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Gram Staining Procedure: a Practical Guide to Bacterial Taxonomy

Learn the differential staining technique, distinguishing between Gram-positive and Gram-negative bacteria. Follow step-by-step instructions on preparing a heat-fixed smear and conducting the Gram stain procedure. Make observations under a microscope to identify organisms. Improve your understanding of bacterial classification and characteristics.

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Gram Staining Procedure: a Practical Guide to Bacterial Taxonomy

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  1. Practical 2 • Gram staining

  2. Differential staining • Acceptance of stains is an important property of bacteria and is the base divission of bacteria to 2 principal groups Gram positive and Gram negative in taxonomy. • Gram staining

  3. GRAM STAINING PROCEDURE • Prepare a heat fixed smear of the culture you wish to examine • Flood the smear with crystal violet (30 sec. to 2 min) • Quickly and gently wash off excess stain (2 seconds) • Flood the smear with Grams iodine (1 minute) • Decolorize with alcohol (10-20 seconds or until the excess alcohol which flow off the slide is colorless) • Quickly and gently wash off excess stain (2 seconds) • Flood the smear with safranin (carbolfuchsin) (30 sec to 2 min.) • Quickly and gently wash off excess stain (2 seconds) • Blot dry with bibulous paper • Examine your slide under the microscope. Record sketches of the organisms, size, color, morphology, and culture identification.

  4. G+ G-

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