Single-shot, dual-mode, water-immersion microscopy platform for biological applications

Appl Opt. 2018 Jan 1;57(1):A242-A249. doi: 10.1364/AO.57.00A242.

Abstract

A single-shot water-immersion digital holographic microscope combined with broadband (white light) illumination mode is presented. This double imaging platform allows conventional incoherent visualization with phase holographic imaging of inspected samples. The holographic architecture is implemented at the image space (that is, after passing the microscope lens), thus reducing the sensitivity of the system to vibrations and/or thermal changes in comparison to regular interferometers. Because of the off-axis holographic recording principle, quantitative phase images of live biosamples can be recorded in a single camera snapshot at full-field geometry without any moving parts. And, the use of water-immersion imaging lenses maximizes the achievable resolution limit. This dual-mode microscope platform is first calibrated using microbeads, then applied to the characterization of fixed cells (neuroblastoma, breast cancer, and hippocampal neuronal cells) and, finally, validated for visualization of dynamic living cells (hippocampal neurons).

MeSH terms

  • Calibration
  • Equipment Design
  • Holography / instrumentation
  • Holography / methods*
  • Immersion
  • Lighting / methods*
  • Microscopy / instrumentation
  • Microscopy / methods*