Fluorescent　probes　with　superior　two-photon　fluorescence　are　highly　attractive　in　the　field　of　bioimaging.　Herein,　we　report　a　one-pot　hydrothermal　route　to　synthesis　amine,　sulfo　cofunctionalized　graphene　quantum　dots　(GQDs),　which　acts　as　efficient　one-photon　and　two-photon　fluorescent　probes　for　cellular　imaging.　As　synthesized　GQDs　exhibit　ultrastability　due　to　the　edge-site　functionalization　of　amine　and　sulfo　groups.　In　addition,　GQDs　display　attractive　two-photon　fluorescence　properties.　The　two-photon　absorption　cross　section　of　GQDs　reaches　up　to　31,000　GM,　which　substantially　exceeds　that　of　the　majority　of　traditional　fluorescent　materials.　Furthermore,　the　noncytotoxicity　GQDs　exhibit　a　negligible　photothermal　effect　under　808　nm　femtosecond　laser　irradiation,　which　is　suitable　for　long-term　two-photon　imaging　and　observation.　These　findings　open　new　possibilities　for　using　two-photon　fluorescent　GQDs　in　various　biological　applications.