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TABLE OF CONTENTS
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December 2017 Volume 14, Issue 12 |
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| In This Issue Editorial This Month Correspondence Research Highlights Methods in Brief Tools in Brief Technology Feature News and Views Analysis Brief Communications Articles Erratum Corrigendum Application Notes | | Advertisement | | | | The new Mastercycler® X50 Take your fast PCR to the next level with the new Mastercycler X50 and 10 °C/s heating. Eppendorf's new 2D-Gradient allows optimization of the annealing and the denaturation temperature in a single run. This gives you the ideal set of temperatures for your PCR assay — quickly, conveniently, reliably. Your PCR will never be the same. | | | | |
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Inspiration does not obey speed limits. Announcing the ORCA-Lightning from Hamamatsu. Our new 12MP sCMOS camera engineered for high speed and low noise and ready for Lightsheet microscopy. hamamatsucameras.com | | | |
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In This Issue | Top |
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In This Issue |
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Editorial | Top |
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So you're writing a paper p1115 doi:10.1038/nmeth.4532 Writing a clear, accurate and convincing scientific paper is both an art and a skill. It is one well worth mastering. |
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This Month | Top |
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The Author File: Ulrich Elling p1117 Vivien Marx doi:10.1038/nmeth.4507 Improving single-cell data in CRISPR screens, with inspiration from open-air theater performances. |
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Points of Significance: Machine learning: a primer pp1119 - 1120 Danilo Bzdok, Martin Krzywinski and Naomi Altman doi:10.1038/nmeth.4526 Machine learning extracts patterns from data without explicit instructions. |
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Correspondence | Top |
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LiteMol suite: interactive web-based visualization of large-scale macromolecular structure data pp1121 - 1122 David Sehnal, Mandar Deshpande, Radka Svobodová Vařeková, Saqib Mir, Karel Berka et al. doi:10.1038/nmeth.4499 |
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Accessible virtual reality of biomolecular structural models using the Autodesk Molecule Viewer pp1122 - 1123 Aidin R Balo, Merry Wang and Oliver P Ernst doi:10.1038/nmeth.4506 |
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MDsrv: viewing and sharing molecular dynamics simulations on the web pp1123 - 1124 Johanna K S Tiemann, Ramon Guixà-Gonzalez, Peter W Hildebrand and Alexander S Rose doi:10.1038/nmeth.4497 |
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Research Highlights | Top |
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Evolution of an A-base editor Protein evolution yields a programmable base editor that can revert the most common pathogenic point mutations in the genome. | Sequencing for carbohydrates A technique combining mass spectrometry and infrared spectroscopy shows promise for unambiguous sequencing of linear oligosaccharides. | Multiscale 3D view of the genome Ultradeep sequencing of Hi-C data from mouse cells in different developmental stages can address outstanding questions about 3D chromatin function. | Glow-in-the-dark nanoparticles for deep imaging A class of semiconductor polymer particles can be used for afterglow imaging to avoid autofluorescence when imaging deep structures in living animals. | Imaging deep in the cortex and beyond Combining red calcium indicators with an optimized illumination strategy enables neuronal activity to be imaged in deep layers of the mouse cortex and hippocampus. | | | | |
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Methods in Brief | Top |
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Genome editing in the brain | Freestyle fluidics | Measuring cellular mass with a picobalance | Improving the efficiency of cryo-EM |
Tools in Brief | Top |
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Stem cells tip toward totipotency | Tracking cell migration in vivo | Haplobank of mutated mouse stem cell clones | A yTRAP for protein aggregates |
Technology Feature | Top |
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Microscopy: hello, adaptive optics pp1133 - 1136 Vivien Marx doi:10.1038/nmeth.4508 Aberrations hinder clear imaging, but an array of options help to correct much distortion. |
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News and Views | Top |
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SPOTs fill a major gap in RNA quantification pp1137 - 1138 Jinzhou Yuan and Peter A Sims doi:10.1038/nmeth.4515 Direct RNA quantification using barcoded fluorescence hybridization enables accurate and scalable gene expression profiling. See also: Brief Communication by Eng et al. |
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Mass spectrometry imaging goes three dimensional pp1139 - 1140 Klaus Dreisewerd and Joanne Y Yew doi:10.1038/nmeth.4513 Two complementary approaches improve 3D mass spectrometry imaging of biomolecules on 3D surfaces and within tissue sections and single cells. See also: Brief Communication by Kompauer et al. | Article by Passarelli et al. |
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Analysis | Top |
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An objective comparison of cell-tracking algorithms pp1141 - 1152 Vladimír Ulman, Martin Maška, Klas E G Magnusson, Olaf Ronneberger, Carsten Haubold et al. doi:10.1038/nmeth.4473 This analysis describes the results of three Cell Tracking Challenge editions for examining the performance of cell segmentation and tracking algorithms and provides practical feedback for users and developers. |
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Brief Communications | Top |
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Profiling the transcriptome with RNA SPOTs pp1153 - 1155 Chee-Huat Linus Eng, Sheel Shah, Julian Thomassie and Long Cai doi:10.1038/nmeth.4500 RNA SPOTs adapts sequential fluorescence in situ hybridization (seqFISH) for highly accurate and inexpensive transcriptome-scale or targeted RNA quantification in vitro. See also: News and Views by Yuan & Sims |
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Autofocusing MALDI mass spectrometry imaging of tissue sections and 3D chemical topography of nonflat surfaces pp1156 - 1158 Mario Kompauer, Sven Heiles and Bernhard Spengler doi:10.1038/nmeth.4433 A mass spectrometry imaging system combined with a laser triangulation system provides simultaneous topographic and molecular information for 3D biological samples and eliminates height-related artifacts in imperfect tissue sections. See also: News and Views by Dreisewerd & Yew |
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High-throughput, image-based screening of pooled genetic-variant libraries pp1159 - 1162 George Emanuel, Jeffrey R Moffitt and Xiaowei Zhuang doi:10.1038/nmeth.4495 Multiplexed FISH readout of barcoded genotypes in single cells allows pooled screening of large genetic-variant libraries for complex, image-based phenotypes. |
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Inducible and multiplex gene regulation using CRISPR-Cpf1-based transcription factors pp1163 - 1166 Y Esther Tak, Benjamin P Kleinstiver, James K Nuñez, Jonathan Y Hsu, Joy E Horng et al. doi:10.1038/nmeth.4483 Inducible dCpf1-based transcriptional activators achieve tunable, multiplexed target gene expression. |
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Antibodies to biotin enable large-scale detection of biotinylation sites on proteins pp1167 - 1170 Namrata D Udeshi, Kayvon Pedram, Tanya Svinkina, Shaunt Fereshetian, Samuel A Myers et al. doi:10.1038/nmeth.4465 Enrichment of biotinylated peptides using an anti-biotin antibody results in substantially improved biotinylation site identifications by mass spectrometry compared to traditional streptavidin-based biotinylated protein enrichment. |
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Deciphering lipid structures based on platform-independent decision rules pp1171 - 1174 Jürgen Hartler, Alexander Triebl, Andreas Ziegl, Martin Trotzmuller, Gerald N Rechberger et al. doi:10.1038/nmeth.4470 The Lipid Data Analyzer uses sets of decision rules to identify lipid subclasses in high-throughput LC-MS/MS data. |
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Advanced Imaging with Proven Optics Olympus is dedicated to your work, vision, and science. Through innovation and service, we seek to aide in your discoveries, advance your research, and inspire you to explore new possibilities. Our wide range of microscopes are built with the optical excellence and proven application expertise you can depend on. Your Science Matters™ > Learn more about Olympus microscopes | | | |
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Articles | Top |
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The 3D OrbiSIMS[mdash]label-free metabolic imaging with subcellular lateral resolution and high mass-resolving power pp1175 - 1183 Melissa K Passarelli, Alexander Pirkl, Rudolf Moellers, Dmitry Grinfeld, Felix Kollmer et al. doi:10.1038/nmeth.4504 The high spatial resolution of secondary ion mass spectrometry and the high resolving power of the Orbitrap mass spectrometer are combined in a single imaging platform, the 3D OrbiSIMS. The instrument's capabilities for resolving lipids and neurotransmitters in the brain with subcellular spatial resolution, and a drug in a single cell in three dimensions is demonstrated. See also: News and Views by Dreisewerd & Yew |
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Localization-based super-resolution imaging meets high-content screening pp1184 - 1190 Anne Beghin, Adel Kechkar, Corey Butler, Florian Levet, Marine Cabillic et al. doi:10.1038/nmeth.4486 An automated system for data acquisition and analysis enables high-content screening localization microscopy and increases the throughput and information content of super-resolution microscopy methods such as dSTORM, DNA-PAINT and (spt)PALM. |
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CRISPR-UMI: single-cell lineage tracing of pooled CRISPR-Cas9 screens pp1191 - 1197 Georg Michlits, Maria Hubmann, Szu-Hsien Wu, Gintautas Vainorius, Elena Budusan et al. doi:10.1038/nmeth.4466 Combining each sgRNA with a unique molecular identifier in a genome-wide screen increases sensitivity and robustness in both positive and negative selection. |
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Thiol-linked alkylation of RNA to assess expression dynamics pp1198 - 1204 Veronika A Herzog, Brian Reichholf, Tobias Neumann, Philipp Rescheneder, Pooja Bhat et al. doi:10.1038/nmeth.4435 SLAM seq detects s4U incorporation into RNA and, combined with 3′-end RNA-seq, provides insight into RNA turnover. |
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Isolation and 3D expansion of multipotent Sox9+ mouse lung progenitors pp1205 - 1212 Massimo Nichane, Asif Javed, V Sivakamasundari, Monisha Ganesan, Lay Teng Ang et al. doi:10.1038/nmeth.4498 This paper describes methods for the 3D culture of mouse lung progenitor cells that can differentiate in vitro and in vivo along all epithelial lineages. |
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Protein interaction perturbation profiling at amino-acid resolution pp1213 - 1221 Jonathan Woodsmith, Luise Apelt, Victoria Casado-Medrano, Ziya Özkan, Bernd Timmermann et al. doi:10.1038/nmeth.4464 A reverse two-hybrid approach combined with massively parallel programmed mutagenesis and sequencing allows profiling protein interaction determinants at amino-acid resolution, as demonstrated by identifying interaction-disrupting mutations in a cilia protein complex associated with Bardet-Biedl syndrome. |
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Application Notes | Top |
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Hardware triggering: maximizing speed and efficiency for live cell imaging Michael A Davis |
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The new 2D Superresolution mode for ZEISS Airyscan Joseph Huff, Annette Bergter, Jan Birkenbeil, Ingo Kleppe and Ralf Engelmann |
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Corrigendum | Top |
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Corrigendum: Accurate identification of single-nucleotide variants in whole-genome-amplified single cells p1222 Xiao Dong, Lei Zhang, Brandon Milholland, Moonsook Lee, Alexander Y Maslov et al. doi:10.1038/nmeth1217-1222b |
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Erratum | Top |
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Erratum: Probes: FRET sensor design and optimization p1222 Vivien Marx doi:10.1038/nmeth1217-1222a |
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