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VERVIT - Sopet nv Karnemelkstraat 2-4 9060 Zelzate Tel: +32499265783 Belgium Fax: +3293456957 BTW: BE0435096468 [email protected] www.vervit.be www.setis-systems.be
UsermanualSETIS™ bioreactorWehopegivingenoughandclearinformationinthismanual,tohelpyoutouseSETIS™bioreactorswithoutanydifficulty.Ifyourequiremoreinformationdonothesitatetocontactus.Manualversion:May‐2012EnjoySETIS™ !!!!PartsofSETIS™ bioreactor:
‐ culturevessel(1x)‐ mediavessel(1x)‐ screwcapforculturevessel(1x)‐ screwcapformediavessel(1x)‐ siliconegasketforculturevessel’scap(1x)‐ siliconegasketformediavessel’scap(1x)‐ airfilters(2x)‐ siliconetube6/9mm(1x,tofurtherbesectionedinthreepieces,see
below)SterilizationPreparegrowthmediaandplaceitinthemediavessel,whilestandingverticalonitsbackside.Amaximumof3litresisrecommended.Placethesiliconegasketinthecorrespondingscrewcapandclosethevesselhardly.Coverbothconnectors(airconnectorandmediaconnector)withaluminiumfoil.Sterilizemediavesselwithgrowthmediaintheautoclave,preferable121°C@1,2Atmfor20‐25min.Placethevesselsalwaysvertically.Onceremovedfromtheautoclaveandstillhot,trytoscrewagainthecaptoguarantyagoodsealing.
Airconnectors
Mediaconnectors
Culturevessel
Mediavessel
VERVIT - Sopet nv Karnemelkstraat 2-4 9060 Zelzate Tel: +32499265783 Belgium Fax: +3293456957 BTW: BE0435096468 [email protected] www.vervit.be www.setis-systems.be
Theculturevesselissterilizedempty,alsointheautoclave.Placethesiliconegasketinthecorrespondingscrewcapandclosethevesselhardly.Coverbothconnectors(airconnectorandmediaconnector)withaluminiumfoil.Sterilizeculturevesselintheautoclave,preferablewithautoclaveprogramformaterialsat121‐125°C@1,4Atmfor15‐20min.Plasticfoilissometimesrecommendedtobeplacedaroundtheneckofthevesseltocoverthecap/neckjoinoncesterile.Thiscanreducethedustaccumulationwhilestorage.Siliconetube(6/9mm)providedwiththebioreactoriscutinthreepieces.Twopiecesof6‐8cmandonepieceof18‐20cm(mediatransfersiliconetube).Thefirsttwopiecesarethenconnectedtotheairfiltersprovided.IMPORTANT!!!Connectionismadeattheoppositesideofthe“IN”sideofthefilter.Please,controlthe“IN”signengravedinthefilterbody.Thesethreepieces,thetwofilterswiththeirrespectivesiliconetubeattachedandthemediatransfersiliconetube,arepackagedandsterilizedintheautoclaveusingsameconditionsasforculturevessel.
VERVIT - Sopet nv Karnemelkstraat 2-4 9060 Zelzate Tel: +32499265783 Belgium Fax: +3293456957 BTW: BE0435096468 [email protected] www.vervit.be www.setis-systems.be
AssemblyattheLaminarFlowPlantmaterialissubcultureandplacedintheculturevesselasabulk.Forthis,theculturevesselisopenedinthelaminarflow,alwaysfacingtheairflow,andtheplantmaterialisintroduced.Closethecaphardlytoavoidairand/orgrowthmedialeaks.Oncetheplantmaterialisintheculturevessel,placetheculturevesselverticallyonitsbackside.Bringthemediavesselbesideit(alsoverticallyplaced)andfacethetopsideofthemediavesselwiththebottomsideoftheculturevessel.Removealuminiumfoilfromtheairconnectorsofbothcontainersandplacetheairfiltersviathesiliconetube.IMPORTANT!!!Avoidtouchingtheconnectorsandedgeofthesiliconetubewithyourhand.Onceairfiltersareconnected,removethealuminiumfoilfromthemediaconnectorsandplacethesiliconetubeof18‐20cmlength.Thenthebioreactorisalreadyassembled!!
VERVIT - Sopet nv Karnemelkstraat 2-4 9060 Zelzate Tel: +32499265783 Belgium Fax: +3293456957 BTW: BE0435096468 [email protected] www.vervit.be www.setis-systems.be
Forprotocolsthatrequireachangeofmediaaftercertainperiod,asecondmediavesselispreparedwiththerespectivegrowthmediaandsterilizedasindicatedabove.BringtheSETIS™bioreactortothelaminarflowandplaceitvertically.Forplantmaterialsthatarealreadyrootedand/orfixedtothebottomoftheculturevessel,turntheculturevesselveryslowlytowardsitsverticalposition,toavoidmaterialdisplacement.Disconnectthemediatransfersiliconetubeatthesideofthemediaconnectorofthemediavessel.Discardthenthismediavesselwitholdgrowthmedia.Bringthenewmediavesselclosetotheculturevesselandremovethealuminiumfoilfromitsmediaconnector.Then,reconnectthemediatransfersiliconetubetothemediaconnectorofthenewmediavessel.Removealuminiumfoilfromtheairconnectorofthenewmediavesselandplaceanewairfilterviathesiliconetube.IMPORTANT!!!Avoidtouchingtheconnectorsandedgeofthesiliconetubewithyourhand.Insomecases,theairfilterfromtheoldmediavesselcouldbetransferredandreusedinthenewmediavessel.IMPORTANT!!!Workextremelycleantomakethistransfer,otherwisetherisksofcontaminationmightincrease.AttheshelfThetopsideofthemediavesseliscomplementarytothebottomsideoftheculturevessel.Viathesepins,theculturevesselisstackedandslidingisavoided.TransporttheSETIS™bioreactorsvertically,ifatrolleyisused,thiswillavoidmediatoenterthroughtheairconnectorofthemediavessel.Usethehandlersintegratedatthebottomofbothsidesofthemediavesseltocarryitwithyourhands.Useyourthumbstoholdtheculturevessel.Placethebioreactorinaperfectlyhorizontalandhardshelfsurface.Thebioreactorisdesignedwithadesiredangleatthebottomsurfaceoftheculturevesseltoallowself‐drainageafterimmersionisfinished.Thisinclinationcanbealteredbyirregularand/orwavyshelf‐surfaces.Placethebioreactorwiththefrontside(whereallconnectorsandcapsarepresent),facingtheoutsideoftheshelf(corridor).Connectbothfilterswiththeirrespectivecompressedaircircuit.Nowthebioreactorisreadytowork!!!
VERVIT - Sopet nv Karnemelkstraat 2-4 9060 Zelzate Tel: +32499265783 Belgium Fax: +3293456957 BTW: BE0435096468 [email protected] www.vervit.be www.setis-systems.be
Foroptimizedshelfsetup,bioreactorsaredesignedtobeplacesidebyside,andbacktoback,withoutloosinganyshelfspace,withtheadvantageofhavingallconnectorsandcapsattheoutsideoftheshelf.InstallationDesignyourshelfsetupinsuchwayhavingamaximumof15SETIS™bioreactorsinonerowandamaximumof4rowsinparallel.SETIS™bioreactorhasbeendesignedwithaminimumheightdifferencebetweenthemediaconnectorsofbothvessels.Therefore,theairpressurerequiredtoperformanimmersionisverylow.Workingairpressurearebetween0,1–0,5BAR.Preferably0,2BAR.GeneralremarksSiliconegasketsaredesignedtoentertightintotheinnersideofeachcap.Thisistoavoidthatthesiliconegasketfallsfromthecaponcethevesselisopenedundersterileconditions.Toplacethesiliconegasketcorrectly,justpresentitintothecapandscrewittoitscorrespondingvessel.Theedgeofthevessel’sneckwillpushthesiliconegasketuniformlytoitsplace.IMPORTANT!!!Donotplacethesiliconegasketbyslidingyourfingertowardsit.Youwillcreatealoopinthesiliconegasket,whichcangenerateleakslateron.Washthevesselsthoroughlyusingneutraldetergent.Aggressivedetergentscaninducecracksinthevesselsbodywhilesterilization.Rinsethemafterwardswithsoftwater,preferablyRO‐water.Avoidthevesselstogetincontactwithsolventsandoils.Performaperiodiccleaningofyourautoclave.Liquidmediaisidealforthegrowthofendogenousbacteria.Please,controlanddiagnosticverygoodyourstartingmaterialsbeforeyouinoculateaSETIS™bioreactor.SETIS™bioreactorisdesignedtoplacetheculturevesselontopofthemediavessel.Theywillbesolelyconnectedviathemediatransfersiliconetube,throughbothmediaconnectors.Makesurethisconnectionisproperlydone,withoutleakage.Avoidtwistthemediatransfersiliconetube,aswellasavoidbendingit.
