Ratio-Free Detection and Partial Field Illumination Improve Time-Domain Dynamic Full-Field Optical Coherence Tomography Sensitivity for Retinal Organoid Imaging

Abstract

Time domain Dynamic full-field optical coherence tomography (D-FFOCT) is a powerful label-free imaging modality that enables functional visualization of cellular activity in living tissues with subcellular resolution. However, its sensitivity remains a major limitation for imaging highly scattering three-dimensional (3D) biological models such as retinal organoids, where incoherent background and inefficient optical flux distribution reduce dynamic contrast and limit imaging depth. In this work, we introduce a ratio-free optical configuration for time-domain D-FFOCT that enables continuous tuning of the sample-to-reference field ratio while minimizing photon losses and suppressing parasitic reflections. This polarization-based architecture allows optimal redistribution of optical flux according to sample scattering conditions and improves sensitivity under both power-limited and dose-limited conditions. Compared with conventional non-polarizing beam splitter configurations, the proposed approach provides a [Formula]-fold (3 dB) sensitivity improvement through optical optimization alone. In addition, we investigate for the first time the use of partial field illumination (PFI) in time-domain D-FFOCT to reduce incoherent background arising from multiple scattering. In retinal organoids imaged at 120 {micro}m depth, PFI yields up to a 14.5-fold (23.2 dB) increase in dynamic signal sensitivity, while preserving functional contrast. When combined, ratio-free detection and PFI provide a cumulative sensitivity improvement of 20.5-fold (26.2 dB). These gains enable improved visualization of photoreceptor precursor organization, rosette structures, and Muller glial cell dynamics in both 3D retinal organoids and 2D cell cultures. This work establishes a practical framework for sensitivity optimization in D-FFOCT and expands its potential for functional imaging, disease modelling, and live-cell monitoring in complex biological systems. O_FIG O_LINKSMALLFIG WIDTH=195 HEIGHT=200 SRC="FIGDIR/small/719402v1_ufig1.gif" ALT="Figure 1"> View larger version (123K): org.highwire.dtl.DTLVardef@1651e7org.highwire.dtl.DTLVardef@15b42e5org.highwire.dtl.DTLVardef@850180org.highwire.dtl.DTLVardef@25a3cc_HPS_FORMAT_FIGEXP M_FIG C_FIG