Silicon nanomaterials are an important class of nanomaterials with great potential for technologies including energy, catalysis, and biotechnology, because of their many unique properties, including biocompatibility, abundance, and unique electronic, optical, and mechanical properties, among others. Silicon nanomaterials are known to have little or no toxicity due to favorable biocompatibility of silicon, which is an important precondition for biological and biomedical applications. In addition, huge surface-to-volume ratios of silicon nanomaterials are responsible for their unique optical, mechanical, or electronic properties, which offer exciting opportunities for design of high-performance silicon-based functional nanoprobes, nanosensors, and nanoagents for biological analysis and detection and disease treatment. Moreover, silicon is the second most abundant element (after oxygen) on earth, providing plentiful and inexpensive resources for large-scale and low-cost preparation of silicon nanomaterials for practical applications. Because of these attractive traits, and in parallel with a growing interest in their design and synthesis, silicon nanomaterials are extensively investigated for wide-ranging applications, including energy, catalysis, optoelectronics, and biology. Among them, bioapplications of silicon nanomaterials are of particular interest. In the past decade, scientists have made an extensive effort to construct a silicon nanomaterials platform for various biological and biomedical applications, such as biosensors, bioimaging, and cancer treatment, as new and powerful tools for disease diagnosis and therapy. Nonetheless, there are few review articles covering these important and promising achievements to promote the awareness of development of silicon nanobiotechnology. In this Account, we summarize recent representative works to highlight the recent developments of silicon functional nanomaterials for a new, powerful platform for biological and biomedical applications, including biosensor, bioimaging, and cancer therapy. First, we show that the interesting photoluminescence properties (e.g., strong fluorescence and robust photostability) and excellent biocompatibility of silicon nanoparticles (SiNPs) are superbly suitable for direct and long-term visualization of biological systems. The strongly fluorescent SiNPs are highly effective for bioimaging applications, especially for long-term cellular labeling, cancer cell detection, and tumor imaging in vitro and in vivo with high sensitivity. Next, we discuss the utilization of silicon nanomaterials to construct high-performance biosensors, such as silicon-based field-effect transistors (FET) and surface-enhanced Raman scattering (SERS) sensors, which hold great promise for ultrasensitive and selective detection of biological species (e.g., DNA and protein). Then, we introduce recent exciting research findings on the applications of silicon nanomaterials for cancer therapy with encouraging therapeutic outcomes. Lastly, we highlight the major challenges and promises in this field, and the prospect of a new nanobiotechnology platform based on silicon nanomaterials.