A post-processing technique for extending depth of focus in conventional optical microscopy

In this paper, we propose a post-processing technique to obtain optical microscope images with extended depth of focus using a conventional microscope. With the proposed technique, we collect a sequence of images focused at different depths. We then combine the in-focus regions of each acquired frame to compose a single all-in-focus image. That is, a new image with extended depth of focus is obtained. The key to such an algorithm is in selecting the “in-focus” regions from each frame. In this paper, we describe the technique used to identify the in-focus region on every depth slice. Quantitative simulation results are presented where mean absolute error is used as a metric to assess the algorithm performance. Results using real imagery are also presented for subjective evaluation. Based on subjective evaluation and the quantitative simulation results, we believe that the proposed algorithm provides useful depth of focus extension.