Dynamic full-field swept-source optical coherence microscope for cellular-resolution, long-depth, and intratissue-activity imaging

TL;DR

This paper introduces a novel full-field swept-source optical coherence microscope that achieves cellular resolution, long imaging depth, and dynamic intra-tissue activity visualization through computational refocusing and innovative acquisition protocols.

Contribution

It presents a new spatially coherent full-field OCT method with computational refocusing and dynamic imaging capabilities, overcoming depth and activity visualization limitations.

Findings

Achieved 1.4 μm lateral resolution and 6.5 μm axial resolution.

Enabled volumetric dynamic imaging of tissue spheroids.

Demonstrated cellular-resolution imaging over full tissue depth.

Abstract

Optical coherence tomography (OCT) microscope (OCM) uses a high-numerical-aperture objective to achieve cellular-level lateral resolution. However, its practical imaging depth range is limited by the depth of focus (DOF). Although computational refocusing can potentially provide sharp images outside the DOF, signal reduction by the confocal effect still limits the imaging depth in practice in point-scanning OCT. In addition, standard OCT cannot visualize intra-tissue activities. To overcome these limitations, we demonstrated a spatially coherent full-field OCM (SC-FFOCM) with computational refocusing. In addition, a repetitive acquisition protocol was designed to visualize intra-tissue activities (i.e., dynamic OCT imaging). The in-focus lateral resolution is 1.4 um, and the axial resolution is 6.5 um (in air) at full-width at half-maximum intensity. Three-dimensional structure and the dynamic OCT imaging using SC-FFOCM with computational refocusing was applied to human breast adenocarcinoma spheroids (MCF-7 cell line). Volumetric dynamic imaging with cellular-level lateral resolution was demonstrated over the full depth of the spheroid.