DAPI

DAPI
	IUPAC名; 2-(4-Amidinophenyl)-1H-indole-6-carboxamidine
别名	4',6-Diamidino-2-phenylindole
识别
CAS号	28718-90-3
PubChem	2954
ChemSpider	2848
SMILES	[N@H]=C(N)c3ccc(c2cc1ccc(cc1n2)C(=[N@H])N)cc3;
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);
InChIKey	FWBHETKCLVMNFS-UHFFFAOYAH
ChEBI	51231
性质
化学式	C16H15N5
摩尔质量	277.32 g·mol−1
	若非注明，所有数据均出自标准状态（25 ℃，100 kPa）下。

DAPI即4',6-二脒基-2-苯基吲哚(4',6-diamidino-2-phenylindole)，是一種能夠與DNA強力結合的熒光染料，常用於螢光顯微鏡觀測^[1]。因為DAPI可以透過完整的細胞膜，它可以用于活細胞和固定細胞的染色。

历史

1971年DAPI在一项关于制抗锥虫病的药物的研究中第一次被合成于Otto Dann的实验室。虽然它不是一种成功的药物，但更深入的研究表明它在与DNA强力结合时发出更强的荧光，因此在1975年它被用在超速离心分析法中以标记线粒体DNA。与DNA结合时激发的强荧光使它被广泛用于荧光显微镜技术中对DNA的染色。上世纪70年代末证实DAPI可以被用来在植物，微生物，多细胞动物和细菌细胞中追踪DNA，1977年证实它可以被用来定量给细胞中的DNA染色，同时也证实DAPI可在流式细胞术中用作DNA染料^[2]

荧光属性

當DAPI與雙股DNA結合時,在螢光顯微鏡觀察下，DAPI染劑在358 nm (紫外线) 处有一个最大吸收峰，并在461 nm (蓝)处有一个最大发射峰，其發射光的波長範圍含蓋了藍色至青綠色，因此在荧光显微技术中DAPI被紫外线照亮且被蓝或青色滤镜检出。DAPI也可以和RNA結合，但產生的螢光強度不及與DNA結合的結果，其發射光的波長範圍約在400nm左右。^[3]^[4] ^[5]

DAPI的發射光為藍色，且DAPI和荧光素、綠色螢光蛋白（Green fluorescent protein, GFP）或Texas Red染劑（紅色螢光染劑）的發射波長，僅有少部分重疊，研究員可以善用這項特性在單一的樣品上進行多重螢光染色。如果需要十分精确的分析，用光谱去混合(spectral unmixing)可以度量这一影响

已隱藏部分未翻譯内容，歡迎參與翻譯。

Outside of analytical fluorescence light microscopy DAPI is also popular for labeling of cell cultures to detect the DNA of contaminating mycoplasma or virus. The labelled mycoplasma or virus particles in the growth medium fluoresce once stained by DAPI making them easy to detect.^[6]

吸收模型和荧光属性

这一DNA荧光探针（英语：fluorescent probe）最近被含时密度泛函理论（英语：Time-dependent density functional theory）有效地刻画^[7]

已隱藏部分未翻譯内容，歡迎參與翻譯。

This DNA fluorescent probe has recently been effectively modeled ^[7]using the Time-dependent density functional theory, coupled with the IEF version of the Polarizable continuum model. This quantum-mechanical modeling has rationalized the absorption and fluorescence behavior given by minor groove binding and intercalation in the DNA pocket, in term of a reduced structural flexibility and polarization.

活细胞毒性

DAPI可用于固定细胞染色，但是因为活细胞染色对DAPI浓度有严格要求，它很少被用于活细胞染色。^[8]DAPI能快速進入活細胞中與DNA結合^{[來源請求]}，因此DAPI對生物體而言，也被視為一種毒性物質與致癌物。然而在MSDS中它被标记为无毒。^[9]尽管DAPI没有显现出对E.coli的诱变性，^[10]它在机器制造商提供的信息上被认为是一种DNA诱变剂^[5]。由于DAPI可以掺入DNA螺旋中，它很可能具有底层的DNA诱变性，在使用過程中應注意配制、使用與拋棄的處理程序。

替代

以DAPI(蓝),鬼笔环肽(红)染色且使用免疫荧光法通过异硫氰酸荧光素结合抗体(FITC)(绿)的内皮细胞

类似于DAPI技术，赫斯特染色是一种既可以用于活细胞也可以用于固定细胞的蓝色荧光染料，并且可以使用与DAPI相同的机器滤镜设置

参见

DAPI

參考資料

^ 4',6-二脒基-2-苯基吲哚. 来邦网. [2010-01-21].
^ Kapuscinski J. DAPI: a DNA-specific fluorescent probe. Biotech Histochem. September 1995, 70 (5): 220–33. PMID 8580206. doi:10.3109/10520299509108199.
^ Scott Prahl, DAPI. accessed 2009-12-08.
^ Kapuscinski J., [1]. accessed 2013-02-25.
^ ^5.0 ^5.1 Invitrogen, DAPI Nucleic Acid Stain 互联网档案馆的存檔，存档日期2009-03-06.. accessed 2009-12-08.
^ RUSSELL, W. C.; NEWMAN, CAROL; WILLIAMSON, D. H. A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses. Nature. 1975, 253 (5491): 461–462. Bibcode:1975Natur.253..461R. doi:10.1038/253461a0.
^ ^7.0 ^7.1 A. Biancardi; T. Biver; F. Secco; B. Mennucci. An investigation of the photophysical properties of minor groove bound and intercalated DAPI through quantum-mechanical and spectroscopic tools.. Phys. Chem. Chem. Phys. 2013, 15: 4596. Bibcode:2013PCCP...15.4596B. doi:10.1039/C3CP44058C.
^ Zink D, Sadoni N, Stelzer E. Visualizing Chromatin and Chromosomes in Living Cells. Usually for the live cells staining Hoechst Staining is used. DAPI gives a higher signal in the fixed cells compare to Hoechst Stain but in the live cells Hoechst Stain is used.. Methods. 2003, 29 (1): 42–50. PMID 12543070. doi:10.1016/S1046-2023(02)00289-X.
^ http://www.kpl.com/docs/msds/710301.pdf
^ Ohta T, Tokishita S, Yamagata H. Ethidium bromide and SYBR Green I enhance the genotoxicity of UV-irradiation and chemical mutagens in E. coli.. Mutat. Res. 2001, 492 (1–2): 91–7. PMID 11377248. doi:10.1016/S1383-5718(01)00155-3.

[1] 4',6-二脒基-2-苯基吲哚. 来邦网. [2010-01-21].

[Kapuscinski1995-2] Kapuscinski J. DAPI: a DNA-specific fluorescent probe. Biotech Histochem. September 1995, 70 (5): 220–33. PMID 8580206. doi:10.3109/10520299509108199.

[3] Scott Prahl, DAPI. accessed 2009-12-08.

[4] Kapuscinski J., [1]. accessed 2013-02-25.

[DAPI_Nucleic_Acid_Stain-5] 5.0 ^5.1 Invitrogen, DAPI Nucleic Acid Stain 互联网档案馆的存檔，存档日期2009-03-06.. accessed 2009-12-08.

[6] RUSSELL, W. C.; NEWMAN, CAROL; WILLIAMSON, D. H. A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses. Nature. 1975, 253 (5491): 461–462. Bibcode:1975Natur.253..461R. doi:10.1038/253461a0.

[model-7] 7.0 ^7.1 A. Biancardi; T. Biver; F. Secco; B. Mennucci. An investigation of the photophysical properties of minor groove bound and intercalated DAPI through quantum-mechanical and spectroscopic tools.. Phys. Chem. Chem. Phys. 2013, 15: 4596. Bibcode:2013PCCP...15.4596B. doi:10.1039/C3CP44058C.

[8] Zink D, Sadoni N, Stelzer E. Visualizing Chromatin and Chromosomes in Living Cells. Usually for the live cells staining Hoechst Staining is used. DAPI gives a higher signal in the fixed cells compare to Hoechst Stain but in the live cells Hoechst Stain is used.. Methods. 2003, 29 (1): 42–50. PMID 12543070. doi:10.1016/S1046-2023(02)00289-X.

[9] ttp://www.kpl.com/docs/msds/710301.pdf

[10] Ohta T, Tokishita S, Yamagata H. Ethidium bromide and SYBR Green I enhance the genotoxicity of UV-irradiation and chemical mutagens in E. coli.. Mutat. Res. 2001, 492 (1–2): 91–7. PMID 11377248. doi:10.1016/S1383-5718(01)00155-3.

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