Propidium iodide (or PI) is a
fluorescent
Fluorescence is one of two kinds of photoluminescence, the emission of light by a substance that has absorbed light or other electromagnetic radiation. When exposed to ultraviolet radiation, many substances will glow (fluoresce) with color ...
intercalating agent
In biochemistry, intercalation is the insertion of molecules between the planar bases of deoxyribonucleic acid (DNA). This process is used as a method for analyzing DNA and it is also the basis of certain kinds of poisoning.
There are several ...
that can be used to
stain
A stain is a discoloration that can be clearly distinguished from the surface, material, or medium it is found upon. They are caused by the chemical or physical interaction of two dissimilar materials. Accidental staining may make materials app ...
cells and
nucleic acids
Nucleic acids are large biomolecules that are crucial in all cells and viruses. They are composed of nucleotides, which are the monomer components: a 5-carbon sugar, a phosphate group and a nitrogenous base. The two main classes of nucleic a ...
. PI binds to DNA by intercalating between the bases with little or no sequence preference. When in an aqueous solution, PI has a fluorescent excitation maximum of 493 nm (blue-green), and an emission maximum of 636 nm (red). After binding DNA, the
quantum yield
In particle physics, the quantum yield (denoted ) of a radiation-induced process is the number of times a specific event occurs per photon absorbed by the system.
\Phi(\lambda)=\frac
Applications
Fluorescence spectroscopy
The fluorescence ...
of PI is enhanced 20-30 fold, and the excitation/emission maximum of PI is shifted to 535 nm (green) / 617 nm (orange-red). Propidium iodide is used as a DNA stain in
flow cytometry
Flow cytometry (FC) is a technique used to detect and measure the physical and chemical characteristics of a population of cells or particles.
In this process, a sample containing cells or particles is suspended in a fluid and injected into the ...
to
evaluate cell viability or DNA content in
cell cycle analysis, or in microscopy to visualize the nucleus and other DNA-containing organelles. Propidium Iodide is not membrane-permeable, making it useful to differentiate
necrotic,
apoptotic and healthy cells based on membrane integrity. PI also binds to
RNA
Ribonucleic acid (RNA) is a polymeric molecule that is essential for most biological functions, either by performing the function itself (non-coding RNA) or by forming a template for the production of proteins (messenger RNA). RNA and deoxyrib ...
, necessitating treatment with nucleases to distinguish between RNA and DNA staining.
PI is widely used in fluorescence staining and visualization of the plant cell wall.
See also
*
Viability assay
*
Vital stain
*
SYBR Green I
*
Ethidium bromide
References
{{DEFAULTSORT:Propidium Iodide
Flow cytometry
DNA-binding substances
Iodides
Phenanthridine dyes
Staining dyes