Analysis of murine BM

HistologicanalysisofmurineBMisanecessarycomplementtoflowcytometricorinvitroanalysis.Techniquestodothisarewellestablishedinhumanhematopathology.AlongtraditioninexperimentalhematologyfocusesonfemurandtibiaasthesourceofBM.However,thethicknessofthecorticalboneandtheoftenpreferreduseofthelargestbone,thefemur,forcellcultureorflowleavedtothepathologistahardspecimenwithlittlemarrow.

ThespineisanexcellentsourceforBManalysisandisnotusedforanyothertypeofanalysis.

Procedure:

Onceananesthetizedmouseissacrified,correctlyfixedtoaboardandopenedup,removeallthevisceralorgansandmostoftheretroperitonealtissue.Withascalpel,makeadeepincisionintheleft(A)andright(B)psoasmuscles,asclosetothespinalprocessesasyoucan.Thencutthespineatthelevelofthelastribs(C)andthenatthepelvis(D).Nowyoucanremovethewholespine,whichwillhaveashinywhitishdorsalsurface,tworoughlateralsideswherethemuscleswereandaventralsidewiththevertrebralbodies.

Youcanfixthespineaccordingtoprotocolandthendecalcifyeitherinaciddecalcifyer(Cal-Ex,FisherDiagnostics,CS510-10D)orEDTA.Afterdecalcification,slicethespinelenghtwisesagittallyacrossthevertebralbodiesandembedthetwohalveswiththecutsidefacingthecuttingplane.Youwillhavesectionswithamplemarrow,bonecartilageandperispinalmuscletoevaluate.