This volume surveys the latest in bioorthogonal chemistry, from foundational methods to cutting-edge biomedical applications. The ability to perform "new-to-nature" chemical reactions within living cells and organisms is revolutionizing how we study biological processes and has enabled the development of powerful new biomedical tools. With over two decades of progress in bioorthogonal chemistry, this timely multi-author volume provides a comprehensive overview of the field and serves as a key reference for both researchers and newcomers. It includes a historical perspective, chapters on the various methodologies for conducting live-compatible transformations, and reviews that showcase the exciting biological and biomedical applications. Each contribution is thematically homogeneous, and experimental descriptions of the more relevant methodologies are also included.