GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 1. Gather all equipment and supplies
Gram stain
Microscope slides
Heat source
Sterile swab
Culture or specimen
Microscope
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 2. Label the slide with the patient’s initials or the specimen number.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 3. Choose an isolated colony off of the agar plate and obtain bacteria with a sterile swab.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 4. Place the swab on the microscope slide and spread the colonies in a circular motion.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 5. Heat fix the microorganisms to the slide by placing the bottom of the slide to heat for approximately 30 seconds.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 6. Place slide on staining tray or hold with forceps above the sink.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 7. Flood the surface of the slide with Crystal Violet stain and let sit for one minute.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 8. Rinse the slide with distilled water.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 9. Flood the slide with Gram’s Iodine and time for one minute.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 10. Rinse the slide with distilled water.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 11. Flood the slide with Gram’s decolorizer and time for 30 seconds.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 12. Rinse the slide with distilled water.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 13. Flood the slide with the counterstain, Safranin, and let sit for one minute.
GRAM STAIN PROCEDURE : GRAM STAIN PROCEDURE 14. Rinse the slide with distilled water.
GRAM STAIN PROCEDURE : 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.