AbstractNanozymes refer to nanomaterial‐based enzyme mimics. Most nanozyme assays use chromogenic molecules as substrates that can produce an absorbance change in the visible region upon reaction. For certain applications, such as imaging, fluorescence and luminescence signals are required. Fluorescence detection is also advantageous for sensor development due to its low background signal and high sensitivity. A few substrates, such as Amplex Red and luminol, allow luminescence generation with many oxidative nanozymes. In addition, these substrates can work at neutral pH, useful for intracellular applications. In this review, we summarize luminescence signalling‐based nanozyme systems. We begin with the properties of commonly used substrates. A few specific nanozymes are highlighted, including metal nanoparticles, metal oxide nanomaterials, carbon‐based nanomaterials, and metal–organic frameworks. Finally, a few future research opportunities are discussed.