Differential Stain- Acid Fast Staining Flashcards
Purpose of Acid Fast Staining
used to detect groups of microbes that do not readily take up dyes (usually Mycobacterium species) (usually bacterias that stain Gram Positive)
What is special about cell walls of mycobacterium that typically undergo acid fast staining?
Their cell walls contains high levels of mycolic acids (wax like fatty acids) that prevent uptake of dyes and require harsher methods
Once mycobacterium are stained,
they are resistant to decolorization (acid fast).
Acid fast technique is great for
diagnosing pathogenic species like mycobacterium, TB and leprosy
Acid Fast/ Ziel-Neelson Stains
Primary Stain is red carbolfuchsin , rinse w distilled water, DECOLORIZING agent w/ acid alcohol applied (REMOVES CARBOLFUCHSIN from all cells except acid fast) , Counterstain Methylene Blue is used. If acid fast, cell is red. If not, stains blue
Acid fast technique and time (5-1-1min)
Carbol fuchsin for 5 mins, rinse w/ distilled water, Add decolorizer (acid-alochol) for 1 min, rinse w/ distilled water, Add methylene bkue for 1 mins, Rinse, Blot dry
Although Acid Fast bacteria stain Gram Positive, the difference in their cell wall is that
it contains mycolic acid (waxy lipid) as its primary component. this wax like component makes it difficult to stain
Primary Stain used to penetrate wax like mycolic acids in Acid Fast Staining
Carbol Fuchsin
If cell retains carbol fuchsin, it is considered
Acid Fast
If cell is decolorized by acid alcohol, and picks up counterstain of methylene blue
it is Acid fast negative
Primary Stain, Mordant, Decolorizer, Counter stain in Acid Fast procedure are
Carbol Fuchsin, “Heat”, Acid-Alcohol, Methylene Blue