Infectious Disease Book

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Gram StainAka: Gram Positive, Gram Negative

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  1. Indications: Evaluation of Infection
    1. Acute Urethritis
    2. Pelvic Inflammatory Disease
    3. Pneumonia
  2. Specimens
    1. Infected body fluids
    2. Aspirated abscesses
    3. Exudates
  3. Technique
    1. Thin smear of material spread on slide
      1. Allow slide to air dry
    2. Fix material onto the slide
      1. Place side on warmer at 70 degrees Celsius or
      2. Pass slide through Bunsen Burner flame 3-4 times
    3. Overlay the slide with crystal violet for 1 minute
    4. Wash slide thoroughly with water
    5. Overlay slide with Gram Iodine for 1 minute
    6. Wash slide thoroughly with water
    7. Flood slide surface with decolorizer (acetone-Alcohol)
      1. Perform until no violet color washes off slide
      2. May require up to 10 seconds
    8. Wash slide completely with water
    9. Overlay smear with safranin counterstain for 1 minute
    10. Wash slide thoroughly with water
      1. Allow excess water to run off slide
    11. Allow slide to air dry or blot with bibulous paper
    12. Examine under high power (100x) oil immersion
  4. Interpretation
    1. Gram Positive (Stains dark blue to purple)
      1. Gram Positive Cocci
    2. Gram Negative (Stains pink to red)
      1. Gram Negative Rod
      2. Gram Negative Coccobacilli
  5. References
    1. Gulick (1983) Med Clin North Am 67:39

Bacterial stain, routine (C0200966)

Definition (NCI)This is a method (named after the Danish bacteriologist, Hans Christian Gram) for staining and identifying bacteria. It is based on the differential retention of a crystal violet-iodine complex within the cell membrane. Gram-positive organisms will appear blue-black or purple while Gram-negative organisms will appear red.
ConceptsLaboratory Procedure (T059)
EnglishGram stain, Gram stain method, Gram Staining Method
Spanishmétodo de tinción de Gram, metodo de tincion de Gram, tinción de Gram, tincion de Gram
CreditsDerived from the NIH UMLS (Unified Medical Language System)



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