AutomatedDrug InjectionPortsEnableKinetic,FunctionalData4druginjectionportsperwell·Addinhibitors,stimulants,substrates,andcompounds·Injectionsaredefined bytheuser360320280c240200120R2110MEminuteSeahorseBioscienceBOSTON·COPENHAGEN·SHANGHAI
Automated Drug Injection Ports Enable Kinetic, Functional Data • 4 drug injection ports per well • Add inhibitors, stimulants, substrates, and compounds • Injections are defined by the user
SSSSMitochondrialGlycolysisGhucoseHRespirationATPOxygenConsumptionRateOGlycolysisExtracellularAcidificationRateRespirationOCRECARATPFCCPAntimycinAOligomycin10mM2-DG&RotenoneOligomycinGlucose455040203580SpareGlycolyti30Respiratory40ReserveCapacity2530Maximal2050RespirationATV1520ProdurtoiGlyoolysisLaa1080540vticdoidibcatinNon010120304050607080900100110108020XFGlycolysisStressTestXFCellMitoStressTestSeahorse BioscienceBOSTON·COPENHAGEN·SHANGHAI
Oxygen Consumption Rate OCR Glycolysis XF Glycolysis Stress Test XF Cell Mito Stress Test Mitochondrial Respiration Extracellular Acidification Rate ECAR
Glycolysis Stress Test--Analyze Metabolic Reprogramming in Primary Immune Cells2-DGGiucoseOtigamycin100Control.LPS7550250-00204060800Time (min)SeahorseBioscienceBOSTON·COPENHAGEN·SHANGHAT
Glycolysis Stress Test -Analyze Metabolic Reprogramming in Primary Immune Cells
SSSSMitochondrialGlycolysisGlucoseH中Respiration?ATPOxygenConsumptionRateOGlycolysisExtracellularAcidificationRateRespirationOCRECARATPFCCPAntimycinAOligomycin10mM&Rotenone502080SpareRespiratory40Capacity00Maximal50RespirationATV20ProdurtoiRase80H40001001101020A6080XFGlycolysisStressTestXFCellMitoStressTestSeahorseBioscienceBOSTONCOPENHAGEN·SHANGHAI
Oxygen Consumption Rate OCR Glycolysis XF Glycolysis Stress Test XF Cell Mito Stress Test Mitochondrial Respiration Extracellular Acidification Rate ECAR
SSSSMitochondrial Electron Transport ChainⅡIIIIVVComplex:12H+2H+2x2H+脚+Aoo04FlavoproteinVllbND1IronOUTprotien8sCytdIyvIbaeeFrdbFrdcInnerCytEQEemitochondrialmembrane1.3.99.1YvoVIeATPCore1.3.5.1Quinone1.9.3.2CortND3poolvllesynthase0IN1.10.2.2SFumarateOHHOSuccinateeeH-SuccinatedehydrogenaseO3.6.1.363.6.1.34hromecoxidaseCytochromebc1complex3.6.1.35HHNAD*NADHdehydrogenaseADPATPNADHKrebsPeterS,RabinwitchafterMandavillietal,MutationResearch509(2002)127-15)CycleISeahorse BioscienceBOSTONCOPENHAGEN·SHANGHAI
Krebs Cycle e - e - e - e - H+H+ H+H+ Cyt C e - e - O H+H+ 2 H+H+H2O ADP ATP Cyt C e - O2 Q H+`H+ NADH NAD+ H+ H+ H+ H+H+ H+ Cyt C H+` e - e - e - IN OUT Peter S. Rabinwitch after Mandavilli et al, Mutation Research 509 (2002) 127–151 Mitochondrial Electron Transport Chain 2OH- OH-