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Ziehl-Neelsen Technique.
also know as the Hot stain.
application of heat during staining for the greater penetration of the primary stain which is the Carbolfushin.
Kinyoun techinique.
also know as the Cold stain.
Acid fast organisms can also be visualized by Flourescence Microscopy using specific flourescent dyes such as Auramine and Rhodamine.
:D
-iampol
Wiki User
∙ 14y ago
Wiki User
∙ 13y agoSome bacteria contain a waxy lipid, mycolic acid, in there cell wall. This lipid makes the cells more durable and is commonly associated with pathogens. Acid fast cell walls are so durable that the stain (carbol fuschin) must be driven into the cells with heat. The cells are then decolorized with acid-alcohol, all other cells will decolorize with this strong solvent, but acid fast bacteria will not. Other cells are then counterstained with methylene blue
Wiki User
∙ 9y agoAcid-fast organisms are characterized by wax-like cell walls. Because the cell wall is so resistant to most compounds, acid-fast organisms require a special staining technique.
1. The primary stain used in acid-fast staining, carbolfuchsin, is lipid-soluble and contains phenol, which helps the stain penetrate the cell wall. (violet or purple).
2. This is further assisted by the addition of heat.
3. The smear is then rinsed with a very strong decolorizer, which strips the stain from all non-acid-fast cells but does not permeate the cell wall of acid-fast organisms.
4. The decolorized non-acid-fast cells then take up the counterstain. (safranin- red).
Gram positive bacteria will stain purple.
Gram negative bacteria will stain red/pink.
When doing this stain all steps are done because you are looking to identify the microbe. You may have guessed what it is but this will help nail it down along with its' shape.
Wiki User
∙ 10y agoThe cold methods of acid fast staining refers to the procedure for staining tubercle bacilli.
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In 1877 Robert Koch thought preparing permanently stained slides would be valuable Why was his assessment correct?
A method was needed to separate these different bacteria. he used permanently staining methods to begin the process of distinguishing microbes and identifying pathogens. These same techniques are essential in studying all microorganism.
What is a primary stain?
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.
Why are the steps in gram staining so carefully standardized?
Gram's stain remains one of the most valuable methods we have for identifying isolates accurately and rapidly. Despite our long-standing familiarity with this method, it still warrants careful attention every step of the way--from preparation and QC of reagents to staining and interpretation. I think one of the main reasons would to avoid contamination.
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