Naderer Christoph, Krobath Heinrich, Sivun Dmitry, Gvindzhiliia Georgii, Klar Thomas A, Jacak Jaroslaw
School of Medical Engineering and Applied Social Sciences, University of Applied Sciences Upper Austria Garnisonstraße 21 4020 Linz Austria
Institute of Theoretical Physics, Johannes Kepler University Linz Altenberger Straße 69 4040 Linz Austria.
RSC Appl Interfaces. 2023 Oct 7;1(1):110-121. doi: 10.1039/d3lf00125c. eCollection 2024 Jan 17.
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.
我们展示了新开发的缓冲系统，该系统在单分子水平上显著提高了光化学诱导表面修饰的效率。含有顺磁阳离子和促进自由基氧生成的物质的缓冲液，通过对单荧光标记的寡核苷酸或生物素进行光漂白，促进其在多光子光刻结构化的二维和三维丙烯酸酯支架上的激光辅助蛋白质吸附（LAPAP）。单分子荧光显微镜已被用于量化光绘效率。我们使用量子力学计算确定了花青、香豆素和罗丹明类荧光团成员的特定阳离子相互作用位点。我们表明，在保持激发参数不变的情况下，我们的缓冲系统可使花青荧光团的LAPAP效率提高多达三倍。
