We developed a new endogenous approach to reveal subcellular metabolic contrast in fresh ex vivo tissues taking advantage of the time dependence of the full field optical coherence tomography interferometric signals. This method reveals signals linked with local activity of the endogenous scattering elements which can reveal cells where other OCT based techniques fail or need exogenous contrast agents. We benefit from the micrometric transverse resolution of full-field OCT to image intracellular features. We used this time dependence to identify different dynamics at the millisecond scale on a wide range of organs in normal or pathological conditions.
We applied quantitative dynamic full-field optical coherence tomography (D-FFOCT) to image retinal explants which are a platform for investigating retinal development, pathophysiology, and cellular therapies. In contrast to histologic analysis and immuno-fluorescence imaging in which multiple specimens fixed at different times are used to reconstruct tissue morphology under various conditions, D-FFOCT imaging can provide repeated images and analysis of the same in vitro tissue with a typical contrast linked to metabolic information and achieve 500nm resolution. We are able to reconstruct three-dimensional volume and follow-up cellular activity in the same plane during several hours.
We applied dynamic FFOCT to imaging of Retinal Pigment Epithelium (RPE) cell culture. After 3D registration we were able to reconstruct 3D volumes.
Thouvenin O, Boccara C, Fink M, Sahel J, Paques M, Grieve K.
Invest Ophthalmol Vis Sci. 2017;58:4605–4615. DOI:10.1167/iovs.17-22375