02/26/2024 Gram- Staining

 

02/26/2024

Introduction: 

We observed Friday’s inoculated plates and noticed contamination. Since it got contaminated, we (Brian) inoculated the plates with Deinococcus Sonorensis again. Alongside Carime and Brian, we gram-strained Caeni and D. Sono. I learned and properly gram-stained the bacteria and took pictures of our view. Around 02:30 PM, Emme and Brian gave a presentation on the research they have completed on the biochemical characteristics tests. 

Method: 

Gram-Staining Steps

1. Wash and dry hands. 

2. Make sure you have all your materials.

3. With the wax pencil, draw a circle in the middle of the slide to place the bacteria inside the circle. You also want to label the slide and place a clip to the end of the slide so that you are able to hold it over the fire. 

4. Turn on the bunsen burner. 

5. Hold the ‘rod’ over the fire for a couple of seconds (rod must be red) to sterilize. 

6. Your plate must be flipped over to easily access the bacteria and place onto the slide. 

7. Once the rod cools down, you will obtain some bacteria by using the small circle at the end of the rod. 

8. The bacteria will then be transferred from the plate to the slide making sure it stays within the circle. Make sure to close the plate as soon as possible, time is crucial. 

9. You will heat-fix the slide with the bacteria in the circle by placing the slide on top of the flame moving it back and forth like six times. 

10.  Turn off the bunsen burner. 

11.  The following steps will be taken in minute increments. 

12.  Crystal Violet: Place 1-2 drops in the circle for a minute. After a minute, use distilled water to clean off the solution but not directly. 

13.  Grams Iodine: Place 1-2 drops in the circle for a minute. After a minute, use distilled water to clean off the solution but not directly. 

14.  Ethanol: Place a couple of drops (not directly of the circle) at an angle of the slide to ‘wash’ the slide and proceed to clean it off with distilled water (not directly). 

15.  Safranin: Place 1-2 drops in the circle for a minute. After a minute, use distilled water to clean off the solution but not directly. 

16.  Take off the clip and dry the slide with bibulous paper by pressing down the paper in between and pat dry. 

17.  To observe the slide, you will get the pink lens cleaner liquid, lens paper, oil, and microscope. 

18.  Finally, place the slide on the microscope and you should be ready to view the gram stain. 

Results:

Caeni Deinococcus Sonorensis 

Discussion: 

Caeni has more colonies formed together in comparison to D. Sono. D. Sono. seems to be smaller and isolated from each other. 

Conclusion: 

Moving forward, I will be gram-staining independently to view bacteria and compare/contrast them with the bacteria we know and have access to.