Gram stain

Today my partner and I preformed a gram stain with our unknown bacteria!! This specific test is used to indicate whether a bacteria is gram negative or gram positive. During this procedure we followed the steps of the simple stain by heat-fixing the bacteria to our slide with an inoculating loop then placing the slide on a staining rack. Next we flooded the slide with crystal violet, and let sit for 20 seconds. We then removed the crystal violet with water and covered the smear with Gram’s iodine for a minute. After rinsing the slide of the iodine we decolorized the bacteria with acetone until the color stopped running. Finally we applied safranin to the smear and waited a minute before rinsing it off the excess. We let the slide dry and proceeded to analyze our results under a microscope. 

The end product of a gram stain depends on the bacteria’s cell wall. If the bacteria were not decolorized during the decolorizing step with acetone and remained a blue/purplish color, the bacteria are said to be gram positive. The gram positive bacteria have a thick cell wall which makes it difficult to remove the crystal violate. If the crystal violate was removed during the decolorizing process, and turned red from the safranin, the bacteria is said to be gram negative. Gram negative bacteria have a thin cell wall which makes it easier to remove the crystal violet. 

Our end result turned out to be redish/purplish! Therefore, our unknown bacteria is gram negative with a thin cell wall.  Our bacteria is beginning to reveal itself through each lab experiment. As of today, the unknown bacteria is a gram negative bacilli/coccobacilli, little guy.