Mycobacterium tuberculosis is an obligate species in the family Mycobacterium and the causative agent of most cases of tuberculosis
For Mycobacterium you will use the Acid-fast staining technique. There are two different methods of stainging: 1) Ziehl-Neelsen Method and 2) Kinyoun Method.1) The Ziel-Neelsen method uses a primary stain of Carbol Fuchsin dye that must be steam treated, rinsed with acid alcohol wash, and a secondary stain of Methylene Blue.2) The Kinyoun Method uses a primary stain of Kinyoun Carbol Fuchsin dye that is not steam treated. An acid alcohol wash is applied and a secondary dye of Brilliant Green. This technique is called "cold staining".The mycolic acid within the Mycobacterium cell membrane has a high affinity for the Carbol Fuchsin dyes.
No. safranin is the classic stain used in gram staining. Concentrated Carbol Fushin is mainly used for the ZN staining procedure to stain organisms such as Vibrio cholerae and Cryptosporidium. Diluted Carbol Fushin can however be used as a replacement counterstain for Safranin in the gram stain.
Leo Carbol was born in 1910.
Leo Carbol died in 1991.
If your talking about Acid Fast staining (aka Ziehl-Neelsen staining), after the addition of the primary stain (carbol fuchsin) heat is applied in order to facilitate proper staining. Bacteria such as Mycobacterium contain large amounts of lipid substances within their cell wall called mycolic acids. These acids resist staining by ordinary methods such as gram staining (where heat is not applied after primary staining). On application of heat, the stain carbol fuschin penetrates the cell wall and stains the cells pink. Following the secondary staining (methylene blue) the acid fast positive cells appear pink while others are stained blue. Endospore staining is yet another staining technique where heat is applied after primary staining (malachite green). In this case the spores are impermeable to stains, hence heating favours proper permeation of stain. Endospores appear green while vegetative cells appear red (secondary stain saffranine). Not all staining procedures involve applying heat after primary staining. However, heat is applied before even beginning the staining procedure. This is called heat fixing, where the cells are fixed so that they are not washed away during the subsequent washing steps.
Both processes use 2 stains. The Gram staining process uses crystal violet as the primary stain and safranin as the secondary stain. Acid-fast staining uses carbol fuchsin as the primary and methylene blue as the secondary.
Primary stain: carbol fuchsin will leave the acid fast cells red, as it can penetrate the lipoidal (thick waxy) shell of the acid fast shell. Counterstain: methylene blue will leave the non-acid fast cells blue. So you should have red and blue.
The hot method uses heat to force the primary stain (carbol fuchsin) into the cell wall, while the cold method uses a detergent to cause the stain to penetrate the wall.
No. It is basic. No. It is basic.
One thing that endospore stains have in common with the acid fast stain is that heat primary stain penetration. Another thing that endospore stains have in common with acid fast stains are counterstain.
to break down the cell wall
methylene blue crystal violet carbol fuchsin