Shenzhen Key Laboratory of Photonics and Biophotonics, Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, China.
Biosensors (Basel). 2022 Jul 20;12(7):539. doi: 10.3390/bios12070539.
The resolution of optical microscopes is limited by the optical diffraction limit; in particular, the axial resolution is much lower than the lateral resolution, which hinders the clear distinction of the three-dimensional (3D) structure of cells. Although stimulated emission depletion (STED) superresolution microscopy can break through the optical diffraction limit to achieve 3D superresolution imaging, traditional 3D STED requires high depletion laser power to acquire high-resolution images, which can cause irreversible light damage to biological samples and probes. Therefore, we developed an ultralow laser power 3D STED superresolution imaging method. On the basis of this method, we obtained lateral and axial resolutions of 71 nm and 144 nm, respectively, in fixed cells with 0.65 mW depletion laser power. This method will have broad application prospects in 3D superresolution imaging of living cells.
光学显微镜的分辨率受到光学衍射极限的限制；特别是轴向分辨率远低于横向分辨率，这阻碍了对细胞三维（3D）结构的清晰区分。虽然受激发射损耗（STED）超分辨率显微镜可以突破光学衍射极限实现 3D 超分辨率成像，但传统的 3D STED 需要高损耗激光功率来获取高分辨率图像，这可能对生物样本和探针造成不可逆转的光损伤。因此，我们开发了一种超低激光功率 3D STED 超分辨率成像方法。在此方法的基础上，我们用 0.65 mW 的损耗激光功率在固定细胞中获得了 71nm 和 144nm 的横向和轴向分辨率。该方法将在活细胞的 3D 超分辨率成像中具有广阔的应用前景。
