Department of Applied Physics, Osaka University, Osaka, Japan.
Advanced Photonics and Biosensing Open Innovation Laboratory, AIST-Osaka University, Osaka, Japan.
Nat Methods. 2024 May;21(5):889-896. doi: 10.1038/s41592-024-02236-3. Epub 2024 Apr 5.
The background light from out-of-focus planes hinders resolution enhancement in structured illumination microscopy when observing volumetric samples. Here we used selective plane illumination and reversibly photoswitchable fluorescent proteins to realize structured illumination within the focal plane and eliminate the out-of-focus background. Theoretical investigation of the imaging properties and experimental demonstrations show that selective plane activation is beneficial for imaging dense microstructures in cells and cell spheroids.
在观察体积样本时,离焦平面的背景光会阻碍结构光照明显微镜的分辨率提高。在这里,我们使用选择性平面照明和可可逆光转换荧光蛋白在焦平面内实现结构照明,并消除离焦背景。对成像特性的理论研究和实验验证表明,选择性平面激活有利于对细胞和细胞球体中的密集微观结构进行成像。