微生物学微生物学 (22).pptVIP

Afterlearningthissection,youshouldbeabletoDifferentiatethestreak-plate,spread-plate,andpour-platemethodsforisolatingpurecultures

Thestreak-platemethodandsimilarmethodsforisolatingpureculturesweredevelopedbytheRobertKoch.Theuseofthesemethodstransformedmicrobiology,andwithin20yearsoftheirdevelopment,mostpathogensresponsibleformajorbacterialdiseaseshadbeenisolatedandidentified.

Kochreasonedthatifcellsfromamixtureofmicrobescouldbespatiallyisolatedfromeachother,eachcellwouldgiverisetoacompletelyseparatecolony-amacroscopicallyvisibleclusterofmicroorganismsinoronasolidmedium.Becauseeachcolonyarisesfromasinglecell,eachcolonyrepresentsapureculture.

StreakplateInthistechnique,cellsaretransferredtotheedgeofanagarplatewithaninoculatingloop.InoculatingloopStreakthecellsoverthesurfaceAdilutionprocessSeparatecolonies

SpreadPlateandPourPlateSpread-plateandpour-platetechniquesaresimilarinthattheybothdiluteasampleofcellsbeforeseparatingthemspatially.Theydifferinthatthespreadplatespreadsthecellsonthesurfaceoftheagar,whereasthepourplateembedsthecellswithintheagar.Bothmethodscanbeusedtodeterminethenumberofmicroorganismsinasample.

Forthespreadplate,asmallvolumeofadilutedmixturecontainingaround25to250cellsistransferredtothecenterofanagarplateandspreadevenlyoverthesurfacewithasterilebentrod.Thedispersedcellsdevelopintoisolatedcolonies.Isolatedcolonies

Thepourplateisextensivelyusedwithbacteria,archaea,andfungi.Itisparticularlyusefulwhensamplingaheterogeneouspopulationofmicrobes,someofwhichmightproducecoloniesthatspreadoveranagarsurfaceifisolatedbythestreak-plateorspread-platemethods.

Inthepour-platemethod,theoriginalsampleisdilutedseveraltimestoreducethemicrobialpopulationsufficientlytoobt