Technical Data #8574a / 2013.07.21

Acridine Orange is a stain used for demonstrating bacteria in normally sterile specimens or when bacteria are present in low quantity.



per liter

Acridine Orange

0,1 gram

Acetate Buffer, 0,5M

1,0 Liter

pH: 3.7 +/- 0,3 à 25o C

This medium is for IN VITRO diagnostic use only.

Store stain at room temperature. Do not use if there is evidence of precipitate, signs of deterioration or if the expiration date has passed.


  1. Prepare a smear on a clean glass slide and allow to air dry.
  2. Fix smear with methanol for 2 minutes and allow to air dry.
  3. Flood slide with Acridine Orange Stain for 2 minutes.
  4. Rinse thoroughly with tap water and allow to dry.
  5. View slide under a high dry objective of a fluorescent microscope.

Bacteria and yeasts cells stained a brilliant orange and background may be black, light green or yellow. Green, yellow or red debris may be observed.


Granules from disintegrated leukocytes may be mistaken for cocci at lower magnifications, but may be distinguished at higher magnifications. The Acridine Orange is a screening procedure, Positive smears must be Gram-stained directly over the Acridine Orange slide.


  1. Burdash, N.M., J.P. Manos, E.R. Bannister and A.L. Welborn. 1983 Acridine Orange Staining and Radiometric Detection of Microorganisms in Blood Cultures. J. Clin. Microbiol. 17:463-465.
  2. Greenwood, J.R. and Kirk-Hillaire. 1981 Evaluation of Acridine Orange Stain for Detection of Trichomonas vaginalis in Vaginal Specimens. J. Clin. Microbiol. 14:699.
  3. Lauer, B.A., L.B. Reller and S. Mirrett. 1981 Comparaison of Acridine Orange and Gram Stains for Detection of Microorganisms in Cerebrospinal Fluid and Other Clinical Specimens. J. Clin. Microbiol. 14:201-205.
  4. Mac Carthy, L.R. and J.E. Senne. 1980 Evaluation of Acridine Orange for Detection of Microorganisms in Blood Culture. J. Clin. Microbiol. 11:281-285.


Acridine Orange Stain 250 ml 8574




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