Automated extended depth of focus digital holographic microscopy using electrically tunable lens

Abstract

A combination of electrically tunable lens with a microscope objective lens (MO) for multifocal plane imaging capability is proposed. The method aims at extending the depth of focus of the MO by changing its axial range and field of view. The combination is implemented in a common path off-axis digital holographic microscopy configuration experimentally and it achieves an axial scanning range of 363.5 μm. It utilizes image plane holography to obtain in-focus images throughout the axial range. The proposed method demonstrates its application in quantitative phase imaging of the USAF 1951 test chart and flowing red blood cells at different axial depths. An axial depth variation of 0.76 μm–0.81 μm is obtained between two consecutive focused planes. The results validate and postulate the advantage of the proposed method over traditional extended depth-of-focus algorithms. The quantitative and qualitative results exploit the possibility of using the proposed method further for localization and tracking of flowing microscopic samples.