Microbial single-cell omics has revolutionized the study of uncultured microorganisms, offering unparalleled insights into their genetic composition, functional potential, and ecological roles. This guide highlights the key advancements in single-cell genomics, transcriptomics, proteomics, and metabolomics as featured in this book. It explores innovative techniques that enhance genome recovery, boost throughput, and lower costs. Additionally, new activity-based methods allow precise targeting of specific microbial populations, while the integration of multi-omics workflows represents a major leap forward. These breakthroughs are set to transform our understanding of microbial diversity and function, with profound implications for ecology, biomedicine, and biotechnology.