Nonlinear structured illumination microscopy by surface plasmon enhanced stimulated emission depletion.
Nonlinear structured illumination microscopy by surface plasmon enhanced stimulated emission depletion.
复制标题
DOI：
10.1364/oe.19.024783
复制
发表时间：
2011-11
期刊：
影响因子：
3.8
通讯作者：
Han Zhang;Ming Zhao;Leilei Peng
中科院分区：
文献类型：
--
作者：
Han Zhang;Ming Zhao;Leilei Peng
非线性结构照明显微镜（SIM）理论上在全视场范围内具有无限分辨率。然而，在实际信噪比和有限的光子预算下，非线性SIM的性能很大程度上取决于非线性效应的行为。饱和SIM （SSIM）由于其强烈的光漂白作用，在生物领域的应用并不理想。与SSIM相比，受激辐射耗尽（STED） SIM具有更高的灵敏度、更高的分辨率和更小的光毒性。然而，所需的激光功率，以支持一个强大的全场STED效应是无法实现与目前的激光技术。我们通过实验证明，表面等离子体共振（SPR）将近表面STED效应增强了8倍，因此在具有SPR增强的全内反射显微镜模式下，STED- sim是可行的。仿真分析预测，SPR增强的二维STED足够强，可以实现非线性SIM的30纳米分辨率和单分子灵敏度的高速成像。STED-SIM超分辨显微镜方法为在体外或活细胞基膜上观察单分子过程提供了一种解决方案。
Nonlinear structured illumination microscopy (SIM) in theory has unlimited resolution over a full field of view. However under a realistic signal-to-noise ratio and a limited photon budget, the performance of nonlinear SIM strongly depends on the behavior of the nonlinear effect. Saturated SIM (SSIM) is not ideal in biological applications due to its strong photobleaching. Stimulated emission depletion (STED) SIM will have high sensitivity, higher resolution and less photo toxicity than SSIM. However, the laser power necessary to support a strong full-field STED effect is not attainable with current laser technology. We experimentally proved that surface plasmon resonance enhances (SPR) near surface STED effect by a factor of 8, and therefore STED-SIM is feasible in the total internal reflection microscopy mode with SPR enhancement. Simulation analysis predicts that SPR enhanced 2D STED is strong enough for nonlinear SIM to achieve high-speed imaging at 30-nm resolution and single molecule sensitivity. The STED-SIM superresolution microscopy method would provide a solution for observing single molecule processes in vitro or on the basal membrane of live cells.