Hydrogen　peroxide　(H2O2)　is　a　small　molecule　of　reactive　oxygen　species,　mainly　produced　by　cell　mitochondria,　which　plays　an　important　role　in　physiological　processes.　Excess　of　H2O2　in　normal　cells　could　causes　a　variety　of　diseases.　For　that　reason,　it　is　very　significant　to　emerge　a　detection　method　with　high-precision　for　H2O2.　Herein,　we　reported　a　structurally　novel　fluorescent　probe,　para-nitrophenyl　oxoacetyl　rhodamine　(RhB-NIR)　for　the　detection　of　H2O2　levels　in　living　cells.　The　results　show　that　the　Stokes　shift　for　RhB-NIR　is　140　nm,　much　larger　than　those　for　other　reported　rodamine　derivatives.　RhB-NIR　shows　satisfactory　specificity　and　high　sensitivity　for　the　detection　of　H2O2.　Excellent　linear　relationship　with　H2O2　within　a　certain　concentration　range　(0-10　mu　M)　was　displayed,　and　the　limit　of　detection　(LOD)　is　61　nM.　More　importantly,　RhB-NIR　realized　fluorescence　imaging　of　H2O2　in　living　cells.　In　addition,　RhB-NIR　also　shows　great　potential　for　targeting　mitochondria　in　cell　colocalization　experiments,　in　which　the　Pearson＇s　coefficient　is　0.91　using　Rhodamine　123　as　a　reference.　This　probe　will　be　a　promising　tool　for　detecting　H2O2　in　the　future　research.