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The steps in Gram staining are:
1. crystal violet added to the smear
2. iodine, the mordant (this fixes the violet)
3. a decolorizer made of acetone and alcohol
4. safranin, the counterstain
If the cell is Gram +, the decolorizer can not remove the violet. If it is Gram -, the decolorizer can remove the violet and the cell can be then colored with the dye, safranin.
Bacteria are grouped in 4 groups by Gram stain:
Gram-positive, the cell wall retains crystal Violet.
Gram-negative, the cell wall does not retain crystal Violet.
Graham not reactive, no staining whatsoever.
Graham variable, uneven staining.
Wiki User
∙ 10y ago
Wiki User
∙ 10y agoThe four reagents are: Crystal violet, Gram's iodine, Decolorizing solution (an alcohol-based solution) and Safranin (or basic fuchsin).
Wiki User
∙ 12y agolist the steps of the gram staining procedure
Wiki User
∙ 13y agoCrystal violet, Gram's iodine, alcohol, and safranin
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How we can use acetocarmine to stain the onion root tip cells?
take the root tip (about 1 cm )and put it in dil. HCL solution for ten min. take the tip out and put in acetocarmine stain for 1min. take out the root put it in water bto remove extra stain .
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How can you differentiate gram positive and gram negative bacteria?
The process of gram staining is simple. 1)smear bacteria from pure culture onto slide, heat fix 2)flood with crystal violet (1min) 3)Add iodine (1 min) 4)acid/alcohol wash (1 min) 5)Flood with safranine (1min) 6)Air dry and examine. These times are for clinical microbiology and experimental methods employ optimal and more precise times (but overall its pretty close). Down side of this method is that you must smear bacteria onto the slide and fix it by heating the underside of the slide with a bunsen burner. if they are pink then you have gram negative (Gram's stain didnt stick) if its purple then its gram positive(Gram's stain did stick) This is due to the peptidoglycan layers. Gram negative bacteria have only a thin layer of peptidoglycan as part of the cell membrane/wall where Gram positive have a very think peptidoglycan layer. Source(s): Medical Microbiology
How do scientists distinguish between gram-positive and gram-negative bacteria?
The process of gram staining is simple. 1)smear bacteria from pure culture onto slide, heat fix 2)flood with crystal violet (1min) 3)Add iodine (1 min) 4)acid/alcohol wash (1 min) 5)Flood with safranine (1min) 6)Air dry and examine. These times are for clinical microbiology and experimental methods employ optimal and more precise times (but overall its pretty close). Down side of this method is that you must smear bacteria onto the slide and fix it by heating the underside of the slide with a bunsen burner. if they are pink then you have gram negative (Gram's stain didnt stick) if its purple then its gram positive(Gram's stain did stick) This is due to the peptidoglycan layers. Gram negative bacteria have only a thin layer of peptidoglycan as part of the cell membrane/wall where Gram positive have a very think peptidoglycan layer. Source(s): Medical Microbiology
How do scientists distinguish between Gram- positive and Gram-negative bacteria?
The process of gram staining is simple. 1)smear bacteria from pure culture onto slide, heat fix 2)flood with crystal violet (1min) 3)Add iodine (1 min) 4)acid/alcohol wash (1 min) 5)Flood with safranine (1min) 6)Air dry and examine. These times are for clinical microbiology and experimental methods employ optimal and more precise times (but overall its pretty close). Down side of this method is that you must smear bacteria onto the slide and fix it by heating the underside of the slide with a bunsen burner. if they are pink then you have gram negative (Gram's stain didnt stick) if its purple then its gram positive(Gram's stain did stick) This is due to the peptidoglycan layers. Gram negative bacteria have only a thin layer of peptidoglycan as part of the cell membrane/wall where Gram positive have a very think peptidoglycan layer. Source(s): Medical Microbiology
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Which basic dyes were used to stain the smears?
1 Gram stainDeveloped by the Danish physician"Hans Christian Gram"Most commonly employed.Important of all the diagnostic staining techniques.By this method bacteria can be recognised as Gram-positive (blue-black) if they retain the primary dye complex of methyl violet and iodine in the face of attempted decolourization or as Gram-negative (red) if decolourisation occurs as shown by the cell accepting the counter stain.Procedure:Flood slide with crystal -10 seconds.washFlood with Gram's iodine - 10 seconds.washCarefully decolorize with 95% ethanolwashFlood with safranin (pink color) - 10 seconds.washAir dry,2 ZEIHL-NEELSEN'S METHOD (Acid-Fast Stain)ACID FAST Os Contains waxlike lipoidal material affecting staining quality.•Carbolfuchsin is primary stain.•Acid fast organisms resist decolorization with acid alcohol.•After decolorization, methyelene blue is added to organisms to counterstain any material that is not acid fast.•It is used for examination of Mycobacterium.•The principle of staining is depend on the resistance of this type of bacteria to decolarization by acid alcohol, because the cell wall contain waxy material (mycolic acid) which prevent the removal of carbol fuchsin from the cell.Staining ProcedureFlood the slide with strong carbol fuchsin and heat until steam rises (but do not boil).After 3-4 min apply more heat until rises again; do not let the stain dry on the slide.About 5-7 min after the first application of heat washes the slide thoroughly under running water.Decolourize in acid-alcohol until all traces of red have disappeared from the film. Decolourization should not be attempted in one stage; there should be intermittent washings in water and re-application of acid-alcohol.Wash well with water and counter stain with Methylene blue for 1 min.Wash and stand end to drain.Acid-fast organisms are red, other organisms are blue.
What is the technique for gram staining test?
the gram staining procedure involves four basic steps; 1.the smear is first flooded wit the primary stain crystal violet dye.in this case the primary stain is the first dye applied in any multicelled staining procedure and it stains all the cells. 2.the smear is rinsed with water to remove any excess crystal violet and then its flooded wit a dilute solution of iodine called grams iodine.iodine acts as a mordant.i.e. the substance that increases the interaction and affinity of cellular components for a dye.this is done so that the cells can stain more strongly. in this case the iodine complex thereby decreases the solubility of the dye within the cells 3.the stained smear is rinsed again and then 95% alcohol or a mixture of alcohol and acetol is briefly added.this solvent acts as decolorizing agents and they readily remove the dye iodine complex from gram negative but not from gram positive bacteria. 4.a counter stain is then applied to import a contrasting color to the now colorless gram negative bacteria.for this purpose,the red dye safrannin is used.this dye stains gram negative as well as gram positive bacteria but because the gram positive are already stained purple,it impacts little difference
How we can use acetocarmine to stain the onion root tip cells?
take the root tip (about 1 cm )and put it in dil. HCL solution for ten min. take the tip out and put in acetocarmine stain for 1min. take out the root put it in water bto remove extra stain .
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