GRAM STAIN PROCEDURE 1. Gather all equipment and supplies –Gram stain –Microscope slides...

GRAM STAIN PROCEDUREGRAM STAIN PROCEDURE

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

2. Label the slide with the patient’s initials or the specimen number.

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

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

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

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

GRAM STAIN PROCEDUREGRAM STAIN PROCEDURE7. Flood the surface of the slide with

Crystal Violet stain and let sit for one minute.

8. Rinse the slide with distilled water.

9. Flood the slide with Gram’s Iodine and time for one minute.

11. Flood the slide with Gram’s decolorizer and time for 30 seconds.

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

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

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