Sensitivity Enhancement of Dynamic Full-Field Optical Coherence Tomography Using Ratio-Free Detection and...

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 2-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 μm 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 cellular-scale visualization in retinal organoids, including cell-resolved imaging within rosette regions, as well as improved detection of intracellular dynamics in Müller glial cell cultures. This work establishes a practical framework for sensitivity optimization in D-FFOCT and expands its potential for functional imaging, disease modeling, and live-cell monitoring in complex biological systems.