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

49 Citations (Scopus)

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

Fingerprint

Atomic Force Microscopy
Proteins
Silanes
Nucleoproteins
DNA
Divalent Cations
mica
Osmolar Concentration
Nucleic Acids
silatrane

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

Cite this

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

Mica functionalization for imaging of dna and protein-dna complexes with atomic force microscopy. / Luda, Shlyakhtenko; Alexander, A. Gall; Lyubchenko, Yuri L.

Cell Imaging Techniques: Methods and Protocols. Vol. 931 Humana Press Inc., 2012. p. 295-312 (Methods in Molecular Biology; Vol. 931).

Research output: Chapter in Book/Report/Conference proceedingChapter

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