微生物学微生物学 (29).pptVIP

Afterlearningthissection,youshouldbeableto:Evaluatedirectcellcounts,viablecountingmethods,andcellmassmeasurementsfordeterminingpopulationsize,Explainwhyplatecountresultsareexpressedintermsofcolonyformingunits(CFU).

Therearemanywaystomeasuremicrobialgrowthtodeterminegrowthrateconstantsandgenerationtimes.Eitherpopulationnumberormassmaybefollowedbecausegrowthleadstoincreasesinboth.DirectMeasurementofCellNumbersViableCountingMethods

DirectMeasurementofCellNumbersCountingchamberConsistofspeciallydesignedslidesandcoverslips;thespacebetweentheslideandcoverslipcreatesachamberofknowndepth.OnthebottomofthechamberisanetchedgridthatfacilitatescountingthecellsThenumberofmicroorganismsinasamplecanbecalculatedbytakingintoaccountthechambersvolumeandanydilutionsmadeofthesamplebeforecounting.

Onedisadvantageofusingcountingchambersisthattodeterminepopulationsizeaccurately,themicrobialpopulationmustberelativelylargeandevenlydispersedbecauseonlyasmallvolumeofthepopulationissampled.

ViableCountingMethodsSeveralplatingmethodscanbeusedtodeterminethenumberofviablemicrobesinasample.Thesearereferredtoaseitherviablecountingmethodsorplatecountsbecausetheycountonlythosecellsthatareabletoreproducewhencultured.Twocommonlyusedproceduresarethespread-plateandthepour-platetechniques.

Whenusingthesemethodstodeterminepopulationsize,thesamplesshouldyieldbetween25and250coloniesperplateforaccuratecounting.Oncethenumberofcoloniesisknown,theoriginalnumberofviablemicroorganismsinthesamplecanbecalculatedfromthatnumberandthesampledilution.Forexample,if1.0milliliterofasolutiondilutedbyafactorof1×106yielded150colonies(i.e.,1.5×102colonies),thentheoriginalsamplecontainedaround1.5×108cellspermilliliter.

CFUBecause