The main production area of HClO and NO is the mitochondria and has modulatory effects on multiple human diseases. Simultaneous detection of signaling molecules such as HClO and NO is an important approach for exploring the complex relationship between HClO and NO in mitochondria. However, most probes can detect only one species or are unable to complete the monitoring of HClO and NO in the NIR channel. There are only few reports on reasonable tools that can simultaneously monitor the presence of HClO and NO in the NIR channel. In this work, to solve this difficulty, a POSS-assisted NIR fluorescent probe with dual-response was rationally devised and developed. The probe Mito-Cy possessed high specificity and responsiveness to HClO and NO in spectral experiments. Notably, the probe exhibited excellent responsiveness and sensitivity to HClO and NO in living cells and the zebrafish model.