loading . . . Supramolecular in situ assemblage and its biological applications Water-soluble macrocycles, including cyclodextrins (CDs), cucurbit[n]urils (CB[n]s), sulfonated calixarenes (CAs), and pillararenes (PAs), can selectively bind organic anions or cations for in situ assemblage to construct supramolecular assemblies. These assemblies can also achieve spatially-confined luminescence or encapsulate drugs, demonstrating their wide applications in biology and medicine, as well as other fields. This review discusses biological applications of supramolecular in situ assemblies constructed by water-soluble macrocyclic derivatives, including macrocyclic hosts modified by polymers, functionalized guests, and macrocyclic hosts, and noncovalent supramolecular polymers. With further development of supramolecular in situ assemblies, intelligent systems with pH, light, and enzyme responsiveness can be widely integrated in macrocyclic cavities with spatially-confined luminescence for cell imaging, targeted drug delivery, and in situ drug formation. http://dlvr.it/TLLxWC