Yeh Chia-Hua, Chen Szu-Yu
Appl Opt. 2015 Mar 20;54(9):2309-17. doi: 10.1364/AO.54.002309.
Conventional structured illumination microscopy (SIM) with wide-field illumination is an applicable tool to provide resolution enhancement. And yet its applications in thick specimens are still full of challenges. By combing the structured illumination concept with two-photon excitation, a laser scanning two-photon structured illumination microscope (LSTP-SIM) was constructed to gain ∼1.42-fold lateral resolution enhancement in contrast to two-photon fluorescence microscopy. With a point-scanning geometry, an acoustic-optical modulator was used to modulate temporally the excitation intensity in order to produce the structured illumination pattern. The theoretical models of image formation and image reconstruction were clearly established. Simulation and experiments were both performed to show the capability of this system to enhance the lateral resolution. Combined with the inherent optical sectioning power of the two-photon excitation, LSTP-SIM would have the potential for applications in optically-thick specimens.
具有宽场照明的传统结构照明显微镜（SIM）是一种可用于提高分辨率的工具。然而，其在厚标本中的应用仍然充满挑战。通过将结构照明概念与双光子激发相结合，构建了一种激光扫描双光子结构照明显微镜（LSTP-SIM），与双光子荧光显微镜相比，其横向分辨率提高了约1.42倍。采用点扫描几何结构，使用声光调制器对激发强度进行时间调制，以产生结构照明图案。清晰地建立了图像形成和图像重建的理论模型。通过模拟和实验都证明了该系统提高横向分辨率的能力。结合双光子激发固有的光学切片能力，LSTP-SIM在光学厚标本中具有应用潜力。
