Nasse Michael J, Woehl Jörg C
Department of Chemistry and Biochemistry, Laboratory for Surface Studies, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin 53211, USA.
J Opt Soc Am A Opt Image Sci Vis. 2010 Feb 1;27(2):295-302. doi: 10.1364/JOSAA.27.000295.
In this paper we present a rigorous and general theoretical model for the illumination point spread function of a confocal microscope that correctly reproduces the optical setup. The model uses vectorial theory and assumes that monochromatic light with a Gaussian intensity distribution (such as from a laser or a single-mode fiber) is focused by a microscope objective with high numerical aperture and passes through stratified media on its way to the sample. This covers the important practical case of illumination through up to three layers, which is the situation most commonly encountered in biological microscopy (immersion oil, glass coverslip, aqueous sample medium). It also accounts for objectives that are corrected for a certain coverslip thickness and refractive index but operated under non-design conditions. Furthermore, illumination with linearly, circularly, or elliptically polarized light is covered by introducing a Babinet-Soleil compensator into the beam path. The model leads to a set of analytical equations that are readily evaluated. Two-dimensional intensity distributions for particular cases of interest are presented and discussed.
在本文中，我们提出了一种用于共聚焦显微镜照明点扩散函数的严格且通用的理论模型，该模型能正确再现光学装置。该模型采用矢量理论，并假设具有高斯强度分布的单色光（如来自激光或单模光纤）由高数值孔径的显微镜物镜聚焦，并在其到达样品的途中穿过分层介质。这涵盖了通过多达三层进行照明的重要实际情况，这是生物显微镜中最常见的情况（浸油、玻璃盖玻片、水性样品介质）。它还考虑了针对特定盖玻片厚度和折射率进行校正但在非设计条件下操作的物镜。此外，通过在光路中引入巴比涅 - 索列尔补偿器，涵盖了线偏振光、圆偏振光或椭圆偏振光的照明情况。该模型导出了一组易于评估的解析方程。给出并讨论了感兴趣的特定情况下的二维强度分布。
