Abstract:
:Fluorescence microscopy of living cells enables visualization of the dynamics and interactions of intracellular molecules. However, fluorescence live-cell imaging is limited by photobleaching and phototoxicity induced by the excitation light. Here we describe controlled light-exposure microscopy (CLEM), a simple imaging approach that reduces photobleaching and phototoxicity two- to tenfold, depending on the fluorophore distribution in the object. By spatially controlling the light-exposure time, CLEM reduces the excitation-light dose without compromising image quality. We show that CLEM reduces photobleaching sevenfold in tobacco plant cells expressing microtubule-associated GFP-MAP4 and reduces production of reactive oxygen species eightfold and prolongs cell survival sixfold in HeLa cells expressing chromatin-associated H2B-GFP. In addition, CLEM increases the dynamic range of the fluorescence intensity at least twofold.
journal_name
Nat Biotechnoljournal_title
Nature biotechnologyauthors
Hoebe RA,Van Oven CH,Gadella TW Jr,Dhonukshe PB,Van Noorden CJ,Manders EMdoi
10.1038/nbt1278subject
Has Abstractpub_date
2007-02-01 00:00:00pages
249-53issue
2eissn
1087-0156issn
1546-1696pii
nbt1278journal_volume
25pub_type
杂志文章abstract::Foxtail millet (Setaria italica), a member of the Poaceae grass family, is an important food and fodder crop in arid regions and has potential for use as a C(4) biofuel. It is a model system for other biofuel grasses, including switchgrass and pearl millet. We produced a draft genome (∼423 Mb) anchored onto nine chrom...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.2195
更新日期:2012-05-13 00:00:00
abstract::Methods used to sequence the transcriptome often produce more than 200 million short sequences. We introduce StringTie, a computational method that applies a network flow algorithm originally developed in optimization theory, together with optional de novo assembly, to assemble these complex data sets into transcripts...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.3122
更新日期:2015-03-01 00:00:00
abstract::The scale and capabilities of single-cell RNA-sequencing methods have expanded rapidly in recent years, enabling major discoveries and large-scale cell mapping efforts. However, these methods have not been systematically and comprehensively benchmarked. Here, we directly compare seven methods for single-cell and/or si...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/s41587-020-0465-8
更新日期:2020-06-01 00:00:00
abstract::The treatment of asthma and allergic rhinitis using unique, humanized anti-IgE monoclonal antibodies with very particular binding specificities is now supported by the results of multiple phase II and III human clinical studies. The therapeutic efficacy of this approach is attributable to several pharmacological mecha...
journal_title:Nature biotechnology
pub_type: 杂志文章,评审
doi:10.1038/72601
更新日期:2000-02-01 00:00:00
abstract::Developments in methods for identifying antigens from infectious agents and cancers has provided exciting new opportunities in prevention and treatment through vaccination. In many of these situations, however, traditional immunization techniques do not stimulate protective immunity because they fail to fully mobilize...
journal_title:Nature biotechnology
pub_type: 杂志文章,评审
doi:10.1038/3469
更新日期:1998-11-01 00:00:00
abstract::Structural genomics has the ambitious goal of delivering three-dimensional structural information on a genome-wide scale. Yet only a small fraction of natural proteins are suitable for structure determination because of bottlenecks such as poor expression, aggregation, and misfolding of proteins, and difficulties in s...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt732
更新日期:2002-09-01 00:00:00
abstract::We describe tethered conformation capture (TCC), a method for genome-wide mapping of chromatin interactions. By performing ligations on solid substrates rather than in solution, TCC substantially enhances the signal-to-noise ratio, thereby facilitating a detailed analysis of interactions within and between chromosomes...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.2057
更新日期:2011-12-25 00:00:00
abstract::In the version of this article initially published, Betalutin was described as directing a radioisotope to CD45-expressing B cells; in fact, it directs it to CD37-expressing B cells. The error has been corrected in the HTML and PDF versions of the article. ...
journal_title:Nature biotechnology
pub_type: 杂志文章,已发布勘误
doi:10.1038/s41587-019-0030-5
更新日期:2019-02-01 00:00:00
abstract::An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...
journal_title:Nature biotechnology
pub_type: 杂志文章,已发布勘误
doi:10.1038/s41587-020-0510-7
更新日期:2020-05-01 00:00:00
abstract::The basics of determining valuation will help you to understand what criteria matter most for investors. ...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:
更新日期:2004-08-01 00:00:00
abstract::The epicardium and its derivatives provide trophic and structural support for the developing and adult heart. Here we tested the ability of human embryonic stem cell (hESC)-derived epicardium to augment the structure and function of engineered heart tissue in vitro and to improve efficacy of hESC-cardiomyocyte grafts ...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/s41587-019-0197-9
更新日期:2019-08-01 00:00:00
abstract::The efficiency of both phage display in Escherichia coli and periplasmic expression of recombinant proteins may be limited by the same periplasmic folding steps. To search for E. coli factors that improve the efficiency of both procedures, a library of E. coli proteins was coexpressed in a phagemid vector that contain...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt0498-376
更新日期:1998-04-01 00:00:00
abstract::An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...
journal_title:Nature biotechnology
pub_type: 杂志文章,已发布勘误
doi:10.1038/s41587-020-0457-8
更新日期:2020-03-01 00:00:00
abstract::Through sequential generations of random mutagenesis and screening, we have directed the evolution of an esterase for deprotection of an antibiotic p-nitrobenzyl ester in aqueous-organic solvents. Because rapid screening directly on the desired antibiotic (loracarbef) nucleus p-nitrobenzyl ester was not feasible, the ...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt0496-458
更新日期:1996-04-01 00:00:00
abstract::Reengineering the receptor footprints of adeno-associated virus (AAV) isolates may yield variants with improved properties for clinical applications. We generated a panel of synthetic AAV2 vectors by replacing a hexapeptide sequence in a previously identified heparan sulfate receptor footprint with corresponding resid...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.1599
更新日期:2010-01-01 00:00:00
abstract::Genome editing with targeted nucleases and DNA donor templates homologous to the break site has proven challenging in human hematopoietic stem and progenitor cells (HSPCs), and particularly in the most primitive, long-term repopulating cell population. Here we report that combining electroporation of zinc finger nucle...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.3408
更新日期:2015-12-01 00:00:00
abstract::The functional state of a cell is largely determined by the spatiotemporal organization of its proteome. Technologies exist for measuring particular aspects of protein turnover and localization, but comprehensive analysis of protein dynamics across different scales is possible only by combining several methods. Here w...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.2281
更新日期:2012-06-24 00:00:00
abstract::An approach to the systematic identification and quantification of the proteins contained in the microsomal fraction of cells is described. It consists of three steps: (1) preparation of microsomal fractions from cells or tissues representing different states; (2) covalent tagging of the proteins with isotope-coded af...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1001-946
更新日期:2001-10-01 00:00:00
abstract::Cells isolated from the embryonic, neonatal, and adult rodent central nervous system divide in response to epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF-2), while retaining the ability to differentiate into neurons and glia. These cultures can be grown in aggregates termed neurospheres, which conta...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/88158
更新日期:2001-05-01 00:00:00
abstract::Programmable DNA cleavage using CRISPR-Cas9 enables efficient, site-specific genome engineering in single cells and whole organisms. In the research arena, versatile CRISPR-enabled genome editing has been used in various ways, such as controlling transcription, modifying epigenomes, conducting genome-wide screens and ...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.3659
更新日期:2016-01-01 00:00:00
abstract::Targeting of secreted and cell-surface proteins to the cell membrane is mediated by a short hydrophobic stretch of amino acids, termed the signal sequence. We have developed a method that detects signal sequences in cDNA fragments based on their ability to redirect a constitutively active mutant of a cytokine receptor...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/8666
更新日期:1999-05-01 00:00:00
abstract::Escherichia coli-based artificial chromosomes have become important tools for physical mapping and sequencing in various genome projects. The lack of a general method to modify these large bacterial clones, however, has limited their utility in functional studies. We developed a simple method to modify bacterial artif...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt0997-859
更新日期:1997-09-01 00:00:00
abstract::Small-interfering RNAs (siRNAs) assemble into RISC, the RNA-induced silencing complex, which cleaves complementary mRNAs. Despite their fluctuating efficacy, siRNAs are widely used to assess gene function. Although this limitation could be ascribed, in part, to variations in the assembly and activation of RISC, downst...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1404
更新日期:2008-05-01 00:00:00
abstract::A wealth of molecular interaction data is available in the literature, ranging from large-scale datasets to a single interaction confirmed by several different techniques. These data are all too often reported either as free text or in tables of variable format, and are often missing key pieces of information essentia...
journal_title:Nature biotechnology
pub_type: 杂志文章,评审
doi:10.1038/nbt1324
更新日期:2007-08-01 00:00:00
abstract::The identification of co-regulated genes and their transcription-factor binding sites (TFBS) are key steps toward understanding transcription regulation. In addition to effective laboratory assays, various computational approaches for the detection of TFBS in promoter regions of coexpressed genes have been developed. ...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt802
更新日期:2003-04-01 00:00:00
abstract::Rice is a staple crop that has undergone substantial phenotypic and physiological changes during domestication. Here we resequenced the genomes of 40 cultivated accessions selected from the major groups of rice and 10 accessions of their wild progenitors (Oryza rufipogon and Oryza nivara) to >15 × raw data coverage. W...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.2050
更新日期:2011-12-11 00:00:00
abstract::Current DNA methylation assays are limited in the flexibility and efficiency of characterizing a large number of genomic targets. We report a method to specifically capture an arbitrary subset of genomic targets for single-molecule bisulfite sequencing for digital quantification of DNA methylation at single-nucleotide...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt.1530
更新日期:2009-04-01 00:00:00
abstract::RNA-interference (RNAi) is a potent mechanism, conserved from plants to humans for specific silencing of genes, which holds promise for functional genomics and gene-targeted therapies. Here we show that bacteria engineered to produce a short hairpin RNA (shRNA) targeting a mammalian gene induce trans-kingdom RNAi in v...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1211
更新日期:2006-06-01 00:00:00
abstract::The human genome encodes approximately 100,000 different genes, and at least partial sequence information for nearly all will be available soon. Sequence information alone, however, is insufficient for a full understanding of gene function, expression, regulation, and splice-site variation. Because cellular processes ...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1296-1675
更新日期:1996-12-01 00:00:00
abstract::Plant satellite RNAs generally reduce the level of helper virus accumulation and attenuate the disease symptoms induced by the helper virus that they depend upon for replication and packaging. As such, satellite RNAs could be used as biocontrol agents to reduce the level of disease in field crops, either by the applic...
journal_title:Nature biotechnology
pub_type: 杂志文章
doi:10.1038/nbt1096-1264
更新日期:1996-10-01 00:00:00