Is propidium iodide a fluorescent dye?
Propidium iodide (PI) is a popular red-fluorescent nuclear and chromosome counterstain. Since propidium iodide is not permeant to live cells, it is also commonly used to detect dead cells in a population.
How does propidium iodide stain DNA?
Propidium iodide (PI) is a cell-impermeant DNA binding dye that can be used to stain cells and nucleic acids. PI binds to DNA by intercalating between the bases with a stoichiometry of one dye per 4-5 base pairs of DNA. As a membrane impermeant dye, PI is generally excluded from viable cells.
What is the excitation and emission spectra of propidium iodide?
Propidium Iodide (PI) binds nucleic acids by intercalation in the bases. When bound to nucleic acid it has an excitation maxima ~535 nm and an emission maxima ~615 nm. On a flow cytometer PI is typically excited by 488 or 561 nm and can be detected in a 610/20 bandpass.
Why does propidium iodide staining indicate apoptosis?
Annexin V and PI are double-staining probes for apoptosis by detecting the externalization of phosphatidylserine and membrane integrity. The apoptotic cells externalize their phosphatidylserine early in apoptosis when the cell membrane is still intact.
What is the color of propidium iodide?
red
Propidium iodide (PI) is a popular red-fluorescent nuclear and chromosome counterstain. Since propidium iodide is not permeant to live cells, it is also commonly used to detect dead cells in a population. PI binds to DNA by intercalating between the bases with little or no sequence preference.
What is propidium iodide staining?
Propidium iodide (or PI) is a fluorescent intercalating agent that can be used to stain cells and nucleic acids. Propidium iodide is used as a DNA stain in flow cytometry to evaluate cell viability or DNA content in cell cycle analysis, or in microscopy to visualize the nucleus and other DNA-containing organelles.
Is propidium iodide cell permeable?
Propidium iodide (or PI) is a fluorescent intercalating agent that can be used to stain cells and nucleic acids. Propidium Iodide is not membrane-permeable, making it useful to differentiate necrotic, apoptotic and healthy cells based on membrane integrity.
Is propidium iodide cytotoxic?
A propidium iodide fluorescence assay (PIA) was developed to characterize the in vitro growth of human tumor cell lines as well as to test the cytotoxic activity of standard compounds. Propidium iodide (PI) was used as a dye which penetrates only damaged cellular membranes.
Does propidium iodide stain apoptotic cells?
Propidium iodide is a membrane-impermeable fluorescent DNA stain. It is used to stain apoptotic cells for flow cytometry, in situ hybridization, and immunohistochemistry applications Belloc et al (1994). It can also be used to differentiate between apoptotic and necrotic cell death.
What happens to phosphatidylserine during apoptosis?
Exposure of phosphatidylserine on the outer leaflet of the plasma membrane is a surface change common to many apoptotic cells. Phosphatidylserine exposure has several potential biological consequences, one of which is recognition and removal of the apoptotic cell by phagocytes.
Where does propidium iodide stain localize in the cell?
Where does the propidium iodine stain in the cell? Nucleus of the cell.
How does propidium iodide work in flow cytometry?
Propidium iodide (PI) is a membrane impermeant dye that is generally excluded from viable cells. It binds to double stranded DNA by intercalating between base pairs. PI is excited at 488 nm and, with a relatively large Stokes shift, emits at a maximum wavelength of 617 nm.
What is propidium iodide used to detect?
Propidium iodide (PI) is a popular red-fluorescent nuclear and chromosome counterstain. Since propidium iodide is not permeant to live cells, it is also commonly used to detect dead cells in a population.
Can propidium iodide enter eukaryotic cells?
Propidium iodide is cell impermeant, and will only enter dead cells. If the eukaryotic cells are dead, they will label with propidium iodide as well. If the eukaryotic cells are alive, propidium iodide will not be able to enter and thus will not label the bacteria inside, whether the bacteria are alive or dead.
What is the fluorescence emission maximum of a bound dye?
Once the dye is bound, its fluorescence is enhanced 20- to 30-fold, the fluorescence excitation maximum is shifted ∼30–40 nm to the red and the fluorescence emission maximum is shifted ∼15 nm to the blue, resulting in an excitation maximum at 535 nm and fluorescence emission maximum at 617 nm.
What is the excitation/emission maxima of Pipi?
PI binds to DNA by intercalating between the bases with little or no sequence preference. In aqueous solution, the dye has excitation/emission maxima of 493/636 nm.