Auramine Phenol, kasutusvalmis

Product information

Auramine Phenol is a fluorochrome and intended for use in the detection of Acid Fast Bacteria such as M. tuberculosis and S. griseus in clinical specimens.  The standard staining method is as follows:

i) Prepare the smear of material to be examined on a glass slide, dry and heat fix using gentle heat. Alternately the smear may be fixed in Methanol.
ii) Apply Auramine Phenol to the slide for 10 minutes.
iii) Wash off excess stain by rinsing the slide in water. Shake off any excess water. 
iv) Apply Auramine Differentiator for 10 minutes. Apply a change of Differentiator after 5 minutes. 
v) Wash off the Differentiator by rinsing the slide in water. Shake off any excess water.
vi) Apply Potassium Permanganate (or Thiazine Red) and counterstain for 30 seconds.
vii) Remove excess stain by rinsing the slide in water. Shake off any excess water. 
viii) Blot the slide gently with clean blotting paper, and either air dry or dry using gentle heat.
ix) Examine the stained slide under the fluorescent microscope using oil immersion objective x 100 under oil.

When the Auramine Phenol staining procedure is performed correctly acid fast organisms (AFB) appear as bright luminous rods under fluorescent microscopy. Organisms and cells and the background that are decolourised by the Differentiator and take up the counterstain will appear as a dark background. Care must be taken when using fluorescent microscopy to differentiate between organisms and non-specific fluorescence.
NB. The fluorescence displayed by the AFB can be weakened if the counterstain is left on the slide for too long. 

Control Results
M. tuberculosis HR37 Rv NCTC 7416 should appear as acid fast bacilli (AFB positive) and appear as bright luminous rods.  S. griseus NCTC 7807 should appear as non-acid fast bacilli (AFB negative) and should appear as purple or red in colour.