Schrader M, Bahlmann K, Giese G, Hell S W
High Resolution Optical Microscopy Group, Max Planck Institute for Biophysical Chemistry, D-37070 Göttingen, Germany.
Biophys J. 1998 Oct;75(4):1659-68. doi: 10.1016/S0006-3495(98)77608-8.
By combining the wavefronts produced by two high-aperture lenses, two-photon 4Pi-confocal microscopy allows three-dimensional imaging of transparent biological specimens with axial resolution in the 100-140-nm range. We reveal the imaging properties of a two-photon 4Pi-confocal microscope as applied to a fixed cell. We demonstrate that a fast, linear point deconvolution suffices to achieve axially superresolved 3D images in the cytoskeleton. Furthermore, we describe stringent algorithms for alignment and control of the two lenses. We also show how to compensate for the effects of a potential refractive index mismatch of the mounting medium with respect to the immersion system.
通过结合两个高孔径透镜产生的波前，双光子4Pi共聚焦显微镜能够对透明生物标本进行三维成像，轴向分辨率在100 - 140纳米范围内。我们揭示了双光子4Pi共聚焦显微镜应用于固定细胞时的成像特性。我们证明，快速、线性点去卷积足以在细胞骨架中实现轴向超分辨三维图像。此外，我们描述了用于两个透镜对准和控制的严格算法。我们还展示了如何补偿安装介质相对于浸没系统可能存在的折射率失配的影响。
