Fluorescent silicon nanoparticles (SiNPs), a type of ultrasmall zero‐dimensional metal‐free silicon‐containing nanomaterials, are featured by excellent biocompatibility, good photoluminescence stability, and easy surface modification property. SiNPs have been widely applied in bioimaging, photoelectric devices, sensing, cancer therapies, and anti‐counterfeiting. In this minireview, the recent advancements of SiNPs in imaging and killing microorganisms (i. e., bacteria, bacterial spores, and fungi) and bacterial biofilms are summarized. Moreover, the synthesis and modification of these SiNPs are mentioned. Finally, concise summary and outlook related to the review topic are provided, and the future challenges are briefly discussed.