You won't be able to positively distinguish gram negative from gram positive organisms. Crystal violet is used first so that gram positive organisms will take up the dye and this color is not affected by safrinin later on. Gram negative organisms will lose the purple coloring during the decolorizer step, therefore need to be counterstained so that you can view the organisms.
You can use acetone or alcohol to remove crystal violet(decolorizer).
It's one of the differentiating dyes that shows a bacterium is gram positive by staying purple at the end of the staining process.
Its the primary stain of the procedure. IT stains the Gram positive organisms
Acetone is used for decolouring method, washing away the iodine-crystal violet complex formed in gram negative bacteria. so that the gram negative bacteria can be then stained with safranin or fuchsin.
It removes the crystal violet (primary stain) from gram negative bacteria. It does not remove Crystal violet as easily from gram positive bacteria, because the highly peptidoglycan walls of gram positive bacteria interact with crystal violet and iodine to form a strong bond (CV-Iodine complex). So, gram positive cells hold on to the stain instead of letting it wash away.
It allows the primary stain crystal violet to remain in the cell instead of being washed out. Due to the larger size of the crystal violet molecule, when the ethanol is applied (the decolorizer) the stain will not be washed out of the Gram (+) positive cells.
The steps in Gram staining are:1. crystal violet added to the smear2. iodine, the mordant (this fixes the violet)3. a decolorizer made of acetone and alcohol4. safranin, the counterstainIf 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.
It is crystal violet & stains all cells purple.
If using a gram stain, they will turn Crystal Violet.
Its the primary stain of the procedure. IT stains the Gram positive organisms
Its the primary stain of the procedure. IT stains the Gram positive organisms
In Gram staining procedure during bacterial staining , iodine forms a complex with crystal violet stain which stains Gram positive bacteria blue to violet .
Acetone is used for decolouring method, washing away the iodine-crystal violet complex formed in gram negative bacteria. so that the gram negative bacteria can be then stained with safranin or fuchsin.
It removes the crystal violet (primary stain) from gram negative bacteria. It does not remove Crystal violet as easily from gram positive bacteria, because the highly peptidoglycan walls of gram positive bacteria interact with crystal violet and iodine to form a strong bond (CV-Iodine complex). So, gram positive cells hold on to the stain instead of letting it wash away.
In a gram stain the primary stain is crystal violet. Iodine then sets that dye into the gram positive cells while alcohol washes out the crystal violet from the gram negative cells. Then safranin, which is the counterstain in a gram stain, is used to dye the rest of the bacteria. This is the example I can give you of why a counterstain does not change the look in all the cells. Though safranin stains all the cells, the gram positive cells that were dyed purple from crystal violet don't look pink - only the gram negative do.
It allows the primary stain crystal violet to remain in the cell instead of being washed out. Due to the larger size of the crystal violet molecule, when the ethanol is applied (the decolorizer) the stain will not be washed out of the Gram (+) positive cells.
If iodine is not applied, both the gram-positive and gram-negative stains will appear to be gram-negative. The iodine acts as a mordant that helps to fix the crystal violet stain in the gram-positive bacteria, making them appear purple. Without iodine, the crystal violet stain can be easily washed out of both gram-positive and gram-negative bacteria, resulting in a pink or red color.
The steps in Gram staining are:1. crystal violet added to the smear2. iodine, the mordant (this fixes the violet)3. a decolorizer made of acetone and alcohol4. safranin, the counterstainIf 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.
Well if you stained with only crystal violet, then they would all be violet! If you do a Gram Stain the right way, you end up getting Gram positive being violet or purple and Gram Negative being red or pink.