Abstract
We describe recent technological progress in multimodal en face full-field optical coherence tomography that has allowed detection of slow and fast dynamic processes in the eye. We show that by combining static, dynamic and fluorescence contrasts we can achieve label-free high-resolution imaging of the retina and anterior eye with temporal resolution from milliseconds to several hours, allowing us to probe biological activity at subcellular scales inside 3D bulk tissue. Our setups combine high lateral resolution over a large field of view with acquisition at several hundreds of frames per second which make it a promising tool for clinical applications and biomedical studies. Its contactless and non-destructive nature is shown to be effective for both following in vitro sample evolution over long periods of time and for imaging of the human eye in vivo.
Highlights
Optical coherence tomography (OCT) is used in the biomedical field to image microstructures in tissue using mostly the endogenous backscattering contrast [1, 2]
The automated plane locking procedure to correct for axial drift means that we are able to repeatedly acquire 3D stacks beginning at the exact same plane, making 3D time-lapse imaging possible and ensuring perfect coincidence of multimodal Full-field optical coherence tomography (FFOCT)/dynamic FFOCT (D-FFOCT) stacks
As a result of the improvements in sample stability and time-lapse acquisition offered by the inverted setup, time-lapse imaging over periods of hours, in conjunction with the fast dynamic signal which creates the D-FFOCT contrast in a single acquisition, can be reliably performed
Summary
Optical coherence tomography (OCT) is used in the biomedical field to image microstructures in tissue using mostly the endogenous backscattering contrast [1, 2]. New multimodal techniques, based either on OCT or FFOCT, have enabled the study of the static 3D morphology of a sample and of the dynamics of its scatterers [7, 8] by measuring phase-sensitive temporal fluctuations of the backscattered light. We have demonstrated that low order geometrical aberrations do not affect the point spread function (PSF) but mostly reduce the signal to noise ratio (SNR) when using interferometry with a spatially incoherent source [18, 19] This allows us to achieve resolution of cone photoreceptors in vivo in the human retina without adaptive optics [19]. An approach that combines fluorescence labeling for live cells with static and dynamic FFOCT is presented
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