Jump to content

DAPI: Difference between revisions

From Wikipedia, the free encyclopedia
Browse history interactively
← Previous editNext edit →
Content deleted Content added
VisualWikitext
No edit summary
No edit summary
Line 39: Line 39:
Apart from labelling cell nuclei, the most popular application of DAPI is in detection of [[mycoplasma]] or [[virus]] DNA in [[cell culture]]s.
Apart from labelling cell nuclei, the most popular application of DAPI is in detection of [[mycoplasma]] or [[virus]] DNA in [[cell culture]]s.


Though DAPI does not enter live cells at physiological pH, it does enter live cells in certain conditions binding tightly to DNA, it is labeled non-toxic in it's MSDS<ref>[http://www.kpl.com/docs/msds/710301.pdf]</ref> and shown to have no mutagenicity <ref>{{cite journal |author=Ohta T, Tokishita S, Yamagata H. |title=Ethidium bromide and SYBR Green I enhance the genotoxicity of UV-irradiation and chemical mutagens in E. coli. |journal=Mutat Res. |volume=492 |issue=1-2 |pages=91–7 |year=2001 |pmid=11377248 }}</ref>. Care should be taken in its handling and disposal.
Though DAPI does not enter live cells at physiological pH, it does enter live cells in certain conditions binding tightly to DNA, it is labeled non-toxic in it's MSDS<ref>[http://www.kpl.com/docs/msds/710301.pdf]</ref> and though was not shown to have mutagenicity to ''E. coli'' <ref>{{cite journal |author=Ohta T, Tokishita S, Yamagata H. |title=Ethidium bromide and SYBR Green I enhance the genotoxicity of UV-irradiation and chemical mutagens in E. coli. |journal=Mutat Res. |volume=492 |issue=1-2 |pages=91–7 |year=2001 |pmid=11377248 }}</ref>, it is labelled as a known mutagen in manufacturer information <ref> Invitrogen, [http://probes.invitrogen.com/media/pis/mp01306.pdf DAPI Nucleic Acid Stain]. accessed 2009-12-08.</ref>. Care should be taken in its handling and disposal.


[[File:DAPIMitoTrackerRedAlexaFluor488BPAE.jpg|250px|left|thumb|Bovine Pulmonary Artery Endothelial cell nuclei stained with DAPI and imaged on a fluorescence microscope.]]The [[Hoechst stain]]s are similar to DAPI in that they are also blue-fluorescent DNA stains which are compatible with both live- and fixed-cell applications.
[[File:DAPIMitoTrackerRedAlexaFluor488BPAE.jpg|250px|left|thumb|Bovine Pulmonary Artery Endothelial cell nuclei stained with DAPI and imaged on a fluorescence microscope.]]The [[Hoechst stain]]s are similar to DAPI in that they are also blue-fluorescent DNA stains which are compatible with both live- and fixed-cell applications.

Revision as of 12:05, 26 April 2010

DAPI
Names
IUPAC name
2-(4-amidinophenyl)-1H -indole-6-carboxamidine
Identifiers
3D model (JSmol)
ChemSpider
  • InChI=1/C16H15N5/c17-15(18)10-3-1-9(2-4-10)13-7-11-5-6-12(16(19)20)8-14(11)21-13/h1-8,21H,(H3,17,18)(H3,19,20)
    Key: FWBHETKCLVMNFS-UHFFFAOYAH
  • [N@H]=C(N)c3ccc(c2cc1ccc(cc1n2)C(=[N@H])N)cc3
Properties
C16H15N5
Molar mass 277.324
Except where otherwise noted, data are given for materials in their standard state (at 25 °C [77 °F], 100 kPa).

DAPI or 4',6-diamidino-2-phenylindole is a fluorescent stain that binds strongly to DNA. It is used extensively in fluorescence microscopy. Since DAPI will pass through an intact cell membrane, it may be used to stain both live and fixed cells, though it passes through the membrane less efficiently in live cells and therefore doesn't stain as well.

For fluorescence microscopy, DAPI is excited with ultraviolet light. When bound to double-stranded DNA its absorption maximum is at 358 nm and its emission maximum is at 461 nm. (This emission is fairly broad, and appears blue/cyan.)[1] DAPI will also bind to RNA, though it is not as strongly fluorescent. Its emission shifts to around 500 nm when bound to RNA. [2]

DAPI (magenta) bound to the minor groove of DNA (green and blue). From PDB: 1D30​.

DAPI's blue emission is convenient for microscopists who wish to use multiple fluorescent stains in a single sample. There is fluorescence overlap between DAPI and green-fluorescent molecules like fluorescein and green fluorescent protein (GFP), or red-fluorescent stains like Texas Red, but using spectral unmixing or taking images sequentially can get around this.

Apart from labelling cell nuclei, the most popular application of DAPI is in detection of mycoplasma or virus DNA in cell cultures.

Though DAPI does not enter live cells at physiological pH, it does enter live cells in certain conditions binding tightly to DNA, it is labeled non-toxic in it's MSDS[3] and though was not shown to have mutagenicity to E. coli [4], it is labelled as a known mutagen in manufacturer information [5]. Care should be taken in its handling and disposal.

Bovine Pulmonary Artery Endothelial cell nuclei stained with DAPI and imaged on a fluorescence microscope.

The Hoechst stains are similar to DAPI in that they are also blue-fluorescent DNA stains which are compatible with both live- and fixed-cell applications.

References

  1. ^ Invitrogen, DAPI Nucleic Acid Stain. accessed 2009-12-08.
  2. ^ Scott Prahl, DAPI. accessed 2009-12-08.
  3. ^ [1]
  4. ^ Ohta T, Tokishita S, Yamagata H. (2001). "Ethidium bromide and SYBR Green I enhance the genotoxicity of UV-irradiation and chemical mutagens in E. coli". Mutat Res. 492 (1–2): 91–7. PMID 11377248.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  5. ^ Invitrogen, DAPI Nucleic Acid Stain. accessed 2009-12-08.
Retrieved from "https://en.wikipedia.org/w/index.php?title=DAPI&oldid=358391454"