-
[
Methods Mol Biol,
1999]
The use of antibodies to visualize the distribution and subcellular localization of gene products powerfully complements genetic and molecular analysis of gene function in Caenorhabditis elegans. Double and triple staining protocols are particularly useful for several reasons. First, colonization of proteins either within tissues or at a subcellular level can be examined. Second, costaining with stage-specific or tissue-specific markers can define the timing and tissue specificity of antigen expression. For these types of studies it is useful to be able to collect data from multiple fluorescence wavelengths simultaneously. A confocal microscope equipped with a krypton/argon laser can simultaneously detect up to three different antigens. Using a confocal microscope it is also possible to collect a series of optical sections through a sample that allows observation of changes in distribution of the antigen in different focal planes of the tissue or cell.
-
[
Biol Proced Online,
2019]
Background: to develop a new estimation procedure that was based on a relationship between area and volume of a larger sample. Results: suspensions. Reactive oxygen concentration was measured as an example of method application and to allow comparison between methods. Conclusion: The proposed method is accurate, facile and reproducible, requiring simple, inexpensive materials that make it an excellent alternative to the DM manual counting procedure. Although the DM is faster, its estimates are not as accurate or as precise as those of the new proposed method.
-
[
Methods Mol Biol,
2020]
Reproductive senescence occurs in a wide range of species with mechanistic aspects that are conserved from Caenorhabditis elegans to humans. Genetic and environmental factors can influence fertility and reproductive output can impact rates of aging. The C. elegans Bristol N2 strain commonly used in laboratories is hermaphroditic, producing a defined number of sperm during larval development before switching exclusively to oogenesis. Here we show a method of assaying both oocyte and sperm quality from a single population of animals.
-
[
Methods Mol Biol,
2019]
This chapter describes methods for the analysis of autophagy proteins in C. elegans aging. We discuss the strains to be considered, the methods for the delivery of double-stranded RNA, and the methods to measure autophagy levels, autophagic flux, and degradation by autophagy.
-
[
Methods Mol Biol,
2000]
The complete description of its nearly invariant cell lineage and the growing availability of cloned genes and markers for the cell lineage make Caenorhabditis elegans particularly favorable for mosaic analysis, and the literature is rich in examples that prove the usefulness of this approach. Because genetic mosaic analysis in C. elegans has recently been reviewed by Herman who developed many of the techniques, this review will be more concerned with the recent technical advances rather than with an extensive background of the approach. First we shall present a brief summary of the principles of mosaic analysis as it is typically performed in C. elegans. This will be followed by a description of markers that indicate mosaicism and by a discussion of a hypothetical analysis of an essential gene.
-
[
Electrophoresis,
2018]
Spatial microgravity is a significant factor affecting and causing physiological changes of organisms in space environment. On-site assessment of the damage associated to microgravity is very important for future long-term space exploration of mankind. In this paper, a new microfluidic device for analyzing the damage of microgravity on Caenorhabditis elegans (C. elegans) has been developed. This device is mainly composed of a microfluidic chip, a dual imaging module, and an imaging acquisition and processing module, which are integrated into a compact system. The microfluidic chip is designed as a platform for monitoring C. elegans, which is captured in an imaging region through a suction structure in the microfluidic chip. A dual imaging module is designed to obtain the images of bright field and fluorescence of C. elegans. The behaviors of C. elegans are analyzed based on the dual-mode imaging of bright field and fluorescence to assess the degree of damage due to microgravity. A comparative study using a commercial microscope is also conducted to demonstrate the unique advantage of the developed system under the simulated microgravity. The results show that the developed system can evaluate the damage of C. elegans under microgravity accurately and conveniently. Furthermore, this device has compact size and weight, easy operation, and low-cost, which could be highly advantageous for on-site evaluation of the damage to microorganisms under microgravity in a space station. This article is protected by copyright. All rights reserved.
-
[
Methods Mol Biol,
2000]
Complete or partial embryonic cell lineages are available for several animal model systems. In the case of the nematode Caenorhabditis elegans, the entire embryonic cell lineage has been determined and is largely invariant. This makes cell lineage analysis a potentially useful tool for assessing mutant phenotypes in C. elegans. Indeed, lineage analysis of some mutants has shown that one cell can be transformed into a different cell resulting in duplication or absence of certain tissues...
-
[
Methods Mol Biol,
1999]
-
[
Methods Mol Biol,
2019]
Mitochondrial selective autophagy (mitophagy) is a critical cellular process for mitochondrial homeostasis and survival both under basal and stress conditions. Distinct cell types display different requirements for mitochondrial turnover depending on their metabolic status, differentiation state, and environmental cues. This points to the necessity of developing novel tools for real-time, tissue-specific assessment of mitophagy. Caenorhabditis elegans is an invaluable model organism for this kind of analysis providing a platform for simultaneous monitoring of mitophagy in vivo in different tissues and cell types, during development, stress conditions, and/or throughout life span. In this chapter we describe three versatile, noninvasive methods, developed for monitoring in vivo early and late mitophagic events in body wall muscles and neuronal cells of C. elegans. These procedures can be readily used and/or provide insights into the generation of novel imaging methods to investigate further the role of mitophagy at the organismal level under normal and pathological conditions.
-
[
Methods Mol Biol,
2021]
Meiosis is a specialized reductional cell division responsible for the formation of gametes and the generation of genetic diversity. A fundamental feature of the meiotic process is the initiation of homologous recombination (HR) by the programmed induction of DNA double-strand breaks (DSBs). Caenorhabditis elegans is a powerful experimental organism, which is used to study meiotic processes mainly due to the germline that allows for visualization of sequential stages of meiosis. C. elegans meiosis-programed DSBs are resolved through HR; hence, the germline provides a suitable model to study DSB repair. Classically direct procedures to detect and study intermediate steps in DSB repair by HR in the nematode rely on germline immunofluorescence against the strand exchange protein RAD-51.