Summary: Propidium iodide (PI) Staining. PI is an intercalating agent and a fluorescent molecule with a molecular mass of 668.4 Da, when bound to nucleic acids, excitation maximum at 535 nm and the emission maximum at 617 nm. PI is used as a DNA stain for both flow cytometry, to evaluate cell viability or DNA content in cell cycle analysis and microscopy to visualize the nucleus and other DNA containing organelles. Since PI also binds to RNA, it is necessary to treat cells with RNase to distinguish between RNA and DNA staining. PI is a membrane impermeant dye that is generally excluded from viable cells. Therefore, for DNA staining with PI, cells must be fixed or permeabilized to allow entry of the dye. Due to this property, PI staining of unpermeabilized cells followed by flow cytometry can be used to distinguish dead and live cells.
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Protocols
- Cell Cycle Analysis by DNA Content (Propidium Iodide) (UCSD Cancer Center Flow Cytometry Core Facility)
Classic cell cycle analysis by PI staining with example graphics
http://cancer.ucsd.edu/flow/Protocols/DNA%20Conten...
Added: Sun Aug 01 2010, Hits: 5089, Reviews: 0 Write review Cached - Cell Cycle Analysis by Propidium Iodide (PI) Staining
(Core Facility Cell Sorting, Universität: Universität zu Lübeck)
http://www.cellsorting.uni-luebeck.de/Dokumente/pr...
Added: Sat Feb 16 2013, Hits: 1340, Reviews: 0 Write review - Propidium iodide staining of cells to assess DNA cell cycle (Abcam)
Cell cycle analysis by quantitation of DNA content was one of the earliest applications of flow cytometry. The DNA of mammalian, yeast, plant or bacterial cells can be stained by a variety of DNA binding dyes. The premise with these dyes is that they are stoichiometric i.e. they bind in proportion to the amount of DNA present in the cell. In this way cells that are in S phase will have more DNA than cells in G1. They will take up proportionally more dye and will fluoresce more brightly until they have doubled their DNA content. The cells in G2 will be approximately twice as bright as cells in G1.
http://www.abcam.com/index.html?pageconfig=resourc...
Added: Sat Feb 16 2013, Hits: 1101, Reviews: 0 Write review Cached -
DNA Content Detection for Cell Cycle Analysis
(Cell Sorter Facility, Yale University)
http://info.med.yale.edu/immuno/cytometry/docs/DNA...
Added: Mon Feb 23 2009, Hits: 5473, Reviews: 0 Write review Cached -
FACS Cell Cycle Staining using DAPI
(Flow Cytometry Core Facility, University of Chicago)
http://iacf.uchicago.edu/Protocols/DAPICellCyclePr...
Added: Sat Feb 21 2009, Hits: 4127, Reviews: 0 Write review Cached -
Fixation and DNA Staining for Cell Cycle Analysis
(U of Florida Flow Cytometry Core Laboratory)
This method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to enter the cells.
http://www.biotech.ufl.edu/FlowCytometry/dna_fix.h...
Added: Tue May 14 2002, Hits: 1574, Reviews: 0 Write review Cached -
Flow Cytometric Analysis of Cell Cycle
(Hannun/Obeid Lab)
Protocol for fixing and staining
http://www.musc.edu/bcmb/ceramide/protocols/0010.h...
Added: Tue May 14 2002, Hits: 5617, Reviews: 0 Write review Cached -
Flow Cytometry of Fibroblast Nuclei for DNA content
(Fero Lab, Fred Hutchinson Cancer Research Center)
http://labs.fhcrc.org/fero/Protocols/MEFnuclei.htm...
Added: Sat Jan 31 2009, Hits: 1734, Reviews: 0 Write review Cached -
PI Labeling Method for FACS Analysis
(Chen Lab, UCSF)
http://bts.ucsf.edu/chen/Protocol/PI%20Labeling%20...
Added: Sun Oct 25 2009, Hits: 6139, Reviews: 0 Write review -
Propidium Iodide Staining for DNA Ploidy
(Bissell Lab, Berkely Lab)
http://www.lbl.gov/LBL-Programs/lifesciences/Bisse...
Added: Sat Feb 21 2009, Hits: 3819, Reviews: 0 Write review -
Propidium Iodide Staining of Dead Cells For FACS Analysis
(UCLA Flow Cytometry Core)
http://cyto.mednet.ucla.edu/Protocols/pi.htm
Added: Sat Jan 31 2009, Hits: 7110, Reviews: 0 Write review Cached