DMSO effects on FRET to dye-labeled DNA in conjugated polymer-based DNA detection

Mijeong Kang; Boram Kim; Han Young Woo

doi:10.1117/12.874398

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11 February 2011 DMSO effects on FRET to dye-labeled DNA in conjugated polymer-based DNA detection

Mijeong Kang, Boram Kim, Han Young Woo

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Proceedings Volume 7908, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VIII; 790809 (2011) https://doi.org/10.1117/12.874398
Event: SPIE BiOS, 2011, San Francisco, California, United States

Abstract

Solvent effects were studied in fluorescence resonance energy transfer (FRET) from a cationic polyfluorene copolymer (FHQ, FPQ) to a fluorescein (Fl)-labeled oligonucleotide (ssDNA-Fl). Upon addition of dimethyl sulfoxide (DMSO), optical properties of the polymers and the probe dye were substantially modified. And the FRET-induced Fl emission was measured by directly exciting the polymer within the complex, polymer/ssDNA-Fl. The FRET signal was successfully modulated with changing the DMSO content. In the case of FHQ, the FRET-induced Fl emission was seriously quenched in phosphate buffer solution (PBS), while a salient FRET signal was observed in a 80 vol% DMSO/PBS mixture (36.8 time higher than that in PBS). The FPQ-sensitized FRET signal was also 3.8-fold amplified by the presence of DMSO. That result is from the decrease of hydrophobic interactions between the polymer and ssDNA-Fl, which induces the weaker polymer/ssDNA-Fl complexation with longer intermolecular separation. The gradual decrease in Fl PL quenching with increasing the DMSO content was investigated by measuring the Stern- Volmer quenching constants (3.3-4.2 × 10⁶ M^-1 in PBS, 0.56-1.1 x 10⁶ M^-1 in 80 vol% DMSO) in PBS/DMSO mixtures. The substantially reduced PL quenching would amplify the resulting FRET Fl signal. This approach suggests a simple way of modifying the fine-structure of polymer/ssDNA-Fl and improving the detection sensitivity in conjugated polymer-based FRET bioassays.

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Mijeong Kang, Boram Kim, and Han Young Woo "DMSO effects on FRET to dye-labeled DNA in conjugated polymer-based DNA detection", Proc. SPIE 7908, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VIII, 790809 (11 February 2011); https://doi.org/10.1117/12.874398

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