Abstract
A simple, controllable and effective sample preparation method was established for atomic force microscopy (AFM) imaging of individual DNA molecules in aqueous solution. Firstly, magnesium ion (Mg2+) at a concentration of 5.0–10.0 mM as a positively charged bridge was transferred onto mica to immobilize DNA molecules. Then Mg2+-modified mica was used to investigate DNA molecules in any buffer without magnesium ion by AFM. AFM images demonstrated that DNA molecules can be successfully observed in solution with good resolution, reproducibility, and stability. Further, this DNA sample preparation method makes AFM successful to investigate DNA molecular interaction in situ and DNA/chitosan complex in gene delivery.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Arscott PG, Bloomfield VA (1992) Scanning tunneling microscopy of nucleic acids. Methods Enzymol 211:491–502
Bustamante C, Keller D, Yang G (1993) Scanning force microscopy of nucleic acids and nucleoprotein assemblies. Curr Opin Struct Biol 3:363–372
Bustamante C, Erie DA, Keller D (1994) Biochemical and structural applications of scanning force microscopy. Curr Opin Struct Biol 4:750–760
Engel A (1991) Biomolecular imaging with the atomic force microscope. Annu Rev Biophys 20:79–108
Hansma HG, Hoh J (1994) Biomolecular imaging with the atomic force microscope. Annu Rev Biophys Biochem Struct 23:115–139
Shlyakhtenko LS, Gall AA, Weimer JJ, Hawn DD, Lyubchenko YL (1999) Atomic force microscopy imaging of DNA covalently immobilized on a functionalized mica substrate. Biophys J 77:568–576
Shlyakhtenko LS, Potaman VN, Sinden RR, Lyubchenko YL (1998) Structure and dynamics of supercoil-stabilized DNA cruciforms. J Mol Biol 280:61–72
Golan R, Pietrasanta LI, Hsieh W, Hansma HG (1999) DNA toroids: stages in condensation. Biochemistry 38:14069–14076
Haberland A, Zaitsev S, Waldöfner N, Erdmann B, Böttger M, Henke W (2009) Structural appearance of linker histone H1/siRNA complexes. Mol Biol Rep 36:1083–1093
Lucius H, Haberland A, Zaitsev S, Dallüge R, Schneider M, Böttger M (2001) Structure of transfection-active histone H1/DNA complexes. Mol Biol Rep 28:157–165
Böttger M, von Mickwitz CU, Scherneck S, Lindigkeit R (1984) Interaction of histone H1 with superhelical DNA. Conformational studies and influence of ionic strength. Mol Biol Rep 10:3–8
Shen XC, Zhang ZL, Zhou B, Peng J, Xie M, Zhang M, Pang DW (2008) Visible light-induced plasmid DNA damage catalyzed by a CdSe/ZnS-photosensitized nano-TiO2 film. Environ Sci Technol 42:5049–5054
Shu W, Shu YT, Shi HB, Fei LX, Lei W, Ming QG (2008) An easy method to discover cell membrane antigen with atomic force microscopy. Mol Biol Rep 35:557–561
Nicolini C, Carrara S, Mascetti G (1997) High order DNA structure as inferred by optical fluorimetry and scanning calorimetry. Mol Biol Rep 24:235–246
Hansma HG, Vesenka J, Siegerist C, Kelderman G, Morrett H, Sinsheimer RL, Elings V, Bustamante C, Hansma PK (1992) Reproducible imaging and dissection of plasmid DNA under liquid with the atomic force microscope. Science 256:1180–1183
Hansma HG (2001) Surface biology of DNA by atomic force microscopy. Annu Rev Phys Chem 52:71–92
Mou J, Czajkowsky DM, Zhang Y, Shao Z (1995) High-resolution atomic-force microscopy of DNA: the pitch of the double helix. FEBS Lett 371:279–282
Acknowledgments
This work was supported by the National Natural Science Foundation of China (Grant No. 20833006 and 20621502).
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Shen, XC., Bao, L., Zhang, ZL. et al. A simple and effective sample preparation method for atomic force microscopy visualization of individual DNA molecules in situ. Mol Biol Rep 38, 965–969 (2011). https://doi.org/10.1007/s11033-010-0190-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-010-0190-3