Femtosecond Laser Pulse Excitation of DNA-Labeled Gold Nanoparticles: Establishing a Quantitative Local Nanothermometer for Biological Applications
Femtosecond Laser Pulse Excitation of DNA-Labeled Gold Nanoparticles: Establishing a Quantitative Local Nanothermometer for Biological Applications
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DOI：
10.1021/acsnano.0c02899
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发表时间：
2020-07-28
期刊：
影响因子：
17.1
通讯作者：
Diaz, Sebastian A.
中科院分区：
文献类型：
--
作者：
Hastman, David A.;Melinger, Joseph S.;Diaz, Sebastian A.
飞秒激光脉冲激发等离子体纳米粒子-生物分子复合物是一种很有前途的局部加热生物材料的方法。研究表明，飞秒光脉冲可以调节DNA或蛋白质的活性，当连接到等离子体纳米粒子;然而，在随后的生物功能的精度仍然在很大程度上是不确定的。具体而言，局部生物分子“经历”的温度仍然未知。我们使用55 nm的金纳米粒子（AuNPs）显示双链（ds）DNA研究如何，对于不同的熔化温度的dsDNA，激光脉冲能量通量和本体溶液温度影响局部DNA变性的速率。一个通用的“模板”单链DNA连接到AuNP表面，和三个染料标记的探针链，不同的长度和熔解温度，杂交到它创建三个单独的dsDNA-AuNP生物缀合物。染料标记的探针链用于定量给定数量的光脉冲后DNA释放的速率和量，然后将其与dsDNA变性温度相关联，得到定量纳米温度计。通过改变脉冲能量通量、DNA解链温度和周围浴温，在8-53 ℃的生物学相关范围内，局部DNA变性率可以被调节三倍以上。与修改后的解离方程为这个系统量身定制的，一个“感测”的温度参数被提取和模拟金纳米粒子的温度曲线进行比较。确定这种系统中的实际生物反应可以让研究人员设计精确的纳米级光热加热源。
Femtosecond (fs) laser pulsed excitation of plasmonic nanoparticle (NP)-biomolecule conjugates is a promising method to locally heat biological materials. Studies have demonstrated that fs pulses of light can modulate the activity of DNA or proteins when attached to plasmonic NPs; however, the precision over subsequent biological function remains largely undetermined. Specifically, the temperature the localized biomolecules "experience" remains unknown. We used 55 nm gold nanoparticles (AuNPs) displaying double-stranded (ds) DNA to examine how, for dsDNA with different melting temperatures, the laser pulse energy fluence and bulk solution temperature affect the rate of local DNA denaturation. A universal "template" single-stranded DNA was attached to the AuNP surface, and three dye-labeled probe strands, distinct in length and melting temperature, were hybridized to it creating three individual dsDNA-AuNP bioconjugates. The dye-labeled probe strands were used to quantify the rate and amount of DNA release after a given number of light pulses, which was then correlated to the dsDNA denaturation temperature, resulting in a quantitative nanothermometer. The localized DNA denaturation rate could be modulated by more than threefold over the biologically relevant range of 8-53 degrees C by varying pulse energy fluence, DNA melting temperature, and surrounding bath temperature. With a modified dissociation equation tailored for this system, a "sensed" temperature parameter was extracted and compared to simulated AuNP temperature profiles. Determining actual biological responses in such systems can allow researchers to design precision nanoscale photothermal heating sources.