Abstract：
:Postembryonic development is an important process of organismal maturation after embryonic growth. Despite key progress in recent years in understanding embryonic development via fluorescence time-lapse microscopy, comparatively less live-cell imaging of postembryonic development has been done. Here we describe a protocol to image larval development in the nematode Caenorhabditis elegans. Our protocol describes the construction of fluorescent transgenic C. elegans, immobilization of worm larvae and time-lapse microscopy analysis. To improve the throughput of imaging, we developed a C. elegans triple-fluorescence imaging approach with a worm-optimized blue fluorescent protein (TagBFP), green fluorescent protein (GFP) and mCherry. This protocol has been previously applied to time-lapse imaging analysis of Q neuroblast asymmetric division, migration and apoptosis, and we show here that it can also be used to image neuritogenesis in the L1 larvae. Other applications are also possible. The protocol can be completed within 3 h and may provide insights into understanding postembryonic development.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Chai Y,Li W,Feng G,Yang Y,Wang X,Ou Gdoi
10.1038/nprot.2012.128subject
Has Abstractpub_date
2012-12-01 00:00:00pages
2090-102issue
12eissn
1754-2189issn
1750-2799pii
nprot.2012.128journal_volume
7pub_type
杂志文章相关文献
Nature Protocols文献大全abstract：:Understanding chemical reactivity through the use of state-of-the-art computational techniques enables chemists to both predict reactivity and rationally design novel reactions. This protocol aims to provide chemists with the tools to implement a powerful and robust method for analyzing and understanding any chemical ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-019-0265-0
更新日期：2020-02-01 00:00:00
abstract：:Cyclodepsipeptides are cyclic peptides in which at least one amide link on the backbone is replaced with an ester link. These natural products present a high structural diversity that corresponds to a broad range of biological activities. Therefore, they are very promising pharmaceutical candidates. Most of the cyclod...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.116
更新日期：2016-10-01 00:00:00
abstract：:Carbohydrate microarrays have received considerable attention as an advanced technology for the rapid analysis of carbohydrate-protein interactions. This protocol provides detailed procedures for the preparation of carbohydrate microarrays by immobilizing hydrazide-conjugated carbohydrates on epoxide-derivatized glass...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.373
更新日期：2007-01-01 00:00:00
abstract：:Although lung transplant is a life-saving therapy for some patients, primary graft dysfunction (PGD) is a leading cause of mortality and morbidity soon after a transplant. Ischemia reperfusion injury is known to be one of the most critical factors in PGD development. PGD is by definition an acute lung injury syndrome ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-018-0019-4
更新日期：2018-08-01 00:00:00
abstract：:Post-translational protein nitration has attracted interest owing to its involvement in cellular signaling, effects on protein function and potential as biomarker of nitroxidative stress. We describe a procedure for enriching nitropeptides for mass spectrometry (MS)-based proteomics that is a simple and reliable alter...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2014.052
更新日期：2014-04-01 00:00:00
abstract：:Post-translational modifications alter protein structure, affecting activity, stability, localization and/or binding partners. Antibodies that specifically recognize post-translationally modified proteins have a number of uses including immunocytochemistry and immunoprecipitation of the modified protein to purify prot...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2014.017
更新日期：2014-02-01 00:00:00
abstract：:The point mutational spectrum over nearly any 75- to 250-bp DNA sequence isolated from cells, tissues or large populations may be discovered using denaturing capillary electrophoresis (DCE). A modification of the standard DCE method that uses cycling temperature (e.g., +/-5 degrees C), CyDCE, permits optimal resolutio...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.79
更新日期：2008-01-01 00:00:00
abstract：:The genomic instability of stem cells in culture, caused by their routine in vitro propagation or by their genetic manipulation, is deleterious both for their clinical application and for their use in basic research. Frequent evaluation of the genomic integrity of stem cells is thus required, and it is usually perform...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2013.051
更新日期：2013-05-01 00:00:00
abstract：:High-throughput transcriptional analysis has unveiled a myriad of novel RNAs. However, technical constraints in RNA sequencing library preparation and platform performance hamper the identification of rare transcripts contained within the RNA repertoire. Herein we present targeted-RNA directional sequencing (TARDIS), ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2015.120
更新日期：2015-12-01 00:00:00
abstract：:During biosynthesis on the ribosome, an elongating nascent polypeptide chain can begin to fold, in a process that is central to all living systems. Detailed structural studies of co-translational protein folding are now beginning to emerge; such studies were previously limited, at least in part, by the inherently dyna...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.101
更新日期：2016-08-01 00:00:00
abstract：:Mouse embryonic stem cells (mESCs) are key tools for genetic engineering, development of stem cell-based therapies and basic research on pluripotency and early lineage commitment. However, successful derivation of germline-competent embryonic stem cell lines has, until recently, been limited to a small number of inbre...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2014.030
更新日期：2014-03-01 00:00:00
abstract：:Desorption electrospray ionization (DESI) allows the direct analysis of ordinary objects or pre-processed samples under ambient conditions. Among other applications, DESI is used to identify and record spatial distributions of lipids and drug molecules in biological tissue sections. This technique does not require sam...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.11
更新日期：2008-01-01 00:00:00
abstract：:Interactions of proteins with DNA mediate many critical nuclear functions. Chromatin immunoprecipitation (ChIP) is a robust technique for studying protein-DNA interactions. Current ChIP assays, however, either require large cell numbers, which prevent their application to rare cell samples or small-tissue biopsies, or...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.68
更新日期：2008-01-01 00:00:00
abstract：:This protocol measures externalization of aminophospholipids (APLs) to the outside of the plasma membrane using mass spectrometry (MS). APL externalization occurs in numerous events, and it is relevant for transplant medicine, immunity and cancer. In this protocol, externalized APLs are chemically modified by using a ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2013.163
更新日期：2014-01-01 00:00:00
abstract：:The colorimetric bio-barcode assay is a red-to-blue color change-based protein detection method with ultrahigh sensitivity. This assay is based on both the bio-barcode amplification method that allows for detecting miniscule amount of targets with attomolar sensitivity and gold nanoparticle-based colorimetric DNA dete...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.201
更新日期：2007-01-01 00:00:00
abstract：:This protocol outlines steps for optimizing the transfection of adherent primary mammalian cells using the readily available off-the-shelf cationic polymer, 25-kDa branched polyethylenimine (bPEI25). Transfection efficiency of cationic polymers varies among cell lines and is highly dependent on the conditions and envi...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.038
更新日期：2012-04-19 00:00:00
abstract：:The use of anisotropic NMR data, such as residual dipolar couplings (RDCs) and residual chemical shift anisotropies (RCSAs), has emerged as a powerful technique for structural characterization of organic small molecules. RDCs typically report the relative orientations of different 1H-13C bonds; RCSAs report the relati...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-018-0091-9
更新日期：2019-01-01 00:00:00
abstract：:Blood circulating endothelial cells (CECs) and circulating hematopoietic progenitor cells (CPCs) represent two cell populations that are thought to play important roles in tissue vascularization. CECs and CPCs are currently studied as surrogate markers in patients for more than a dozen pathologies, including heart dis...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.111
更新日期：2007-01-01 00:00:00
abstract：:Top-down proteomics (TDP) by mass spectrometry (MS) is a technique by which intact proteins are analyzed. It has become increasingly popDesalting and concentrating GELFrEEular in translational research because of the value of characterizing distinct proteoforms of intact proteins. Compared to bottom-up proteomics (BUP...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-018-0085-7
更新日期：2019-01-01 00:00:00
abstract：:The fabrication of 3D tissues retaining the original functions of tissues/organs in vitro is crucial for optimal tissue engineering and regenerative medicine. The fabrication of 3D tissues also contributes to the establishment of in vitro tissue/organ models for drug screening. Our laboratory has developed a fabricati...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.027
更新日期：2012-04-05 00:00:00
abstract：:We describe a simple and versatile scheme to prepare a series of poly(ethylene glycol)-based bidentate ligands that permit strong interactions with colloidal semiconductor nanocrystals (quantum dots, QDs) and gold nanoparticles (AuNPs) alike and promote their dispersion in aqueous solutions. These ligands are synthesi...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.243
更新日期：2009-01-01 00:00:00
abstract：:Most currently available colorectal cancer (CRC) mouse models are not suitable for studying progression toward the metastatic stage. Recently, establishment of tumor organoid lines, either from murine CRC models or patients, and the possibility of engineering them with genome-editing technologies, have provided a larg...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2017.137
更新日期：2018-02-01 00:00:00
abstract：:Herein we describe a protocol that uses hollow-fiber flow field-flow fractionation (FFF) coupled with multiangle light scattering (MALS) for hydrodynamic size-based separation and characterization of complex protein aggregates. The fractionation method, which requires 1.5 h to run, was successfully modified from the a...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2015.009
更新日期：2015-01-01 00:00:00
abstract：:In the past decade, Drosophila has emerged as an ideal model organism for studying the genetic components of sleep as well as its regulation and functions. In fruit flies, sleep can be conveniently estimated by measuring the locomotor activity of the flies using techniques and instruments adapted from the field of cir...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.041
更新日期：2012-04-26 00:00:00
abstract：:The protein called 'small ubiquitin-like modifier' (SUMO) is post-translationally linked to target proteins at the ɛ-amino group of lysine residues. This 'SUMOylation' alters the behavior of the target protein, a change that is utilized to regulate diverse cellular processes. Understanding the target-specific conseque...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2015.095
更新日期：2015-09-01 00:00:00
abstract：:The rodent malaria parasite Plasmodium chabaudi chabaudi shares many features with human malaria species, including P. falciparum, and is the in vivo model of choice for many aspects of malaria research in the mammalian host, from sequestration of parasitized erythrocytes, to antigenic variation and host immunity and ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2011.313
更新日期：2011-04-01 00:00:00
abstract：:A big challenge in proteomics is the identification of cell-type-specific proteomes in vivo. This protocol describes how to label, purify and identify cell-type-specific proteomes in living mice. To make this possible, we created a Cre-recombinase-inducible mouse line expressing a mutant methionyl-tRNA synthetase (L27...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-018-0106-6
更新日期：2019-02-01 00:00:00
abstract：:Here, we describe a simple protocol for the design and construction of a laser-guided direct writing (LGDW) system able to micropattern the self-assembly of liver sinusoid-like structures with micrometer resolution in vitro. To the best of our knowledge, LGDW is the only technique able to pattern cells "on the fly" wi...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.386
更新日期：2006-01-01 00:00:00
abstract：:This protocol describes a method combining phase-contrast and fluorescence microscopy, Raman spectroscopy and optical tweezers to characterize the germination of single bacterial spores. The characterization consists of the following steps: (i) loading heat-activated dormant spores into a temperature-controlled micros...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2011.307
更新日期：2011-05-01 00:00:00
abstract：:A detailed protocol for the synthesis of core/shell semiconductor nanocrystal, their encapsulation into phospholipid micelles, their purification and their coupling to a controlled number of small molecules is given. The protocol for the core/shell quantum dot (QD) CdSe/CdZnS synthesis has been specifically designed w...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.351
更新日期：2007-01-01 00:00:00
