Long‐lived and highly efficient room temperature phosphorescence (RTP) materials are in high demand for practical applications in lighting and display, security signboards, and anti‐counterfeiting. Achieving RTP in aqueous solutions, near‐infrared (NIR) phosphorescence emission and NIR‐excited RTP are crucial for applications in bio‐imaging, but these goals pose significant challenges. Supramolecular self‐assembly provides an effective strategy to address the above problems. This review focuses on the recent advances in the enhancement of RTP via supramolecular self‐assembly, covering four key aspects: small molecular self‐assembly, cocrystals, the self‐assembly of macrocyclic hosts and guests, and multi‐stage supramolecular self‐assembly. This review not only highlights progress in these areas but also underscores the prominent challenges associated with developing supramolecular RTP materials. The resulting strategies for the development of high‐performance supramolecular RTP materials are discussed, aiming to satisfy the practical applications of RTP materials in biomedical science.This article is protected by copyright. All rights reserved