Description | Thecausesandeffectsofneuronaldegenerationareofmajorinteresttoawidevarietyofneuroscientists.Parallelingthisgrowinginterestisanincreasingnumberofmethodsapplicabletothedetectionofneuronaldegeneration.Fluoro-Jade®Cstainsalldegeneratingneuronsregardlessofspecificinsultormechanismofcelldeath.Fluoro-JadeCexhibitsthegreatestsignaltobackgroundratio,aswellasthehighestresolution.Thistranslatestoastainofmaximalcontrastandaffinityfordegeneratingneurons.Thismakesitidealforlocalisingnotonlydegeneratingnervecellbodiesbutalsodistaldendrites,axonsandterminals.ThedyeishighlyresistanttofADIngandiscompatIBLewithvirtuallyallhistologicalprocessingandstainingprotocols. |
Applications | TheFluoro-JadeC"ReadytoDilute"(RTD)StainingKitprovidesaneasytouseassortmentofFluoro-JadeC,DAPI,sodiumhydroxideandpotassiumpermanganateinliquidform.Followingourdetailedprotocol,Fluoro-JadeClabelleddegeneratingneuronsarevisualisedwithbluelightexcitationwhileDAPIcounterstainedcellnucleiarevisualisedwithultra-violetIllumination.TheFluoro-JadeCStainingKitcanbeusedonallkindsofpreservedtissues,includingfresh-frozen,paraformaldehydeorformalinfixed,andformalinfixed,paraffin-embeddedtissues. |
Comments | MATERIALSPROVIDED SodiumHydroxide,SolutionA(Dilute1:10priortouse)-40mL PotassiumPermanganate,SolutionB(Dilute1:10priortouse)-40mL Fluoro-JadeC,SolutionC(Dilute1:10priortouse)-40mL DAPI,SolutionD(AddtodilutedFluoro-JadeC)-40mL EQUIPMENTANDREAGENTSNEEDED Gelatincoatedmicroscopeslides Stainingdishes/Coplinjars Coverslips DPXmountingmedia Slidewarmer Convectionoven Distilledwater Ethanol Xylene NUMBEROFSLIDESPROCESSED: Theactualnumberofslidesprocessedbythiskitwilldependlargelyuponthevesselthatisusedtoincubatetheslides.IfusingastandardCoplinJar,itscapacityis50mLandtypicallyholds5slidesperjar.Ifusingsuchadevice,then80-100slidesstainedper50mlofworkingsolution(or,5mlofstocksolution)couldbeprocessedinoneday.NotethediluteddyeisNOTstableandwillnotstoreovernight.Itisbesttousefreshlydiluteddyeeachtimeanexperimentalbatchisstarted. FinalworkingconcentrationsofFJC:0.0001% FinalworkingconcentrationofKMnO4:0.06% |
Specificity | Degeneratingneurons,andneuronaldegeneration.Thereisnospecificstaininginnormalhealthybrain. |
Cross-reactivity | Note:SomeresearchersundersomeconditionsreportbloodvesselstainingwithFluoroJade.ThismaybebecauseFluoroJadeisananalogueofeosin(whichstainsbloodcells).Ingeneral,goodperfusionandpreparationofthetissueshouldhelppreventbloodvesselstainingbutitmaynotbepossibletoeliminateitentirely.Inourexperienceitisgenerallypossibletodistinguishneuronalfrombloodvesselsstainingbyeye. |
Form | ThereagentsintheFluoroJadekitareallsuppliedinaliquidformatandareready-to-dilute. |
Appearance | FJCvisualizationisaccomplishedusingbluelightora488nmLaser.ExcitationPeak:495nmEmissionPeak:521nmFiltersystemforvisualizing:Fluorescein/FITC |
Reconstitution | Dilutesolutionsasdirectedintheprotocolinstructions.Sometimessmallprecipitatesmaybepresentinthestockordilutedsolutions.Completemixingofthedilutedsolutionsusuallydissolvestheprecipitates.Theprecipitates,ifnotremoved,donotusuallycauseanydifficultiesifthewashingstepsarefollowedasinstructed.Optional:ForentirelycleansolutionsBiosensisrecommendsfilteringthedilutedsolutionthoughethanolandNaOHcompatiblesyringeorvacuumfilterdevicespriortocontactwithtissueslides. |
Storage | Theunopenedkitcanbestoredforupto6monthsat2-8ºCafterthedateofreceipt.Thekitandcomponentsshouldbestoredprotectedfromlight.DilutedFJCdyesolutionsarenotstableandshouldbeusedwithin4hoursofmaking.Theotherdilutedsolutionscanbereusedandstoredforupto48hoursifrefrigeratedandprotectedfromlight.Bestresultsrequirefreshlydilutedsolutions.Werecommendusingaseptictechniqueswhenhandlingthereagentstoavoidbacterialgrowthandcontamination. TheFJCReadytodilutekitisshippedambientandstableatroomtemperatureduringtransport.Refrigerateuponarrival,donotfreeze. |
ExpiryDate | Unopenedkit6monthsat2-8ºCprotectedfromlight.SeeStorageinstructionsforworkingsolutionsrecommendations. |
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ReagentKitprotocol | TR-100-FJ_January2017_protocol.pdf |
MSDS | MSDS_TR-100-FJ_as_at_May2017.pdf |
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