Lee Changwon, Carney Randy P, Hazari Sidhartha, Smith Zachary J, Knudson Alisha, Robertson Christopher S, Lam Kit S, Wachsmann-Hogiu Sebastian
Center for Biophotonics, University of California, Davis, Sacramento, CA 95817, USA.
Nanoscale. 2015;7(20):9290-7. doi: 10.1039/c5nr01333j. Epub 2015 May 5.
Thin silver film coated nanobowl Surface Enhanced Raman Spectroscopy (SERS) substrates are used to capture exosomes in solution for SERS measurements that can provide biochemical analysis of intact and ruptured exosomes. Exosomes derived via Total Exosome Isolation Reagent (TEIR) as well as ultracentrifugation (UC) from the SKOV3 cell line were analyzed. Spectra of exosomes derived via TEIR are dominated by a signal characteristic for the TEIR kit that needs to be subtracted for all measurements. Differences in SERS spectra recorded at different times during the drying of the exosome solution are statistically analyzed with Principal Component Analysis (PCA). At the beginning of the drying process, SERS spectra of exosomes exhibit peaks characteristic for both lipids and proteins. Later on during the drying process, new SERS peaks develop, suggesting that the initially intact exosome ruptures over time. This time-dependent evolution of SERS peaks enables analysis of exosomal membrane contents and the contents inside the exosomes.
涂有薄银膜的纳米碗表面增强拉曼光谱(SERS)底物用于捕获溶液中的外泌体以进行SERS测量,该测量可提供完整和破裂外泌体的生化分析。分析了通过总外泌体分离试剂(TEIR)以及超速离心(UC)从SKOV3细胞系中获得的外泌体。通过TEIR获得的外泌体光谱主要由TEIR试剂盒的信号特征主导,所有测量都需要减去该信号特征。使用主成分分析(PCA)对在外泌体溶液干燥过程中不同时间记录的SERS光谱差异进行统计分析。在干燥过程开始时,外泌体的SERS光谱显示出脂质和蛋白质的特征峰。在干燥过程后期,出现了新的SERS峰,表明最初完整的外泌体随着时间的推移而破裂。SERS峰的这种随时间的演变能够分析外泌体膜内容物和外泌体内部的内容物。