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A miniature portable tomograph allows the imaging of awake, behaving rats with positron emission tomography while simultaneously measuring their behavior. This method for noninvasive whole-brain imaging can link studies of brain activity with particular behaviors.
RNAi screening combined with automated microscopy of Drosophila melanogaster cells reveal genetic interactions and allow the reconstruction of signaling pathways.
Light sheet microscopy using a scanned Bessel beam in combination with structured illumination or two-photon excitation reduces photobleaching and phototoxicity, improves axial resolution and allows isotropic three-dimensional imaging. The authors demonstrate performance of the method via fast volumetric subcellular imaging of several dynamic processes in single living cells.
The combination of an ultrahigh-resolution dual optical trap with a confocal microscope allowed single-fluorophore detection of labeled oligonucleotide binding and simultaneous measurement of angstrom-scale changes in DNA tether extension.
Confined photoactivation of photoactivatable mCherry using two-photon illumination with line-scanning temporal focusing in combination with three-dimensional localization algorithms allows three-dimensional super-resolution microscopy of cellular features at <50 nm lateral and <100 nm axial resolution and depths greater than 8 μm.
A genetic multicolor cell-labeling technique for Droshophila melanogaster, Flybow, is described and applied to the study of neural circuits. This method implements a variant of the mouse Brainbow strategy in combination with specific neuronal targeting using the Gal-4–upstream activating sequence system to select for membrane-tethered fluorescent proteins. Also in this issue, Hampel et al. report a similar strategy, Drosophila Brainbow, to select for epitope-tagged proteins detectable via immunofluorescence.
A genetic multicolor cell-labeling technique for Droshophila melanogaster, Drosophila Brainbow, is described and applied to the study of neural circuits. This method implements a variant of the mouse Brainbow strategy in combination with specific neuronal targeting using the Gal-4–upstream activating sequence system to select for epitope-tagged proteins detectable with immunofluorescence. Also in this issue, Hadjieconomou et al. develop a similar strategy, Flybow, to select for membrane-tethered fluorescent proteins.
By methylating the phosphate groups of PtdIns(3,4,5)P3 researchers can load this lipid more efficiently into a mass spectrometer and thus this lipid can be quantified in the presence of an internal synthetic standard.
An optogenetic illumination system based on the use of a digital micromirror device and video tracking software is reported, which allows real-time light delivery with high spatial resolution to specified targets in freely moving Caenorhabditis elegans. Also in this issue, Stirman et al. report a similar illumination system using a liquid crystal display projector. Both methods allow optogenetic perturbation of a variety of neural circuits in the behaving worm.
An optogenetic illumination system based on the use of a liquid crystal display projector and video tracking software is reported, which allows real-time multispectral light delivery with high spatial resolution to specified targets in freely moving Caenorhabditis elegans. Also in this issue, Leifer et al. report a similar illumination system using a digital micromirror device. Both methods allow optogenetic perturbation of a variety of neural circuits in the behaving worm.
By individually replacing 16 yeast genes encoding ABC transporters by GFP, mating and selecting for strains with accumulated mutations the authors create a Green Monster, a strain with deletions in all 16 genes.