Super-resolution Surface-Enhanced Raman Scattering Imaging of Single Particles in Cells
Super-resolution Surface-Enhanced Raman Scattering Imaging of Single Particles in Cells
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DOI：
10.1021/acs.analchem.0c01864
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发表时间：
2020-07-07
影响因子：
7.4
通讯作者：
Schultz, Zachary D.
中科院分区：
文献类型：
--
作者：
de Albuquerque, Carlos Diego Lima;Schultz, Zachary D.
定位和识别细胞中分子相互作用的能力对于理解蛋白质功能和分子生物学具有重要意义。功能化的金属纳米颗粒已经被用作蛋白质追踪和药物递送的探针，因为它们能够携带治疗剂和容易功能化的表面。在这项工作中，我们提出了一个超分辨率的表面增强拉曼散射（Sers）的成像和跟踪膜受体与肽功能化的金纳米星（AuNS）的方法。结肠癌细胞中的α（v）β（3）整联蛋白受体被成功地靶向，并使用附接有高亲和力氨基酸序列甘氨酸-甘氨酸-天冬氨酸-苯丙氨酸氨基转移酶（RGDFC）的AuNS成像。RGDFC肽与整联蛋白受体的相互作用提供了明亮且波动的Sers信号，可以用定位显微镜算法进行分析。此外，观察到的Sers光谱用于确认蛋白质-肽相互作用。功能化和裸AuNS的实验说明特异性和非特异性结合事件。以类似于6 nm的定位精度监测特异性结合。观察到的空间分辨率与紧密结合有关，这通过AuNS-RGDFC的4.4 x 10（-11）cm（2）/s测量的较慢的扩散系数与裸AuNS的7.8 x 10（10）cm（2）/s相比得到证实。不同焦平面的超分辨率Sers图像显示了内化颗粒的证据，并暗示了对细胞表面蛋白质取向的见解。我们的工作证明了超分辨率Sers成像可以探测细胞中的膜受体相互作用，提供化学信息和空间分辨率，具有在生命科学和生物医学中广泛应用的潜力。
The ability to locate and identify molecular interactions in cells has significant importance for understanding protein function and molecular biology. Functionalized metallic nanoparticles have been used as probes for protein tracking and drug delivery because of their ability to carry therapeutic agents and readily functionalized surfaces. In this work, we present a super-resolution surface-enhanced Raman scattering (SERS) approach for imaging and tracking membrane receptors interacting with peptide-functionalized gold nanostars (AuNS). The alpha(v)beta(3) integrin receptors in colon cancer cells are successfully targeted and imaged using AuNS with the high-affinity amino acid sequence arginine-glycine-aspartic acid-phenylalaninecysteine (RGDFC) attached. The RGDFC peptide interaction with the integrin receptor provides a bright and fluctuating SERS signal that can be analyzed with localization microscopy algorithms. Additionally, the observed SERS spectrum is used to confirm protein-peptide interaction. Experiments with functionalized and bare AuNS illustrate specific and nonspecific binding events. Specific binding is monitored with a localization precision of similar to 6 nm. The observed spatial resolution is associated with tight binding, which was confirmed by the slower diffusion coefficient measured from 4.4 x 10(-11) cm(2)/s for the AuNS-RGDFC compared to 7.8 x 10(10) cm(2)/s for the bare AuNS. Super-resolution SERS images at different focal planes show evidence of internalized particles and suggest insights into protein orientation on the surface of cells. Our work demonstrates super-resolution SERS imaging to probe membrane receptor interactions in cells, providing chemical information and spatial resolution with potential for diverse applications in life science and biomedicine.