Please use this identifier to cite or link to this item:
http://hdl.handle.net/1942/8438
Title: | Topographical and functional characterization of the ssDNA probe layer generated through EDC-mediated covalent attachment to nanocrystalline diamond using fluorescence microscopy | Authors: | VERMEEREN, Veronique WENMACKERS, Sylvia DAENEN, Michael HAENEN, Ken WILLIAMS, Oliver AMELOOT, Marcel VAN DE VEN, Martin WAGNER, Patrick MICHIELS, Luc |
Issue Date: | 2008 | Publisher: | AMER CHEMICAL SOC | Source: | LANGMUIR, 24(16). p. 9125-9134 | Abstract: | The covalent attachment method for DNA on nanocrystalline diamond (NCD), involving the introduction of COOH functionalities on the surface by photoattachment of 10-undecenoic acid (10-UDA), followed by the 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC)-mediated coupling to NH2-labeled ssDNA, is evaluated in terms of stability, density, and functionality of the resulting biological interface. This is of crucial importance in DNA biosensor development. The covalent nature of DNA attachment will infer the necessary stability and favorable orientation to the ssDNA probe molecules. Using confocal fluorescence microscopy, the influence of buffer type for the removal of excess 10-UDA and ssDNA, the probe ssDNA length, the probe ssDNA concentration, and the presence of the COOH-linker on the density and functionality of the ssDNA probe layer were investigated. It was determined that the most homogeneously dense and functional DNA layer was obtained when 300 pmol of short ssDNA was applied to COOH-modified NCD samples, while H-terminated NCD was resistant for DNA attachment. Exploiting this surface functionality dependence of the DNA attachment efficiency, a shadow mask was applied during the photochemical introduction of the COOH-functionalities, leaving certain regions on the NCD H-terminated. The subsequent DNA attachment resulted in a fluorescence pattern corresponding to the negative of the shadow mask. Finally, NCD surfaces covered with mixtures of the 10-UDA linker molecule and a similar molecule lacking the COOH functionality, functioning as a lateral spacer, were examined for their suitability in preventing nonspecific adsorption to the surface and in decreasing steric hindrance. However, purely COOH-modified NCD samples, patterned with H-terminated regions and treated with a controlled amount of probe DNA, proved the most efficient in fulfilling these tasks. | Notes: | Hasselt Univ & Transnatl Univ Limburg, Biomed Res Inst, Sch Life Sci, B-3590 Diepenbeek, Belgium. Hasselt Univ & Transnatl Univ Limburg, Inst Mat Res, Sch Life Sci, B-3590 Diepenbeek, Belgium. IMEC VZW, Div IMOMEC, B-3590 Diepenbeek, Belgium. | Document URI: | http://hdl.handle.net/1942/8438 | ISSN: | 0743-7463 | e-ISSN: | 1520-5827 | DOI: | 10.1021/la800946v | ISI #: | 000258377900110 | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2009 |
Appears in Collections: | Research publications |
Show full item record
SCOPUSTM
Citations
29
checked on Sep 5, 2020
WEB OF SCIENCETM
Citations
29
checked on Oct 11, 2024
Page view(s)
74
checked on Sep 7, 2022
Google ScholarTM
Check
Altmetric
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.