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Q: What is the most important reagent in gram staining?
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Which step is most crucial or most likely to cause poor results in the gram stain?

Gram staining is a type of differential staining in which two types of bacteria are differentiated on the basis of their cell wall either gram positive or gram negative although all the steps in gram staining are crucial, the most important step the most crucial step in the performance of the Gram staining procedure is the decolorization step which is the Acid-Alcohol (3% HCl and 95% Ethanol) and must be timed correctly; the crystal violet stain will be removed from both Gram-positive and negative cells if the decolorizing agent is left on too long (a matter of seconds).


What is commonly found in a cell membrane?

Peptidoglycan, being the most important element for gram staining, cell differentiation....etc...


What is the most common stain?

Gram staining


If at the end of gram staining both the gram positive and gram negative bacteria appear red what is the most likely explanation?

Contamination or old culture.Type your answer here...


Why must young cultures be used when doing staining?

Young cultures must be used when doing a gram stain to get more accurate results. The cell was is the part of the bacterial cell that is most involved with gram staining because it holds the crystal violet.


What is the importance of gram staining?

Gram staining (or Gram's method) is an empirical method of differentiating bacterial species into two large groups (Gram-positive and Gram-negative) based on the chemical and physical properties of their cell walls. It is a first step to determine the identity of a particular bacterial sample. Gram stains are performed on body fluid or biopsy when infection is suspected. It yields results much more quickly than culture, and is especially important when infection would make an important difference in the patient's treatment and prognosis.


Why are the steps in gram staining so carefully standardized?

Gram's stain remains one of the most valuable methods we have for identifying isolates accurately and rapidly. Despite our long-standing familiarity with this method, it still warrants careful attention every step of the way--from preparation and QC of reagents to staining and interpretation. I think one of the main reasons would to avoid contamination.


What is the most crucial step in the gram stain procedure?

The most critical step of gram staining is the decolorization step as crystal violet stain will be removed from both G+ve & G-ve cells if the decolorizing agent(e.g alchohol ) is left on too long.


What is the purpose of staining an unknown organism?

By doing differential stains on an unknown organism, you can learn more about that organism. One of the most helpful stains would be the Gram stain. The gram stain will differentiate from Gram positive and Gram negative cells, narrowing your bacteria down a lot. Other stains include: Acid-Fast stain, Capsule stain, Endospore stain, and PHB stain.


Why must fresh bacterial cultures be used in a Gram stain?

Why must young cultures be used when doing a Gram stain Young cultures must be used so the crystal violet can stick to the cell walls of Gram positive bacteria. The cell walls break down in old cultures and the staining process is not accurate


What part of bacterial cell is most involve with gram staining and why?

In the gram staining process, gram positive bacteria appear to be purple because their cell walls, which contain a large concentration of peptidoglycans, are strongly dyed. Gram negative bacteria appear pink because their walls asborb less dye. This occurs becayse there is a smaller concentration of peptidoglycans and an additional lipid layer surrounds the cell wall. ANSWER The exact mechanism of action of this staining technique is not clearly understood. However, it is known that differences in the biochemical composition of bacterial cell walls parallel differences in their Gram-stain reactions. Gram-positive bacterial walls are rich in tightly linked peptidoglycans (protein-sugar complexes) that enable cells to resist decolorization. Gram-negative bacterial walls have a high concentration of lipids (fats) that dissolve in the decolorizer (alcohol, acetone, or a mixture of these) and are washed away with the crystal violet. The decolorizer thus prepares gram-negative organisms for the counterstain.


What are the different techniques used in Biology?

Gram staining: This is to determine if a bacterial cell is Gram positive or negative. This uses Crystal violet dye, Gram's iodine as a mordant, Ethyl Alcohol as a decolorization medium, and Safranin as a secondary dye. Spore staining: Primary dye is Malachite green, then slide is placed over boiling beaker, cooled, rinsed with water, then Safranin is used as a counter stain. This test is used to show whether a bacteria is a spore former. Acid fast staining: Primary dye is Carbolfuchsin, heated over beaker like the spore stain, acid alcohol is used as the decolorizing agent, and Methylyene Blue is used as a counter stain. This is used to show bacteria with acid-fast walls, which have a thick waxy lipid around them. These are the most commonly used staining techniques with Bacteria.