Of the many metabolic pathways found in nature, fatty acid biosynthesis plays vital roles in cellular biology. Beyond its typically role in cellular energy metabolism, fatty acids integrate and interact with signaling pathways, contribute to post-translational modifications of proteins (palmitoylation), and influence the membrane structure and function. Additionally, the valorization of fatty acids and their derivatives (e.g. esters, alkanes, olefins, ketones, alcohols, polyesters) drives further interest in metabolic engineering. They are involved manufacturing highly valuable products such as biofuels, detergents, cosmetics, lubricants, and pharmaceutical ingredients. The cellular lipids and fatty acids compositions are quite diverse among various biological systems. This makes fast and accurate quantification of lipids and fatty acids to be a difficult task especially when the target molecules are complex and rare low-abundance lipids and fatty acids species. In addition, high-throughput methods for fast screening of the cells producing lipids or fatty acids is highly desirable. This collection aims to cover the recent advances in methods developed for the analysis of lipids and fatty acids, from the sample preparation, separation, identification, and quantification.