This review aims to describe some recent and novel applications of stable isotope tracer technology to study the metabolism of C-13-polyunsaturated fatty acids. Stable isotope methodology has existed for several decades, and in that sense, it is not novel per se. However, in the past 10 years, developments in the fields of isotope ratio mass spectrometry, nuclear magnetic resonance (NMR) spectroscopy, and biological approaches to labelling tracer compounds with stable isotopes have provided new opportunities in fatty acid research. Sample preparation for isotope ratio mass spectrometry has been converted from a manual combustion method to an on-line or continuous flow method, making it much more versatile and easier to use. Similarly, C-13-NMR spectroscopy has recently developed as a remarkably useful method for monitoring metabolic steps and pathways bath in vivo and at the molecular level. Coincident with these improvements in instrumentation, the commercial availability of numerous uniformly C-13-labelled compounds has made these studies more affordable. The application of some of these developments to questions in the field of polyunsaturated fatty acid metabolism is described.