Acridine orange
Encyclopedia
Acridine orange is a nucleic acid
selective fluorescent cationic dye
useful for cell cycle determination. It is cell-permeable, and interacts with DNA
and RNA
by intercalation
or electrostatic attractions respectively. When bound to DNA, it is very similar spectrally to fluorescein
, with an excitation
maximum at 502 nm
and an emission maximum at 525 nm (green). When it associates with RNA, the excitation maximum shifts to 460 nm (blue) and the emission maximum shifts to 650 nm (red). Acridine orange will also enter acidic compartments such as lysosomes and become protonated and sequestered. In these low pH
conditions, the dye will emit orange light when excited by blue light. Thus, acridine orange can be used to identify engulfed apoptotic cells, because it will fluoresce upon engulfment. The dye is often used in epifluorescence microscopy
.
Acridine orange is prepared from coal tar
and creosote
oil.
Acridine orange can be used in conjunction with ethidium bromide
to differentiate between viable, apoptotic and necrotic cells. Additionally, Acridine orange may be used on blood samples to fluoresce bacterial DNA, aiding in clinical diagnosis of bacterial infection once serum and debris have been filtered.
Acridine orange can be used in the assessment of sperm chromatin
quality.
Nucleic acid
Nucleic acids are biological molecules essential for life, and include DNA and RNA . Together with proteins, nucleic acids make up the most important macromolecules; each is found in abundance in all living things, where they function in encoding, transmitting and expressing genetic information...
selective fluorescent cationic dye
Dye
A dye is a colored substance that has an affinity to the substrate to which it is being applied. The dye is generally applied in an aqueous solution, and requires a mordant to improve the fastness of the dye on the fiber....
useful for cell cycle determination. It is cell-permeable, and interacts with DNA
DNA
Deoxyribonucleic acid is a nucleic acid that contains the genetic instructions used in the development and functioning of all known living organisms . The DNA segments that carry this genetic information are called genes, but other DNA sequences have structural purposes, or are involved in...
and RNA
RNA
Ribonucleic acid , or RNA, is one of the three major macromolecules that are essential for all known forms of life....
by intercalation
Intercalation (chemistry)
In chemistry, intercalation is the reversible inclusion of a molecule between two other molecules . Examples include DNA intercalation and graphite intercalation compounds.- DNA intercalation :...
or electrostatic attractions respectively. When bound to DNA, it is very similar spectrally to fluorescein
Fluorescein
Fluorescein is a synthetic organic compound available as a dark orange/red powder soluble in water and alcohol. It is widely used as a fluorescent tracer for many applications....
, with an excitation
Excited state
Excitation is an elevation in energy level above an arbitrary baseline energy state. In physics there is a specific technical definition for energy level which is often associated with an atom being excited to an excited state....
maximum at 502 nm
Nanometre
A nanometre is a unit of length in the metric system, equal to one billionth of a metre. The name combines the SI prefix nano- with the parent unit name metre .The nanometre is often used to express dimensions on the atomic scale: the diameter...
and an emission maximum at 525 nm (green). When it associates with RNA, the excitation maximum shifts to 460 nm (blue) and the emission maximum shifts to 650 nm (red). Acridine orange will also enter acidic compartments such as lysosomes and become protonated and sequestered. In these low pH
PH
In chemistry, pH is a measure of the acidity or basicity of an aqueous solution. Pure water is said to be neutral, with a pH close to 7.0 at . Solutions with a pH less than 7 are said to be acidic and solutions with a pH greater than 7 are basic or alkaline...
conditions, the dye will emit orange light when excited by blue light. Thus, acridine orange can be used to identify engulfed apoptotic cells, because it will fluoresce upon engulfment. The dye is often used in epifluorescence microscopy
Fluorescence microscope
A fluorescence microscope is an optical microscope used to study properties of organic or inorganic substances using the phenomena of fluorescence and phosphorescence instead of, or in addition to, reflection and absorption...
.
Acridine orange is prepared from coal tar
Coal tar
Coal tar is a brown or black liquid of extremely high viscosity, which smells of naphthalene and aromatic hydrocarbons. Coal tar is among the by-products when coal iscarbonized to make coke or gasified to make coal gas...
and creosote
Creosote
Creosote is the portion of chemical products obtained by the distillation of a tar that remains heavier than water, notably useful for its anti-septic and preservative properties...
oil.
Acridine orange can be used in conjunction with ethidium bromide
Ethidium bromide
Ethidium bromide is an intercalating agent commonly used as a fluorescent tag in molecular biology laboratories for techniques such as agarose gel electrophoresis. It is commonly abbreviated as "EtBr", which is also an abbreviation for bromoethane...
to differentiate between viable, apoptotic and necrotic cells. Additionally, Acridine orange may be used on blood samples to fluoresce bacterial DNA, aiding in clinical diagnosis of bacterial infection once serum and debris have been filtered.
Acridine orange can be used in the assessment of sperm chromatin
Chromatin
Chromatin is the combination of DNA and proteins that make up the contents of the nucleus of a cell. The primary functions of chromatin are; to package DNA into a smaller volume to fit in the cell, to strengthen the DNA to allow mitosis and meiosis and prevent DNA damage, and to control gene...
quality.