OTHERTOOLSANDRESOURCES
CHEMICALSTRUCTURE
3,5-difluoro-4-hydroxybenzylideneimidazolinone(DFHBI)
MOLECULARFORMULA:
C12H10F2N2O2
MOLECULARWEIGHT:
252.22
CASNAME/NUMBER:
4H-Imidazol-4-one,5-[(3,5-difluoro-4-hydroxyphenyl)methylene]-3,5-dihydro-2,3-dimethyl-,5(Z)- /1241390-29-3
FLUORESCENCESPECTRA
AbsorptionandfluorescenceemissionspectraofDFHBIinpH7.4buffer.
PRESENTATION: Lyophilizeddye
QUALITYASSURANCE: Productsareanalyzedby 1HNMRandLC-MSandprovidedatpurityof>95%byHPLC.
USAGESTATEMENT:Thisproductisintendedforresearchuseonlyandarenottobeusedforanyotherpurpose,whichincludesbutisnotlimitedto,unauthorizedcommercialuses,invitrodiagnosticuses,exvivoorinvivotherapeuticusesoranytypeofconsumptionorapplicationtohumansoranimals.Duetothehighlyspecificnatureoffluorophoresandaptamers,wecannotpredictorbeheldresponsIBLewithrespecttohowLucernaTM productswillbehaveinitscustomers"systems.ResearchersusingLucernaTMproductsshouldconductoptimizationstudiestoachievetheoptimalresultpossiblefortheirintendedapplication.
DFHBIIMAGES
HEK293Tcellsweretransfectedwitheither5S-controlaptameror5S-SpinachTM plasmidsandincubatedwith20µMDFHBI.ImagesarephaseoverlaywithHoechst-stainednuclei(blue)andSpinachTM fluorescence(green).
SpinachTM RNArobustlyactivatesthefluorescenceofDFHBI. EachtubecontainedtheindicatedsolutionandwasirrADIatedat465nm.
REFERENCEFORTHEPRODUCT:
RNAmimicsofgreenfluorescentprotein.PaigeJS,WuKY,JaffreySR. Science.2011Jul29;333(6042):642-6.
THISPRODUCTWASCITEDIN:
RNAsignalamplifiercircuitwithintegratedfluorescenceoutput.AkterFandYokobayashiY.ACSSynthBiol.2015May15.
Design,synthesis,andapplicationofSpinachmolecularbeaconstriggeredbystranddisplacement.BhadraSandEllingtonAD.MethodsEnzymol.2015Jan15.
Live-cellimagingofmammalianRNAswithSpinach2.StrackRLandJaffreySR.MethodsEnzymol.2015Jan6.
MonitoringmRNAandproteinlevelsinbulkandinmodelvesicle-basedartificalcells.vanNiesP,CantonAS,Nourianz,DanelionC.MethodsEnzymol.2015Jan6.
UsingSpinachaptamertocorrelatemRNAandproteinlevelsinEscherichiacoli.PothoulakisGandEllisT.MethodsEnzymol.2015Jan5.
Invitroanalysisofriboswitch-Spinachaptamerfusionsasmetabolite-sensingfluorescentbiosensors.KellenbergerCAandHammondMC.MethodsEnzymol.2014Dec19.
TheSpinachRNAAptamerasaCharacterizationToolforSyntheticBIOLOGy. PothoulakisG,CeroniF,ReeveB,EllisT. ACSSynBiol.2013Sep13.
AsuperfoldingSpinach2revealsthedynamicnatureoftrinucleotiderepeat-containingRNA.StrackRL,DisneyMD,JaffreySR. NatMehtods.2013Dec;10(12):1219-24.
Genepositionmorestronglyinfluencescell-freeproteinexpressionfromoperonsthanT7transcriptionalpromoterstrength.ChizzoliniF,ForlinM,CecchiD,MansySS. ACSSynBiol.2013Dec9.
LICENSING:
Lucernahastheexclusiverightstoallpatentscoveringtheuseofthisproduct. AllcommercialusersshouldcontactLucernaforauselicense.
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这些染料都非常成熟,光毒和淬灭都很低,当然要考虑到你采集图像时的显微镜参数。
比如calcein,常用的示踪,绿光(虽然这些颜色只是根据光谱加上去的伪彩),但要考虑你的实验过程中结合细胞结构,是否会伴随calcein的泄漏,就是荧光降低。
dri,还可以看膜啊
cfda也不错
bcef虽是ph指示,但你试验时不仅可观察细胞,还可看细胞ph变化,也行。
当然你或许还要结合其他方法,如细胞免疫化学等手段去双染或多染,都需要综合考虑染料之间特性。单独染一个染料,有点浪费,不如多染,数据和图像也好看些。现在流行细胞成像。
想请问一下,DAPI这个染料到底有没有膜通透性,我通过百度搜索查询关于DAPI染料的,基本上是说它能透过细胞膜对活细胞和死细胞均能染上蓝色;但是也有人说DAPI只可以透过死细胞膜,不能对活细胞进行染色,用以区分活死细胞,到底哪个是对的啊,蒙了!!!!!!
For principle,look at this site: