Mica functionalization for imaging of dna and protein-dna complexes with atomic force microscopy

Shlyakhtenko Luda, A. Gall Alexander, Yuri L Lyubchenko

Research output: Chapter in Book/Report/Conference proceedingChapter

53 Scopus citations

Abstract

Surface preparation is a key step for reliable and reproducible imaging of DNA and protein-DNA complexes with atomic force microscopy (AFM). This article describes the approaches for chemical functionalization of the mica surface. One approach utilizes 3-aminopropyl-trietoxy silane (APTES), enabling one to obtain a smooth surface termed AP-mica. This surface binds nucleic acids and nucleoprotein complexes in a wide range of ionic strengths, in the absence of divalent cations and in a broad range of pH. Another method utilizes aminopropyl silatrane (APS) to yield an APS-mica surface. The advantage of APS-mica compared with AP-mica is the ability to obtain reliable and reproducible time-lapse images in aqueous solutions. The chapter describes the methodologies for the preparation of AP-mica and APS-mica surfaces and the preparation of samples for AFM imaging. The protocol for synthesis and puri fication of APS is also provided. The applications are illustrated with a number of examples

Original languageEnglish (US)
Title of host publicationCell Imaging Techniques: Methods and Protocols
PublisherHumana Press Inc.
Pages295-312
Number of pages18
Volume931
ISBN (Print)9781627030557
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume931
ISSN (Print)10643745

Keywords

  • AFM
  • Atomic force microscopy
  • DNA structure and dynamics
  • Mica functionalization
  • Protein-DNA complexes
  • Silanes
  • Silatranes
  • Surface chemistry

ASJC Scopus subject areas

  • Medicine(all)
  • Molecular Biology
  • Genetics

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    Luda, S., Alexander, A. G., & Lyubchenko, Y. L. (2012). Mica functionalization for imaging of dna and protein-dna complexes with atomic force microscopy. In Cell Imaging Techniques: Methods and Protocols (Vol. 931, pp. 295-312). (Methods in Molecular Biology; Vol. 931). Humana Press Inc.. https://doi.org/10.1007/978-1-62703-56-4_14