Noninvasive monitoring of kidney elimination of engineered nanoparticles at high temporal and spatial resolution will not only significantly advance our fundamental understandings of nephrology at nano scale but also render engineered nanoparticles new functionalities in early detection of kidney disease, which is influencing more than 10% of population worldwide. Taking advantage of strong NIR absorption of the well‐defined Au25(SG)18 nanocluster, we successfully used photoacoustic (PA) imaging to in‐situ visualize its transport through the aorta to the renal parenchyma and its subsequent filtration into the renal pelvis at a temporal resolution down to 1s. High temporal and spatial resolution imaging of Au25(SG)18 kidney elimination allows us for the first time to accurately quantify glomerular filtration rate (GFR) of individual kidneys in normal and pathological conditions using PA method, broadening biomedical applications of engineered nanoparticles in preclinical kidney research as anew tool for interrogating kidney functions at high temporal resolution and the anatomic level.