user manual setis™ bioreactor

VERVIT - Sopet nv Karnemelkstraat 2-4 9060 Zelzate Tel: +32499265783 Belgium Fax: +3293456957 BTW: BE0435096468 info@setis-systems.be www.vervit.be www.setis-systems.be

UsermanualSETIS bioreactorWehopegivingenoughandclearinformationinthismanual,tohelpyoutouseSETISbioreactorswithoutanydifficulty.Ifyourequiremoreinformationdonothesitatetocontactus.Manualversion:May2012EnjoySETIS !!!!PartsofSETIS bioreactor: culturevessel(1x) mediavessel(1x) screwcapforculturevessel(1x) screwcapformediavessel(1x) siliconegasketforculturevesselscap(1x) siliconegasketformediavesselscap(1x) airfilters(2x) siliconetube6/9mm(1x,tofurtherbesectionedinthreepieces,seebelow)SterilizationPreparegrowthmediaandplaceitinthemediavessel,whilestandingverticalonitsbackside.Amaximumof3litresisrecommended.Placethesiliconegasketinthecorrespondingscrewcapandclosethevesselhardly.Coverbothconnectors(airconnectorandmediaconnector)withaluminiumfoil.Sterilizemediavesselwithgrowthmediaintheautoclave,preferable121C@1,2Atmfor2025min.Placethevesselsalwaysvertically.Onceremovedfromtheautoclaveandstillhot,trytoscrewagainthecaptoguarantyagoodsealing.

Airconnectors

MediaconnectorsCulturevessel Mediavessel

VERVIT - Sopet nv Karnemelkstraat 2-4 9060 Zelzate Tel: +32499265783 Belgium Fax: +3293456957 BTW: BE0435096468 info@setis-systems.be www.vervit.be www.setis-systems.be

Theculturevesselissterilizedempty,alsointheautoclave.Placethesiliconegasketinthecorrespondingscrewcapandclosethevesselhardly.Coverbothconnectors(airconnectorandmediaconnector)withaluminiumfoil.Sterilizeculturevesselintheautoclave,preferablewithautoclaveprogramformaterialsat121125C@1,4Atmfor1520min.Plasticfoilissometimesrecommendedtobeplacedaroundtheneckofthevesseltocoverthecap/neckjoinoncesterile.Thiscanreducethedustaccumulationwhilestorage.Siliconetube(6/9mm)providedwiththebioreactoriscutinthreepieces.Twopiecesof68cmandonepieceof1820cm(mediatransfersiliconetube).Thefirsttwopiecesarethenconnectedtotheairfiltersprovided.IMPORTANT!!!ConnectionismadeattheoppositesideoftheINsideofthefilter.Please,controltheINsignengravedinthefilterbody.Thesethreepieces,thetwofilterswiththeirrespectivesiliconetubeattachedandthemediatransfersiliconetube,arepackagedandsterilizedintheautoclaveusingsameconditionsasforculturevessel.

VERVIT - Sopet nv Karnemelkstraat 2-4 9060 Zelzate Tel: +32499265783 Belgium Fax: +3293456957 BTW: BE0435096468 info@setis-systems.be www.vervit.be www.setis-systems.be

AssemblyattheLaminarFlowPlantmaterialissubcultureandplacedintheculturevesselasabulk.Forthis,theculturevesselisopenedinthelaminarflow,alwaysfacingtheairflow,andtheplantmaterialisintroduced.Closethecaphardlytoavoidairand/orgrowthmedialeaks.Oncetheplantmaterialisintheculturevessel,placetheculturevesselverticallyonitsbackside.Bringthemediavesselbesideit(alsoverticallyplaced)andfacethetopsideofthemediavesselwiththebottomsideoftheculturevessel.Removealuminiumfoilfromtheairconnectorsofbothcontainersandplacetheairfiltersviathesiliconetube.IMPORTANT!!!Avoidtouchingtheconnectorsandedgeofthesiliconetubewithyourhand.Onceairfiltersareconnected,removethealuminiumfoilfromthemediaconnectorsandplacethesiliconetubeof1820cmlength.Thenthebioreactorisalreadyassembled!!

VERVIT - Sopet nv Karnemelkstraat 2-4 9060 Zelzate Tel: +32499265783 Belgium Fax: +3293456957 BTW: BE0435096468 info@setis-systems.be www.vervit.be www.setis-systems.be

Forprotocolsthatrequireachangeofmediaaftercertainperiod,asecondmediavesselispreparedwiththerespectivegrowthmediaandsterilizedasindicatedabove.BringtheSETISbioreactortothelaminarflowandplaceitvertically.Forplantmaterialsthatarealreadyrootedand/orfixedtothebottomoftheculturevessel,turntheculturevesselveryslowlytowardsitsverticalposition,toavoidmaterialdisplacement.Disconnectthemediatransfersiliconetubeatthesideofthemediaconnectorofthemediavessel.Discardthenthismediavesselwitholdgrowthmedia.Bringthenewmediavesselclosetotheculturevesselandremovethealuminiumfoilfromitsmediaconnector.Then,reconnectthemediatransfersiliconetubetothemediaconnectorofthenewmediavessel.Removealuminiumfoilfromtheairconnectorofthenewmediavesselandplaceanewairfilterviathesiliconetube.IMPORTANT!!!Avoidtouchingtheconnectorsandedgeofthesiliconetubewithyourhand.Insomecases,theairfilterfromtheoldmediavesselcouldbetransferredandreusedinthenewmediavessel.IMPORTANT!!!Workextremelycleantomakethistransfer,otherwisetherisksofcontaminationmightincrease.AttheshelfThetopsideofthemediavesseliscomplementarytothebottomsideoftheculturevessel.Viathesepins,theculturevesselisstackedandslidingisavoided.TransporttheSETISbioreactorsvertically,ifatrolleyisused,thiswillavoidmediatoenterthroughtheairconnectorofthemediavessel.Usethehandlersintegratedatthebottomofbothsidesofthemediavesseltocarryitwithyourhands.Useyourthumbstoholdtheculturevessel.Placethebioreactorinaperfectlyhorizontalandhardshelfsurface.Thebioreactorisdesignedwithadesiredangleatthebottomsurfaceoftheculturevesseltoallowselfdrainageafterimmersionisfinished.Thisinclinationcanbealteredbyirregularand/orwavyshelfsurfaces.Placethebioreactorwiththefrontside(whereallconnectorsandcapsarepresent),facingtheoutsideoftheshelf(corridor).Connectbothfilterswiththeirrespectivecompressedaircircuit.Nowthebioreactorisreadytowork!!!

VERVIT - Sopet nv Karnemelkstraat 2-4 9060 Zelzate Tel: +32499265783 Belgium Fax: +3293456957 BTW: BE0435096468 info@setis-systems.be www.vervit.be www.setis-systems.be

Foroptimizedshelfsetup,bioreactorsaredesignedtobeplacesidebyside,andbacktoback,withoutloosinganyshelfspace,withtheadvantageofhavingallconnectorsandcapsattheoutsideoftheshelf.InstallationDesignyourshelfsetupinsuchwayhavingamaximumof15SETISbioreactorsinonerowandamaximumof4rowsinparallel.SETISbioreactorhasbeendesignedwithaminimumheightdifferencebetweenthemediaconnectorsofbothvessels.Therefore,theairpressurerequiredtoperformanimmersionisverylow.Workingairpressurearebetween0,10,5BAR.Preferably0,2BAR.GeneralremarksSiliconegasketsaredesignedtoentertightintotheinnersideofeachcap.Thisistoavoidthatthesiliconegasketfallsfromthecaponcethevesselisopenedundersterileconditions.Toplacethesiliconegasketcorrectly,justpresentitintothecapandscrewittoitscorrespondingvessel.Theedgeofthevesselsneckwillpushthesiliconegasketuniformlytoitsplace.IMPORTANT!!!Donotplacethesiliconegasketbyslidingyourfingertowardsit.Youwillcreatealoopinthesiliconegasket,whichcangenerateleakslateron.Washthevesselsthoroughlyusingneutraldetergent.Aggressivedetergentscaninducecracksinthevesselsbodywhilesterilization.Rinsethemafterwardswithsoftwater,preferablyROwater.Avoidthevesselstogetincontactwithsolventsandoils.Performaperiodiccleaningofyourautoclave.Liquidmediaisidealforthegrowthofendogenousbacteria.Please,controlanddiagnosticverygoodyourstartingmaterialsbeforeyouinoculateaSETISbioreactor.SETISbioreactorisdesignedtoplacetheculturevesselontopofthemediavessel.Theywillbesolelyconnectedviathemediatransfersiliconetube,throughbothmediaconnectors.Makesurethisconnectionisproperlydone,withoutleakage.Avoidtwistthemediatransfersiliconetube,aswellasavoidbendingit.