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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.