Dynamic full-field optical coherence tomography (D-FFOCT) has recently emerged as a label-free imaging tool, capable of resolving cell types and organelles within 3D live samples, whilst monitoring their activity at tens of milliseconds resolution. Here, a D-FFOCT module design is presented which can be coupled to a commercial microscope with a stage top incubator, allowing non-invasive label-free longitudinal imaging over periods of minutes to weeks on the same sample. Long term volumetric imaging on human induced pluripotent stem cell-derived retinal organoids is demonstrated, highlighting tissue and cell organisation as well as cell shape, motility and division. Imaging on retinal explants highlights single 3D cone and rod structures. An optimal workflow for data acquisition, postprocessing and saving is demonstrated, resulting in a time gain factor of 10 compared to prior state of the art. Finally, a method to increase D-FFOCT signal-to-noise ratio is demonstrated, allowing rapid organoid screening.
Label-free live optical imaging of dynamic cellular and subcellular features has been made possible in recent years thanks to the advances made in optical imaging techniques, including dynamic optical coherence tomography (D-OCT) methods. These techniques analyze the temporal fluctuations of an optical signal associated with the active movements of intracellular organelles to obtain an ensemble metric recapitulating the motility and metabolic state of cells. They hence enable visualization of cells within compact, static environments and evaluate their physiology. These emerging microscopies show promise, in particular for the three-dimensional evaluation of live tissue samples such as freshly excised biopsies and 3D cell cultures. In this review, we compare the various techniques used for dynamic OCT. We give an overview of the range of applications currently being explored and discuss the future outlook and opportunities for the field.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.