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Differential staining . Types of staining

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Differential staining . Types of staining

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Microbiology
pharmaceutical microbilogy
grams staining
differential staining
types of staining
b pharm 3rd sem
types of staining
detail explination of the grams staining
staining techniques
identification of bacteria

Microbiology
pharmaceutical microbilogy
grams staining
differential staining
types of staining
b pharm 3rd sem
types of staining
detail explination of the grams staining
staining techniques
identification of bacteria

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Differential staining . Types of staining

  1. 1. GRAM STAINING Differential Staining
  2. 2. Principle : The gram staining is differential staining it involves the differentiation of gram –ve and +ve bacteria . The primary stain crystal violet forms complex with iodine leads formation of crystal violet magnesium ribo nucleate iodine and crystal violet iodine complex, these are forms in gram +ve and -ve bacteria respectively. In gram +ve bacteria the colour of primary stain is retained and in –ve bacteria colour of primary stain was not retained after adding decolourising agent. Gram –ve bacteria counter stained by methyl red and carbolfuschin forms red or pink colour at end.
  3. 3. Reagents used in Grams staining 1.Primary stain: crystal violet 2.Mordant stain: Grams iodine 3.Decolourising agent: Ethanol 4.Counter stain: Methyl red or carbolfuschin
  4. 4. 1.Primary stain : Carbolfuschin The carbolfuschin is the primary stain forms crystal violet magnesium ribonucleate iodine complex in grams +ve bacteria, it retained after the adding decolourising agent . The colour of primary stain was retained because gram +ve bacteria containing waxy material on cell wall, these resistance to entry of decolourising agent.
  5. 5. 2. Mordant: Iodine The iodine forms complex with crystal violet that is water insoluble compound. 3. Decolourising agent The decolourising agent decolourises complex formed by primary stain and mordant iodine solution.
  6. 6. 4. Counter stain: methyl red The counter stain used differentiate the gram +ve and gram –ve bacteria. It It stains only the gram –ve bacteria and cell of gram +ve bacteria was impermeable to the entry of methyl red through it. It was added after decolourisation. Note: Some type of grams staining involves counter stain as carbolfuschin in place of methyl red
  7. 7. Procedure involved in Grams staining 1. Smear making 2. Heat fixing 3. Primary staining 4. Decolourising 5. Counter staining 6. Obeserve under microscope Report: 1. If the colour of the primary stain violet is left in side the cell wall of bacteria reported as Gram +ve bacteria 2. If the colour of primary stain does not retains and appears red or pink colour.

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