<jats:p>The dynamics of physical parameters in cells is strongly related to life phenomena; thus, a method to monitor and visualize them on a single-organelle scale would be useful to reveal unknown biological processes. We demonstrate real-time nanometre-scale T1-weighted imaging using a fluorescent nanodiamond. We explored optically detected magnetic resonance (ODMR) contrast at various values of interval laser pulse (τ), showing that sufficient contrast is obtained by appropriate selection of τ. By this method, we visualized nanometre-scale pH changes using a functionalized nanodiamond whose T1 has a dependence on pH conditions.</jats:p>