Shaevitz Joshua W, Fletcher Daniel A
Department of Integrative Biology, University of California, Berkeley 94720, USA.
J Opt Soc Am A Opt Image Sci Vis. 2007 Sep;24(9):2622-7. doi: 10.1364/josaa.24.002622.
We present a technique to systematically measure the change in the blurring function of an optical microscope with distance between the source and the coverglass (the depth) and demonstrate its utility in three-dimensional (3D) deconvolution. By controlling the axial positions of the microscope stage and an optically trapped bead independently, we can record the 3D blurring function at different depths. We find that the peak intensity collected from a single bead decreases with depth and that the width of the axial, but not the lateral, profile increases with depth. We present simple convolution and deconvolution algorithms that use the full depth-varying point-spread functions and use these to demonstrate a reduction of elongation artifacts in a reconstructed image of a 2 microm sphere.
我们提出了一种技术，用于系统地测量光学显微镜的模糊函数随光源与盖玻片之间距离（深度）的变化，并展示其在三维（3D）反卷积中的效用。通过独立控制显微镜载物台和光镊捕获的珠子的轴向位置，我们可以记录不同深度处的3D模糊函数。我们发现，从单个珠子收集到的峰值强度随深度降低，并且轴向（而非横向）轮廓的宽度随深度增加。我们提出了简单的卷积和反卷积算法，这些算法使用完整的随深度变化的点扩散函数，并使用它们来证明在2微米球体的重建图像中伸长伪影的减少。
