• IntroductiontoImmunocytochemistry(HouseEarInstitute)Abriefoverviewofcommonavailablemethods.
• BrDUImmunocytochemistryusingperoxidaseandDABforrattissues(ContributedbyShyamLaxminarayan)BrDU(5-Bromo-2-Deoxyuridine)isasyntheticThymidineanalogwhichincorporatesintosinglestrandedDNAduringtheS-PhaseoftheCellCycle.AntibodiesagainstBrDUcanbeusedalongwithadetectionsystem(suchasPeroxidase/DAB)toimmunostainforBrDUinmammaryglandsectionstorevealthedistributionofactivelydividingcells,hencethe%ofproliferatingcells.
• EstrogenReceptor/ProgesteroneReceptorImmunocytochemistry(ContributedbyShyamLaxminarayan)Protocolspecificallyfordetectingestrogen/progesteronereceptorbyimmunocytochemistry.
• Immunocytohistochemistry(HowellLab)Proceduresforcellplating,fixing,antibodyincubationandmounting

·ImmunocytochemicalIdentificationofProliferatingCellsinVascularTissue(anti-BrdU);VectorRedDetection(WilcoxLab)

·ImmunocytocheimicalIdentidicationofProliferatingCellsinVascularTissue(anti-BrdU);DiaminobenzADIne(DAB)BrownDetection(WilcoxLab)

• Immunohisto-andImmunocyto-chemisty-EnzymaticProtocol(R&D)Generalandniceprotocolforimmunocytoandimmunohistochemistryincludingsamplepreparation,stainingandtroubleshooting
• Immunocytochemisty-FluorescenceProtocol(R&D)GeneralprotocolincludingdetailsonstainingcellinsUSPensionandonslide
• EmbeddinginResinsforImmunocytochemistry(HouseEarInstitute)Describestheuseofthespecializedandnotsospecializedresins.
• FreezeSubstitutionforEmbeddinginResins(HouseEarInstitute)Asimple,inexpensivewayofembeddinginresinusingfreezesubstitutionmethods.
• UsingResinSectionsforImmunofluorescence(HouseEarInstitute)AprotocolforapplyingantibodieswithfluorescentMarkersontosectionsofresin-embeddedmaterial.Thethinsectionshelpincreaseresolution.
• PreparationofColloidalGoldProbes(HouseEarInstitute)Seehoweasily,andinexpensively,theseexpensiveprobescanbeprepared.Excellentinformationiflargevolumesofgoldprobesarerequired.
• SilverEnhancementofColloidalGold(HouseEarInstitute)Howtoprecipitatesilverontosmallcolloidalgoldparticlessothatthesignalcanbedetectedbylightmicroscopy.
• ProblemswithImmunolabeling(HouseEarInstitute)Whenlabelingcellsorsectionswithantibodiesorotheraffinitymarkers,theresultsarenoteasilyinterpreted.Lookhereforsomehelp.
• TheUseofAffinityCytochemistrytoDetectLigandsBoundtoTargetCells(ChildsLab)Thisprotocolisusedtodetectneuropeptidesandgrowthfactorsboundtotargetsitesonpituitarycells.Pituitarycellsaregrowninculturefor1-5days(timedependentontheexperimentaldesign).Thecellsareexposedtophysiologicalconcentrationsofbiotinylatedligandsfor1minto4h,dependingonthetypeofexperiment.Afterexposure,thecellsarefixedforlightorelectronmicroscopicdetectionofthesiteofbinding.
• Preparingcoatedcoverslips(HoshiLab)Somecellssticktocoveslipsbetteriftheyarecoatedwithpolyaminoacidsorcollagen.Poly-(D,L,D/L)-lysineiscommonlyusedtotreatcoverslips.

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