编辑“︁DAPI”︁

{{other uses}}

{{noteTA
|1=zh-hans:荧光; zh-hant:螢光;
}}
{{chembox
| Watchedfields = changed
| verifiedrevid = 443556310
| ImageFile=DAPI.svg
| ImageSize=200px
| ImageFile1=DAPI Space Fill.png
| ImageSize1=200px
| IUPACName=2-(4-Amidinophenyl)-1''H''-indole-6-carboxamidine
| OtherNames=4',6-Diamidino-2-phenylindole
|Section1={{Chembox Identifiers
| 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
| SMILES1 = [H]/N=C(/c1ccc(cc1)c2cc3ccc(cc3[nH]2)/C(=N/[H])/N)\N
| ChEMBL_Ref = {{ebicite|correct|EBI}}
| ChEMBL = 48217
| StdInChI_Ref = {{stdinchicite|correct|chemspider}}
| StdInChI = 1S/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)
| StdInChIKey_Ref = {{stdinchicite|correct|chemspider}}
| StdInChIKey = FWBHETKCLVMNFS-UHFFFAOYSA-N
| CASNo_Ref = {{cascite|correct|??}}
| CASNo=28718-90-3
| ChemSpiderID_Ref = {{chemspidercite|correct|chemspider}}
| ChemSpiderID = 2848
| PubChem=2954
| ChEBI_Ref = {{ebicite|correct|EBI}}
| ChEBI = 51231
| SMILES = [N@H]=C(N)c3ccc(c2cc1ccc(cc1n2)C(=[N@H])N)cc3
}}
|Section2={{Chembox Properties
| C=16 | H=15 | N=5
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| MeltingPt=
| BoilingPt=
| Solubility=
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|Section3={{Chembox Hazards
| MainHazards=
| FlashPt=
| AutoignitionPt =
  }}
}}

'''DAPI'''即'''4',6-二脒基-2-苯基吲哚'''（4',6-'''d'''i'''a'''midino-2-'''p'''henyl'''i'''ndole），是一種能夠與[[DNA]]強力結合的[[螢光]][[染料]]，常用於螢光[[顯微鏡]]觀測<ref>{{cite web|url=http://www.labome.cn/综述/试剂/7|title=4',6-二脒基-2-苯基吲哚|accessdate=2010-01-21|work=来邦网|archive-date=2010-01-24|archive-url=https://web.archive.org/web/20100124100535/http://www.labome.cn/%E7%BB%BC%E8%BF%B0/%E8%AF%95%E5%89%82/7|dead-url=no}}</ref>。因為DAPI可以透過完整的[[細胞膜]]<ref name="膜穿透性">{{cite journal|author2=王黎|author3=周清|author4=杨艳|author5=陈剑|author1=徐锦堂|title=CM_Dil与DAPI联合标记人羊膜上皮细胞的可行性研究_王黎.pdf|doi=10.13389/j.cnki.rao.2013.11.001|url=http://kns.cnki.net/KCMS/detail/detail.aspx?dbcode=CJFQ&dbname=CJFD2013&filename=XKJZ201311001&uid=WEEvREcwSlJHSldTTEYzVDhsN3d4K24xZ0JTUVh5cnNodGZxcGVBRWpKdz0=$9A4hF_YAuvQ5obgVAqNKPCYcEjKensW4IQMovwHtwkF4VYPoHbKxJw!!&v=MTI1NjlEaDFUM3FUcldNMUZyQ1VSTE9mWXVkcEZDam5WNzNBUFNiQmRMRzRIOUxOcm85RlpZUjhlWDFMdXhZUzc=|year=2013|journal=|access-date=2019-05-25|archive-date=2019-09-24|archive-url=https://web.archive.org/web/20190924221557/http://kns.cnki.net/KCMS/detail/detail.aspx?dbcode=CJFQ&dbname=CJFD2013&filename=XKJZ201311001&uid=WEEvREcwSlJHSldTTEYzVDhsN3d4K24xZ0JTUVh5cnNodGZxcGVBRWpKdz0=$9A4hF_YAuvQ5obgVAqNKPCYcEjKensW4IQMovwHtwkF4VYPoHbKxJw!!&v=MTI1NjlEaDFUM3FUcldNMUZyQ1VSTE9mWXVkcEZDam5WNzNBUFNiQmRMRzRIOUxOcm85RlpZUjhlWDFMdXhZUzc=|dead-url=no}}</ref>，它可以用于活[[細胞]]和固定細胞的[[染色_(生物學) |染色]]。

==历史==
1971年DAPI在一项关于制抗{{le|锥虫病|Trypanosomiasis}}的药物的研究中由Otto Dann的实验室首次合成，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[[离心机|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]]<ref>{{cite journal |author=M Cassan |author2=E Boy |author3=F Borne |author24=J C Patte |title=Regulation of the lysine biosynthetic pathway in Escherichia coli K-12: isolation of a cis-dominant constitutive mutant for AK III synthesis. |journal=J Bacteriol. |date=1975 Aug |pmid=238953 |url=https://www.ncbi.nlm.nih.gov/pmc/articles/PMC235741/ |access-date=2019-05-13 |archive-date=2019-09-24 |archive-url=https://web.archive.org/web/20190924221558/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC235741/ |dead-url=no }}</ref>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。<ref name="Kapuscinski1995">{{cite journal |author=Kapuscinski J |title=DAPI: a DNA-specific fluorescent probe |journal=Biotech Histochem |volume=70 |issue=5 |pages=220–33 |date=September 1995 |pmid=8580206 |doi= 10.3109/10520299509108199|url=}}</ref>

==荧光属性==
===发射与吸收波长===
在[[螢光顯微鏡]]觀察下，與雙股DNA結合的DAPI染劑在358&nbsp;nm（[[紫外线]]）处有一个最大吸收峰，并在461&nbsp;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400&nbsp;nm左右。<ref>Scott Prahl, [http://omlc.ogi.edu/spectra/PhotochemCAD/html/dapi(H2O).html DAPI] {{Wayback|url=http://omlc.ogi.edu/spectra/PhotochemCAD/html/dapi(H2O).html |date=20131029185614 }}. accessed 2009-12-08.</ref><ref>Kapuscinski J., [https://www.ncbi.nlm.nih.gov/pubmed/1696951] {{Wayback|url=https://www.ncbi.nlm.nih.gov/pubmed/1696951 |date=20180422202658 }}. accessed 2013-02-25.</ref><ref name="DAPI Nucleic Acid Stain">Invitrogen, [http://probes.invitrogen.com/media/pis/mp01306.pdf DAPI Nucleic Acid Stain] {{Wayback|url=http://probes.invitrogen.com/media/pis/mp01306.pdf |date=20090306113851 }}. accessed 2009-12-08.</ref>

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，可以使用光谱去混合（spectral unmixing）技術計算此一影响。
[[File:1D30 DNA DAPI.png|250px|thumb|DAPI 结合于DNA小沟(蓝和绿). From {{PDB|1D30}}.]]
===吸收模型===
2013年，有研究者以{{tsl|en|Time-dependent density functional theory|含时密度泛函理论}}與[[等電位聚焦]]的{{link-en|连续极化介质模型|Polarizable continuum model}}描述了DAPI與DNA結合與發出螢光的機制，此一量子力学模型解释了DAPI與DNA結合後吸收與发射光譜的現象，與DNA的結合會使DAPI分子因位向限制導致幾何彈性降低，且周圍環境的[[極性]]甚低會導致DAPI分子的極性降低。<ref name="model">{{cite journal |author1=A. Biancardi |author2=T. Biver |author3=F. Secco |author4=B. Mennucci |title=An investigation of the photophysical properties of minor groove bound and intercalated DAPI through quantum-mechanical and spectroscopic tools. |doi=10.1039/C3CP44058C  |journal=Phys. Chem. Chem. Phys. |year=2013 |volume=15 |page=4596|bibcode=2013PCCP...15.4596B }}</ref>

==用途==
除了用于分析荧光显微法以外，DAPI也经常在[[细胞培养]]中被用于追踪侵染的[[支原体]]或[[病毒]]。在[[培养基]]上培养的[[支原体]]或[[病毒]]颗粒被DAPI染色后会发出荧光而易于追踪。<ref>{{cite journal|last=RUSSELL|first=W. C.|author2=NEWMAN, CAROL |author3=WILLIAMSON, D. H. |title=A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses|url=https://archive.org/details/sim_nature-uk_1975-02-06_253_5491/page/460|journal=Nature|date=1975|volume=253|issue=5491|pages=461–462|doi=10.1038/253461a0|bibcode=1975Natur.253..461R}}</ref>

DAPI也经常在[[流式细胞术]]中用于区分不同[[细胞周期]]时相的细胞。<ref>{{cite journal|author=Ashraf M. Khalil|title=B Cell Signal Transduction in Germinal Center B Cells is Short-Circuited by Increased Phosphatase Activity|journal=Science|date=2012-06-01|url=https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3777391/|accessdate=2019-05-11|archive-date=2021-05-26|archive-url=https://web.archive.org/web/20210526011148/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3777391/}}</ref><ref>{{cite journal |last1=Kerenyi |first1=Marc A |last2=Shao |first2=Zhen |last3=Hsu |first3=Yu-Jung |last4=Guo |first4=Guoji |last5=Luc |first5=Sidinh |last6=O'Brien |first6=Kassandra |last7=Fujiwara |first7=Yuko |last8=Peng |first8=Cong |last9=Nguyen |first9=Minh |last10=Orkin |first10=Stuart H |title=Histone demethylase Lsd1 represses hematopoietic stem and progenitor cell signatures during blood cell maturation |journal=eLife |date=2013-06-18 |volume=2 |issue=2013; 2 |doi=10.7554/eLife.00633 |pmid=23795291 |url=https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3687337/ |accessdate=2019-05-11 |archive-date=2019-09-24 |archive-url=https://web.archive.org/web/20190924221559/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3687337/ |dead-url=no }}</ref>

==活细胞安全性==
===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。<ref>{{cite journal |vauthors=Zink D, Sadoni N, Stelzer E |title=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. |doi=10.1016/S1046-2023(02)00289-X|journal=Methods |volume=29 |issue=1 |pages=42–50 |year=2003 |PMID=12543070}}</ref>虽然DAPI能快速進入活細胞中與DNA結合<ref name="膜穿透性" />，但是[[材料安全性数据表|MSDS]]<ref>{{Cite web |url=http://www.kpl.com/docs/msds/710301.pdf |title=存档副本 |accessdate=2018-04-18 |archive-date=2016-03-03 |archive-url=https://web.archive.org/web/20160303211317/http://www.kpl.com/docs/msds/710301.pdf |dead-url=yes }}</ref>和其他文献<ref name="膜穿透性" />认为DAPI没有毒性。

有研究表明，大剂量的DAPI可以导致小鼠中毒死亡,LD<sub>50</sub>= 基质 37.3 mg/kg。<ref>{{cite journal|author=Slieger RF et al.|title=DAPI对鼠疟的治疗作用.pdf|doi=10.13389/j.cnki.rao.2013.11.001|url=http://kns.cnki.net/KCMS/detail/detail.aspx?dbcode=CJFQ&dbname=CJFD7984&filename=GWJC198104015&uid=WEEvREcwSlJHSldTTEYzVDhsN3d4K24xZ0JTUVh5cnNodGZxcGVBRWpKdz0=$9A4hF_YAuvQ5obgVAqNKPCYcEjKensW4IQMovwHtwkF4VYPoHbKxJw!!&v=MTAxOTdiL0lJanJCYmJLeEZ0RE1xNDlFWVlSOGVYMUx1eFlTN0RoMVQzcVRyV00xRnJDVVJMT2ZZdWRwRkNqblU=|year=1980|access-date=2019-05-25|archive-date=2019-09-24|archive-url=https://web.archive.org/web/20190924221559/http://kns.cnki.net/KCMS/detail/detail.aspx?dbcode=CJFQ&dbname=CJFD7984&filename=GWJC198104015&uid=WEEvREcwSlJHSldTTEYzVDhsN3d4K24xZ0JTUVh5cnNodGZxcGVBRWpKdz0=$9A4hF_YAuvQ5obgVAqNKPCYcEjKensW4IQMovwHtwkF4VYPoHbKxJw!!&v=MTAxOTdiL0lJanJCYmJLeEZ0RE1xNDlFWVlSOGVYMUx1eFlTN0RoMVQzcVRyV00xRnJDVVJMT2ZZdWRwRkNqblU=|dead-url=no}}</ref>
===潜在的诱变性===
DAPI被視為一種[[致癌物]]。尽管DAPI没有显现出对[[大腸桿菌]]的诱变性<ref>{{cite journal |vauthors=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 |doi=10.1016/S1383-5718(01)00155-3}}</ref>，在机器制造商提供的信息上，DAPI被认为是一种DNA诱变剂<ref name="DAPI Nucleic Acid Stain" />。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==
[[File:FluorescentCells.jpg|250px|right|thumb|以DAPI（蓝）與[[鬼笔环肽]]（红）染色的內皮細胞，另外綠色為與使用與[[异硫氰酸荧光素]]結合的抗體進行{{link-en|免疫荧光法|Immunofluorescence}}的結果]]
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<ref>{{cite journal|last=Latt|first=SA|author2=Stetten, G |author3=Juergens, LA |author4=Willard, HF |author5=Scher, CD |title=Recent developments in the detection of deoxyribonucleic acid synthesis by 33258 Hoechst fluorescence.|journal=Journal of Histochemistry and Cytochemistry|date=July 1975|volume=23|issue=7|pages=493–505|pmid=1095650|doi=10.1177/23.7.1095650}}</ref>，并且可以使用与DAPI相同的机器滤镜设置。

==参见==
*DAPI在[[PDB]]上的[https://pdbj.org/mine/search?query=DAPI 搜索结果]{{Wayback|url=https://pdbj.org/mine/search?query=DAPI |date=20190505155953 }}

==參考資料==
{{reflist|2}}
{{Fluorophore}}

[[Category:染色着色剂]]
[[Category:荧光染料]]
[[Category:DNA结合物质]]
[[Category:吲哚]]
[[Category:脒]]
[[Category:联芳烃]]