MaterialsRequiredbutNotDelivered | 1.PrecisionPipettes:sufficientforaliquotingcells. 2.Harvestingbuffer:EDTAortrypsinbasedcelldetachmentbuffer,orothercelldetachmentformulationsasoptimizedbyindividualinvestigators.Chemicon"sready-to-usenon-mammaliandetachmentsolution,Accutase?(Cat.No.SCR005)canalsobeused. 3.Tissueculturegrowthmediumappropriateforsubjectcells,suchasDMEMcontaining5%FBS. 4.QuenchingMedium:serum-freemedium,suchasDMEM,MEMetccontaining5%BSA.Mustcontaindivalentcations(Mg2+,Ca2+)sufficientforquenchingEDTAinharvestingbuffer. 5.SterilePBSorHBSStowashcells. 6.Distilledwater 7.(Optional)Chemoattractantorpharmacologicalagentaddedtoculturemedium. 8.Lowspeedcentrifugeandtubesforcellharvesting. 9.CO2incubatorappropriateforsubjectcells. 10.Hemocytometerorothermeansofcountingcells. 11.TrypanblueorequivalentviABIlitystain. 12.Microplatereader(540-570nmdetection)orspectrophotometer. 13.Sterilecellculturehood 14.(Optional)Graduatedocular(calibrated),orautomatedmethodforcountingstainedcellsonamembrane. 15.Shaker |