GRAM STAIN PROCEDURE

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GRAM STAIN PROCEDURE

• 1. Gather all equipment and supplies
• Gram stain
• Microscope slides
• Heat source
• Sterile swab
• Culture or specimen
• Microscope

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GRAM STAIN PROCEDURE

• 2. Label the slide with the patients initials
  or the specimen number.

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GRAM STAIN PROCEDURE

• 3. Choose an isolated colony off of the agar
  plate and obtain bacteria with a sterile swab.

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GRAM STAIN PROCEDURE

• 4. Place the swab on the microscope slide and
  spread the colonies in a circular motion.

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GRAM STAIN PROCEDURE

• 5. Heat fix the microorganisms to the slide by
  placing the bottom of the slide to heat for
  approximately 30 seconds.

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GRAM STAIN PROCEDURE

• 6. Place slide on staining tray or hold with
  forceps above the sink.

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GRAM STAIN PROCEDURE

• 7. Flood the surface of the slide with Crystal
  Violet stain and let sit for one minute.

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GRAM STAIN PROCEDURE

• 8. Rinse the slide with distilled water.

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GRAM STAIN PROCEDURE

• 9. Flood the slide with Grams Iodine and time
  for one minute.

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GRAM STAIN PROCEDURE

• 10. Rinse the slide with distilled water.

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GRAM STAIN PROCEDURE

• 11. Flood the slide with Grams decolorizer and
  time for 30 seconds.

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GRAM STAIN PROCEDURE

• 12. Rinse the slide with distilled water.

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GRAM STAIN PROCEDURE

• 13. Flood the slide with the counterstain,
  Safranin, and let sit for one minute.

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GRAM STAIN PROCEDURE

• 14. Rinse the slide with distilled water.

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GRAM STAIN PROCEDURE

• 15. Blot the slide and read with the oil
  immersion lens of the microscope. Look for
  Gram-negative and Gram-positive bacteria.