Caenorhabditis elegans: Cell Biology and Physiology

High-Resolution Imaging of Cellular Processes in Caenorhabditis elegans

I. Microscope Stand

II. Objective Lenses and Resolution

III. Transmitted Light Imaging

IV. Fluorescence Imaging

V. Detectors

VI. Confocal Imaging

VII. Deconvolution Imaging

VIII. Whole Worm (Adult and Larval Stages)

IX. Embryos

Immunofluorescence Microscopy

I. Introduction

II. Preparing Specimens for Immunolocalization Studies

III. Preparation of Antibodies to Nematode Antigens

IV. Microscopy: Necessary Equipment and Imaging Tips

V. Concluding Remarks

VI. Recipes

Fluorescent Protein Methods: Strategies and Applications

I. Introduction

II. Methods

III. Adopting New Fluorescent Proteins for Use in C. elegans

Modern Electron Microscopy Methods for C. elegans

I. Introduction

II. Modern Electron Microscopes

III. Alternate Immersion Fixation Techniques

IV. Serial Thin Sectioning

V. ImmunoEM Protocols

VI. Alternate Methods to Post Stain Thin Sections

VII. Electron Tomography

VIII. Emerging EM Technologies

IX. Computer-Based Analytical Tools

X. Landmarks for Recognizing Tissues

Acknowledgments

Culture and Manipulation of Embryonic Cells

I. Introduction: Uses and Limitations of the System

II. Embryo Devitellinization and Blastomere Isolation

III. Culture Methods

IV. Drug Treatments

V. Radioactive Labeling

VI. Fixation, Cytochemistry, and Electron Microscopy

VII. Solutions and Culture Media

Laser Microsurgery in Caenorhabditis elegans

I. Overview

II. Identifying Cells in C. elegans

III. Laser Ablation Theory and Apparatus

IV. Laser Killing of Cells

V. Assessing Damage to the Operated Cell

VI. Unintended Damage

VII. Laser Cutting of Nerve Fibers

VIII. Related Methods

Acknowledgments

Methods in Cell Biology: Analysis of Cell Polarity in C. elegans Embryos

I. Introduction

II. Methods for Analyzing Gene Function

III. Analyzing Cell Polarity: Protein and Organelle Localization

IV. Dynamics of Polarity Establishment

V. Summary

Analysis of Membrane-Bound Organelles

I. Introduction

II. Visualizing Organelles

III. Functional Studies

IV. Protein Extract and Organelle Isolation

V. Summary

Acknowledgments

C. elegans Cell Cycle Analysis

I. Introduction

II. Types of C. elegans cell cycles

III. Using the C. elegans Lineage to Follow Cell Divisions

IV. Methods of Developmental Synchronization

V. Quantification of DNA Content

VI. Detection of Different Cell Cycle Phases

VII. Common Cell Cycle Phenotypes and Potential Interpretations

Methods for Studying Programmed Cell Death in C. elegans

I. Introduction

II. Methods

III. Conclusions

Acknowledgments

Methods for Studying the DNA Damage Response in the Caenorhabdatis elegans Germ Line

I. Introduction

II. Assay Development

III. Assays

IV. Materials

V. Discussion

Acknowledgements

Analysis of Aging in Caenorhabditis elegans

I. Introduction

II. Aging Pathways and Their Mechanism of Action

III. Life-Span Analysis

IV. Analysis and Interpretation of Lifespan Data

V. Measurement of Age-Related Changes

VI. Conclusion

Acknowledgments

Analyses of C. elegans Fat Metabolic Pathways

I. Introduction

II. Staining of Fixed Animals and Histochemical Methods

III. Biochemical Methods

IV. Biochemical Measurements of Metabolic Rates

V. Visualization of Fat Depots and Fat Metabolism Using Vital Dyes

VI. Visualizing Hypodermal, Intestinal, and Gonad Lipid Stores using Nile Red

VII. Fatty Acid Uptake

VIII. Raman Scattering-Based Methods of Lipid Imaging in C. elegans

IX. Summary

Acknowledgments

Electrophysiological Methods for Caenorhabditis elegans Neurobiology

I. Introduction

II. Recording Currents from Identified Neurons

III. Recording Synaptic Events from Identified Neurons

IV. Recording Currents and Synaptic Events at the Neuromuscular Junction

V. Materials

VI. Discussion

VII. Summary

Acknowledgments