OTHERTOOLSANDRESOURCES
CHEMICALSTRUCTURE
(Z)-4-(3,5-difluoro-4-hydroxybenzylidene)-2-methyl-1-(2,2,2-trifluoroethyl)-1H-imidazol-5(4H)-one (DFHBI-1T)
MOLECULARFORMULA:
C13H9F5N2O2
MOLECULARWEIGHT:
320.21
CASNAME/NUMBER:
4H-Imidazol-4-one,5-[(3,5-difluoro-4-hydroxyphenyl)methylene]-3,5-dihydro-2-methyl-3-(2,2,2-trifluoroethyl)-,(5Z)/1539318-36-0
FLUORESCENCESPECTRA
AbsorptionandfluorescenceemissionspectraofSpinach2TM/DFHBI-1TinpH7.4buffer.
PRESENTATION: Lyophilizeddye
QUALITYASSURANCE: Productsareanalyzedby 1HNMRandLC-MSandprovidedatpurityof>95%byHPLC.
USAGESTATEMENT: Thisproductisintendedforresearchuseonlyandarenottobeusedforanyotherpurpose,whichincludesbutisnotlimitedto,unauthorizedcommercialuses,invitrodiagnosticuses,exvivoorinvivotherapeuticusesoranytypeofconsumptionorapplicationtohumansoranimals.Duetothehighlyspecificnatureoffluorophoresandaptamers,wecannotpredictorbeheldresponsIBLewithrespecttohowLucernaTM productswillbehaveinitscustomers"systems.ResearchersusingLucernaTMproductsshouldconductoptimizationstudiestoachievetheoptimalresultpossiblefortheirintendedapplication.
LICENSING:CornellUniversityhasfiledpatentstitled"RNAsequencesthatswitchonthefluorescenceofsmallmoleculesandtheiruseindetectionofRNAinvitroandincells"and"novelmethodsforRNAdetectionandquantification." AllcommercialusersshouldcontactCornellUniversityforauselicense.
DFHBI-1TIMAGES
A. Live-cellimagingofaCOS7cellexpressingCGG60-Spinach2TM inthepresenceofeither20µMDFHBIorDFHBI-1T. Imagesofthesamecellwereacquiredusinga100msecexposurewithaGFPfilterset. B. Quantificationofaverage DFHBI-1Tbrightnessof10cellsexpressingCGG60-SpinachTM normalizedtothebrightnessofDFHBI.
REFERENCEFORTHEPRODUCT:
SongW,StrackRL,SvensenN,JaffreySR.2014. Plug-and-PlayFluorophoresExtendtheSpectral PropertiesofSpinach. JACS136(4):1198-1201.
THISPRODUCTWASCITEDIN:
SvensenN,JaffreySR.2016. FluorescentRNAaptamersasatooltostudyRNA-modifyingenzymes. CellChemBiol23(3):415-25.
AwSS,Tang,MX,TeoYN,CohenSM.2016.Aconformation-inducedfluorescencemethodformicroRNAdetection. NucleicAcidsRes44(10):e92.
AlamKK,TawiahKD,LichteMF,Porciani,BurkeDH.2017.AfluorescentsplitaptamerforvisualizingRNA-RNAassemblyinvivo. ACSSynthBiol6(9):1710-1721.
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: