第1页：Experiment one
Experiment one
- Basic methods of microbiological laboratory
- Bacterial culture
- Staining of bacteria
- Use of oil-immersion microscope
- Observation of several special bacteria
- Isolation and culture of bacteria(streakplate culture)
- Pyogenic cocci
- Pathogenic enterobacteria
第2页：Bacterial Culture and Growth
Bacterial Culture and Growth
- PURPOSE :To obtain information about bacterial biologicalcharacteristics
- METHODS :streak plate method isolation and cultureagar slope methodliquid medium culture pure culturestab cultureanaerobic culture
第3页：Streak Plate Method
Streak Plate Method
- PURPOSE:Isolation and culture of bacteria growingtogether in a specimen
- MATERIALS:l.MixedbrothcultureofEscherichiacoliandstaphylococcusaureus.2. Nutrient agar plate.
第4页：Streak Plate Method
Streak Plate Method
- PROCEDURE:
- Flameyourinoculatingloopuntilthewireglowsred.
- Allowthelooptocoolandgetaloopfulofthesuspensionofsample.
- Pickupyourplateandstreakthesurface,Flametheloopbeforestreakingnextsection.When
第5页：Streak Plate Method
Streak Plate Method
- PROCEDURE:
- Coverthepetridish,inverttheplate.Sterilizetheloop,labelyourname,dateetal.
- Incubatetheplateat37℃for18-24hours.
- Observe the bacterialcolonies.
第6页：Streak Plate Method
Streak Plate Method
- RESULT:observe the location and characteristicsof the bacterial colonies:SizeShape:circular, irregular, spreadingColorDensity:transparent, or opaqueElevationMarginHemolysisSurface:
第7页：Streptococcus pyogenes
Streptococcus pyogenes
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第8页：Agar Slope Method
Agar Slope Method
- MATERIALS :1. Agar slope2. Colonies on agar plate
- PROCEDURE :1.Withtheflame-sterilizedwireinoculatingloop,transferasmallamountofbacteriafromthecolonyonagarplate.Thenstreakontheagarslope.2.
第9页：Agar Slope Method
Agar Slope Method
- RESULTS :Therearemanysimilarwetcoloniesonthesurface.Iftherearesomeotherforms,itindicatesculturesampleisnotpure.
第10页：Liquid Medium Culture
Liquid Medium Culture
- MATERIALS：1. Peptone water2.Coloniesonagarplate
- PROCEDURE：1.Flame-sterilizethewireinoculatingloop.2.Insertthewireloopcontainingasmallamountofbacteriaintotheliquidculturerobe.3.Scratchthewalloftube
第11页：Liquid Medium Culture
Liquid Medium Culture
- PROCEDURE：4. Flame the mouth of the tube and reinsert thecotton plug. Flame-sterilize the wire loop.5.Labelthetube,incubateat37℃for24hours6. Observe the result.
- RESULTS:turbid,flocculent,pellicle
第12页：Stab Culture
Stab Culture
- METHODS:1.Flame-sterilizeinoculatingneedle.2.Inserttheneedlewithasmallbacteriatothecenteroftheculture,becarenottotouchthebottomofthetube,thendrawitoutinthesameway.3.Flamethemouthofthetube
第13页：Stab Culture
Stab Culture
- RESULTS:Motilebacteriawillmigratefromthelineofinoculationtoformadiffuseturbidityinthesurroundingmedium;nonmobilebacteriawillgrowonlyalongthelineofinoculation.
第14页：Staining of Bacteria
Staining of Bacteria
- PURPOSE :To make bacteria more easily observableTo acquaint you with Gram stain
- MATERIALS:
- Simplestain
- Gramstain
- Acid-faststain
- Specialstain
- Sporestain
- Capsulestain
- Flagellastain
- Metachromaticgranulesstain.
第15页：Gram stain
Gram stain
- purpose:differentiating bacteria
- MATERIALS :
- Slant cultures of and Escherichia coli andS.aureus (18 to 24 hours old)
- Crystal violet, iodine solution, 95% alcohol,safranin
- Microscope slides
第16页：Gram stain
Gram stain
- PROCEDURE:
- Smear:size of a dime to form athin film
- Dry :air dry
- Fix:through the warm air abovethe flame two or three times.
第17页：Gram stain
Gram stain
- PROCEDURE:
- Stain
- primary stain: crystal violet，60 seconds ,Wash with water
- mordant stain: iodine solution , 60 seconds ,Wash with water
- decolorize: 95% alcohol, 20 secondsWash with water
- Counterstain: safranin 30 secondsWash with water
第18页：Gram stain
Gram stain
- results:
- typicalshapesandcolorsofthebacteria.
- GrampositivebacteriaarevioletcoloredandGramnegativecellsareredcolored.
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第20页：Use of The Oil-immersion
Use of The Oil-immersion
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第21页：study morphology
study morphology
- study morphology
- structure
- staining characteristics
- motility of different microorganisms.Use of The Oil-immersionMicroscope
第22页：MATERIALS
MATERIALS
- Microscope,immersionoil,lenspaper,glassslides,andcoverslips.
- Preparedstainedslidesofseveraltypesofbacteria.
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第27页：Types of objective lense
Types of objective lense
- 1.thelow-powerobjectivelensethe10×or16-millimetre(mm)
- 2.thehigh-dryonethe40×,50×,or4mmobjective
- 3.theoilimmersionthe90×,100×,or1.8mmobjective.
第28页：PROCEDURE
PROCEDURE
- Carefully place the microscope on the desk andexamine it
- preparation:selecttheobjectivelenseMaketheirisdiaphragmopenandthetopofthecondenseratthelevelofthestage.
- placeadropofimmersionoilonthecoverglassofthemicroslide
- lowertheoilimmersionobjectivetubewhileuntilitengagesthedrop
第29页：Precautions
Precautions
- whenloweringthetube,mustlookatthemicroscopefromtheside
- Donotallowchemicalstocontactthemicroscope
- Cleanthemechanicalpartswithgauze;theoilcanbewipedoffwithlenspaper.
- Toremoveimmersionoilfromtheopticalglassparts,wipewithlenspapermoistenedwithxylol.Discardthelenspaper.Wiperapidlytopreventinjurytoopticalglass
第30页：focusupwardslowlyuntiltheimageappear
focusupwardslowlyuntiltheimageappear
- focusupwardslowlyuntiltheimageappear.
- Completefocuswiththefineadjustment
- Cleanthemicroscopewhenthisexperimentisended
- Leavethemicroscopewiththelowestpowerobjectiveintheworkingposition
- Removeimmersionoilfrompreparedpracticeslides.Returnthesetoinstructor.
- Placeaplasticcoveroverthemicroscopeorplaceitinitscase.
第31页：Laboratory Diagnosis of
Laboratory Diagnosis of
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第32页：Colonialcharacteristicobservation
Colonialcharacteristicobservation
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