Citation: QIN Ge-Ge, LI Wen-Hui, XU Jia-Chao, KOU Xiao-Long, ZHAO Rong, LUO Fang, FANG Xiao-Hong. Development of Integrated Atomic Force Microscopy and Fluorescence Microscopy for Single-Molecule Analysis in Living Cells[J]. Chinese Journal of Analytical Chemistry, ;2017, 45(12): 1813-1823. doi: 10.11895/j.issn.0253-3820.171313 shu

Development of Integrated Atomic Force Microscopy and Fluorescence Microscopy for Single-Molecule Analysis in Living Cells

QIN Ge-Ge ^1,2 ,
LI Wen-Hui ^1,2 ,
XU Jia-Chao ^1,2 ,
KOU Xiao-Long ^1,2 ,
ZHAO Rong ^1,2 ,
LUO Fang ^1,2 ,
FANG Xiao-Hong ^1,2,,

1.
Key Laboratory of Molecular Nanostructure and Nanotechnology, CAS Research/Education Center for Excellence in Molecular Sciences, Institute of Chemistry, Chinese Academy of Science, Beijing 100190, China
2.
University of Chinese Academy of Sciences, Beijing 100049, China

Corresponding author: FANG Xiao-Hong, xfang@iccas.ac.cn
Received Date: 9 October 2017
Accepted Date: 30 October 2017

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 21735006, 91413119)the National Natural Science Foundation of China 21735006the National Natural Science Foundation of China 91413119

Figures(5)

Atomic force microscope (AFM) and fluorescence microscope (FM) have been emerging as two most commonly used tools for single-molecule study in living cells. Combining the advantages of two microscopes, the development of the integrated AFM-FM technique with high spatiotemporal resolution and multi-function has attracted increasing interest. In this review, the principles of AFM single-molecular force spectroscopy and single-molecule fluorescence imaging were briefly discussed, and the recent advances in the integrated AFM-FM instrumentation were summarized. Subsequently based on our own research in the investigation of ligand-receptors interactions with the integrated AFM-FM technique, its applications in live-cell single-molecule imaging and characterization were introduced.
- Atomic force microscopy,
- Fluorescence microscopy,
- Single-molecule fluorescence imaging,
- Single-molecule force spectroscopy,
- Single-molecule analysis,
- Review
1. [1]
  YU Jun-Ping, LIN Yi, JIANG Ya-Xin, ZHAO Xin-Sheng, FANG Xiao-Hong. Single-molecule Fluorescence Imaging and Single Molecular Force Spectroscopy in the Study of Single Molecular Behavior In Living Cells. Beijing:Science Press, 2005:13
2. [2]
  Müller D J, Dufrêne Y F. Nature Nanotech., 2008, 3(5):261-269 doi: 10.1038/nnano.2008.100
3. [3]
  Gahlmann A, Moerner W E. Nat. Rev. Microbiol., 2014, 12(1):9-22
4. [4]
  Hinterdorfer P, Dufrêne Y F. Nat. Methods, 2006, 3(5):347-355 doi: 10.1038/nmeth871
5. [5]
  Fotiadis D, Liang Y, Filipek S, Saperstein D A, Engel A, Palczewski K. Nature, 2003, 421(6919):127-128 doi: 10.1038/421127a
6. [6]
  Stroh C, Wang H, Bash R, Ashcroft B, Nelson J, Gruber H, Lohr D, Lindsay S M, Hinterdorfer P. Proc. Natl. Acad. Sci. USA, 2004, 101(34):12503-12507 doi: 10.1073/pnas.0403538101
7. [7]
  CHENG Ming, LI Nan, FANG Xiao-Hong. Sci. China-Chem., 2012, (12):1663-1671
8. [8]
  Sun Y, Li N, Fang X. Prog. Biophys. Mol. Biol., 2015, 118(3):95-102 doi: 10.1016/j.pbiomolbio.2015.04.009
9. [9]
  Wang S, Su J H, Beliveau B, Bintu B, Moffitt J, Wu C, Zhuang X. Science, 2016, 353(6299):598-602 doi: 10.1126/science.aaf8084
10. [10]
  Wang C, Han B, Zhou R, Zhuang X W. Cell, 2016, 165(4):990-1001 doi: 10.1016/j.cell.2016.04.040
11. [11]
  Lu H P, Xun L, Xie X S. Science, 1998, 282(5395):1877-1882 doi: 10.1126/science.282.5395.1877
12. [12]
  Binnig G, Quate C F, Gerber C. Phys. Rev. Lett., 1986, 56(9):930-933 doi: 10.1103/PhysRevLett.56.930
13. [13]
  Seelert H, Poetsch A, Dencher N A, Engel A, Stahlberg H, Müller D J. Nature, 2000, 405(6785):418-419 doi: 10.1038/35013148
14. [14]
  Kufer S K, Puchner E M, Gumpp H, Liedl T, Gaub H E. Science, 2008, 319(5863):594-596 doi: 10.1126/science.1151424
15. [15]
  Shi X, Zhang X, Xia T, Fang X H. Nanomedicine, 2012, 7(10):1625-1637 doi: 10.2217/nnm.12.130
16. [16]
  Benoit M, Gabriel D, Gerisch G, Gaub H E. Nat. Cell. Biol., 2000, 2(6):313-317 doi: 10.1038/35014000
17. [17]
  Lee S, Mandic J, Van Vliet K J. Proc. Natl. Acad. Sci. USA, 2007, 104(23):9609-9614 doi: 10.1073/pnas.0702668104
18. [18]
  Horton M, Charras G, Lehenkari P. J. Recept Signal Transduct Res., 2002, 22(1-4):169-190 doi: 10.1081/RRS-120014594
19. [19]
  Xiao J, Dufrêne Y F. Nat. Microbiol., 2016, 1(11):16186 doi: 10.1038/nmicrobiol.2016.186
20. [20]
  Yang Y, Xia T, Zhang W, Fang X. Chin. Sci. Bull., 2011, 56(11):1063-1067 doi: 10.1007/s11434-011-4415-1
21. [21]
  Kaksonen M, Toret C P, Drubin D G. Cell, 2005, 123(2):305-320 doi: 10.1016/j.cell.2005.09.024
22. [22]
  Dyachok O, Isakov Y, Sågetorp J, Tengholm A. Nature, 2006, 439(7074):349-352 doi: 10.1038/nature04410
23. [23]
  Bougourd S, Marrison J, Haseloff J. Plant J. Cell Mol. Biol., 2000, 24(4):543-550 doi: 10.1046/j.1365-313x.2000.00892.x
24. [24]
  Erickson J R, Joiner M A, Guan X, Kutschke W, Yang J, Oddis C V, Bartlett R K, Lowe J S, O'Donnell S, Aykinburns N. Cell, 2008, 133(3):462-474 doi: 10.1016/j.cell.2008.02.048
25. [25]
  Laskowski P R, Pfreundschuh M, Stauffer M, Ucurum Z, Fotiadis D, Müller D J. ACS Nano, 2017, 11(8):8292-8301 doi: 10.1021/acsnano.7b03456
26. [26]
  Hell S W, Wichmann J. Opt. Lett., 1994, 19(11):780-782 doi: 10.1364/OL.19.000780
27. [27]
  Willig K I, Rizzoli S O, Westphal V, Jahn R, Hell S W. Nature, 2006, 440(7086):935-939 doi: 10.1038/nature04592
28. [28]
  Betzig E, Patterson G H, Sougrat R, Lindwasser O W, Olenych S, Bonifacino J S, Davidson M W, Lippincottschwartz J, Hess H F. Science, 2006, 313(5793):1642-1645 doi: 10.1126/science.1127344
29. [29]
  Rust M J, Bates M, Zhuang X W. Nat. Methods, 2006, 3(10):793-795 doi: 10.1038/nmeth929
30. [30]
  York A G, Ghitani A, Vaziri A, Davidson M W, Shroff H, Nat. Methods, 2011, 8(4):327-333 doi: 10.1038/nmeth.1571
31. [31]
  Shaw J E, Oreopoulos J, Wong D, Hsu J C Y, Yip C M. Surf. Interface Anal., 2010, 38(11):1459-1471
32. [32]
  Alsteens D, Newton R, Schubert R, Martinezmartin D, Delguste M, Roska B, Müller D J. Nat. Nanotech., 2016, 12(2):177-183 doi: 10.1038/nnano.2016.228
33. [33]
  Yu J, Yuan J, Zhang X, Liu J, Fang X. Chin. Sci. Bull., 2013, 58(33):4045-4050 doi: 10.1007/s11434-013-6011-z
34. [34]
  Fukuda S, Uchihashi T, Iino R, Okazaki Y, Yoshida M, Igarashi K, Ando T. Rev. Sci. Instrum., 2013, 84(7):073706 doi: 10.1063/1.4813280
35. [35]
  Timmel T, Schuelke M, Spuler S. Microsc. Microanal., 2014, 20(2):514-520 doi: 10.1017/S1431927613014098
36. [36]
  Axelrod D. Method Cell Biol., 1989, 30(11):245-270
37. [37]
  Mathur A B, Truskey G A, Reichert W M. Biophys. J., 2000, 78(4):1725-1735 doi: 10.1016/S0006-3495(00)76724-5
38. [38]
  Gumpp H, Stahl S W, Strackharn M, Puchner E M, Gaub H E. Rev. Sci. Instrum., 2009, 80(10):063704
39. [39]
  Shibata M, Yamashita H, Uchihashi T, Kandori H, Ando T. Nat. Nanotech., 2010, 5(3):208-212 doi: 10.1038/nnano.2010.7
40. [40]
  Colom A, Casuso I, Rico F, Scheuring S. Nat. Commun., 2013, 4:2155
41. [41]
  Hillner P E, Walters D A, Lal R, Hansma H G, Hansma R. Microsc. Microanal., 1995, 1(3):127-130 doi: 10.1017/S1431927695111277
42. [42]
  Meyer G, Amer N M. Appl. Phys. Lett., 1988, 53(12):1045-1047 doi: 10.1063/1.100061
43. [43]
  Kassies R, Ko V D W, Lenferink A, Hunter C N, Olsen J D, Subramaniam V, Otto C. J. Microsc., 2005, 217(1):109-116 doi: 10.1111/jmi.2005.217.issue-1
44. [44]
  Ebenstein Y, Mokari T, Banin U. Appl. Phys. Lett., 2002, 80(21):4033-4035 doi: 10.1063/1.1482785
45. [45]
  Müller D J, Schabert F A, Büldt G, Engel A. Biophys. J., 1995, 68(5):1681-1686 doi: 10.1016/S0006-3495(95)80345-0
46. [46]
  Harke B, Chacko J V, Haschke H, Canale C, Diaspro A. Optical Nanoscopy, 2012, 1(1):1-6 doi: 10.1186/2192-2853-1-1
47. [47]
  Chacko J V, Canale C, Harke B, Diaspro A. Plos One, 2013, 8(6):e66608 doi: 10.1371/journal.pone.0066608
48. [48]
  Monserrate A, Casado S, Flors C. Chemphyschem, 2014, 15(4):S1 647-650
49. [49]
  Chacko J V, Zanacchi F C, Diaspro A. Cytoskeleton, 2013, 70(11):729-740 doi: 10.1002/cm.v70.11
50. [50]
  Odermatt P D, Shivanandan A, Deschout H, Jankele R, Nievergelt A P, Feletti L, Davidson M W, Radenovic A, Fantner G E. Nano Lett., 2015, 15(8):4896-4904 doi: 10.1021/acs.nanolett.5b00572
51. [51]
  Weghuber J, Sunzenauer S, Plochberger B, Brameshuber M, Haselgrübler T, Schütz G J. Anal. Bioanal.Chem., 2010, 397(8):3339-3347 doi: 10.1007/s00216-010-3854-x
52. [52]
  Yu J, Sun S, Jiang Y, Ma X, Chen F, Zhang G, Fang X. Polymer, 2006, 47(7):2533-2538 doi: 10.1016/j.polymer.2005.12.087
53. [53]
  Yu J, Wang Q, Shi X, Ma X, Yang H, Chen Y G, Fang X. J. Phys. Chem. B, 2007, 111(48):13619-13625 doi: 10.1021/jp0758667
54. [54]
  Yang H, Yu J, Fu G, Shi X, Xiao L, Chen Y, Fang X, He C. Exp. Cell Res., 2007, 313(16):3497-3504 doi: 10.1016/j.yexcr.2007.08.001
55. [55]
  Andre G, Deghorain M, Bron P A, van Swam I I, Kleerebezem M, Hols P, Dufrene Y F, ACS. Chem. Biol., 2011, 6(4):366-376 doi: 10.1021/cb1003509
56. [56]
  Yang Y, Xu Y, Xia T, Chen F, Zhang C, Liang W, Lai L, Fang X. Chem. Commun., 2011, 47(19):5440-5442 doi: 10.1039/c1cc10778j
57. [57]
  Yang Y, Wolfram J, Shen H, Fang X, Ferrari M. Eur. J. Med. Chem., 2012, 58:390-395 doi: 10.1016/j.ejmech.2012.10.028
58. [58]
  Zhang W, Yuan J, Yang Y, Xu L, Wang Q, Zuo W, Fang X, Chen Y G. Cell Res., 2010, 20(11):1216-1223 doi: 10.1038/cr.2010.105
59. [59]
  Zhang W, Jiang Y, Wang Q, Ma X, Xiao Z, Zuo W, Fang X, Chen Y G. Proc. Natl. Acad. Sci. USA, 2009, 106(37):15679-15683 doi: 10.1073/pnas.0908279106
60. [60]
  Ye Z, Li N, Zhao L, Sun Y, Ruan H, Zhang M, Yuan J, Fang X. Sci. Bull., 2016, 61(8):632-638 doi: 10.1007/s11434-016-1043-9
61. [61]
  Xiao H, Ma X, Feng W, Fu Y, Lu Z, Xu M, Shen Q, Zhu Y, Zhang Y. Cardiovasc. Res., 2010, 87(3):504-513 doi: 10.1093/cvr/cvq066
62. [62]
  Xiao H, Zhang J, Xu Z H, Li J Y, Xu Z, Feng Y, Zhang M, Liu J, Chen R, Shen J, Wu J, Lu Z, Fang X, Li J, Zhang Y. Sci. Rep., 2016, 6(6):28597
63. [63]
  Li Y, Shi X L, Liu H L, Yi S Q, Zhang X J, Fang X H. Sci. China-Chem., 2010, 53(4):752-758 doi: 10.1007/s11426-010-0111-2
64. [64]
  Shi X, Xu L, Yu J, Fang X. Exp. Cell Res., 2009, 315(16):2847-2855 doi: 10.1016/j.yexcr.2009.05.023
65. [65]
  Wei Y, Zhang X, Xu L, Yi S, Li Y, Fang X, Liu H. Sci. China Life Sci., 2012, 55(10):891-897 doi: 10.1007/s11427-012-4383-y
66. [66]
  Liu J, Zhang X, Wang X, Li X, Li J, Fang X. Sci. Bull., 2016, 61(15):1-8
67. [67]
  Valle-Delgado J J, Urbán P, Fernàndez-Busquets X. Nanoscale, 2013, 5(9):3673-3680
68. [68]
  Zhang L, Yang F, Cai J Y, Yang P H, Liang Z H. Biosens.Bioelectron., 2014, 56(1):271-277
69. [69]
  Alsteens D, Garcia M C, Lipke P N, Dufrene Y F. Proc. Natl. Acad. Sci. USA, 2010, 107(48):20744-20749 doi: 10.1073/pnas.1013893107
70. [70]
  He K, Luo W, Zhang Y, Liu F, Liu D, Xu L, Qin L, Xiong C, Lu Z, Fang X. ACS Nano, 2010, 4(6):3015-3022 doi: 10.1021/nn1002198
71. [71]
  He K, Shi X, Zhang X, Dang S, Ma X, Liu F, Xu M, Lyu Z, Han D, Fang X. Cardiovasc. Res., 2011, 92(1):39-47 doi: 10.1093/cvr/cvr189
72. [72]
  Dixon I M, Davies J J. Cardiovasc. Res., 2011, 92(1):5-6 doi: 10.1093/cvr/cvr216
73. [73]
  Liu Q, Zhang X, Zhao Y, Lin J, Shu C, Wang C, Fang X. Environ. Sci. Technol., 2013, 47(13):7490-7498 doi: 10.1021/es4010224
1. [1]
  Jingwen Wang , Minghao Wu , Xing Zuo , Yaofeng Yuan , Yahao Wang , Xiaoshun Zhou , Jianfeng Yan . Advances in the Application of Electrochemical Regulation in Investigating the Electron Transport Properties of Single-Molecule Junctions. University Chemistry, 2025, 40(3): 291-301. doi: 10.12461/PKU.DXHX202406023
2. [2]
  Chunmei GUO , Weihan YIN , Jingyi SHI , Jianhang ZHAO , Ying CHEN , Quli FAN . Facile construction and peroxidase-like activity of single-atom platinum nanozyme. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1633-1639. doi: 10.11862/CJIC.20240162
3. [3]
  Fei Xie , Chengcheng Yuan , Haiyan Tan , Alireza Z. Moshfegh , Bicheng Zhu , Jiaguo Yu . d带中心调控过渡金属单原子负载COF吸附O₂的理论计算研究. Acta Physico-Chimica Sinica, 2024, 40(11): 2407013-. doi: 10.3866/PKU.WHXB202407013
4. [4]
  Yang WANG , Xiaoqin ZHENG , Yang LIU , Kai ZHANG , Jiahui KOU , Linbing SUN . Mn single-atom catalysts based on confined space: Fabrication and the electrocatalytic oxygen evolution reaction performance. Chinese Journal of Inorganic Chemistry, 2024, 40(11): 2175-2185. doi: 10.11862/CJIC.20240165
5. [5]
  Pingwei Wu . Application of Diamond Software in Simplex Teaching. University Chemistry, 2024, 39(3): 118-121. doi: 10.3866/PKU.DXHX202311043
6. [6]
  Shiyang He , Dandan Chu , Zhixin Pang , Yuhang Du , Jiayi Wang , Yuhong Chen , Yumeng Su , Jianhua Qin , Xiangrong Pan , Zhan Zhou , Jingguo Li , Lufang Ma , Chaoliang Tan . 铂单原子功能化的二维Al-TCPP金属-有机框架纳米片用于增强光动力抗菌治疗. Acta Physico-Chimica Sinica, 2025, 41(5): 100046-. doi: 10.1016/j.actphy.2025.100046
7. [7]
  Wenyan Dan , Weijie Li , Xiaogang Wang . The Technical Analysis of Visual Software ShelXle for Refinement of Small Molecular Crystal Structure. University Chemistry, 2024, 39(3): 63-69. doi: 10.3866/PKU.DXHX202302060
8. [8]
  Yong Shu , Xing Chen , Sai Duan , Rongzhen Liao . How to Determine the Equilibrium Bond Distance of Homonuclear Diatomic Molecules: A Case Study of H₂. University Chemistry, 2024, 39(7): 386-393. doi: 10.3866/PKU.DXHX202310102
9. [9]
  Jiaxun Wu , Mingde Li , Li Dang . The R eaction of Metal Selenium Complexes with Olefins as a Tutorial Case Study for Analyzing Molecular Orbital Interaction Modes. University Chemistry, 2025, 40(3): 108-115. doi: 10.12461/PKU.DXHX202405098
10. [10]
  Yuchen Zhou , Huanmin Liu , Hongxing Li , Xinyu Song , Yonghua Tang , Peng Zhou . 设计热力学稳定的贵金属单原子光催化剂用于乙醇的高效非氧化转化形成高纯氢和增值产物乙醛. Acta Physico-Chimica Sinica, 2025, 41(6): 100067-. doi: 10.1016/j.actphy.2025.100067
11. [11]
  Rui Li , Huan Liu , Yinan Jiao , Shengjian Qin , Jie Meng , Jiayu Song , Rongrong Yan , Hang Su , Hengbin Chen , Zixuan Shang , Jinjin Zhao . 卤化物钙钛矿的单双向离子迁移. Acta Physico-Chimica Sinica, 2024, 40(11): 2311011-. doi: 10.3866/PKU.WHXB202311011
12. [12]
  Siyi ZHONG , Xiaowen LIN , Jiaxin LIU , Ruyi WANG , Tao LIANG , Zhengfeng DENG , Ao ZHONG , Cuiping HAN . Targeting imaging and detection of ovarian cancer cells based on fluorescent magnetic carbon dots. Chinese Journal of Inorganic Chemistry, 2024, 40(8): 1483-1490. doi: 10.11862/CJIC.20240093
13. [13]
  Jiarong Feng , Yejie Duan , Chu Chu , Dezhen Xie , Qiu'e Cao , Peng Liu . Preparation and Application of a Streptomycin Molecularly Imprinted Electrochemical Sensor: A Suggested Comprehensive Analytical Chemical Experiment. University Chemistry, 2024, 39(8): 295-305. doi: 10.3866/PKU.DXHX202401016
14. [14]
  Wenliang Wang , Weina Wang , Sufan Wang , Tian Sheng , Tao Zhou , Nan Wei . “Schrödinger Equation – Approximate Models – Core Concepts – Simple Applications”: Constructing a Logical Framework and Knowledge Graph of Atom and Molecule Structures. University Chemistry, 2024, 39(8): 338-343. doi: 10.3866/PKU.DXHX202312084
15. [15]
  Jinlong YAN , Weina WU , Yuan WANG . A simple Schiff base probe for the fluorescent turn-on detection of hypochlorite and its biological imaging application. Chinese Journal of Inorganic Chemistry, 2024, 40(9): 1653-1660. doi: 10.11862/CJIC.20240154
16. [16]
  Jin Tong , Shuyan Yu . Crystal Engineering for Supramolecular Chirality. University Chemistry, 2024, 39(3): 86-93. doi: 10.3866/PKU.DXHX202308113
17. [17]
  Zehua Zhang , Haitao Yu , Yanyu Qi . 多重共振TADF分子的设计策略. Acta Physico-Chimica Sinica, 2025, 41(1): 2309042-. doi: 10.3866/PKU.WHXB202309042
18. [18]
  Wen-Bing Hu . Systematic Introduction of Polymer Chain Structures. University Chemistry, 2025, 40(4): 15-19. doi: 10.3866/PKU.DXHX202401014
19. [19]
  Yuhui Yang , Jintian Luo , Biao Zuo . A Teaching Approach to Polymer Surface and Interface in Undergraduate Polymer Physics Courses. University Chemistry, 2025, 40(4): 126-130. doi: 10.12461/PKU.DXHX202408056
20. [20]
  Laiying Zhang , Yinghuan Wu , Yazi Yu , Yecheng Xu , Haojie Zhang , Weitai Wu . Innovation and Practice of Polymer Chemistry Experiment Teaching for Non-Polymer Major Students of Chemistry: Taking the Synthesis, Solution Property, Optical Performance and Application of Thermo-Sensitive Polymers as an Example. University Chemistry, 2024, 39(4): 213-220. doi: 10.3866/PKU.DXHX202310126

Get Citation

PDF

XML

Metrics

PDF Downloads(34)
Abstract views(1717)
HTML views(352)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142