embryonic stem cells. Also in this issue, a paper by Carey et al. presents related tools.
Mouse strains in which three or four cellular reprogramming factors are expressed from a defined genomic locus are presented. They will enable studies of reprogramming in multiple cell types as well as facilitate comparisons between induced pluripotent stem cells and embryonic stem cells. Also in this issue, a paper from Stadtfeld et al. presents related tools.
By combining a protein complementation assay with a transcriptional reporter assay based on short expressed oligonucleotide tags (EXTs), the authors monitor tyrosine kinase receptor dimerization in conjunction with effector recruitment and downstream signaling.
An efficient system for the reversion and modification of mouse gene trap alleles is presented. It is applicable to available collections of gene trap embryonic stem cell lines.
Selected reaction monitoring (SRM) is a powerful mass spectrometry technology to reliably detect selected protein targets, even those at very low abundance, but requires tedious assay development for each protein of interest. High-throughput SRM assay development is now possible by using crude synthetic peptide libraries without purification to represent each protein target.
A polarity-sensitive annexin-based biosensor called pSIVA becomes strongly fluorescent only after reversibly binding to the plasma membrane. pSIVA allows live-cell imaging of the apoptotic process in degenerating neurons in vitro and in vivo.
Single-molecule sequencing of poly(A)-tailed chromatin immunoprecipitated DNA proves equal in sensitivity and accuracy to amplification-based sequencing technologies and allows analysis of samples sizes as small as 50 pg.
Staining with a mitochondrial dye permits high-purity isolation of cardiomyocytes from embryonic and induced pluripotent stem cells of several species, without genetic modification.
A degradation pathway found in plants, dependent on the hormone auxin, can be transplanted and harnessed to induce rapid and reversible target protein degradation in both yeast and animal cells.
Microsources positioned with holographic optical tweezers can establish a highly localized, three-dimensional chemical gradient that allows the manipulation of polarization and migration in single cells.
Tissue-specific expression of microRNA sponges allows precise regulation of microRNA activity in living flies. The authors investigate the role of miR-8 in the formation of neuromuscular junctions in detail.
Methods for automated fluorescence imaging allow high-throughput examination of reporter expression patterns in zebrafish embryos. They are applied to mapping promoter-enhancer interactions in this organism.
An improved version of the GCaMP genetically encoded calcium indicator, called GCaMP3, has higher calcium affinity and increased baseline fluorescence, dynamic range and stability. GCaMP3 performs better than existing genetically encoded calcium indicators in several assays and organisms, including in vivo imaging of neuronal signaling in worms, flies and mice.
Neuronal stimulation with channelrhodopsin-2 is combined with calcium fluorescence imaging to study neural connections in intact Caenorhabditis elegans.
Fusion of the genetically-encoded calcium indicator GCaMP2 to synaptophysin localizes the sensor to neuron presynaptic terminals and conveys linear responsiveness over a wider range of spike frequencies. The sensor allowed measurement of synaptic activity caused by spiking as well as graded voltage signals during in vivo imaging in zebrafish.
A combination of scattering interferometry and single-molecule fluorescence microscopy allows visualization of both the position and orientation of single Simian virus 40 particles on lipid bilayers and provides evidence of viral interaction with receptors in membrane nanodomains.
A combination of forward and reverse two hybrid screening allows systematic identification of 'edgetic' or edge-specific alleles, which encode proteins that have lost a single physical interaction but for which other interactions remain unperturbed.
