Cell culture has emerged as an important research method for studying various types of primary cells, including neurons and glial cells. This method has been especially instrumental in assessing intracellular Ca 2+ dynamics of neural cells. The invertebrate model organism Caenorhabditis elegans has been extensively used in neurobiology to study wide-spread issues ranging from gene regulation to behavior. We present some of the basic morphological characteristics of the four C. elegans glial cells residing in the cephalic sensilla of the worm, followed by a description of cell culturing methods for these glial cells. We describe the combined genetic and fluorescence microscopy approaches for identification of C. elegans glial cells in culture and assessment of their cytosolic Ca 2+ dynamics. © 2012 Springer Science+Business Media, LLC.