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A. B. C. D. E. F. HANGING-DROP METHOD USE OF SEMI-SOLID MEDIA STAINING OF FLAGELLA SEROLOGICAL TEST FLUORESCENT TEST SWARMING PHENOMENA
2. MORPHOLOGY AND STAINING 3 GENERAL WAYS A. WET MOUNT B. HANGING DROP C. FIXED STAINING
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PROCESS OF ARTIFICIAL COLORING OF MICROORGANISM WITH DYES OR REAGENTS IN ORDER TO FACILITATE MORPHOLOGICAL IDENTITY AND ARRANGEMENT OF BACTERIA UNDER THE MICROSCOPE.
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SIMPLE STAIN
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ONE PARTICULAR STAIN OR DYE RESULTING INTO ONE COLOR ALSO KNOWN AS DIRECT STAINING AN AQUEOUS OR ALCOHOLIC SOLUTION OF A SINGLE DYE.
2. DIFFERENTIAL STAIN
TWO OR MORE DYES OR STAINS ARE USUALLY EMPLOYED COMPOSITION : 1. PRIMARY STAIN 2. MORDANT 3. DECOLORIZER 4. SECONDARY OR COUNTER STAIN EX. : GRAMS STAIN, ACID FAST STAIN
REAGENTS :
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2. 3. 4.
THE DIFFERENCE IN THE COMPOSITION OF BACTERIAL CELL WALL COMPOSITION Gram positive organisms: with thick peptidoglycan layer and teichoic acid cross linkages. Gram negative organisms : thin layer of peptidoglycan.
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5. Bacillus and Clostridium ARE SPORE FORMING ORGANISMS 6. THE HIGHER FORMS OF ORGANISM INCLUDING Actinomyces, Streptomyces, yeast and mold ARE GRAM POSITIVE 7. SPIRAL ORGANISMS ARE NOT STAINABLE EXCEPT FOR SOME WHICH ARE GRAM NEGATIVE.
PRINCIPLE : USED FOR SUBSET OF BACTERIA WHOSE CELL WALL CONTAINS LONG CHAIN FATTY ACID. ACID FAST : ORGANISMS WITH MYCOLIC ACID IN THEIR CELL WALL. NON-ACID FAST : ORGANISMS WITHOUT MYCOLIC ACID IN THEIR CELL WALL.
METHODS : ZIEHL NEELSEN METHOD KINYOUN METHOD (phenol) PAPPENHEIMS METHOD ( alcohol/Rosolic acid) BAUMGARTENS METHOD (diluted alc.fuchsin) RHODAMINE AURAMINE METHOD
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CAPSULAR STAINS
1. HISSS COPPER SULFATE METHOD
- CAPSULATED ORGANISMS APPEAR AS DARK PURPLE BODY WITH A FAINT BLUE CAPSULE AROUND IT
2. GINS METHOD - BACTERIA WILL BE STAINED BUT THE CAPSULE IS UNSTAINED WITH THEIR MARGIN DELINEATED BY THE INK. 3. WELCHS METHOD - CAPSULE STAINS PALE VIOLET 4. WADSWORTHS METHOD -BACTERIA ( BLUE ), Capsule ( PINKISH )
LOEFFLERS ALKALINE METHYLENE BLUE ( LAMB ) 2. NEISSER STAIN - BACILLI APPEAR ENTIRELY BROWN OR SHOW DARK BLUE ROUND BODY AT BOTH ENDS. 3. ALBERTS METHOD - GRANULES APPEAR BLUE-BLACK 4. LJUBINSKY METHOD
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LEIFSON METHOD
2. 3. 4. 5. 6. - FLAGELLA ( RED ), BACTERIAL CELL ( BLUE ) SILVER STAIN FOR FLAGELLA FISHER CONN CASARES GILS METHOD LOFFLERS METHOD VAN ERMENGENS METHOD
* FLAGELLAR STAIN USED A SPECIAL MORDANT SUCH AS TANNIC ACID THAT SWELLS, COATS, FORMS A PRECIPITATE WITH THE FLAGELLA.
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- SPIROCHETES/BACTERIA (WHITE IN DARK FIELD) -USED FOR ORGANISMS WITH POOR STAINING PROPERTIES 2. NIGROSIN - USED FOR STUDYING SPIROCHETES 3. RELIEF STAIN ( DORNER ) - CELLS ARE UNSTAINED AGAINST THE DARK BACKGROUND.