Depth enhancement of 3D microscopic living-cell image using incoherent fluorescent digital holography

Abstract

Multilayer images of living cells are typically obtained using confocal or multiphoton microscopy. However, limitations on the distance between consecutive scan layers hinder high-resolution three-dimensional reconstruction, and scattering strongly degrades images of living cell components. Consequently, when overlapping information from different layers is focused on a specific point in the camera, this causes uncertainty in the depiction of the cell components. We propose a method that combines the Fresnel incoherent correlation holography and a depth-of-focus reduction algorithm to enhance the depth information of three-dimensional cell images. The proposed method eliminates overlap between light elements in the different layers inside living cells and limitations on the interlayer distance, and also enhances the contrast of the reconstructed holograms of living cells.

Keywords: Digital holography, holographic display; image recognition; microscopy; phase modulation; spatial light modulator.