Application of Super-Sampling to Microscopy Images Produces Image Resolution below Optical Diffraction Limit

Abstract

Image Phase Alignment Super-Sampling (ImPASS) is a computational imaging algorithm for converting a sequence of displaced low-resolution images into a single high-resolution image. The method consists of a unique combination of Phase Correlation image registration and SeDDaRA blind deconvolution. The method has previously been validated in simulations and applied successfully to images captured in a laboratory setting. As discussed here, the performance of ImPASS surpasses similar methods that provide quantitative results. ImPASS is applied for the first time to images taken by a widefield microscope, requiring no customization other than a translation stage, to determine if this approach can subceed the diffraction limit for this application. The 80-frame image sets had as targets a slide with a slice of Porcine Cornea, and a standard US Air Force resolution chart, providing quantitative and quantitative assessments. The sets were up-sampled by a factor of eight, aligned, combined, and processed. The measurement revealed that image resolution improved by a factor of 2.68 and subceeded the diffraction limit by a factor of 1.79.