During Gram's stain

Options

A. all bacteria whether Gram (+) ve or (-) ve, take crystal violet stain.
B. only Gram+ ve bacteria take crystal violet stain.
C. only Gram -ve bacteria take crystal violet stain.
D. Gram ( + )ve bacteria lose this stain after alcohol treatment and take red stain of safranin

Answer

### Step-by-Step Solution for the Question: "During Gram's stain"

1. **Understanding Gram Staining**:
- Gram staining is a laboratory technique used to classify bacteria into two main groups: Gram-positive and Gram-negative. This classification is based on the structure of their cell walls.

2. **Cell Wall Composition**:
- **Gram-positive bacteria** have a thick peptidoglycan layer in their cell wall, which allows them to retain the crystal violet stain used in the procedure.
- **Gram-negative bacteria** have a thinner peptidoglycan layer and an outer membrane, which does not retain the crystal violet stain.

3. **Staining Process**:
- The first step involves applying **crystal violet stain** to the bacterial sample. Both Gram-positive and Gram-negative bacteria initially take up this stain, appearing purple.

4. **Decolorization**:
- The next step is the addition of **alcohol** (either ethanol or acetone).
- In this step, Gram-positive bacteria retain the purple color due to their thick peptidoglycan layer, while Gram-negative bacteria lose the purple color and become colorless (decolorization).

5. **Counterstaining**:
- After decolorization, a **counterstain** called **safranin** is applied.
- Gram-positive bacteria remain purple, while Gram-negative bacteria take up the safranin and appear pink or red.

6. **Conclusion**:
- The final result of the Gram staining process is that Gram-positive bacteria appear purple, and Gram-negative bacteria appear pink/red. This differentiation is crucial for identifying bacterial species and determining appropriate treatments.