Multiplane differential phase contrast imaging using asymmetric illumination in volume holographic microscopy

J Biomed Opt. 2020 Nov;25(12):123704. doi: 10.1117/1.JBO.25.12.123704.

Abstract

Significance: Differential phase contrast (DPC) is a well-known imaging technique for phase imaging. However, simultaneously acquiring multidepth DPC images is a non-trivial task. We propose simultaneous multiplane DPC imaging using volume holographic microscopy (VHM).

Aim: To design and implement a new configuration of DPC-VHM for multiplane imaging.

Approach: The angularly multiplexed volume holographic gratings (AMVHGs) and the wavelength-coded volume holographic gratings (WC-VHGs) are used for this purpose. To obtain asymmetric illumination for DPC images, a dynamic illumination system is designed by modifying the regular Köhler illumination using a thin film transistor panel (TFT-panel).

Results: Multidepth DPC images of standard resolution chart and biosamples were used to compare imaging performance with the corresponding bright-field images. An average contrast enhancement of around three times is observed for target resolution chart by DPC-VHM. Imaging performance of our system is studied by modulation transfer function analysis, which suggests that DPC-VHM not only suppresses the DC component but also enhances high-frequency information.

Conclusions: Proposed DPC-VHM can acquire multidepth-resolved DPC images without axial scanning. The illumination part of the system is adjustable so that the system can be adapted to bright-field mode, phase contrast mode, and DPC mode by controlling the pattern on the TFT-panel.

Keywords: diffraction gratings; holography; illumination design; imaging systems; microscopy; volume gratings.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Diagnostic Tests, Routine
  • Holography*
  • Lighting
  • Microscopy*
  • Microscopy, Phase-Contrast