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基于点扫描的超分辨显微成像进展

赵光远 郑程 方月 匡翠方 刘旭

基于点扫描的超分辨显微成像进展

赵光远, 郑程, 方月, 匡翠方, 刘旭
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  • 光学显微镜一直推动着现代科学技术的发展.随着科学的进步,对显微成像分辨率的要求在生物、材料等领域日渐凸显,而常规宽场显微成像一直面临着成像分辨率衍射受限的问题.1968年出现的共聚焦显微镜作为点扫描显微镜的开端第一次实现了远场下成像分辨率的突破,它具有层切性好、信噪比高等优点.在1994年出现的受激辐射荧光损耗显微镜将显微成像能力突破到2.8 nm左右,并成为目前效果最佳、应用较广泛的超分辨显微技术.荧光差分显微和饱和荧光吸收竞争等点扫描技术具有无荧光染剂限制、饱和光强低、光路简单等优势,并且能取得1/6波长的分辨能力,进而在超分辨显微领域仍有着发挥空间.Airyscan技术作为以上方法的补充可以弥补点扫描系统中由于探测小孔半径减小而带来的信号丢失,从而提高成像信噪比和分辨率,但阵列探测器成本较高.上述点扫描显微镜通过改变照明或者探测的方式实现了分辨率突破.本文详细讨论了点扫描超分辨方法的原理、成像效果及面临的瓶颈,并分析了点扫描超分辨显微镜在应用和技术上的趋势.
      通信作者: 匡翠方, cfkuang@zju.edu.cn
    • 基金项目: 国家重点基础研究发展计划(批准号:2015CB352003)、国家重点研发计划(批准号:2016YFF0101400)、国家自然科学基金(批准号:61335003,61377013,61378051,61427818)、浙江省自然科学基金(批准号:LR16F050001)和中央高校基本科研业务费资助的课题.
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    Abbe E 1873 Archiv fr Mikroskopische Anatomie 9 413

    [2]

    Stephenson J W 1877 Monthly Microsc. J. 17 82

    [3]

    Rayleigh L 1874 Philos. Mag. Ser. 47 81

    [4]

    Houston W V 1927 Phys. Rev. 29 478

    [5]

    Kirz J, Jacobsen C, Howells M 1995 Q. Rev. Biophys. 28 33

    [6]

    Petrň M, Hadravsky M, Egger M D, Galambos R 1968 J. Opt. Soc. Am. A 58 661

    [7]

    Hell S W, Wichmann J 1994 Opt. Lett. 19 780

    [8]

    Betzig E, Patterson G H, Sougrat R, Lindwasser O W, Olenych S, Bonifacino J S, Davidson M W, Lippincott-Schwartz J, Hess H F 2006 Science 313 1642

    [9]

    Lindwasser O W, Olenych S, Bonifacino J S, Davidson M W, Lippincott-Schwartz J, Hess H F 2006 Science 313 1642

    [10]

    Rust M J, Bates M, Zhuang X 2006 Nat. Methods 3 793

    [11]

    Douglass K M, Sieben C, Archetti A, Lambert A, Manley S 2016 Nat. Photon. 10 705

    [12]

    Shechtman Y, Weiss L E A, Backer S, Lee M Y, Moerner W E 2016 Nat. Photon. 10 590

    [13]

    Gustafsson M G 2000 J. Microsc. 198 82

    [14]

    Heintzmann R, Cremer C G 1999 Proceedings of SPIE-The International Society for Optical Engineering 3568 1399

    [15]

    Mudry E, Belkebir K, Girard J, Savatier J, Moal E L, Nicoletti C, Allain M, Sentenac A 2012 Nat. Photon. 6 312

    [16]

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    [17]

    Webb R H 1996 Rep. Prog. Phys. 59 427

    [18]

    Sheppard C J, Wilson T 1981 J. Microsc. 124 107

    [19]

    Wilson T 2011 J. Microsc. 154 143

    [20]

    Ellinger P 2008 Biol. Rev. 15 323

    [21]

    Brakenhoff G J, Ht V D V, Spronsen E A, Nanninga N 1989 J. Microsc. 153 151

    [22]

    Brakenhoff G J, Blom P, Barends P 1979 J. Microsc. 117 219

    [23]

    Borlinghaus R T, Kappel C 2016 Nat. Methods 13

    [24]

    Kuang C, Li S, Liu W, Hao X, Gu Z, Wang Y, Ge J, Li H, Liu X 2013 Sci. Rep. 3 1441

    [25]

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    [26]

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    [27]

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    [28]

    Xie H, Liu Y, Jin D, Santangelo P J, Xi P 2013 J. Opt. Soc. Am. A 30 1640

    [29]

    Hao X, Kuang C, Wang T, X Liu 2010 J. Opt. 12 115707

    [30]

    Hao X, Kuang C, Li Y, Liu X 2012 J. Optics 14 045702

    [31]

    Zhang C, Li H, Wang S, Zhao W, Feng X, Wang K, Wang G, Bai J 2016 J. Laser Micro Nanoen. 11 290

    [32]

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    [33]

    Hao X, Kuang C, Gu Z, Li S 2012 Commun. Photon. Conference 1 3

    [34]

    Hell S W, Kroug M 1995 Appl. Phys. B 60 495

    [35]

    Keller J 2006 Ph. D. Dissertation (Heidelberg: Heidelberg University)

    [36]

    Wildanger D, Patton B R, Schill H, Marseglia L, Hadden J P, Knauer S, Schnle A, Rarity J G, O'Brien J L, Hell S W 2011 Adv. Mater. 24 OP309

    [37]

    Gigan S 2017 Nat. Photon. 11 14

    [38]

    Zhang P, Goodwin P M, Werner J H 2014 Opt. Express 22 12398

    [39]

    Yu W, Ji Z, Dong D, Yang X, Xiao Y, Gong Q, Xi P, Shi K 2015 Laser Photon. Rev. 10 147

    [40]

    Patton B R, Burke D, Owald D T J, Bewersdorf Gould J, Booth M J 2016 Opt. Express 24 8862

    [41]

    Wang Y, Hao X, Liu X 2013 Opt. Engineer. 52 093107

    [42]

    Wildanger D, Rittweger E, Kastrup L, Hell S W 2008 Opt. Express 16 9614

    [43]

    Winter F R, Loidolt M, Westphal V, Butkevich A N, Gregor C, Sahl S J, Hell S W 2017 Sci. Rep. 7 46492

    [44]

    Reuss M, Engelhardt J, Hell S W 2010 Opt. Express 18 1049

    [45]

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    [46]

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    [47]

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    [48]

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    Hanne J, Falk H J, Grlitz F, Hoyer P, Engelhardt J, Sahl S J, Hell S W 2015 Nat. Commun. 6 7127

    [50]

    Gao P, Prunsche B, Zhou L, Nienhaus K, Nienhaus G U 2017 Nat. Photon. 11 163

    [51]

    Bordenave M D, Balzarottt F, Stefani F D, Hell S W 2016 J. Phys. D: Appl. Phys. 49 365102

    [52]

    Liu Y, Lu Y, Yang X, Zheng X, Wen S, Fan W, Vidal X, Zhao J, Liu D, Zhou Z 2017 Nature 543 229

    [53]

    Yang X, Xie H, Alonas E, Liu Y, Chen X, Santangelo P J, Ren Q, Xi P, Jin D 2016 Light-Sci. Appl. 5 e16134

    [54]

    Danzl J G, Sidenstein S C, Gregor C, Urban N T, Ilgen P, Jakobs S, Hell S W 2016 Nat. Photon. 10 122

    [55]

    Gttfert F, Pleiner T, Heine J, Westphal V, Grlich D, Sahl S J, Hell S W 2017 Proc. Natl. Acad. Sci. USA 114 2125

    [56]

    Zhao G, Kabir M M, Toussaint K C, Kuang C, Zheng C, Yu Z, Liu X 2017 Optica 4 633

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    [71]

    Zhao G, Rong Z, Zheng C, Liu X, Kuang C 2016 J. Innov. Opt. Heal. Sci. 9 1793

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    [73]

    Sheppard C J, Mehta S B, Heintzmann R 2013 Opt. Lett. 38 2889

    [74]

    Huff J 2015 Nat. Methods 12

    [75]

    Mller C B, Enderlein J 2010 Phys. Rev. Lett. 104 198101

    [76]

    Roth S, Sheppard C J, Kai W, Heintzmann R 2013 Opt. Nanoscopy 2 1

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    [79]

    Sheppard C J, Roth S, Heintzmann R, Castello M, Vicidomini G, Chen R, Chen X, Diaspro A 2016 Opt. Express 24 27280

    [80]

    Kuang C, Ma Y, Zhou R, Zheng G, Fang Y, Xu Y, Liu X, So P T 2016 Phys. Rev. Lett. 117 028102

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    Ge B, Wang Y, Huang Y, Kuang C, Fang Y, Xiu P, Rong Z, Liu X 2016 Opt. Lett. 41 2013

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    [83]

    Wang P, Slipchenko M N, Mitchell J, Yang C, Potma E O, Xu X, Cheng J X 2013 Nat. Photon. 7 449

    [84]

    Jin N, Rahmat-Samii Y 2007 IEEE Trans. Antenn. Propag. 55 556

    [85]

    Rogers E T, Lindberg J, Roy T, Savo S, Chad J E, Dennis M R, Zheludev N I 2012 Nat. Mater. 11 432

    [86]

    van Putten E G, Akbulut D, Bertolotti J, Vos W L, Lagendijk A, Mosk A P 2011 Phys. Rev. Lett. 106 193905

    [87]

    Park J H, Park C, Yu H S, Park J, Han S, Shin J, Ko S H, Nam K T, Cho Y H, Park Y K 2013 Nat. Photon. 7 454

    [88]

    Fang Z, Zhu X 2013 Adv. Mater. 25 253840

    [89]

    Zhang W, Fang Z, Zhu X 2016 Chem. Rev. 117 5095

    [90]

    Diekmann R, Helle O I, Oie C I, McCourt P, Huser T R, Schttpelz M, Ahluwalia B S 2017 Nat. Photon. 11 322

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    Roider C, Ritsch-Marte M, Jesacher A 2016 Opt. Lett. 41 3825

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基于点扫描的超分辨显微成像进展

  • 1. 浙江大学光电科学与工程学院, 现代光学仪器国家重点实验室, 杭州 310027
  • 通信作者: 匡翠方, cfkuang@zju.edu.cn
    基金项目: 

    国家重点基础研究发展计划(批准号:2015CB352003)、国家重点研发计划(批准号:2016YFF0101400)、国家自然科学基金(批准号:61335003,61377013,61378051,61427818)、浙江省自然科学基金(批准号:LR16F050001)和中央高校基本科研业务费资助的课题.

摘要: 光学显微镜一直推动着现代科学技术的发展.随着科学的进步,对显微成像分辨率的要求在生物、材料等领域日渐凸显,而常规宽场显微成像一直面临着成像分辨率衍射受限的问题.1968年出现的共聚焦显微镜作为点扫描显微镜的开端第一次实现了远场下成像分辨率的突破,它具有层切性好、信噪比高等优点.在1994年出现的受激辐射荧光损耗显微镜将显微成像能力突破到2.8 nm左右,并成为目前效果最佳、应用较广泛的超分辨显微技术.荧光差分显微和饱和荧光吸收竞争等点扫描技术具有无荧光染剂限制、饱和光强低、光路简单等优势,并且能取得1/6波长的分辨能力,进而在超分辨显微领域仍有着发挥空间.Airyscan技术作为以上方法的补充可以弥补点扫描系统中由于探测小孔半径减小而带来的信号丢失,从而提高成像信噪比和分辨率,但阵列探测器成本较高.上述点扫描显微镜通过改变照明或者探测的方式实现了分辨率突破.本文详细讨论了点扫描超分辨方法的原理、成像效果及面临的瓶颈,并分析了点扫描超分辨显微镜在应用和技术上的趋势.

English Abstract

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