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|ImageFile=Propidium iodide.png | ImageFile=Propidium iodide.svg
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| InChI = 1/C27H33N4.2HI/c1-4-31(3,5-2)17-9-16-30-26-19-22(29)13-15-24(26)23-14-12-21(28)18-25(23)27(30)20-10-7-6-8-11-20;;/h6-8,10-15,18-19,29H,4-5,9,16-17,28H2,1-3H3;2*1H/q+1;;/p-1 | InChI = 1/C27H33N4.2HI/c1-4-31(3,5-2)17-9-16-30-26-19-22(29)13-15-24(26)23-14-12-21(28)18-25(23)27(30)20-10-7-6-8-11-20;;/h6-8,10-15,18-19,29H,4-5,9,16-17,28H2,1-3H3;2*1H/q+1;;/p-1
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⚫ | PubChem=104981
| UNII = TP416O228T
⚫ | SMILES = CC(C)(CC)CCC1c2cc(ccc2c3ccc(cc3c1c4ccccc4)N)N..
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'''Propidium iodide''' (or PI) is an ] and a ] ] with a ] of 668.4 ] that can be used to ] ]. When excited with 488&nbsp;nm wavelength light, it fluoresces red. Propidium iodide is used as a DNA stain for both ] to evaluate cell viability or DNA content in ]<ref>http://www.biolegend.com/propidium-iodide-solution-2651.html</ref> and microscopy to visualise the nucleus and other DNA containing organelles. It can be used to differentiate ], ] and normal ].<ref>{{cite journal |author=Lecoeur H |title=Nuclear apoptosis detection by flow cytometry: influence of endogenous endonucleases |journal=Exp. Cell Res. |volume=277 |issue=1 |pages=1–14 |year=2002 |pmid=12061813 |doi=10.1006/excr.2002.5537}}</ref> '''Propidium iodide''' (or '''PI''') is a ] ] that can be used to ] ] and ]. 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 ] of PI is enhanced 20-30 fold, and the excitation/emission maximum of PI is shifted to 535 nm (green) / 617 nm (orange-red).<ref>{{cite web |url=https://www.thermofisher.com/us/en/home/life-science/cell-analysis/fluorophores/propidium-iodide.html |title=Propidium Iodide |author=<!--Not stated--> |date=2019-11-14 |publisher=Thermo Fisher Scientific |access-date=2019-11-14 }}</ref> Propidium iodide is used as a DNA stain in ] to ] or DNA content in ], <ref>{{cite web|url=http://www.biolegend.com/propidium-iodide-solution-2651.html|title=Propidium Iodide Solution - BioLegend|access-date=10 January 2015|archive-url=https://web.archive.org/web/20150210095844/http://www.biolegend.com/propidium-iodide-solution-2651.html|archive-date=10 February 2015|url-status=dead}}</ref> or in microscopy to visualize the nucleus and other DNA-containing organelles. Propidium Iodide is not membrane-permeable, making it useful to differentiate ], ] and healthy cells based on membrane integrity.<ref>{{cite journal |author=Lecoeur H |title=Nuclear apoptosis detection by flow cytometry: influence of endogenous endonucleases |journal=Exp. Cell Res. |volume=277 |issue=1 |pages=1–14 |year=2002 |pmid=12061813 |doi=10.1006/excr.2002.5537}}</ref><ref>{{cite web|title=Propidium Iodide|url=https://www.thermofisher.com/order/catalog/product/P1304MP|publisher=ThermoFisher}}</ref> PI also binds to ], necessitating treatment with nucleases to distinguish between RNA and DNA staining.<ref>{{cite journal|vauthors=Suzuki T, Fujikura K, Higashiyama T, Takata K|date=1 January 1997|title=DNA staining for fluorescence and laser confocal microscopy|journal=J. Histochem. Cytochem.|volume=45|issue=1|pages=49–53|doi=10.1177/002215549704500107|pmid=9010468|doi-access=free}}</ref> PI is widely used in fluorescence staining and visualization of the plant cell wall.<ref name=Bidhendi2020>{{cite journal|last1=Bidhendi|first1=AJ|last2=Chebli|first2=Y|last3=Geitmann|first3=A|title=Fluorescence Visualization of Cellulose and Pectin in the Primary Plant Cell Wall|journal=Journal of Microscopy|volume=278 |issue=3 |pages=164–181|date=May 2020|doi=10.1111/jmi.12895|pmid=32270489|s2cid=215619998}}</ref><br />

Propidium Iodide is the most commonly used dye to quantitatively assess DNA content.<ref>Cancer Research UK. 2004. Cell Cycle Analysis - Propidium Iodide. http://science.cancerresearchuk.org/sci/facs/facs_major_apps/cell_cycle_analysis/propidium_iodide/?version=1</ref>{{Citation needed| reason=Better reference needed|date=May 2010}}

Propidium iodide (PI) binds to DNA by intercalating between the bases with little or no sequence preference and with a stoichiometry of one dye per 4–5 base pairs of DNA. PI also binds to ], necessitating treatment with nucleases to distinguish between RNA and DNA staining.<ref>{{cite journal |doi=10.1177/002215549704500107 |author=Suzuki T, Fujikura K, Higashiyama T, Takata K |title=DNA staining for fluorescence and laser confocal microscopy |journal=J. Histochem. Cytochem. |volume=45 |issue=1 |pages=49–53 |date=1 January 1997|pmid=9010468 |url=http://www.jhc.org/cgi/pmidlookup?view=long&pmid=9010468 }}</ref> Once the dye is bound to nucleic acids, its fluorescence is enhanced 20- to 30-fold, the fluorescence excitation maximum is shifted ~30–40&nbsp;nm to the red and the fluorescence emission maximum is shifted ~15&nbsp;nm to the blue. Although its molar absorptivity (extinction coefficient) is relatively low, PI exhibits a sufficiently large ] to allow simultaneous detection of nuclear DNA and ]-labeled antibodies, provided the proper optical filters are used. PI is suitable for ], ], ], and ].

PI is membrane impermeant and generally excluded from viable cells. PI is commonly used for
identifying dead cells in a population and as a counterstain in multicolor fluorescent techniques.<ref>{{cite journal |author=Moore A, Donahue CJ, Bauer KD, Mather JP |title=Simultaneous measurement of cell cycle and apoptotic cell death |journal=Methods Cell Biol. |volume=57 |issue= |pages=265–78 |year=1998 |pmid=9648110 |doi=10.1016/S0091-679X(08)61584-8}}</ref> The counterstaining protocols below are compatible with a wide range of cytological labeling techniques—direct or indirect antibody-based detection methods, mRNA in situ hybridization, or staining with fluorescent reagents specific for cellular structures. These protocols can be modified for tissue staining.

A typical use of propidium iodide in plant biology is to stain the cell wall. Especially useful for '']'' seedling root tissue observed by ], it increases visibility of the outlines of cells in the root tip. This red fluorescent background is useful to determine the sub-localization of a gene of interest expressed as a ] fusion.

Also, propidium iodide is used as a stain in animal cells. For example, in ''Apodemus sylvaticus,'' more commonly known as the ], it can be used to indicate the location of the nuclear region by emitting its characteristic red fluorescence.<ref>Moore, Harry et al., Exceptional sperm cooperation in Wood Mouse.Nature 418, 174-177 (2002).</ref>


== See also == == See also ==
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==References== ==References==
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