Supercritical angle fluorescence biosensor for the detection of molecular interactions on cellulose-modified glass surfaces

Cellulose films have been proposed as a convenient substrate for producing flat and homogeneous surface coatings. Additionally, amino-labelled cellulose species, like aminopropyltrimethylsilylethercellulose (ATMSC), are excellent support matrices for covalent binding of biomolecules, with low probe density and prevention of non-specific adsorption of unbound analyte molecules. Due to ATMSC films fulfil important requirements as substrate for analyse techniques of surface-tethered proteins and nucleic acids, we consequently report a new preparation for DNA-functionalised surfaces. Single-stranded DNA molecules are covalently coupled to cellulose-coated glass cover slips to interact with complementary free Cy5-labelled oligonucleotides in solution. Hybridisation efficiencies at the new substrate and at standard surface coatings are determined by detection of the surface-generated fluorescence. In order to discriminate against the fluorescence from unbound oligonucleotides the detection volume was restricted to the surface by collecting supercritical angle fluorescence (SAF). Thus, it is demonstrated that cellulose films are utilised to investigate DNA-hybridisation reactions highly sensitive