In order to understand related pathogenesis of some diseases and design new intracellular drug delivery systems, investigation of pH change in living cells in real time is important. In this paper, a new style of fluorescent silicon nanoparticles (SiNPs) as a pH-sensitive probe and for the visualization of the pH changes in cells was designed and prepared using 4-aminophenol as a reducing agent and N-aminoethyl-γ-aminopropyltrimethyl as a silicon source by a one-pot hydrothermal method. It was particularly noteworthy that the fluorescence intensity emitted from the SiNPs positively correlated with the pH value of solutions, making the SiNPs a viable probe used for sensitive sensing of pH. At the same time, a response of the probe to the pH was found in 5.0-10.0, and the SiNPs have an excellent biocompatibility (e.g., ∼74% of cell viability was remained after treatment for 24 h at 500 μg/mL of the SiNPs). The proposed method that could display the change in pH of live cells provided an effective means for visually diagnosing diseases related to intracellular pH.