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Crystal violet iodine complex is a chemical compound formed when crystal violet dye interacts with iodine. This complex is commonly used in microbiology as a stain for bacterial cells, allowing them to be visualized under a microscope. The crystal violet iodine complex binds to the peptidoglycan layer of bacterial cell walls, imparting a purple color to the cells.

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Why is Iodine used in Gram staining?

Iodine is used in Gram staining as a mordant, which helps to bind the crystal violet dye to the cell wall of bacteria. This mordant-iodine complex forms larger complexes with the crystal violet dye, making it difficult for the dye to be washed away during the decolorization step. This allows for differentiation of Gram-positive and Gram-negative bacteria based on their ability to retain the crystal violet dye.


What is the secondary stain in gram stain procedure?

It is crystal violet & stains all cells purple.


What happens if grams iodine is not applied after staining with crystal violet?

If the iodine in the Gram's stain was omitted, the crystal violet which enters the cell will not be able to form the Crystal Violet Iodine (CVI) complex. The CVI complex molecule has a larger size than that of Crystal violet molecule and hence Gram +ve bacteria are able to retain the stain. If iodine were omitted, even Gram+ve cells would get decolorised by Alcohol treatment and would be seen as Gram-ve cells.


If iodine step was amitted how would gram negative cells appear?

Iodine work as modrant to crystal violet if the iodine step will be emitted then the crystal violet will not be able to show its effect and all cells will appear pink when counter stain with saffranin


What is the action of iodine in grams stain?

Iodine is used to bind the Crystal Violet to the Gram Positive microbes.


What is the use of Iodine in gram stains?

Iodine is used in Gram staining to form a complex with crystal violet, which helps to trap the dye within the cell wall of Gram-positive bacteria. This step is important in differentiating between Gram-positive and Gram-negative bacteria based on their ability to retain the crystal violet dye.


Does the age of the culture affect the gram staining reaction?

Yes, the age of the culture may influence the results of the stain. With the current theory behind gram staining, it is thought that in gram-positive bacteria, the crystal violet and iodine combine to form a larger molecule that precipitates out within the cell. The alcohol/acetone mixture then causes dehydration of the multilayered peptidoglycan in the gram-positive call wall, thus decreasing the space between the molecules and causing the cell wall to trap the crystal violet-iodine complex within the cell. In the case of gram-negative bacteria, the alcohol/acetone mixture, being a lipid solvent, dissolves the outer membrane of the gram-negative cell wall (and may also damage the cytoplasmic membrane to which the peptidoglycan is attached). The thin layer of peptidoglycan is unable to retain the crystal violet-iodine complex and the cell is decolorized. It is important to note that gram-positivity (the ability to retain the purple crystal violet-iodine complex) is not an all-or-nothing phenomenon but a matter of degree. There are several factors that could result in a gram-positive organism staining gram-negatively: 1. The method and techniques used. Overheating during heat fixation, over decolorization with alcohol, and even too much washing with water between steps may result in gram-positive bacteria losing the crystal violet-iodine complex. 2. The age of the culture. Cultures more than 24 hours old may lose their ability to retain the crystal violet-iodine complex. 3. The organism itself. Some gram-positive bacteria are more able to retain the crystal violet-iodine complex than others.


Does the age of the culture affect the gram stain?

Yes, the age of the culture may influence the results of the stain. With the current theory behind gram staining, it is thought that in gram-positive bacteria, the crystal violet and iodine combine to form a larger molecule that precipitates out within the cell. The alcohol/acetone mixture then causes dehydration of the multilayered peptidoglycan in the gram-positive call wall, thus decreasing the space between the molecules and causing the cell wall to trap the crystal violet-iodine complex within the cell. In the case of gram-negative bacteria, the alcohol/acetone mixture, being a lipid solvent, dissolves the outer membrane of the gram-negative cell wall (and may also damage the cytoplasmic membrane to which the peptidoglycan is attached). The thin layer of peptidoglycan is unable to retain the crystal violet-iodine complex and the cell is decolorized. It is important to note that gram-positivity (the ability to retain the purple crystal violet-iodine complex) is not an all-or-nothing phenomenon but a matter of degree. There are several factors that could result in a gram-positive organism staining gram-negatively: 1. The method and techniques used. Overheating during heat fixation, over decolorization with alcohol, and even too much washing with water between steps may result in gram-positive bacteria losing the crystal violet-iodine complex. 2. The age of the culture. Cultures more than 24 hours old may lose their ability to retain the crystal violet-iodine complex. 3. The organism itself. Some gram-positive bacteria are more able to retain the crystal violet-iodine complex than others.


What is dark violet iodine crystals.. is it a chemical change or not?

In order for something to be considered a change some sort of change must occur. A crystal is an object, not a type of change. Dark violet iodine crystals are pieces of relatively pure elemental iodine.


Function of Gram's iodine?

Iodine is used as a mordant in the gram staining procedure to make large crystals when it is used with crystal violet dye. In gram positive cell walls those crystals get stuck and wont get washed off with the alcohol. In gram negative cell walls the crystals are washed out.


Can grams iodine be used before primary stain?

Probably not. The CVI complex formed between the iodine and crystal violet will be trapped in the lipid layer and not the peptidoglycan layer, so you would probably end up over decolorizing. This would cause a lot of false gram negatives. Probably not. The CVI complex formed between the iodine and crystal violet will be trapped in the lipid layer and not the peptidoglycan layer, so you would probably end up over decolorizing. This would cause a lot of false gram negatives.


How does the Gram stain work What is happening as each reagent is used?

The positive turn purple and the negative turns red-colored. The positive is purple because the stain is able to pass through the thick peptoglycan wall where as the negative is red/pink because the stain can't get through the thick lipid layer (Membrane) to get to the thin peptoglycan layer.