微生物学微生物学 (21).docVIP

MeasurementofMicrobialPopulationSize

Evaluatedirectcellcounts,viablecountingmethods,andcellmassmeasurementsfordeterminingpopulationsize

Explainwhyplatecountresultsareexpressedintermsofcolonyformingunits(CFU)

Therearemanywaystomeasuremicrobialgrowthtodeterminegrowthrateconstantsandgenerationtimes.Eitherpopulationnumberormassmaybefollowedbecausegrowthleadstoincreasesinboth.Herethemostcommonlyemployedtechniques

DirectMeasurementofCellNumbers

Themostobviouswaytodeterminemicrobialnumbersisbydirectcountsusingacountingchamber(e.g.,Petroff-Haussercountingchamberandhemacytometers).

Thisapproachiseasy,inexpensive,andrelativelyquick.Alwaysemployedinthelab.

Italsogivesinformationaboutthesizeandmorphologyofmicroorganisms.Countingchambersconsistofspeciallydesignedslidesandcoverslips;thespacebetweentheslideandcoverslipcreatesachamberofknowndepth.

Onthebottomofthechamberisanetchedgridthatfacilitatescountingthecells(figure7.33).Thenumberofmicroorganismsinasamplecanbecalculatedbytakingintoaccountthechambersvolumeandanydilutionsmadeofthesamplebeforecounting.

Figure7.33ThePetroff-HausserCountingChamber.

(a)Sideviewofthechambershowingthecoverglassandthespacebeneathitthatholdsabacterialsuspension.

(b)Atopviewofthechamber.Thegridislocatedinthecenteroftheslide.

(c)Anenlargedviewofthegrid.Thebacteriainseveralofthecentralsquaresarecounted,usuallyatX400toX500magnification.Theaveragenumberofbacteriainthesesquaresisusedtocalculatetheconcentrationofcellsintheoriginalsample.

Onedisadvantageofusingcountingchambersisthattodeterminepopulationsizeaccurately,themicrobialpopulationmustberelativelylargeandevenlydispersedbecauseonlyasmallvolumeofthepopulationissampled.

ViableCountingMethods

Severalplatingme