Hybridization of Deoxyribonucleic Acid and Immobilization of Green Fluorescent Protein on Nanostructured Organosilane Templates

Abstract

We propose a novel process for preferential immobilization of deoxyribonucleic acid (DNA) and green fluorescent protein (GFP) onto organosilane self-assembled monolayer (SAM) templates. One of the advantages of using the organosilane SAM as the template is that it is electron-beam sensitive and, by utilizing the SAM as an alternative resist film, we can make nanopatterns on a molecular scale because the proximity effect is negligible owing to the film's thinness. An other advantage is that the organosilane SAM is repellent to both DNA and GFP. Thus, the patterned SAM can be utilized as the passivation film covering the outside of the pattern while DNA and GFP are immobilized within the pattern. We investigate two kinds of organosilane SAMs for the template. One is n-octadecyltrimethoxysilane (ODS) SAM, and the other is 1H,1H,2H,2H-perfluorodecyltriethoxysilane (FDS) SAM. Our results indicate that the ODS SAM is more repellent to DNA than the FDS SAM and is suitable for DNA immobilization, while the FDS SAM template is suitable for GFP immobilization via biotin-avidin coupling because the FDS SAM surface prevents the nonspecific adsorption of streptavidin. Although the nonspecific adsorption of DNA and GFP on a SAM is also recognized, by controlling both the concentration and the incubation time, we can immobilize DNA and GFP preferentially onto nanopatterns of 100 nm diameter.

Application of Organosilane Monolayer Template to Quantitative Evaluation of Cancer Cell Adhesive Ability

“…The above treatment of the SiO 2 surface with ODS and APTES has been wellcharacterized to form a SAM of ODS 24,25) and to functionalize the surface with amino groups, [26][27][28] respectively.…”

“…Finally, the substrate was immersed in a 2% solution of 3aminopropyltriethoxysilane (APTES) in 95% ethanol/water for 30 min at room temperature, and was sonicated in ethanol for 5 min. The above treatment of the SiO 2 surface with ODS and APTES has been wellcharacterized to form a SAM of ODS 24,25) and to functionalize the surface with amino groups, [26][27][28] respectively.…”

Detecting nuclear spins in an organosilane monolayer using nitrogen-vacancy centers for analysis of precursor self-assembly on diamond surface

“…Such surface modification is useful for controlling surface properties, such as wettability in liquids and immobilizing other molecules on the monolayer with covalent bonds. [17][18][19] In this work, several types of organosilane monolayers were deposited on the diamond surface. Nuclear spins in the monolayer were detected by using shallow NV centers as the spin probe.…”

In situ synthesis and direct immobilization of ssDNA on electron beam patterned hydrogen silsesquioxane

“…In "positive-tone" electron beam lithography, patterning defines the clear areas for silane chemistry and subsequent DNA immobilization. 7,8 Similarly, this can also be achieved through the use of induced charge trapping produced by electron beam exposure. 9 In "negative-tone" electron beam patterning a DNA coated substrate is directly exposed to electron beam radiation to destroy DNA by the exposure, thus defining areas that are inaccessible to DNA hybridization, resulting in nanopatterned features surrounded by areas of DNA sequence.…”