The use of 13C and 15N labeled precursors in combination with adequate analytical tools makes it possible to study metabolic pathways in cultured neural cells. The most commonly used precursors are 13C labeled glucose, lactate, glutamate and acetate. For a dynamic evaluation of intermediary metabolism of cell cultures, incubation with 13C containing substrates followed by nuclear magnetic resonance spectroscopy (NMRS) and mass spectrometry (MS) is excellent. NMRS can be used on cell extracts or living cells if a sufficient quantity of labeled atoms is present. MS is the more sensitive of the two methods but often it requires derivatization and separation of the components before analysis. The review provides descriptions of the basic and practical aspects of culturing neural cells, incubation and superfusion experiments and NMRS and MS analyses. It focuses on the analytical tools and the use of primary cultures of neurons and astrocytes for the elucidation of metabolic interactions between neurons and astrocytes.