Chemical Microbiology, Part B, Volume 665, the latest release in the Methods of Enzymology series, highlights new advances in the field, including comprehensive chapters on the Application of Antibiotic-derived Fluorescent Probes to Bacterial Studies, Metabolomic approaches for enzyme function and pathway discovery in bacteria, Adding a diazo-transfer reagent to culture to generate secondary metabolite probes for click chemistry, Customized Peptidoglycan Surfaces to Investigate Innate Immune Recognition via Surface Plasmon Resonance, Development and application of highly sensitive labeling reagents for amino acids, Bacterial Cell Wall Modification with a Glycolipid Substrate, and much more.