It is generally accepted that microRNAs (miRNAs) play a crucial role in gene expression regulation and that their aberrant expression is intimately linked with pathologies, most notably cancer. There is thus significant interest in detecting and quantifying these important regulators. Herein, we report the fluorescence imaging of miRNAs within a few hours using a nucleic-acid templated Staudinger reaction. A good correlation between the level of miRNAs and the fluorescence intensity was observed across different cell lines. This method was shown to also be applicable for suspended cells with fluorescence quantification by flow cytometry.