MiniChamber for Regulating Gaseous Environment During Culture

1.ThebasicunitisconstructedfromamediafiltrationunitsuchasCorning’s115mlfiltersystem(catalognumbers430944-430947).

2.Tygontubing(3.2mmi.d.x6.4mmo.d.)withastopcockfastenedtotheendisattachedtothehoseconnector.

3.A00sizerubberstopperisinsertedintothepourspoutofthefiltersystem.

4.Themembraneinthefiltersystemispuncturedinseveralplacestofacilitatemovementofgases.

5.Cultureplates(uptoa60mmpetridishplate)aregentlypositionedontopofthemembrane.Iftheplateislarge,itisdifficulttomaneuvertheplateintothechamberwithfingers.Inthiscase,acradleunderneaththeplatecanbeconstructedwithtapetofacilitateplacementofthedish.

6.ThelidofthefiltersystemistapedsecurelywithscotchtapeandParafilmisplacedaroundthelidtomakethechamberairtight.

7.Gasesareinjectedfor3minthroughthepourspoutfromahoseconnectedtothegascylinderwhilethestopcockwasintheopenposition.

8.Afterinjectionofgasesiscompleted,therubberstopperisfastenedtothepourspoutandthestopcockismovedtotheclosedposition.

9.Topreventcontaminationduringinjectionofgases,afilter(0.22mm)isattachedtothegashose.