Kim Jeongyong, Kim Dae-Chul, Back Seng-Hun
Department of Physics, University of Incheon, Incheon 402-749, Korea.
Microsc Res Tech. 2009 Jun;72(6):441-6. doi: 10.1002/jemt.20689.
The authors present the experimental result of improved lateral resolution in laser confocal microscopy (LCM) by using annular and radially polarized light as the input illumination of an existing LCM. The authors examined the lateral resolution of the LCM by imaging a single fluorescent bead and measuring the lateral width of the single bead profile appearing in the optical image. Compared to no aperture and linearly polarized light, the central peak of the single bead profile narrowed by approximately 40%, being as small as 122 nm in full width at half maximum using 405 nm laser excitation in a reflection imaging. In addition, the authors showed that radial polarization helps to preserve the circular shape of the single bead profile whereas linearly polarized light tends to induce an elongation along the polarization direction.
作者展示了通过使用环形光和径向偏振光作为现有激光共聚焦显微镜(LCM)的输入照明,从而提高其横向分辨率的实验结果。作者通过对单个荧光微珠成像并测量光学图像中单个微珠轮廓的横向宽度,来检测LCM的横向分辨率。与无孔径光和线偏振光相比,在反射成像中使用405nm激光激发时,单个微珠轮廓的中心峰变窄了约40%,半高宽小至122nm。此外,作者还表明,径向偏振有助于保持单个微珠轮廓的圆形,而线偏振光则倾向于沿偏振方向诱导伸长。