Micro Lab Gram-Stains 9/1

Today in lab we did gram-stains on two slides. One of the slides had bacteria out of our broth on it and the other one came from a colony of bacteria off of the streak plate. The steps for making a gram-stain are listed below…

1. Get yourself 2 clean slides
2. Turn on your bunsen burner
3. In order to make your slide from the broth you need to:
- Heat your IN. loop until it is red hot, and then let it cool for a minute
- Dip the loop into the broth being sure to only touch sterile parts of the bottle
- Spread the drop of broth from the loop onto the slide
- Repeat steps if you want more bacteria on there
- Let air dry
In order to make your slide from the streak plate you need to:
- Get a dropper that is filled with water
- Put a small drop of water onto your clean slide
- Heat your IN. loop until it is red hot, then allow to cool for a moment or stick it into sterile auger to cool
- Touch/tap your bacteria colony lightly
- Smear the bacteria onto the water droplet
- Heat the IN. loop in order to clean it
4. Next you need to either heat fix, or methanol fix, your slide.
- If you methanol fix it allow the methanol to sit on it for 2 minutes
5. Dump the extra methanol off of the slide
6. Cover the entire area with crystal violet and allow to sit for 1+ minute(s)
7. Rinse GENTLY and blot the slide
8. Then cover the entire slide with gram iodine for about 30 seconds
9. Dump off excess and rinse GENTLY, blot to dry
10. Decolorize your slide with 5-10 drops of E+OH
11. Rinse GENTLY and blot dry
12. Cover the slide with saframin for 1+minute(s)
13. Finally rinse the slide GENTLY, blot it dry, and take a look at your bacteria!

Good Luck and Have Fun! :)

Unless otherwise stated, the content of this page is licensed under Creative Commons Attribution-ShareAlike 3.0 License