some orgs are too sensitive to be passed thru a flame. if the org cant withstand heat, it will be distorted and it will not yield an accurate representation of its phys properties in vivo when viewed under a mscope. Also, negative staining often involves nigrosine, India ink, etc which creates a thick layer which light cannot penetrate. So if heated, this layer cracks (many thin microscopic cracks) and you would not like that.
Negative stains like nigrosin rely on the acidic properties of the stain to be repelled by the negatively charged bacterial cells, resulting in a contrasting background. Applying heat can disrupt this process by altering the charge distribution on both the bacterial cell and the stain molecules, leading to inaccurate staining results.
Steam heating is used in acid-fast staining to drive the primary stain, carbol fuchsin, into the lipid-rich cell wall of acid-fast bacteria. Heat fixing, on the other hand, is done to adhere the bacterial cells to the slide and denature proteins to make them more permeable to stains, but it is not strong enough to penetrate the tough cell wall of acid-fast bacteria.
Unfortunately, if bleach doesn't fix it them nothing probably will. It may be time to get a new table cloth.
If the bleach has soaked in it will have removed colour from the material. So even removing the bleach if possible, will not fix the stain. You may need to re-dye if possible.
Unfortunately, yellow bleach stains are permanent as the bleach has altered the color of the fabric. You can try masking the stain with fabric dye or fabric markers, or consider adding a decorative patch over the stained area. Additionally, you can try using a color remover to lighten the entire garment to make the stain less noticeable.
Dont remove it, paint over it with gold-colored paint.
since you do not heat fix the slide when you use a negative stain the cells do not shrink or become distorted
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.
You absolutely do not heat fix a blood smear before staining, that is, if you are looking at the blood cells. For bacteria, why wouldn't you culture it first and then heat fix, stain etc. I don't think heat fixing the blood stain would damage the bacterial cells so much as make it hard to differentiate the bacterial cells from the dead, shriveled, ruined blood cells, unless maybe you have like an electron microscope or something.
to fix the color of the stain
If you forget to heat fix your slide your bacterial sample will be lost with the next wash step. So if you are doing a Gram stain when you add the crystal violet the liquid will mix with the bacteria, and when you wash later in the protocol the bacteria will wash away with the stains.
Gram's iodine stain is applied after the culture is stained with the primary stain. It acts as a mordant, fixing the primary stain to the cell wall while lending no additional colour to the cell (i.e. the mordant itself is not a stain). The mordant is only able to fix the stain to Gram-positive bacteria because of the characteristic thick, peptidoglycan coat that they possess. Because the mordant is not able to fix the stain to Gram-negative bacteria (who's coat have a different composition), the crystal violet stain will wash away from Gram-negative bacteria when the decolourizing agent is added.
smear will be washed( no smear will be left on the slide)
Methanol is used to fix the smear during Giemsa staining because it helps to preserve the cellular structure by denaturing proteins and preventing their degradation. It also permeabilizes the cells, allowing for better penetration of the dye and increased staining intensity. Additionally, methanol evaporates quickly, which helps to reduce the staining time and allows for quicker results.
methods that fix different colors on different components of cells.
Passing the bacterial smear through the flame before staining is done to heat-fix the bacteria onto the slide, making them adhere firmly and preventing them from washing off during the staining process. Heat fixing also kills the bacteria, which helps in the preservation of their cellular structures for visualization under the microscope.
It is used to fix because to make the cell inactive or immoblie, but the main purpose is to fix the smear so that when we put stain and then flush it out with water ( or some time with alcohol) the smear should not wash out with dye.
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