When imaging a sample, it is desirable to have the entire area of interest in focus in the acquired image. Typically, microscopes have a limited depth of field (DOF) and this makes the acquisition of such an all-in-focus image difficult. This is a major problem in many microscopic applications and applies equally in the realm of scanning electron microscopy as well. In multifocus fusion, the central idea is to acquire focal information from multiple images at different focal planes and fuse them into one all-in-focus image where all the focal planes appear to be in focus.Large chamber scanning electron microscopes (LC-SEM) are one of the latest members in the SEM family that has found extensive use for nondestructive evaluations. Large objects (~1 meter) can be scanned in micro- or nano-scale using this microscope. An LC-SEM can provide characterization of conductive and non-conductive surfaces with a magnification from 10× to 200,000×. The LC-SEM, as with other SEMs, suffers from the problem of limited DOF making it difficult to inspect a large object while keeping all areas in focus.