| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 484/504 |
| MW | N/A |
| CAS# | N/A |
| Solvent | DMSO |
| Storage | F/D/L |
| Category | MicroBIOLOGy FlowCytometry |
| Related | FluorescenceImaging BiochemicalAssays |
| Spectrum | AdvancedSpectrumViewer |
Note1:Thefollowingprotocolcanbeadaptedformostcelltypes.Theseconditionsrequireadjustmentforeachcelltypeandexperimentalsystem.Growthmedium,celldensity,thepresenceofothercelltypesandfactorsmayinfluencestaining.Residualdetergentonglasswaremayalsoaffectstainingofmanyorganisms,andcausebrightlystainedmaterialtoappearinsolutionswithorwithoutcellspresent.
Note2:UseplastictubeswhendilutingMycoLightTMGreenJJ98andJJ99,becausethedilutedstainadherestoglass.Ingeneral,thebestresultsareobtainedinbuffersthatdonotcontainphosphate.
1.Adherentcellsinculturemaybestainedinsituoncoverslips.PelletcellsinsUSPensionbycentrifugationand
resuspendinbufferedsaltsolutionorwater.
2.DilutetheMycoLightTMGreenJJ98orJJ99withnon-phosphatebuffersuchasHepesbufferorbufferofyourchoice.Add MycoLightTMGreenJJ98.UsingtheconcentrationslistedbelowTable1asaguideline.Ininitialexperiments, itmaybebesttotryseveraldyeconcentrationsovertheentiresuggestedrangetodeterminetheconcentrationthat yieldsoptimalstaining.
Table1.SuggestedconditionsforstainingcellswithMycoLightTMGreenJJ98
Application | Concentration | StainingConditions |
Bacterialcells | 50nM–20μM | Vortextomix,thenincubatefor1–30minutes. |
Eukaryoticcells | 10nM–5μM | Incubatefor10–120minutes. |
Microarrays | 50nMinTEbuffer | Incubatefor5minutes,rinseandthendry. |
3.Stainedeukaryoticcellsgenerallyshowdiffusecytoplasmicstainingaswellasnuclearstaining.Particularly
MycoLightTMGreenJJ98andJJ99intensestainingofintranuclearbodiesarefrequentlyobserved.
| References&Citations |  PrinterFriendlyVersion |
1. WattsMR,JamesG,SultanaY,GinnAN,OuthredAC,KongF,VerweijJJ,IredellJR,ChenSC,LeeR.(2014)Aloop-mediatedisothermalamplification(LAMP)assayforStrongyloidesstercoralisinstoolthatusesavisualdetectionmethodwithSYTO-82fluorescentdye.AmJTropMedHyg,90,306.
2. WeidnerJ,CassensU,GohdeW,WullenweberJ,GreveB.(2013)AnewtriplexrealtimePCRwhichdistinguishesbetweenMRSA,MSSA,andmecAcoagulasenegativestrainsbymeansofmeltingpointanalysisusingSYTO9.ClinLab,59,795.
3. HubbardKS,GutIM,ScheelerSM,LymanME,McNuttPM.(2012)CompatibilityofSYTO13andHoechst33342forlongitudinalimagingofneuronviABIlityandcelldeath.BMCResNotes,5,437.
4. LarrosaM,TruchadoP,EspinJC,Tomas-BarberanFA,AllendeA,Garcia-ConesaMT.(2012)EvaluationofPseudomonasaeruginosa(PAO1)adhesiontohumanalveolarepithelialcellsA549usingSYTO9dye.MolCellProbes,26,121.
5. EischeidAC.(2011)SYTOdyesandEvaGreenoutperformSYBRGreeninreal-timePCR.BMCResNotes,4,263.
6. WlodkowicD,SkommerJ,DarzynkiewiczZ.(2011)RapidquantificationofcellviabilityandapoptosisinB-celllymphomaculturesusingcyanineSYTOprobes.MethodsMolBiol,740,81.
7. SeputieneV,VilkoicaiteA,ArmalyteJ,PavilonisA,SuziedelieneE.(2010)Detectionofmethicillin-resistantStaphylococcusaureususingdoubleduplexreal-timePCRanddyeSyto9.FoliaMicrobiol(Praha),55,502.
8. UllalAJ,PisetskyDS,ReichCF,3rd.(2010)UseofSYTO13,afluorescentdyebindingnucleicacids,forthedetectionofmicroparticlesininvitrosystems.CytometryA,77,294.
9. UdovichJA,BesselsenDG,GmitroAF.(2009)AssessmentofacridineorangeandSYTO16forinvivoimagingoftheperitonealtissuesinmice.JMicrosc,234,124.
10. WlodkowicD,SkommerJ,FaleyS,DarzynkiewiczZ,CooperJM.(2009)DynamicanalysisofapoptosisusingcyanineSYTOprobes:fromclassicaltomicrofluidiccytometry.ExpCellRes,315,1706.
