Due to the low background and deep tissue penetration of near-infrared light, near-infrared fluorescence (NIRF) probes have received much attention in recent years. Cell membrane targeting probes can be used in in vitro and in vivo for living cell imaging, disease diagnosis and tumor targeted therapy. However, most of the currently available cell membrane probes are blue or green emissive fluorophores, NIRF probes are relatively rare. Herein, we designed and prepared a NIRF chimeric peptide probe named C-16-CARK, which could target the tumor cell membrane with longtime retention (more than 4 h). Moreover, the C-16-CARK can heat and disrupt the cell membrane upon high irradiation dose, revealing its dual roles as a cell membrane imaging probe and a photothermal agent. This kind of near-infrared fluorescence probe with cell membrane targeting and retention property provides a well design paradigm for the field of cancer imaging and treatment.