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胶原蛋白、纤连蛋白、玻连蛋白、水性凝胶、低代成纤维细胞
℡ 4000-520-616
℡ 4000-520-616
Advanced BioMatrix/CytoSoft® Imaging 96-well Plate//5258-1EA 8 kPa
产品编号:5258-1EA8kPa
市  场 价:¥3300.00
场      地:美国(厂家直采)
产品分类: 其它>耗材>>
联系QQ:1570468124
电话号码:4000-520-616
邮      箱: info@ebiomall.com
美  元  价:$165.00
品      牌: Advanced BioMatrix
公      司:Advanced BioMatrix ,Inc.
公司分类:
Advanced BioMatrix/CytoSoft® Imaging 96-well Plate//5258-1EA 8 kPa
商品介绍

ProductDescription

ThisCytoSoft®Imaging96-wellproducthasadefinedelasticmodulus(seecertificateofanalysis)withaglassbottomforhighresolutionimagingThethicknessofthesiliconegelisuniformwitha~0.03mmthicklayerofsiliconeineachwell.ThesiliconegelsareactivatedandreadytobindtoapurifiedECM,suchasPureCol®typeIcollagen(#5005)priortocelladdition.

Therigidityofthesubstratetowhichcellsadherecanhaveaprofoundeffectoncellmorphologyandgeneexpression.CytoSoft®productsprovideatooltoculturecellsonsubstrateswithvariousrigiditiescoveringabroadphysiologicalrangeusingathinlayerofbiocompatIBLesilicone.

TheCytoSoft®–Imagingplatehasaglassbottomforhigh-resolutionimagingwherelowautofluorescenceandexceptionalopticalclarityarerequired.The96-wellformatisoptimalforhigh-throughputcellstudiesandscreening.Theplateconsistsofa#1.5thicknessglassbottombondedtoablackpolystyreneframeandincludesalid.Theplatesaresterilizedusingozoneandprovidedwith1plateperpackage.

Onthebottomofeachwell,thereisathinlayerofspeciallyformulatedbiocompatiblesilicone,whoseelasticmodulus(rigidity)iscarefullymeasuredandcertified.ThesurfacesofthegelsinCytoSoft®productsarefunctionalizedtoformcovalentbondswithaminesonproteins.Thechemicalfunctionalizationisstableandthereactiondoesnotrequireacatalyst,facilitatingcoatingofthegelsurfaceswithmatrixproteinsandplatingcells.

Thesiliconesubstratesareopticallyclearandhaveanearzeroauto-florescence.Thelayerofsiliconeineachwellisfirmlybondedtothebottomofthewell.Unlikehydrogels(suchaspolyacrylamidegels),siliconegelsarenotsusceptibletohydrolysis,donotdryorswell,areresilientandresistanttotearingorcracking,andtheirelasticmoduli(rigidities)remainnearlyunchangedduringextendedstoragetimes.

CytoSoft®-Imagingproductsaccommodatelivecellstainingusingmembraneandcellpermeabledyes;fixationofcellsusingcommontechniquessuchasparaformaldexyde;andimmunostainingoffixedcells.

Parameter,Testing,andMethodCytoSoft®Imaging96-WellPlates
SterilizationMethodOzone
PlateSize96-MultiwellBlackPolystyrenePlatewithGlassBottom
QuantityperPackage1Plate
Rigidty(ElasticModulus)0.2,0.5,2,8,16,32,64kPa(exactvaluesprovidedontheCofA)
StorageTemperatureRoomTemperature
ShelfLifeMinimumof6monthsfromdateofreceipt
PlateSurfaceMaterialFunctionalizedSilicone

GrowthAreaperWell

0.32cm2

TypicalWorkingVolumeperWell

40to280μL

CellAttachmentAssay

Pass

DirectionsforUse

DownloadthefullPDFversionorcontinuereADIngbelow:

CoatingProcedure

Note:Usetheserecommendationsasguidelinestodeterminetheoptimalcoatingconditionsforyourculturesystem.

CautionaryNote:Thebottomofa96-wellplatecanbedetachedbyexcessivemechanicalforcesuchascentrifugationordirectcontactwithliquidhandlingtools(tips,Pipettes,etc.).

RemovetheCytoSoft®productfromtheprotectivesleeveinasterilehood.

  1. Prepareextracellularmatrixmaterialbyneutralizinginamine-freebufferpH7.4to7.9(suchas1XDPBS).WedonotrecommendusinggelatinasyourECMprotein.

Note:Pre-warmthecoatingsolutiontoapproximatelyroomtemperaturebeforeuse.

  1. Diluteasneeded,anddispense~150uLofsolutionintoeachwelltofullycoatthesurface.

Note:RecommendeddilutionforPureCol®3mg/mlTypeIcollagenis1:30(~100µg/ml).

Note:Thehydrophobicsurfacerequireslargervolumestocoverthesurfacethandoconventionalplasticdishes

  1. IncubateECMcoatedCytoSoft®atroomtemperature,coveredfor0.5-1hour.
  1. Afterincubation,aspirateanyremainingmaterialandrinsecoatedsurfacesimmediatelytwotimeswithculturemediumorPBS.Leaveabout150uLofmediumperwelltokeepsurfacecovered.

Note:DonotallowtheCytoSoft®surfacetobecomedryoncethesurfacehasbeenwetted.

  1. Coatedsurfacesarereadyforuse.
  2. StandardharvestingproceduresusedforremovingcellsfromculturewarecanbeemployedforharvestingcellsfromtheCytoSoft®productincludinguseoftrypsin,Accutase®andnon-enzymaticcelldetachmentsolutions.

ProductQ&A

Werecommenda60Xobjectivefortheimagingplates.

Theelasticmodulusismeasuredbytrackingbeadsonthegelsurfaceunderawide-fieldfluorescencemicroscopewithoutanyotherspecializedequipment.Themeasurementshavesmallandsimpletoestimateerrorsandtheirresultsareconfirmedbyconventionaltensiletests.AmastercurveisobtainedrelatingthemixingratiosofthetwocomponentsofSylgard184withtheresultingelasticmoduliofthegels.

Usingprewarmedmediawilldecreasegassolubilityandhelppreventbubblesonthesurfaceofthesilicone.

Thesiliconegelcancrackifthesurfacebecomesdry.

No.Cellmatrixproteinsareattachedtothesurfaceofthegelviacovalentbonding.ItisdifficulttoformanewECMlayeraftercelldetachment.Therearenolongeranyreactivegroupsonthesurfaceofthegelaftertheinitialcellculture.

Thechangeintherefractiveindexofthesiliconedistortsthingsalittle,soDICwillbepossiblebutnotperfect.Also,DICisonlypossibleontheglassbottomimagingplates,notthe6-wellplasticplates.

1.41

Thetwomainissuesare:

1.IneffecientcoatingofthematrixproteinsduetolowpHofthecoatingsolution.

2.Insufficientwashofleftovercellmatrixafterthecoatingprocedure.

No.CytosoftmustbecoatedwithanECMproteinsuchasCollagenorFibronectin.

Thesurfaceisdecoratedwithanhydridefunctionalgroups.

Plasmauseisnotrecommended.PlasmawillinduceformationofahardcrustonthesurfaceandwillchangethemechanicalpropertiesoftheCytoSoft®products.

DonotfreezetheCytoSoft®products.

Whenfrozen,thereisagoodchancethatthesiliconesurfacegetshydrolyzedandabsorbsmoisture,whichwouldinactivatethebindingsitesandmaketheproductnot-usable.

Ifyoufrozetheproductanditisstillfrozen,warmtheCytoSoft®upat60Cwiththebagvented.Thatwillminimizethechanceofthemabsorbingmoisture–butthereisstillachancethattheywillnolongerbefunctional.

UsinggelatinforcoatingCytoSoftisoftenproblematicbecausegelatinoftenhaslowmolecularweightimpuritiesthatblockbindingsitesontheactivatedsurfaceofthesilicone.WerecommendusinghighlypurifiedECM"sinstead.

ProductCellAssay

CytoSoft®platescanalsobeusedtoshowhowfibroblastsareabletodiscriminatebetweentheunderlyingstiffness.Thisismanifestedinbothadhesionsizeandstressfibers,asseenbelow.Itappearsthatthecellsonthe8kPastiffnesshavereducedintracellulartensionandincreasedadhesion.

ProductReferences

ReferencesforCytoSoft®Products:

1.Modaresi,Saman,etal.“DecipheringtheRoleofSubstrateStiffnessinEnhancingtheInternalizationEfficiencyofPlasmidDNAinStemCellsUsingLipid-BasedNanocarriers.”Nanoscale,vol.10,no.19,2018,pp.8947–8952.,doi:10.1039/c8nr01516c.

2.Wilson,ChristinaL.InVitroModelsOfBrainForStudyOfMolecularMechanismsInBrainDisorder.TheUniversityofNEBraska-Lincoln,2016.

3.Wilson,C.L.,Hayward,S.L.,&Kidambi,S.(2016).Astrogliosisinadish:Substratestiffnessinducesastrogliosisinprimaryratastrocytes.RSCAdvances,6(41),34447-34457.doi:10.1039/c5ra25916a

4.Prager-KhoutorskyM,LichtensteinA,KrishnanR,RajendranK,MayoA,KamZ,GeigerB,BershadskyAD.Fibroblastpolarizationisamatrix-rigidity-dependentprocesscontrolledbyfocaladhesionmechanosensing.Nat.CellBiol.2011;13:1457–1465.

5.GutierrezE,TkachenkoE,BesserA,SunddP,LeyK,DanuserG,GinsbergMH,GroismanA.HighRefractiveIndexSiliconeGelsforSimultaneousTotalInternalReflectionFluorescenceandTractionForceMicroscopyofAdherentCells.PLoSOne.2011;6:e23807.

6.MerkelR,KirchgessnerN,CesaCM,HoffmannB.Cellforcemicroscopyonelasticlayersoffinitethickness.Biophys.J.2007;93:3314–23.

7.Schellenberg,A.etal.Matrixelasticity,replicativesenescenceandDNAmethylationpatternsofmesenchymalstemcells.Biomaterials35,6351–8(2014).

8.Cesa,C.M.etal.Micropatternedsiliconeelastomersubstratesforhighresolutionanalysisofcellularforcepatterns.Rev.Sci.Instrum.78,034301(2007).

9.Gutierrez,E.&Groisman,A.MeasurementsofElasticModuliofSiliconeGelSubstratewithaMicrofluidicDevice.PlosOne6(2011).

10.Mori,H.,Takahashi,A.,Horimoto,A.,andHara,M.Migrationofglialcellsdifferentiatedfromneurosphere-formingneuralstem/Progenitorcellsdependsonthestiffnessofthechemicallycross-linkedcollagengelsubstrate.NeuroscienceLetters,Vol.555,October(2013)

11.Banerjee,I.,Carrion,K.,Serrano,R.,Dyo,J.,Sasik,R.,Lund,S.etal.Cyclicstretchofembryoniccardiomyocytesincreasesproliferation,growth,andexpressionwhilerepressingTgf-βsignaling.JMolCellCardiol.2015;79:133–144

12.Vertelov,G.etal.Rigidityofsiliconesubstratescontrolscellspreadingandstemcelldifferentiation.Sci.Rep.6,33411;doi:10.1038/srep33411(2016).

13.TkachenkoE,RawsonR,LaE,etal.RigidSubstrateInducesEsophagealSmoothMuscleHypertrophyandEoEFibroticGeneExpression.TheJournalofallergyandclinicalimmunology.2016;137(4):1270-1272.e1.doi:10.1016/j.jaci.2015.09.020.

14.Sao,K.etal.MyosinIIgovernsintracellularpressureandtractionbydistincttropomyosin-dependentmechanisms.MolecularBIOLOGyoftheCell30,1170–1181(2019).

15.Cooper,J.G.etal.SpinalCordInjuryResultsinChronicMechanicalStiffening.JournalofNeurotrauma(2019).doi:10.1089/neu.2019.6540

ProductCertificateofAnalysis

Noresultfor.

ProductVideos

link to library blog - What is CytoSoft<sup>®?
WhatisCytoSoft®?

Video

BDceacb54ca4b15499b4.jpg"alt="linktolibraryblog-HowtoCoatCytoSoft®Plates">
HowtoCoatCytoSoft®Plates

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SafetyandDocumentation

SafetyDataSheet

CertificateofOrigin

ProductDisclaimer

ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.

品牌介绍
美国Advanced BioMatrix(简称ABM) www.advancedbiomatrix.com
Advanced BioMatrix(简称ABM)是美国一家著名的生物公司,获得了Allergan Inc的授权(Allergan用25年时间不断完善胶原蛋白相关的产品的生产工艺),将Allergan的专业和技术用于蛋白生产与检测,致力于为组织工程、细胞分析及细胞增殖等研究领域提供优质稳定的产品。

Advanced BioMatrix不断丰富已有产品线,目前可为三维细胞培养提供各种胶原蛋白、纤连蛋白、玻连蛋白、水性凝胶、不同粘度与分子量的透明质酸以及低代成纤维细胞等。在美国全部产品授权Sigma销售。

Advanced BioMatrix是组织培养,细胞分析和细胞增殖三维(3D)应用的生命科学领域的领导者。 我们的产品被公认为纯度,功能性和一致性的标准。
我们在生产,分离,纯化,冷冻干燥,细胞培养和蛋白质测试,粘附肽,附着因子,底物刚性和其他3D矩阵产品方面拥有丰富的专业知识。 我们的专业技术和知识正在被用来确保我们的产品质量最高,批次之间一致且易于为我们的研究客户使用。

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