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Proceedings Article

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

[+] Author Affiliations
Mijeong Kang, Boram Kim, Han Young Woo

Pusan National Univ. (Korea, Republic of)

Proc. SPIE 7908, Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VIII, 790809 (February 11, 2011); doi:10.1117/12.874398
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From Conference Volume 7908

  • Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VIII
  • Alexander N. Cartwright; Dan V. Nicolau
  • San Francisco, California, USA | January 22, 2011

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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Citation

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 (February 11, 2011); doi:10.1117/12.874398; http://dx.doi.org/10.1117/12.874398


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