Endocytosis is a basic cellular process that describes a form of active transport across the plasma membrane into the cell. The endocytic pathway consists of distinct membrane compartments; internalized molecules are delivered to early endosomes, and some of them are recycled back to the surface, whereas other molecules are sent to late endosomes and lysosomes for degradation. However, little is known about how mitochondria are involved in the endocytic pathway. Here, we report that FM dyes, membrane-impermeant fluorescent lipid probes, can traffic to mitochondria directly from the plasma membrane by clathrin-mediated endocytosis. FM dye entry into mitochondria uses microtubule-dependent active transport, but the mechanism is different from the classical endocytic pathway. Hence, this study reveals a previously unrealized lipid trafficking pathway from the plasma membrane to mitochondria.