Atomic force microscopy imaging and probing of DNA, proteins, and protein-DNA complexes: Silatrane surface chemistry

Yuri L. Lyubchenko, Luda S. Shlyakhtenko, Alexander A. Gall

Research output: Chapter in Book/Report/Conference proceedingChapter

49 Scopus citations

Abstract

Despite their rather recent invention, atomic force microscopes are widely available commercially. AFM and its special modifications (tapping mode and noncontact operation in solution) have been successfully used for topographic studies of a large number of biological objects including DNA, RNA, proteins, cell membranes, and even whole cells. AFM was also successfully applied to studies of nucleic acids and various protein DNA complexes. Part of this success is due to the development of reliable sample preparation procedures. This chapter describes one of the approaches based on chemical functionalization of mica surface with 1-(3-aminopropyl) silatrane (APS). One of the most important properties of APS-mica approach is that the sample can be deposited on the surface in a wide range of ionic strengths, in the absence of divalent cations and a broad range of pH. In addition to imaging of dried sample, APS-mica allows reliable and reproducible time lapse imaging in aqueous solutions. Finally, APS mica is terminated with reactive amino groups that can be used for covalent and ionic attachment of molecules for AFM force spectroscopy studies. The protocols for the preparation of APS, functionalization with APS mica and AFM probes, preparation of samples for imaging in air and in aqueous solutions, and force spectroscopy studies are outlined. All these applications are illustrated with a few examples.

Original languageEnglish (US)
Title of host publicationDNA-Protein Interactions
Subtitle of host publicationPrinciples and Protocols, Third Edition
EditorsTom Moss, Benoit Leblanc
Pages337-351
Number of pages15
DOIs
StatePublished - 2009

Publication series

NameMethods in Molecular Biology
Volume543
ISSN (Print)1064-3745

Keywords

  • Atomic force microscopy (AFM)
  • DNA local structures
  • Force spectroscopy
  • Mica functionalization
  • Protein DNA complexes
  • Silanes
  • Silatranes
  • Surface chemistry

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Lyubchenko, Y. L., Shlyakhtenko, L. S., & Gall, A. A. (2009). Atomic force microscopy imaging and probing of DNA, proteins, and protein-DNA complexes: Silatrane surface chemistry. In T. Moss, & B. Leblanc (Eds.), DNA-Protein Interactions: Principles and Protocols, Third Edition (pp. 337-351). (Methods in Molecular Biology; Vol. 543). https://doi.org/10.1007/978-1-60327-015-1_21