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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.
In a Gram stain, the reagents used are crystal violet (primary stain), iodine (mordant), ethanol or acetone (decolorizer), and safranin (counterstain). The steps involved in a Gram stain include applying crystal violet, rinsing with iodine, decolorizing with alcohol/acetone, and counterstaining with safranin. Gram-positive bacteria retain the crystal violet stain and appear purple, while Gram-negative bacteria lose the crystal violet stain and take up the safranin, appearing pink under the microscope.
The four reagents are: Crystal violet, Gram's iodine, Decolorizing solution (an alcohol-based solution) and Safranin (or basic fuchsin).
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What is the composition of PEA agar?
PEA (Phenylethyl alcohol agar) contains peptone, phenylethanol, agar, and distilled water. Phenylethanol is added as a selective agent to inhibit the growth of Gram-negative bacteria, making PEA agar useful for isolating Gram-positive bacteria.
How can you differentiate gram positive and gram negative bacteria?
Gram positive bacteria have a thicker peptidoglycan layer in their cell wall, which retains the crystal violet stain during the Gram staining process, giving them a purple color. Gram negative bacteria have a thinner peptidoglycan layer and an outer membrane, which does not retain the stain, causing them to appear pink when counterstained with safranin.
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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What is the technique for gram staining test?
The technique for Gram staining involves applying crystal violet stain, followed by iodine, alcohol decolorization, and then a counterstain like safranin. Gram-positive bacteria retain the purple color, while Gram-negative bacteria appear pink after the process. This technique is commonly used to classify bacteria based on differences in cell wall composition.
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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.
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Trace out the algorithm Max Min on a data set containing atleast 8 elements?
Steps to perform MaxMin on a data set (2,4,6,3,8,1,9,7) are:(2,4,6,3) (8,1,9,7)((2,4)(6,3)) ((8,1)(9,7))In sublist (4,6), max is 6 and min is 4. In sublist (8,9), max is 9 and min is 8.Comparing max and min values of sublist (2,4) and sublist (6,3), value of max is 6 and min is 2.Therefore, for sublist (2,4,6,3) max is 6 and min is 2.Similarly, comparing max and min values of sublist (8,1) and sublist (9,7), value of max is 9 and min is 1.Therefore, for sublist (8,1,9,7) max is 9 and min is 1.Finally, comparing max and min values of sublist (2,4,6,3) and sublist (8,1,9,7), value of max is 9 and min is 1. Steps to perform MaxMin on a data set (2,4,6,3,8,1,9,7) are:(2,4,6,3) (8,1,9,7)((2,4)(6,3)) ((8,1)(9,7))In sublist (4,6), max is 6 and min is 4. In sublist (8,9), max is 9 and min is 8.Comparing max and min values of sublist (2,4) and sublist (6,3), value of max is 6 and min is 2.Therefore, for sublist (2,4,6,3) max is 6 and min is 2.Similarly, comparing max and min values of sublist (8,1) and sublist (9,7), value of max is 9 and min is 1.Therefore, for sublist (8,1,9,7) max is 9 and min is 1.Finally, comparing max and min values of sublist (2,4,6,3) and sublist (8,1,9,7), value of max is 9 and min is 1. Steps to perform MaxMin on a data set (2,4,6,3,8,1,9,7) are:(2,4,6,3) (8,1,9,7)((2,4)(6,3)) ((8,1)(9,7))In sublist (4,6), max is 6 and min is 4. In sublist (8,9), max is 9 and min is 8.Comparing max and min values of sublist (2,4) and sublist (6,3), value of max is 6 and min is 2.Therefore, for sublist (2,4,6,3) max is 6 and min is 2.Similarly, comparing max and min values of sublist (8,1) and sublist (9,7), value of max is 9 and min is 1.Therefore, for sublist (8,1,9,7) max is 9 and min is 1.Finally, comparing max and min values of sublist (2,4,6,3) and sublist (8,1,9,7), value of max is 9 and min is 1. sonika aggarwal GNIIT
