Open AccessA silanized mica substrate suitable for high‐resolution fiber FISH analysis by scanning near‐field optical/atomic force microscopy
Author(s) -
Sugiyama Shigeru,
Fukuta Megumi,
Hirose Tamaki,
Ohtani Toshio,
Yoshino Tomoyuki
Publication year - 2010
Publication title -
scanning
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.359
H-Index - 47
eISSN - 1932-8745
pISSN - 0161-0457
DOI - 10.1002/sca.20214
Subject(s) - substrate (aquarium) , near field scanning optical microscope , fluorescence , fluorescence microscope , microscopy , materials science , mica , peptide nucleic acid , resolution (logic) , nanoscopic scale , analytical chemistry (journal) , chemistry , optical microscope , optics , nucleic acid , nanotechnology , scanning electron microscope , chromatography , physics , biochemistry , oceanography , artificial intelligence , computer science , composite material , geology
We applied a novel silanized mica substrate with an extremely flat surface constructed according to Sasou et al . ( Langmuir 19 , 9845–9849 (2003)) to high‐resolution detection of a specific gene on a DNA fiber by scanning near‐field optical/atomic force microscopy (SNOM/AFM). The interaction between the substrate and fluorescence‐dye conjugated peptide nucleic acid (PNA) probes, which causes fluorescence noise signal, was minimal. By using the substrate, we successfully obtained a fluorescence in situ hybridization signal from the ea47 gene on a λphage DNA labeled with an Alexa 532‐conjugated 15‐base PNA probe. As the results, no fluorescence noises were observed, indicating that the surface adsorbed almost none of the PNA probe. The combination of the substrate and SNOM/AFM is an effective tool for visualizing DNA sequences at nanometer‐scale resolution. SCANNING 32: 383–389, 2010. © 2010 Wiley Periodicals, Inc.