Toyouchi Shuichi, Oomachi Seiya, Hasegawa Ryoma, Hayashi Kota, Takagi Yumiko, Tamura Mamoru, Tokonami Shiho, Iida Takuya
Research Institute for Light-induced Acceleration System (RILACS), Osaka Metropolitan University, 1-2 Gakuencho, Nakaku, Sakai, Osaka 599-8570, Japan.
Department of Physics, Graduate School of Science, Osaka Metropolitan University, 1-2 Gakuencho, Nakaku, Sakai, Osaka 599-8570, Japan.
ACS Sens. 2025 Feb 28;10(2):751-760. doi: 10.1021/acssensors.4c02119. Epub 2025 Jan 23.
The unique characteristics of biological structures depend on the behavior of DNA sequences confined in a microscale cell under environmental fluctuations and dissipation. Here, we report a prominent difference in fluorescence from dye-modified single-stranded DNA in a light-induced assembly of DNA-functionalized heterogeneous probe particles in a microwell of several microliters in volume. Strong optical forces from the Mie scattering of microparticles accelerated hybridization, and the photothermal effect from the localized surface plasmons in gold nanoparticles enhanced specificity to reduce the fluorescence intensity of dye-modified DNA to a few %, even in a one-base mismatched sequence, enabling us to clearly highlight the single nucleotide polymorphisms in DNA. Fluorescence intensity was positively correlated with complementary DNA concentrations ranging in several tens fg/μL after only 5 min of laser irradiation. Remarkably, a total amount of DNA in an optically assembled structure of heterogeneous probe particles was estimated between 2.36 ymol (2.36 × 10 mol) and 2.36 amol (2.36 × 10 mol) in the observed concentration range. These findings can promote an innovative production method of nanocomposite structures via biological molecules and biological sensing with simple strategies avoiding genetic amplification in a PCR-free manner.
生物结构的独特特性取决于在环境波动和耗散下限制在微观尺度细胞内的DNA序列的行为。在此,我们报道了在体积为几微升的微孔中,DNA功能化异质探针颗粒的光诱导组装过程中,染料修饰的单链DNA荧光存在显著差异。微粒的米氏散射产生的强大光力加速了杂交,金纳米颗粒中局域表面等离子体的光热效应提高了特异性,即使在单碱基错配序列中,也能将染料修饰DNA的荧光强度降低到百分之几,使我们能够清晰地突出显示DNA中的单核苷酸多态性。仅在激光照射5分钟后,荧光强度就与几十fg/μL范围内的互补DNA浓度呈正相关。值得注意的是,在观察到的浓度范围内,异质探针颗粒光学组装结构中的DNA总量估计在2.36 ymol(2.36×10⁻¹²mol)至2.36 amol(2.36×10⁻¹⁸mol)之间。这些发现可以通过生物分子促进纳米复合结构的创新生产方法,并以简单的策略实现生物传感,无需PCR进行基因扩增。