• Preparea10mlsyringefittedwitha26Gshortneedleandfilledwith5to7mlofmediumand2to3mlofair.Airisimportant.
• PrepareaPasteurPipettewithrubberpipetter.

• EuthanizebyCO₂inhalation.AskyourComparativeMedicineDepartmentforproceduresandfacilities.Makesureyourmouseisdeadbeforestarting.
• Ifyouneedtocollectbloodfromheart,takewithalateralapproachtothethorax,soyoudon"tspillbloodintheperitoneum.
• NicktheaBDominalskinbelowthesternum,takingcarenottonicktheperitoneum.
• Grabthetwosidesofthecutandgentlyripaparttheabdominalskinwithafirmbutgentlemovement,exposingthesternumandthepelvis(Aimage).Becarefulnottoripmajorabdominalvesselsoryouwillgetbloodinperitoneum.
• Grabthesternumandgentlyfilltheabdominalcavitywiththemediumandtheair(Bimage).MakesuretokeeptheneedlefirmandsteadysoyoumakethesmallestpossIBLehole.Removetheneedle.Theperitoneumshouldself-seal.
• Holdthemouseinyourhandandshake(image)itvigorouslyaboutten-fifteentimes.Putitbackonthetable.
• Grabthesternumandsomeperitoneumwithatweezer(Cimage).Youshouldnotloosethegripuntilyouaredone.
• Withaswirlingmovementinserttheair-filledPasteurpipetteintheperitonealcavity(Cinsetimage).
• Expresstheairinthecavity,distendingtheperitoneumandstarttoaspiratethemediumandtheperitonealcells(Cimage).Yoursucessdependsofthedistensionoftheperitoneum.
• Collectallthefluidyouinserted.
• Whilestillholdingthesubsternalperitoneum,openuptheabdominalwalllongitudinally(Dimage)andcollecttheremainingfluid.

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