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STAINING
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Purpose of bacterial staining
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Purpose of bacterial staining
Staining is used to visualize bacteria, which are small and colorless, to study their properties and aid in identification.
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Differential Staining
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Purpose of bacterial staining
Staining is used to visualize bacteria, which are small and colorless, to study their properties and aid in identification.
Simple staining
Uses one stain to visualize bacterial shape and arrangement.
Differential staining
Uses two or more stains to differentiate bacterial species or structures.
Examples of differential stains
Gram stain, acid fast stain, capsule stain, and spore stain.
Gram stain primary stain
Crystal violet stains all cells purple initially.
Gram stain mordant
Iodine increases cells' affinity for crystal violet, forming a complex.
Gram stain decolorizer
95% ethanol dissolves lipids, increasing cell wall porosity and removing stain from some cells.
Gram stain counterstain
Safranin stains cells that lost the primary stain pink.
Gram positive bacteria cell wall
Thick peptidoglycan layer retains crystal violet-iodine complex, staining cells purple.
Gram negative bacteria cell wall
Thin peptidoglycan and outer membrane lose crystal violet during decolorization, staining pink with safranin.
Common shapes of bacteria
Coccus (round), bacillus (rod), and spiral (vibrio, spirillum, spirochete).
Acid fast stain primary stain
Carbol fuchsin stains acid fast bacteria red by penetrating waxy mycolic acid cell walls.
Acid fast stain decolorizer
Acid alcohol removes stain from non-acid fast cells.
Acid fast stain counterstain
Methylene blue stains non-acid fast bacteria blue.
Acid fast bacteria examples
Mycobacterium tuberculosis and Mycobacterium leprae.
Spore stain primary stain
Malachite green stains spores.
Spore stain decolorizer
Water removes stain from vegetative cells.
Spore stain counterstain
Safranin stains vegetative cells pink.
Capsule stain technique
Negative staining with dyes like nigrosin stains background, leaving capsules unstained and visible.
Purpose of capsule stain
Visualizes gelatinous outer layer protecting bacteria from host immune cells.