Question

For microbiology:

1. Consider the following scenario: My work study student accidentally mixed up the safranin and crystal violet stains when filling the reagent bottles. Predict the outcomes when you gram stain each of the bacteria that we utilized in this laboratory.

Answer 1

ANSWER :-

1. According to the case mentioned above, the Crystal violet dye has been mixed up with Safranin dye.
2. To speak about Crystal violet, it is a basic dye which carries a positive charge as it is able to release hydroxy ions which renders them an overall positive charge. This dye is used in initial stages and is coupled with Gram's iodine to form a bigger complex which resides within the membranes of cell.
3. In gram positive bacteria, due to presence of thick peptidoglycan layer, there is more retention of the complex as compared to the gram negative bacteria.
4. Safranin is not water soluble and at the same time is lighter in appearance than the initial strain Crystal violet. So, the Crystal violet dye better penetrates and gets retained within the cell membranes. This retention as discussed above is more prominent in gram positive bacteria due to which they appear purple.
5. So, the basic, water soluble dye Crystal violet and water insoluble dye Safranin show variations in their binding potential and retention due to which chances of errors even when the components are mixed becomes minimized.
6. Although, visualization of the cells depends upon the intensity of complex formed between the Crystal violet and the Gram's iodine.

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