ProductDescription
SphereCol®beadsarecoatedwithVitroCol®humanTypeIcollagenandareidealforgrowingcellsinsUSPension.Thecollagencoatedbeadsprovideanaturalinvivo-likeenvironmenttopromotehighcellgrowthwhileprovidingalargesurfaceareaforcellstoattachwithoptimalsurfaceareatovolumeratios.SphereCol®providesa3Dbio-scaffoldwhichisoptimalinmanycellcultureprocedures.
SphereCol®,humancollagencoatedbeadsiscoatedwithhighlypurifiedTypeIhumancollagenderivedfromahumanfibroblastcellcultureprocess,VitroCol®.Thecollagenprovidesanoptimalcoatingonthebeadstoenhancecellattachment,cellviABIlity,cellproliferationandcellfunction.Thecollagenbeadsrangeinsizefromabout125to250micron.Theproductispackagedina20mlbottleandsterilized.SphereCol®isprovidedinauser-friendlypackagingforuseandstorage.
| Parameter,Testing,andMethod | SphereCol®#5138 |
| BeadShape | Spherical |
| PackageSize | 10grams |
| BeadsperGram | 4.6x105 |
| BeadSizeDistribution | 125-250microns |
RelativeDensityRange | 1.022-1.030 |
| SurfaceAreaperBead | 360cm2/gram |
| CollagenUsedforCoating | VitroCol®HumanTypeICollagen |
| StorageTemperature | RoomTemperature |
| ShelfLife | Minimumof6monthsfromdateofreceipt |
| SterilizationMethod | IrrADIation |
DirectionsforUse
DownloadthefullPDFversionorcontinuereadingbelow:
Note:SphereColisprovidedasasterile,drypowderandthusmustbehandledinanasepticmanner.
Theinstructionsbelowaredesignedtoserveasageneralguidelineforthecultureofa200mlspinnercontainingaSphereCol®,humancollagenbeadsataconcentrationof5cm2/mlataseedingdensityof20,000cells/cm2utilizingalowserumattachmentphase(i.e.,0.05%FBS-containingmedium).
Preparation
- Weighout2.78grams(1000cm2)oftheSphereCol®beadsandaddthemtoasterilized250mlspinnerflask.Note:WeighouttheappropriatemassofSphereCol®beadsintoasteriletube.AddavolumeofsterilePBSormediumsuchthatthemassofbeads(andthus,surfacearea)pervolumeisknown.Aliquotthedesiredmassintoasterilevessel.Youmayremovetheliquidbycarefullydecantingoraspirating.
- Add180mloflowserum-containingcellculturemedium(i.e.,0.05%)tothespinnerflask.Thisallowsfora10mladditionofcellinoculumfollowedbya10mladditionofFBSuponsatisfactorycellattachment.
Acclimation
- Tomaximizecellattachment,themediumandSphereCol®beadsmixtureshouldbeacclimatedtothecultureenvironmentpriortoaddingthecellinoculum.Forexample,placementofthevessel(i.e.,a200mlspinneronastirplate)ina37°C,5%CO2incubatorforaminimumof30minutesallowsfortemperature,gasandpHequilibration.
GenerateCellInoculum
- Harvestcellsusingstandardcellculturetechniquesandreagents.Note:Theexactprocedurerequiredtoproducetheoptimalcellinoculumwillvarybasedonthecelltypeandcellculturesystem.Ideally,auniform,single-cellsuspensionisdesiredtoallowforanevendistributionofcellsacrosstheSphereCol®beadspopulation.
- Uponachievingasatisfactorycellsuspension,transfer20X106cellstoacentrifugetube,spindownandre-suspendin10mllow-serumcellculturemedium.Note:20X106cellsacross1000cm2equatesto20,000cells/cm2
CellAttachmenttoSphereCol®Beads(“AttachmentPhase”)
- Removespinnerflaskfromincubatorandplaceonstirplateunderlaminarflowcabinet.
- Add10mlcellsuspensiontospinnerflask.
- Uponsatisfactorycellattachment,add10mlserumtobringthefinalconcentrationofserumto5%.Note:CellswilltypicallybegintoattachtoSphereCol®beadsoverthefirstfewhoursoftheculture,althoughthiswillvarybasedoncelltypeandcultureconditions.Ideally,theattachmentphaseisconsideredcompleteonce>90%ofcellsareattached,whichcanbeconfirmedmicroscopically(seeMonitoringandMaintainingtheCulture).
AdditionalConsiderations:
- Alow-serumattachmentphaseissometimesrequiredforcellsthatwilleithernotattachatall,ordonotattachinanevenlydistributedmannerinthepresenceofthestandardserumconcentrations.Theoptimalconcentrationofserumduringtheinitialattachmentphase(typicallythefirst1to4hours)mustbedeterminedforeachcellculturesystem.Oftentimesithasbeenfoundtobeverylow(i.e.,0.05%).Uponsatisfactorycellattachment,serummaybeaddedtothedesiredfinalconcentration.
- Anallowancefortheadditionofserumaftercellattachmentmustalsobeaccountedforifperformingalow-serumattachmentstep.
- AnevendistributionofcellsacrosstheSphereCol®beadspopulationiscriticalinmaximizingtheusageofavailablesurfacearea,leadingtomaximalcellyield.
- Agitationspeedisaprocessparameterthatrequiresoptimizationforgoodcellattachmentinspinners.Ataminimum,theagitationrateshouldbesufficienttoevenlysuspendtheSphereCol®beads.Ingeneral,thelowestagitationratethatallowsforgoodcellattachmentandevensuspensionofSphereCol®beadsshouldbechosen,soastolessensheerforcesexerteduponthecellsbythedynamicenvironmentofthespinner.
MonitoringandMaintainingtheCulture
- Theculturemaybemonitoredbycollectingrepresentativesamplesinculturedishes(i.e.,multiwellplates)andvisualizingunderamicroscope.Cellattachmentandspreadingcanbeeasilyobservedat100Xmagnificationatvarioustimepointsatwhichtimeaqualitativeassessmentoftheattachmentandspreadingcanbemade.CellscanbevisualizedattheedgesorcircumferenceoftheSphereCol®beadsasrounded(initialattachmentphase),“gumdrop-shaped”(earlyspreading)orflattened(completelyspread).
- Aswithflatcultureware,mediaexchangesmaybenecessarytomaintainastablesupplyofnutrientsoverthecourseoftheculture.Inalaminarflowhood,allowtheSphereCol®beadstosettletothebottomofthevesselandwithdrawthedesiredvolumeofmediumfromthetop,takingcarenottoremoveanySphereCol®beads.Replacewithanequalvolumeoffresh,warmedculturemedium.
AdditionalConsiderations:
- SeveraltechniquesmaybeusedtoenhancevisualizationofcellsonSphereCol®beadsifneeded:
- Fluorescencetechniques(i.e.,DAPIstainingmethod)
- Acridineorange
- Directvisualizationbyphasemicroscopy
- Asthecellsgrow,SphereCol®beadswillbecomeheavier,andtheagitationrate(inthecaseofaspinnerculture)mayneedtobeincreasedtomaintainauniformsuspension.
HarvestingCells
- WhilestandardcellharvestingreagentsandtechniquesusedforflatculturewaretypicallyworkwellwithSphereCol®beadscultures,harvestconditionswillneedtobeoptimizedforeachcelltypeandcellculturesystemtogetthebestresultspossIBLe.Optimalharvestconditionsinflatculturewareprovideagoodstartingpoint.Ingeneral,theconditionsgentlestonthecellsshouldbeused.
- Removethemediafromthespinner,beingcarefulnottoremovecell-ladenSphereCol®beads.
- WashtheSphereCol®beadswith40mlphosphatebufferedsaline(PBS).Incubateatroomtemperaturefor10minutes.
- AspiratePBS,andadd10mldissociationenzyme(i.e.,trypsin).AllowcellstoincubateintheenzymeuntiltheydissociatefromtheSphereCol®beadssurface,thentrituratetoobtainasinglecellsuspension.SphereCol®beads/cellsmaybeplacedat37°Ctofacilitatedetachment.
- Countcellswithahemocytometerusingtrypanblue.SphereCol®beadsgenerallydonotgetunderthecoverslip,sotheywillnotinterferewiththecount.Keeptrackofallthereagentvolumesusedaswellasthemediavolumeremovedsothatthecellcountcanbeadjustedwiththeappropriatedilution/concentrationfactor.
AdditionalConsiderations:
- Washingisperformedtoaidintheremovaloftrypsininhibitingmediacomponents.SolutionsotherthanPBSarealsowellsuitedforthispurpose.
- Trypsinconditionsshouldbeasgentleaspossible,solongasasingle-cellsuspensionisobtainedinastimelyfashion.Highertrypsinconcentrations,longerincubationtimes,andhighertemperaturesaresomefactorsthatcouldnegativelyimpactcellhealth.
- Somecells,suchasMDCKcells,aredifficulttodissociate,andthereforerequireharshertechniques,suchastheuseof0.25%(5X)Trypsin-EDTA,andincubationat37°Cforover10minutes.ItmayalsobebeneficialtoperformanEDTAwashpriortotrypsinaddition.
- Foranimalcomponentderivedfreesystems,trypsinsubstitutes,suchasTrypLE,canbeusedtochemicallydissociatecells.
- ToseparatedissociatedcellsfromSphereCol®beads,thesamplecanbepassedthroughacellstrainer.
- SphereCol®beads/cellseparationcanalsobeperformedviadifferentialsettlinginaconicaltube.
ProductCellAssay
Todemonstratecellattachment,cellswereseededontoSphereCol®humancollagencoatedbeads.ThephotobelowshowsafluorescentimageofhumanMesenchymalStemCellsattachedtothebeads.DAPIstainednucleiappearblueandPhalloidin-Alexa-488stainingofactinfilamentsisgreen.
ProductReferences
SphereCol®References:
Park,Yonsil,etal."Hepaticdifferentiationofhumanembryonicstemcellsonmicrocarriers."Journalofbiotechnology174(2014):39-48.
Dai,Lin,etal."Inorganic–organicnanocompositeassemblyusingcollagenasatemplateandsodiumtripolyphosphateasabiomimeticanalogofmatrixphosphoprotein."Crystalgrowth&design11.8(2011):3504-3511.
Rafiq,QasimA.,etal."Systematicmicrocarrierscreeningandagitatedcultureconditionsimproveshumanmesenchymalstemcellyieldinbioreactors."Biotechnologyjournal11.4(2016):473-486.
Lin,YoushanMelissa,etal."Criticalattributesofhumanearlymesenchymalstromalcell-ladenmicrocarrierconstructsforimprovedchondrogenicdifferentiation."Stemcellresearch&therapy8.1(2017):93.
Yoon,Junghyo,etal."FabricationoftypeIcollagenmicrocarrierusingamicrofluidic3DT-junctiondeviceanditsapplicationforthequantitativeanalysisofcell–ECMinteractions."Biofabrication8.3(2016):035014.
Rafiq,QasimA."TowardascalableandconsistentmanufacturingprocessfortheproductionofhumanMSCs."CellandGeneTherapyInsights2.1(2016):127-140.
Wang,Zhenxing,etal."Developmentofdemineralizedbonematrix-basedimplantableandbiomimeticmicrocarrierforstemcellexpansionandsingle-steptissue-engineeredbonegraftconstruction."JournalofMaterialsChemistryB5.1(2017):62-73.
Suess,P.M.,Chinea,L.E.,Pilling,D.&Gomer,R.H.ExtracellularPolyphosphatePromotesMacrophageandFibrocyteDifferentiation,InhibitsLeukocyteProliferation,andActsasaChemotacticAgentforNeutrophils.TheJournalofImmunology(2019).doi:10.4049/jimmunol.1801559
Tavassoli,H.etal.Large-scaleproductionofstemcellsutilizingmicrocarriers:ABiomaterialsengineeringperspectivefromacademicresearchtocommercializedproducts.Biomaterials181,333–346(2018).
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ThisproductisforR&Duseonlyandisnotintendedforhumanorotheruses.PleaseconsulttheMaterialSafetyDataSheetforinformationregardinghazardsandsafehandlingpractices.
美国AdvancedBioMatrix（简称ABM） www.advancedbiomatrix.comAdvancedBioMatrix（简称ABM）是美国一家著名的生物公司，获得了AllerganInc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。AdvancedBioMatrix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。AdvancedBioMatrix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。
美国AdvancedBioMatrix是3D组织培养、细胞检测和细胞增殖等领域实验解决方案的佼佼者。AdvancedBioMatrix在分离、纯化、冻干、细胞培养和蛋白检测、多肽粘附、附着因子、基质硬度和其他3Dmatrix 产品开发方面有着丰富的经验。AdvancedBioMatrix的研发经验和专业知识确保其产品可达到最佳质量，并保证产品之间一致性，方便研究客户使用。以下为AdvancedBioMatrix3DMatrices 产品竞争优势：1. 提供高纯度和成分确定的胞外基质；2. 超过1000余篇文献引用PureCol产品，品质非常均一；3. 在3D培养基领域可提供最全面的产品线；4. 唯一可提供特异性刚性有机硅基板的公司（CytoSoft）;5. 唯一可提供可溶性丝纤蛋白的供应商（可运用于多种3D培养）；6. 如果客户首次接触3D胶原凝胶，AdvancedBioMatrix还是唯一的预制胶原蛋白(PureColEZGel)供应商；
以下产品为AdvancedBioMatrix全球畅销品：1.PureCol 牛源I型胶原蛋白 3mg/ml#5005-100ML2.Nutragen牛源I型胶原蛋白 6mg/ml#5010-50ML3.FibriCol 牛源I型胶原蛋白 10mg/ml#5133-20ML4.VitroCol 人源I型胶原蛋白 #5007-20ML5. 弹性蛋白原 #5052-1MG6.ECMSelectArraykitUltra-36#5170-1EA7.CytoSoft（刚性可变的基底，AdvancedBioMatrix最新添加产品5190-7EA）8. 人III型胶原蛋白 #5021-10MG9. 人IV型胶原蛋白 #5022-5MG10.SilkFibroin溶液 #5154-20ML11.Fibronectin#5080-5MG12.Vitronectin#5051-0.1MG
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问题：
1，既然外国公司的原材料这么贵，我就想购买国内厂家的，请各位大侠帮忙提供一下国内哪些厂家的玻璃胚针物廉价美？
2，国内的哪些大学的测试中心，能够提供拉针仪的有偿使用服务的？
具体操作是:局麻下将3～4根电极导管经股静脉、锁骨下静脉送入冠状静脉窦、高位右心房及希氏束、右心室等部位，刺激心房和心室诱发与临床一致的心动过速，定位心动过速起源点，然后将消融用的电极导管送达已定位的起源点并与体外的射频发生器相连。放电后重复电生理检查，若不能诱发心动过速且临床随访无发作，则说明消融成功。
此方法治疗的疾病有:预激综合征和房室结双经路引起的阵发性室上性心动过速、房扑和房颤、室性心动过速及房性心动过速。
脑电波：
脑电波（Electroencephalogram,EEG）是大脑在活动时，大量神经元同步发生的突触后电位经总和后形成的。它记录大脑活动时的电波变化，是脑神经细胞的电生理活动在大脑皮层或头皮表面的总体反映。
脑电波来源于锥体细胞顶端树突的突触后电位。脑电波同步节律的形成还与皮层丘脑非特异性投射系统的活动有关。
脑电波是脑科学的基础理论研究，脑电波监测广泛运用于其临床实践应用中。
电子波
电子波是指电子产品等所发出的电子辐
电子波的害处
电脑及大多数家用电器设备等都是可以产生各种形式不同频率、不同强度的电磁辐射源。
电磁辐射--对人体机理的危害
电场辐射危害人体的机理主要是热效应、非热效应和累积效应等。
热效应：人体70%以上是水，水分子受到电子波辐射后相互摩擦，引起机体升温，从而影响到体内器官的正常工作。
非热效应：人体的器官和组织都存在微弱的电磁场，它们是稳定和有序的，一旦受到外界电场的干扰，处于平衡状态的微弱电场即将遭到破坏，人体也会遭受损伤。
累积效应：热效应和非热效应作用于人体后，对人体的伤害尚未来得及自我修复之前（通常所说的人体承受力---内抗力），再次受到电子波辐射的话，其伤害程度就会发生累积，久之会成为永久性病态，危及生命。对于长期接触电子波辐射的群体，即使功率很小，频率很低，也能诱发体内想不到的病变，应引起警惕。
谢谢啦~
1.在没有开始记录（空跑的状态下）和开始记录时的波形的基线都不在0点而是处于负值，是因为仪器设备设置的问题还是仪器本身有损坏？
2.记录ACC场电的通道50Hz干扰特别大，接地线排干扰后仍然存在，可能是什么问题呢？
3.Brownlee440的Amplifier上有Gain，lowpassfilter，Highpassfilter的设置，这个设置对ACC场电的记录有影响吗？如果记录ACC的场电，一般常用的参数是多少啊？
4.有没有用过这个仪器记录过肌电的前辈，我用A-B模式可以记录到类似Chart5软件记录的肌电波形。可是用clampfit10.2的Analyze--statistics--Measurement--Area分析,分析出来的数值太小，与波形不符。从波形看，明显有强的肌肉收缩，但是数值却没有明显差异。有前辈分析过肌电吗？是否我的分析方法不对？或者是应为问题1中提到的基线不在0点所引起的曲线下面积分析的误差？
感谢！
