Ultrasmall and highly fluorescent gold nanoclusters (Au NCs) have been widely used for the construction of sensing and imaging platforms. Specifically, through a combination of surface functionalization and spectral analysis and/or imaging techniques, effective intracellular detection and imaging are realized. In this review, we summarize the recently adopted intracellular analysis and imaging events with Au NCs-based probes. The synthesis of Au NCs is briefly introduced based on stabilizer selection. The principles and applications of fluorometric intracellular detection systems toward different analytes, including small molecules and biomacromolecules, are presented by turnoff, turn-on, and ratiometric tactics. The cell imaging events are summarized based on conventional imaging and high-resolution imaging techniques, respectively. In the end, this review highlights the challenges of intracellular applications with Au NCs.