Significant progress on upconversion‐nanoparticle (UCNP)‐based probes is witnessed in recent years. Compared with traditional fluorescent probes (e.g., organic dyes, metal complexes, or inorganic quantum dots), UCNPs have many advantages such as non‐autofluorescence, high chemical stability, large light‐penetration depth, long lifetime, and less damage to samples. This article focuses on recent achievements in the usage of lanthanide‐doped UCNPs as efficient probes for biodetection since 2014. The mechanisms of upconversion as well as the luminescence resonance energy transfer process is introduced first, followed by a detailed summary on the recent researches of UCNP‐based biodetections including the detection of inorganic ions, gas molecules, reactive oxygen species, and thiols and hydrogen sulfide.