Bioluminescence　tomography　(BLT)　is　a　novel　in　vivo　technique　in　small　animal　studies,　which　can　reveal　the　molecular　and　cellular　information　at　the　whole-body　small　animal　level.　At　present,　there　is　an　increasing　interest　in　multispectral　bioluminescence　tomography,　since　multispectral　data　acquisition　could　improve　the　BLT　performance　significantly.　In　view　to　the　ill-posedness　of　BLT　problem,　we　develop　an　optimal　permissible　source　region　strategy　to　constrain　the　possible　solution　of　the　source　by　utilizing　spectrum　character　of　bioluminescent　source.　Then　a　linear　system　to　link　the　measured　data　with　the　unknown　light　source　variables　is　established　by　utilizing　the　optimal　permissible　region　strategy　based　on　adaptive　finite　element　analysis.　Furthermore,　singular　value　decomposition　analysis　is　used　for　data　dimensionality　reduction　and　improving　computational　efficiency　in　multispectral　case.　The　reconstructed　speed　and　stability　benefit　from　adaptive　finite　element,　the　permissible　region　strategy　and　singular　value　decomposition.　In　the　numerical　simulation,　the　heterogeneous　phantom　experiment　has　been　used　to　evaluate　the　performance　of　the　proposed　algorithm　with　the　Monte　Carlo　based　synthetic　data.　The　reconstruction　results　demonstrate　the　merits　and　potential　of　our　methodology　for　localizing　bioluminescent　source.　©　2009　SPIE.