Precise measurement of the resolution in light microscopy using Fourier transform.
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.
Vainrub
Arnold
A
Department of Anatomy, Physiology and Pharmacology, College of Veterinary Medicine, Auburn University, Auburn, Alabama 36849, USA. vainrar@auburn.edu
D000465:Algorithms; D002138:Calibration; D019544:Equipment Failure Analysis / Q000379:methods*; D005583:Fourier Analysis; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008853:Microscopy / Q000379:methods* / Q000592:standards*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity