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Antibody Addition and Detection for Staining Caenorhabditis elegans Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4523

Category: C. elegans Protocols: C. elegans Staining Protocols

Extreme care should be used to identify and verify positive reactions, however, because cross-reactions are common. Counterstaining is essential for examining worms by immunofluorescence and is used to identify the exact cell in which an antigen appears. Methods for counterstaining include labeling all cells with a fluorescent dye that is specific for nucleic acids (e.g., DAPI or propidium iodide) and using GFP driven by tissue-specific promoters. - [Read Antibody Addition and Detection for Staining Caenorhabditis elegans Protocol]

Cosuppression in C. elegans Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4318

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Cosuppression is a process in Caenorhabditis elegans that closely resembles RNAi. In contrast to RNAi, however, the cosuppression effect in C. elegans does not spread throughout the animal. Cosuppression in C. elegans can be triggered by highly repetitive transgenes that contain gene constructs. - [Read Cosuppression in C. elegans Protocol]

Detection of si/miRNA in C. elegans by Northern Blot Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4320

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Protocol uses northern blot technology to detect short interfering RNAs (siRNAs) or micro-RNAs (miRNAs) in RNA preparations from Caenorhabditis elegans. - [Read Detection of si/miRNA in C. elegans by Northern Blot Protocol]

Detection of siRNA in C. elegans Using Rnase Protection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4319

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Protocol uses Rnase protection to detect short interfering RNAs (siRNAs) in RNA preparations from Caenorhabditis elegans. SiRNAs can also be detected by northern blot. However, the Rnase protection assay seems to be more sensitive. - [Read Detection of siRNA in C. elegans Using Rnase Protection Protocol]

Fixing Caenorhabditis elegans in Bouin’s Fixative Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4521

Category: C. elegans Protocols: C. elegans Fixing Protocols

Bouin’s fixative is a particularly good choice for worms because it penetrates dense tissues well and is extremely good for fixing antigens. Like all strong fixatives, however, it is unsuitable for some antibody-antigen pairs. In such cases, the length of time in the Bouin’s fixative can be shortened, or paraformaldehyde fixation can be used instead. - [Read Fixing Caenorhabditis elegans in Bouin’s Fixative Protocol]

Fixing Caenorhabditis elegans in Paraformaldehyde Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4522

Category: C. elegans Protocols: C. elegans Fixing Protocols

Common method for fixing worms is to use paraformaldehyde. This method provides a gentler fixation than the Bouin’s method, but often requires the use of collagenase. This method is particularly good for examining adult worms. - [Read Fixing Caenorhabditis elegans in Paraformaldehyde Protocol]

Introduction of Double-Stranded RNA in C. elegans by Feeding Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4317

Category: C. elegans Protocols: C. elegans Genetics Protocols

Protocol describes an easily scalable way of introducing double-stranded RNA (dsRNA) in Caenorhabditis elegans: feeding the nematode with bacteria that express dsRNA. When using an RNase-III-negative Escherichia coli strain (HT115), the efficiency of this method is comparable to the alternative. - [Read Introduction of Double-Stranded RNA in C. elegans by Feeding Protocol]

Introduction of Double-Stranded RNA in C. elegans by Feeding Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4317

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Protocol describes an easily scalable way of introducing double-stranded RNA (dsRNA) in Caenorhabditis elegans: feeding the nematode with bacteria that express dsRNA. When using an Rnase-III-negative Escherichia coli strain (HT115), the efficiency of this method is comparable to the alternative. - [Read Introduction of Double-Stranded RNA in C. elegans by Feeding Protocol]

Introduction of Double-Stranded RNA in C. elegans by Injection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4315

Category: C. elegans Protocols: C. elegans Genetics Protocols

Double-stranded RNA (dsRNA) can be efficiently introduced into Caenorhabditis elegans by microinjection into the gonad, the gut, or the body fluid. The RNAi effect will spread within the nematode, exerting an effect beyond the site of injection. - [Read Introduction of Double-Stranded RNA in C. elegans by Injection Protocol]

Introduction of Double-Stranded RNA in C. elegans by Injection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4315

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Introduction of Double-Stranded RNA in C. elegans by Soaking Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4316

Category: C. elegans Protocols: C. elegans Genetics Protocols

Double-stranded RNA (dsRNA) can be introduced into Caenorhabditis elegans by soaking the animals in a solution of dsRNA. Alternative methods are dsRNA injection (see Introduction of Double-stranded RNA in C. elegans by Injection) and feeding the animals with bacteria that produce dsRNA. - [Read Introduction of Double-Stranded RNA in C. elegans by Soaking Protocol]

Introduction of Double-Stranded RNA in C. elegans by Soaking Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4316

Category: RNAi Technology Protocols: RNAi C. elegans Protocols

Double-stranded RNA (dsRNA) can be introduced into Caenorhabditis elegans by soaking the animals in a solution of dsRNA. Alternative methods are dsRNA injection and feeding the animals with bacteria that produce dsRNA. - [Read Introduction of Double-Stranded RNA in C. elegans by Soaking Protocol]

Isolation of RNA from C. elegans Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4321

Category: C. elegans Protocols: C. elegans Genetics Protocols

Protocol describes an efficient approach to isolate RNA from Caenorhabditis elegans. - [Read Isolation of RNA from C. elegans Protocol]

Live Imaging of Caenorhabditis elegans: Examples - http://www.cshprotocols.org/cgi/content/extract/2006/29/pdb.ip19

Category: C. elegans Protocols

To image early cleavages and chromatin dynamics, it is convenient to use histone H2B fused to GFP or lamin::GFP. Time-lapse movies can be obtained using conventional confocal microscope systems and their included software. Early embryos dissected from transgenic hermaphrodites are placed with egg salts on agar pads. - [Read Live Imaging of Caenorhabditis elegans: Examples]

Live Imaging of Caenorhabditis elegans: Observation of Nematodes and Data Collection - http://www.cshprotocols.org/cgi/content/extract/2006/29/pdb.ip18

Category: Microscopy Protocols: In Vivo Imaging Protocols

Live nematodes can be studied at all levels of microscopic resolution. Data collection includes information on: Photographic Records, Video Microscopy,Multiple-Focal-Plane Time-Lapse Recording Systems and Digital Imaging. - [Read Live Imaging of Caenorhabditis elegans: Observation of Nematodes and Data Collection]

Live Imaging of Caenorhabditis elegans: Observation of Nematodes and Data Collection Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/29/pdb.ip18

Category: C. elegans Protocols

At low magnification, dissection microscopes are useful, whereas at higher magnification and resolution, a wide range of optical methods can be used, including differential interference contrast (DIC) optics, phase contrast, fluorescence, polarized light, confocal, and dark field. - [Read Live Imaging of Caenorhabditis elegans: Observation of Nematodes and Data Collection Protocol]

Live Imaging of Caenorhabditis elegans: Preparation of Samples - http://www.cshprotocols.org/cgi/content/abstract/2006/29/pdb.prot4601

Category: Microscopy Protocols: In Vivo Imaging Protocols

Caenorhabditis elegans, a small (adults are ~1 mm long), free-living soil nematode that feeds on bacteria, is an ideal organism for applying various live microscopy techniques. This protocol describes useful techniques for preparing C. elegans for live microscopic analysis. Details of sample preparation depend on the developmental stage of the worm to be studied. - [Read Live Imaging of Caenorhabditis elegans: Preparation of Samples]

Live Imaging of Caenorhabditis elegans: Preparation of Samples - http://www.cshprotocols.org/cgi/content/abstract/2006/29/pdb.prot4601

Category: C. elegans Protocols

Protocol describes useful techniques for preparing C. elegans for live microscopic analysis. Details of sample preparation depend on the developmental stage of the worm to be studied. - [Read Live Imaging of Caenorhabditis elegans: Preparation of Samples]

Protocol Live Imaging of Caenorhabditis Elegans - http://www.cshprotocols.org/cgi/content/extract/2006/29/pdb.ip19

Category: Microscopy Protocols: In Vivo Imaging Protocols

To image early cleavages and chromatin dynamics, it is convenient to use histone H2B fused to GFP or lamin::GFP. Time-lapse movies can be obtained using conventional confocal microscope systems and their included software. Early embryos dissected from transgenic hermaphrodites are placed with egg salts on agar pads. Chromatin dynamics can be followed easily, and wild-type embryonic cells can be compared with mutants or RNAi-treated embryos. - [Read Protocol Live Imaging of Caenorhabditis Elegans]