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Aladin creations zeal Nielsen method of acid-fast staining technique is used to stain Mycobacterium species including Mycobacterium tuberculosis Mycobacterium ulcer ins and Mycobacterium leprae detection of acid-fast bacilli in stained and acid-washed smears examined microscopically may provide the initial bacteria logic evidence of the presence of mycobacteria in a clinical specimen principle the cell wall of mycobacteria contain high concentration of lipid making them waxy hydrophobic and impermeable to routine stain such as the Gram stain but stained with caramel Fuchs and combined with phenol zeal Nielsen staining technique has three main steps is primary staining to colorization end counter stain the phenol Carville Fuchs and stain is heated to enable the dye to penetrate the waxy mycobacterial cell wall the stain binds to the mycolic acid in the mycobacterial cell wall after staining decolorizing solution is applied this removes the red dye from the background cells tissu