AimToensureallcellcultureproceduresareperformedtoastandardthatwillpreventcontaminationfrombacteria,fungiandmycoplasmaandcrosscontaminationwithothercelllines.

Materials

• Chloros/Preseptsolution(2.5g/l)
• 1%formaldehydebaseddisinfectante.g.Virkon,Tegador
• 70%ethanolinwater(Prod.No.R8382)

Equipment

• Personalprotectiveequipment(sterilegloves,laboratorycoat,safetyvisor)
• MicroBIOLOGicalsafetycABInetatappropriatecontainmentlevel

Procedure

1. Sanitizethecabinetusing70%ethanolbeforecommencingwork.
2. Sanitizeglovesbywashingthemin70%ethanolandallowingtoairdryfor30secondsbeforecommencingwork.
3. Putallmaterialsandequipmentintothecabinetpriortostartingworkaftersanitizingtheexteriorsurfaceswith70%ethanol.
4. Whilstworkingdonotcontaminateglovesbytouchinganythingoutsidethecabinet(especiallyfaceandhair).Ifglovesbecomecontaminatedre-sanitizewith70%ethanolasabovebeforeproceeding.
5. Discardglovesafterhandlingcontaminatedculturesandattheendofallcellcultureprocedures.
6. Equipmentinthecabinetorthatwhichwillbetakenintothecabinetduringcellcultureprocedures(mediabottles,Pipettetipboxes,pipetteaids)shouldbewipedwithtissuesoakedwith70%ethanolpriortouse.
7. Movementwithinandimmediatelyoutsidethecabinetmustnotberapid.Slowmovementwillallowtheairwithinthecabinettocirculateproperly.
8. Speech,sneezingandcoughingmustbedirectedawayfromthecabinetsoasnottodisrupttheairflow.
9. Aftercompletingworkdisinfectallequipmentandmaterialbeforeremovingfromthecabinet.Spraytheworksurfacesinsidethecabinetwith70%ethanolandwipedrywithtissue.Disposeoftissuebyautoclaving.
10. Cellculturediscardinchloros(10,000)ppmmustbekeptinthecabinetforaminimumoftwohours(preferablyovernight)priortodiscardingdownthesinkwithcopiousamountsofwater.
11. PeriodicallycleanthecabinetsurfaceswithadisinfectantsuchasPresept,TegadororVirkonorfumigatethecabinetaccordingtothemanufacturersinstructions.Howeveryoumustensurethatitissafetofumigateyourownlaboratoryenvironmentduetothegenerationofgaseousformaldehyde,consultyouron-siteHealthandSafetyAdvisor.