New buffer systems for photopainting of single biomolecules
Sprache des Titels:
Englisch
Original Kurzfassung:
We present newly developed buffer systems that significantly improve the efficiency of a photochemically
induced surface modification at the single molecule level. Buffers with paramagnetic cations and radical
oxygen promoting species facilitate laser-assisted protein adsorption by photobleaching (LAPAP) of single
fluorescently labelled oligonucleotides or biotin onto multi-photon-lithography-structured 2D and 3D
acrylate scaffolds. Single molecule fluorescence microscopy has been used to quantify photopainting
efficiency. We identify specific cation interaction sites for members of the cyanine, coumarin and
rhodamine classes of fluorophores using quantum mechanical calculations. We show that our buffer
systems provide an up to three-fold LAPAP-efficiency increase for the cyanine fluorophore, while keeping
excitation parameters constant.