| Overview |  PrinterFriendlyVersion |
| Ex/Em(nm) | 485/520 |
| MW | N/A |
| CAS# | N/A |
| Solvent | N/A |
| Storage | F/D/L |
| Category | CellBIOLOGy LabelingCells |
| Related | FluorescenceImaging |
| Spectrum | AdvancedSpectrumViewer |
- PrepareNileGreen™stainingsolution:
- WarmNileGreen™(ComponentA)toroomtemperature.
- PrepareNileGreen™stainingsolutionbydiluting5µLofNileGreen™(ComponentA)to1mLofStainingBuffer(ComponentB).
Note1.50µLofNileGreen™(ComponentA)isenoughforone96-wellplate.AliquotandstoreunusedNileGreen™at<-20=""ºc.=""protect=""it=""from=""light=""and=""avoid=""repeated=""freeze-thaw="">
Note2.TheoptimalconcentrationoftheNileGreen™variesdependingonspecificapplications.ThestainingconditionsmaybemodifiedaccordingtoaparticularcelltypeandthepermeABIlityofthecellsortissuestotheprobe.
- Prepareandstaincells:
- Foradherentcells:
a.Growcellseitherina96-wellblackwall/clearbottomplate(100µL/well/96-well)oroncover-slipsinsideapetridishfilledwiththeappropriateculturemedium.
b.Gentlyaspiratetheculturemedium,andaddequalvolume(suchas100µL/well/96-wellplate)oftheNileGreen™stainingsolution(fromStep1.2).
c.Incubatethecellsina37°C,5%CO2incubatorfor10~30minutes.
d.RemoveNileGreen™stainingsolution(Optional,seeNote1below).
e.ReadFluorescenceat485/520nmwithamicroplatereaderorobservethecellsusingafluorescencemicroscopewithaFITCfilterset. - ForsUSPensioncells:
a.Centrifugethecellsat1000rpmfor5minutestoget1-5×105cellspertube.
b.Resuspendcellsin500μLofNileGreen™workingsolution(fromStep1.2).
c.Incubateatroomtemperatureor37ºCfor10to30min,protectedfromlight.
d.CentrifugetoremovetheNileGreen™workingsolution,andresuspendcellsin500μLofpre-warmedmediumorbufferofyourchoicetoget1-5×105cellspertube(Optional,seeNote1below).
e.MonitorthefluorescenceincreaseusingfluorescencemicroscopewithaFITCfiltersetorflowcytometeratFL1channel.
Note1:SinceNileGreen™hasminimalfluorescenceinaqueousmedia,aspirationofthegrowthmedium(step2.1b)andremovalofNileGreen™stainingsolution(step2.1d)afterstainingisoptional.
Note2:Stainedcellscanbefixedwith3-4%formaldehyde.Inaddition,prefixedcells(3-4%formaldehydefixation)canbestainedwithNileGreen™stainingsolution.
- Foradherentcells:
| References&Citations |  PrinterFriendlyVersion |
1. Nevo-Yassaf,I.;Lovelle,M.;Nahmias,Y.;Hirschberg,K.;Sklan,E.H.,LivecellimagingandanalysisoflipiddropletsbiogenesisinHCVinfectedcells.Methods2017.
2. Makino,A.;Hullin-Matsuda,F.;Murate,M.;Abe,M.;Tomishige,N.;Fukuda,M.;Yamashita,S.;Fujimoto,T.;Vidal,H.;Lagarde,M.;Delton,I.;Kobayashi,T.,Acuteaccumulationoffreecholesterolinducesthedegradationofperilipin2andRab18-dependentfusionofERandlipiddropletsinculturedhumanhepatocytes.MolBiolCell2016,27(21),3293-3304.
3. Wang,C.W.,Lipiddroplets,lipophagy,andbeyond.BiochimBiophysActa2016,1861(8PtB),793-805.
4. Takahashi,H.;Kutasy,B.;Friedmacher,F.;Takahashi,T.;Puri,P.,Expressionofhepaticlipiddropletsisdecreasedinthenitrofenmodelofcongenitaldiaphragmatichernia.PediatrSurgInt2016,32(2),155-60.
5. Schneider,M.R.,Lipiddropletsandassociatedproteinsinsebocytes.ExpCellRes2016,340(2),205-8.
6. Natarajan,S.K.;Rasineni,K.;Ganesan,M.;Feng,D.;McVicker,B.L.;McNiven,M.A.;Osna,N.A.;Mott,J.L.;Casey,C.A.;Kharbanda,K.K.,Structure,functionandmetabolismofhepaticandadiposetissuelipiddroplets:implicationsinalcoholicliverdisease.CurrMolPharmacol2015.
7. Papadopoulos,C.;Orso,G.;Mancuso,G.;Herholz,M.;Gumeni,S.;Tadepalle,N.;Jungst,C.;Tzschichholz,A.;Schauss,A.;Honing,S.;Trifunovic,A.;Daga,A.;Rugarli,E.I.,Spastinbindstolipiddropletsandaffectslipidmetabolism.PLoSGenet2015,11(4),e1005149.
8. Hashemi,H.F.;Goodman,J.M.,Thelifecycleoflipiddroplets.CurrOpinCellBiol2015,33,119-24.
9. Kochan,K.;Maslak,E.;Krafft,C.;Kostogrys,R.;Chlopicki,S.;Baranska,M.,Ramanspectroscopyanalysisoflipiddropletscontent,distributionandsaturationlevelinNon-AlcoholicFattyLiverDiseaseinmice.JBiophotonics2015,8(7),597-609.
10. Billecke,N.;Rago,G.;Bosma,M.;Eijkel,G.;Gemmink,A.;Leproux,P.;Huss,G.;Schrauwen,P.;Hesselink,M.K.;Bonn,M.;Parekh,S.H.,ChemicalimagingoflipiddropletsinmuscletissuesusinghyperspectralcoherentRamanmicroscopy.HistochemCellBiol2014,141(3),263-73.
