Biophysics Department, Biological Faculty, Moscow State University, Moscow, Russian Federation.
Biophys J. 2009 Dec 16;97(12):3206-14. doi: 10.1016/j.bpj.2009.09.029.
The article presents a noninvasive approach to the study of erythrocyte properties by means of a comparative analysis of signals obtained by surface-enhanced Raman spectroscopy (SERS) and resonance Raman spectroscopy (RS). We report step-by-step the procedure for preparing experimental samples containing erythrocytes in their normal physiological environment in a mixture of colloid solution with silver nanoparticles and the procedure for the optimization of SERS conditions to achieve high signal enhancement without affecting the properties of living erythrocytes. By means of three independent techniques, we demonstrate that under the proposed conditions a colloid solution of silver nanoparticles does not affect the properties of erythrocytes. For the first time to our knowledge, we describe how to use the SERS-RS approach to study two populations of hemoglobin molecules inside an intact living erythrocyte: submembrane and cytosolic hemoglobin (Hb(sm) and Hb(c)). We show that the conformation of Hb(sm) differs from the conformation of Hb(c). This finding has an important application, as the comparative study of Hb(sm) and Hb(c) could be successfully used in biomedical research and diagnostic tests.
本文提出了一种非侵入性的方法来研究红细胞的性质，方法是通过比较表面增强拉曼光谱（SERS）和共振拉曼光谱（RS）获得的信号。我们逐步介绍了制备实验样品的步骤，这些样品在含有胶体溶液和银纳米粒子的混合物中包含正常生理环境下的红细胞，并介绍了优化 SERS 条件的步骤，以在不影响活红细胞性质的情况下实现高信号增强。通过三种独立的技术，我们证明在提出的条件下，胶体银纳米粒子溶液不会影响红细胞的性质。据我们所知，我们首次描述了如何使用 SERS-RS 方法来研究完整活红细胞内的两种血红蛋白分子群体：细胞膜下和细胞质血红蛋白（Hb(sm) 和 Hb(c)）。我们表明，Hb(sm)的构象与 Hb(c)的构象不同。这一发现具有重要的应用价值，因为 Hb(sm)和 Hb(c)的比较研究可以成功地用于生物医学研究和诊断测试。
