Optical super-resolution imaging technology, acknowledged by the Nobel Prize in Chemistry for transcending optical diffraction limits, has revolutionised life science research with its groundbreaking observation scale. However, conventional super-resolution fluorescence microscopes face challenges, requiring intricate optical systems that often result in significant phototoxicity and low temporal resolution, limiting their widespread use in biomedical research. Therefore, research teams actively seek alternative fluorescent probes with near-infrared capabilities, high brightness, and resistance to photobleaching, aiming to extend the application of super-resolution microscopy in biomedical research. Rare-earth nanomaterials, renowned for their exceptional physicochemical properties such as anti-Stokes spectral shift, lack of background noise, resistance to photobleaching, photostability, low toxicity, and high imaging penetration, have emerged as stable and superior inorganic fluorescent probes. This review paper provides a brief overview of the luminescent mechanism of upconversion nanoparticles, exploring the primary constraints in achieving photon upconversion in nanostructured materials. Additionally, it highlights the applications and advantages of lanthanide-doped upconversion nanoparticles in super-resolution biological imaging, molecular detection, and other domains. These advantages encompass reducing laser power requirements, addressing technical challenges in coupling, improving laser scanning imaging resolution and speed, and enhancing multiplexing imaging efficiency. This paper concludes by emphasizing significant challenges in particle synthesis, proposing feasible improvement measures, and outlining prospects for future development. It establishes a robust theoretical foundation and provides technical support for the widespread integration of rare-earth nanomaterials in the field of life imaging sciences.