Subcellular organelles play indispensable roles in diverse biological processes by their precise mutual cooperation. Thus, the development of a single fluorescent probe (SF-probe) for simultaneous and discriminable visualization of different organelles and their dynamics during certain bioprocess is significant, yet remains greatly challenging. Herein, for the first time, we rationally prepared a pH-sensitive SF-probe (named HMBI) for the simultaneous two-color visualization of nuclei and mitochondria and monitoring cell apoptosis. HMBI shows remarkable ratiometric fluorescence changes toward pH changes. Due to different pH environments in subcellular organelles, HMBI can image nuclei and mitochondria with green and red emission, respectively. HMBI can monitor drug-induced cell apoptosis with dramatically decreased red emission in mitochondria but almost unchanged green emission in nuclei, and the shrinking and pyknotic nuclei are also observed during cell apoptosis. HMBI possesses tremendous potential in two-color biomedical imaging of the dynamic changes of nuclei and mitochondria in many physiological processes.
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