Tugas Presentasi :

Pemeriksaan Mikroskopis
Acridine orange

▪ Priciple :
▪ Acridine orange is a cell-permeable,nucleic acid selective dye that emits green fluorescence
when bound to dsDNA(at 520 ) and red fluorescence when bound to ssDNA or RNA(at 650 nm)
▪ It is a cationic dye → also enter acidic compartments such as lysosomes which in low pH
conditions, will emit orange light.
▪ Note: RNA is more abundant during cellular growth and may mask the green fluorescence of the
DNA within the cell

▪ Procedur :
▪ Examine first with the 10X objective lens to see the distribution of fluorescing material, and then
with the 40X objective lens to identify Trichomonas vaginalis and to detect yeast cells, bacteria
and parasites( chromatoid bars of Entamoeba histolytica/ dispar).
Uses of Acridine Orange Staining

1. Rapid identification of Trichomonas vaginalis, yeast cells, and clue cells in vaginal smears which can also
use to detect intracellular gonococci, meningococci, and other bacteria particularly in blood cultures.
2. Analyzing mitochondria and lysosomal content by flow cytometer.
3. Applicable for visual detection of DNA or RNA ( nucleic acids) on agarose and polyacrylamide gels.
4. May enumerate the microbial load in a sample because acridine orange binds with the nucleic acid of
both living and dead organisms.
5. Identifying engulfed apoptotic cells, because it will fluoresce upon engulfment.
6. Acts as differential staining for human cells and prokaryotic cell with a fluorescence microscope.
7. To differentiate cells in the live and dead tumor cell groups.
8. Test for Assessment of Human Sperm DNA Integrity
Result interpretation

▪ Trichomonas vaginalis : Orange-red with yellow-green nucleus

▪ Yeast cells : Orange
▪ Bacteria : Orange

Limitations of Acridine Orange Staining

1. Cellular debris within a specimen (WBCs, epithelial cells, and dead bacteria) may
distort the microscopic image.
2. Due to being very sensitive stain, caution should be used when interpreting tests
results.

WBC :Yellow-green
Epithelial cells : Yellow-green
Contoh interpretasi

staphylococcal colonisation (orange microorganisms) from a burn skin specimen

 Referensi

▪ Merlino J. Resistance to the biocidal activity of silver in burn wound dressings - is it a problem?.
Microbiology Australia 31(4):168-170 DOI: 10.1071/MA10168. January 2010

▪ Aryal S. Acridine Orange Stain- Principle, Procedure and Result Interpretation. Microbe notes. August 4,
2017