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Acridine Orange Staining

Acridine Orange contains metachromatic properties, that’s why it is employed in fluorescence microscopy and flow cytometry study of cellular physiology and cell cycle state, including the fluorescent microscopic examination of microorganisms.

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MN Editors

Acridine Orange Stain is used for the differentiation of cellular organelles. When the organisms are treated with Acridine Orange, it binds with the nucleic acid ( either DNA or RNA) by the electrostatic interactions and fluoresces to emit various colors.

Acridine Orange contains metachromatic properties, that’s why it is employed in fluorescence microscopy and flow cytometry study of cellular physiology and cell cycle state, including the fluorescent microscopic examination of microorganisms.

Aim

Acridine Orange (AO) stain is a differential staining method of nucleic acids such as RNA and DNA.

Principle of Acridine Orange Staining

AO is a type of fluorochromatic dye, it binds with the nucleic acids (RNA and DNA) of cells and results deoxyribonucleic acid (DNA) to fluoresce green and ribonucleic acid (RNA) or single stranded DNA to fluoresce orange-red under UV light. In 1942,  Strugge and Hilbrich first introduced this stain.

Requirements

Acridine Orange Staining Procedure

  1. Take a clean grease free glass slide.
  2. Prepare a smear on it and make a thin layer of specimen.
  3. Air-dry the smear
  4. Perform Alcohol fixing by Flooding the smear with methanol.
  5. Air dry the slide
  6. Flood the smear with acridine orange stain and wait for 2 minutes.
  7. Rinse the slide with running tap water.
  8. Allow to air dry.
  9. No slide is ready to observe under a microscope by using oil immersion technique.

Result interpretation

Limitations 

Citation

APA

MN Editors. (April 7, 2021).Acridine Orange Staining. Retrieved from https://microbiologynote.com/acridine-orange-staining/

MLA

MN Editors. "Acridine Orange Staining." Microbiology Note, Microbiologynote.com, April 7, 2021.

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