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PourplateMethod:Principle,Procedure,Uses,and
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Pourplatemethodisusuallythemethodofchoiceforcountingthenumberofcolonyformingbacteriapresentin Join1,005othersubscribers
aliquidspecimen.Inthismethod,fixedamountofinoculum(generally1ml)fromabroth/sampleisplacedinthe
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centerofsterilePetridishusingasterilepipette.Moltencooledagar(approx.15mL)isthenpouredintothePetri
dishcontainingtheinoculumandmixedwell.Afterthesolidificationoftheagar,theplateisinvertedand Subscribe
incubatedat37Cfor2448hours.

Microorganismswillgrowbothonthesurfaceandwithinthemedium.Coloniesthatgrowwithinthemedium

generallyaresmallinsizeandmaybeconfluentthefewthatgrowontheagarsurfaceareofthesamesizeand

appearanceasthoseonastreakplate.Each(bothlargeandsmall)colonyiscarefullycounted(usingmagnifying
colonycounterifneeded).Eachcolonyrepresentsacolonyformingunit(CFU). TripleSugarIronAgar(TSI):
Principle,Procedureand
Interpretation
Thenumberofmicroorganismspresentintheparticulartestsampleisdeterminedusingtheformula:
july16,2013

CFU/mL=CFU*dilutionfactor*1/aliquot
Catalasetest:principle,uses,
procedureandresults
Foraccuratecounts,theoptimumcountshouldbewithintherangeof30300colonies/plate.Toinsurea
october7,2013
countableplateaseriesofdilutionsshouldbeplated.

GramStaining:Principle,Procedure
andResults
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BloodAgar:Composition,
Preparation,UsesandTypesof
Hemolysis
august22,2013

API20ETestSystem:Introduction,
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may6,2015

CATEGORIES

Bacteriology
AnaerobicBacteriology
BacteriologyNote
BiochemicaltestsinMicrobiology
laboratorydiagnosisofBacterialDisease
MCQBacteriology
Thepourplatemethodofcountingbacteriaismoreprecisethanthestreakplatemethod,but,ontheaverage,it StructureofBacterialCells
virulencefactorsofpathogenicbacteria
willgivealowercountasheatsensitivemicroorganismsmaydiewhentheycomecontactwithhot,moltenagar
DifferenceBetween
medium. EmergingInfectiousDiseases
Immunology
Uses: MCQimmunology
serology
Thepourplatetechniquecanbeusedtodeterminethenumberofmicrobes/mLinaspecimen.Ithasthe LifeofBidur
Microbialgenetics
advantageofnotrequiringpreviouslypreparedplates,andisoftenusedtoassaybacterialcontaminationoffood
Microbiology
stuffs.
CultureMediausedinMicrobiology
FunnyMicrobiology
MaterialsandEquipments MCQMicrobiology
MCQMycology
1.Testsample MedalsandawardsinMicrobiology
2.PlateCountAgar(PCA)orNutrientAgar MicrobiologyforBeginners
3.Hotwaterbath45C recentdiscoveryinmicrobiology
4.SterilePetridishes StainingtechniquesinMicrobiology
5.Flame Uncategorized
6.Colonycounterwithmagnifyingglass USMLEMicrobiologypracticequestions
7.Sterilecapped16*150mmtesttubes MolecularBiology
8.Pipettesofvarioussizes(e.g.01,1.0and2.0mL) Mycology
labdiagnosisoffungaldisease

ProcedureofPourplatetechnique Parasitology
labdiagnosisofparasiticDisease
http://microbeonline.com/pourplatemethodprincipleprocedureusesdisadvantages/ 1/4
3/8/2017 PourplateMethod:Principle,Procedure,Uses,and(Dis)Advantagesmicrobeonline
labdiagnosisofparasiticDisease
1.Preparethedilutionofthetestsampleexpectedtocontainbetween30300CFU/mL.(Followserialdilutiontechnique) MCQParasitology
2.InoculatelabeledemptypetridishwithspecifiedmL(0.1or1.0mL)ofdilutedspecimen ParasitologyNote
Principlesofsterilizationanddisinfection
SexuallyTransmittedInfections
Note:forthedetaildescriptionregardinguseofpipette,inoculationofsample,dilutiontechniqueetc,followthe
Virology
reference1.
LabdiagnosisofViralDisease
MCQVirology
Pouringthemoltenagarandincubation StructureofVirus

1.Collectonebottleofsterilemoltenagar(containing15mLofmeltedPlateCountAgaroranyotherstandardculturemedia)
fromthewaterbath(45C).

2.Holdthebottleintherighthandremovethecap
withthelittlefingerofthelefthand.
3.Flametheneckofthebottle.
4.LiftthelidofthePetridishslightlywiththeleft
handandpourthesterilemoltenagarintothe
Petridishandreplacethethelid.
5.Flametheneckofthebottleandreplacethecap.
6.Gentlyrotatethedishtomixthecultureandthe
mediumthoroughlyandtoensurethatthemedium
coverstheplateevenly.Donotsliptheagarover
theedgeofthepetridish.
7.Allowtheagartocompletelygelwithoutdisturbingit,
itwilltakeapproximately10minutes. Pouringthemoltenagarmedium
8.Sealandincubatetheplateinaninvertedpositionat
37Cfor2448hours.

OverviewofPourplatemethodandspreadplatemethod

Results:

After2448hours,countallthecolonies(again:notethattheembeddedcolonieswillbemuchsmallerthanthose
whichhappentoformonthesurface).Amagnifyingcolonycountercanaidincountingsmallembeddedcolonies.

CalculateCFU/mLusingtheformula:CFU/mL=CFU*dilutionfactor*1/aliquot

(thevolumeofdilutedspecimen(aliquot)iseither0.1or1.0mL)

DisadvantagesofPourplatemethod

1.Preparationforpourplatemethodistimeconsumingcomparedwithstreakplate/andorspreadplatetechnique.
2.Lossofviabilityofheatsensitiveorganismscomingintocontactwithhotagar.
3.Embeddedcoloniesaremuchsmallerthanthosewhichhappentobeonthesurface.Thus,onemustbecarefultoscore
thesesothatnoneareoverlooked.
4.Reducedgrowthrateofobligateaerobesinthedepthoftheagar.

Referencesandfurtherreadings

1.BasicPracticalMicrobiologyAManualbySocietyforGeneralMicrobiology(SGM)
http://microbeonline.com/pourplatemethodprincipleprocedureusesdisadvantages/ 2/4
3/8/2017 PourplateMethod:Principle,Procedure,Uses,and(Dis)Advantagesmicrobeonline
1.BasicPracticalMicrobiologyAManualbySocietyforGeneralMicrobiology(SGM)

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3thoughtsonPourplateMethod:Principle,Procedure,Uses,and(Dis)
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Egwuatu REPLY
October16,2016at11:57amEdit

quitegood

michael REPLY
October26,2016at8:45pmEdit

Wouldanaerobicbacteriagrowinapourplateasinoculatedandincubatedabove?reason?

Prachi REPLY
February14,2017at11:03amEdit

Whyuse250mlsampleforwateranalysisthroughmembranefiltermethod

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