Beruflich Dokumente
Kultur Dokumente
MB102b
LABREPORT#3:
NegativestainingofStaphylococcusaureuswith
nigrosin
MuhammedSayedHassan
ID:171741
GROUP(F)
Submittedto:L.A.Suzanneali
NegativestainingofStaphylococcusaureuswith
nigrosin
INTRODUCTION
Thislabcoveredsmearpreparationofstaphylococcusaureusonthe
microscopicslidewiththeheatfixationmethod,Thennegativestainingwith
acidicdye(Nigrosin)andsuccessfullyidentifyingit'sshapeunderthelight
microscope.Thenegativestainingmethoddependsontherepulsionofthe
bacterialcellsurfaceofthenegativechargeddyetocolorizethe
background(microscopicslide).Aimofthelabistonegativestainstaphylococcus
aureusandidentifyit'sshapeunderthelightmicroscope.
MATERIALS
Chemicalagents:
1. Immersionoil
2. Nigrosin(Nigrosin100gm/L,Formalin5ml/Linwater)
3. Distilledwater
4. Ethanol(70%)
Biologicalagents:
1. Staphylococcusaureus
Equipments:
1. LightMicroscope
2. Microscopicslide
3. Aluminiumfoilsheet
4. Inoculationloop
5. Bunsenburner
PROCEDURE
a) Smearingofbacteria
1. 1dropsofwaterwasputonthemicroscopicslide.
2. Theinoculationloopwassterilizedbybunsenburner.
3. 1loopfulwasputonthemicroscopicslide.
4. Thebacteriawassmearedontheslide.
b) Heatfixationofbacteria
5. Thebacteriawasheatedwithbunsenburnerwithindirectflamefixation.
c) Negativestainingoftheslide
6. Themicroscopicslidewasputonthealuminiumfoilsheet.
7. 2dropofdyeweredroppedontheedgeofthemicroscopicslide.
8. Anotherslidewasusedatangleof45tosmearthedyecarefullyonthebacteria
slide.
9. Thentheslidewasdroughtforabout10minutesbeforeexamination.
d) Microscopicexaminationoftheslide
10. 1dropofimmersionoilwasdroppedontheslide.
11. Theslidewasputinthemicroscopicclip.
12. Knobwaszoomeduntilthelenstouchedtheoilconcavesurface.
13. ThentheslidewasexaminedwiththeOillens.
RESULTS
Fig(1): Negative stain of staphylococcus aureus bacteria shown under a light microscope
Using Oil-immersion technique.
DISCUSSION
Abovefig(1)showsthatculturedstaphylococcusaureusunderlightmicroscopeusing
oil-immersionlens.Smearandnegativestainingwassuccessfullyapplied,the
backgroundoftheslidehadthedyebutthebacterialcellsurfacerepulsedthedye
becauseit'sacidicandhasanegativechargesasthesurfaceofthebacterialcell.
CONCLUSION
Theaimofthelapwassuccessfullyachievedwhichisnegativestainingofstaphylococcus
aureususingnigrosinacidicdye.