Cell Motility as Contrast Agent in Retinal Explant Imaging With Full-Field Optical Coherence Tomography

Invest Ophthalmol Vis Sci. 2017 Sep 1;58(11):4605-4615. doi: 10.1167/iovs.17-22375.

Abstract

Purpose: To use cell motility as a contrast agent in retinal explants.

Methods: Macaque and mouse retinal explants were imaged with high resolution full field optical coherence tomography (FFOCT) and dynamic FFOCT, coupled with fluorescence imaging.

Results: Static and dynamic FFOCT create complementary contrast from different structures within a cell. When imaging in vitro samples, static FFOCT detects steep refractive index gradients to reveal stationary structures including fibers, vessels, collagen, and cell contours, while dynamic FFOCT emphasizes metabolic activity of moving structures that are mainly intracellular, thus creating or enhancing contrast in cells that were previously hidden in noise. Dynamic FFOCT enables detection of most of the retinal cell types in the ganglion cell, inner and outer nuclear layers, where static FFOCT contrast is too low in relation to the noise background.

Conclusions: Composite static and dynamic FFOCT provides a new kind of FFOCT image containing valuable information for imaging of retinal explants. It provides label-free en face images of living retinas, with a subcellular resolution. Dynamic FFOCT adds information about cell activity, which is of interest in longitudinal explant studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers
  • Cell Movement / physiology*
  • Contrast Media*
  • Dependovirus / genetics
  • Genetic Vectors
  • Green Fluorescent Proteins / genetics
  • Macaca
  • Mice
  • Mice, Inbred C57BL
  • Microscopy, Fluorescence
  • Recombinant Fusion Proteins
  • Retina / cytology*
  • Retinal Ganglion Cells / cytology*
  • Rhodopsin / genetics
  • Tomography, Optical Coherence / methods*

Substances

  • Biomarkers
  • Contrast Media
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • Rhodopsin