使用傅里叶变换对光学显微镜中的分辨率进行精确测量。

Precise measurement of the resolution in light microscopy using Fourier transform.

作者信息

Vainrub Arnold

机构信息

Department of Anatomy, Physiology and Pharmacology, College of Veterinary Medicine, Auburn University, Auburn, Alabama 36849, USA.

出版信息

Rev Sci Instrum. 2008 Apr;79(4):046112. doi: 10.1063/1.2912954.

DOI:10.1063/1.2912954

PMID:18447570

Abstract

The resolution power of light microscope has been accurately measured (+/-5%) by Fourier transform of various object images and further evaluation of the highest spatial frequency in Fourier spectrum. Any unknown shape plane object with a shape feature's size smaller than the resolution to be measured was shown to provide a reliable resolution test. This simple method gives a direct measurement of the resolution power as defined by Abbe [Archiv. F. Mikroskopische Anat. 9, 413 (1873)]. The results have been justified by comparison to a standard resolution measurement by using calibrated periodic line patterns. Notably, the approach is applicable in super-resolution light microscopy (transmission, reflection, and fluorescence), where calibrated resolution targets do not occur. It was conveniently implemented by using a compact disk as a test object and free IMAGEJ imaging software.

摘要

通过对各种物体图像进行傅里叶变换，并进一步评估傅里叶频谱中的最高空间频率，已精确测量了光学显微镜的分辨率（±5%）。任何形状特征尺寸小于待测分辨率的未知形状平面物体都可用于提供可靠的分辨率测试。这种简单方法可直接测量由阿贝定义的分辨率[《显微镜解剖学档案》9, 413 (1873)]。通过与使用校准周期性线条图案的标准分辨率测量结果进行比较，验证了这些结果。值得注意的是，该方法适用于超分辨率光学显微镜（透射、反射和荧光），在这种显微镜中不存在校准的分辨率目标。使用光盘作为测试对象和免费的IMAGEJ成像软件可方便地实现该方法。