Accurate shape from focus based on focus adjustment in optical microscopy

Optical microscopy allows a magnified view of the sample while decreasing the depth of focus. Although the acquired images from limited depth of field have both blurred and focused regions, they can provide depth information. The technique to estimate the depth and 3D shape of an object from the images of the same sample obtained at different focus settings is called shape from focus (SFF). In SFF, the measure of focus--sharpness--is the crucial part for final 3D shape estimation. The conventional methods compute sharpness by applying focus measure operator on each 2D image frame of the image sequence. However, such methods do not reflect the accurate focus levels in an image because the focus levels for curved objects require information from neighboring pixels in the adjacent frames too. To address this issue, we propose a new method based on focus adjustment which takes the values of the neighboring pixels from the adjacent image frames that have approximately the same initial depth as of the center pixel and then it re-adjusts the center value accordingly. Experiments were conducted on synthetic and microscopic objects, and the results show that the proposed technique generates better shape and takes less computation time in comparison with previous SFF methods based on focused image surface (FIS) and dynamic programming.