Organic　dyes　generally　suffer　from　small　Stokes　shift　that　usually　leads　to　self-quenching　and　-gaining　errors　during　the　fluorescent　imaging　process.　Here,　a　through-bond　energy　transfer　(TBET)　cassette　is　developed　with　large　Stokes　shift　to　pursue　precise　cell　imaging.　The　TBET　is　constructed　by　covalently　conjugated　tetraphenylethene　(acts　as　donor)　and　rhodamine　(acceptor)　through　an　acetylene　bond.　The　constructed　TBET　cassette　distinctly　behaves　as　dual-Stokes　shifts,　including　a　large　pseudo-Stokes　shift　caused　by　energy　transfer,　from　donor＇s　absorption　to　acceptor＇s　emission　(up　to　260　nm)　and　a　smaller　Stokes　shift　of　acceptor　molecules　itself.　Due　to　the　intrinsic　dual-Stokes　shifts,　TBET　cassette　exhibits　specific　dual　distinct　absorbances,　single　shared　emission　properties,　which　can　be　excitated　under　two　different　laser　channels.　By　colocalization　of　the　imaging　readouts　of　these　two　channels,　the　precisely　double　checked　fluorescent　imaging　is　achieved　in　living　cells.