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 organization processes such as rosette formation and mitosis as well as cell shape and motility. 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.

Dynamic optical coherence tomography (DOCT) enhances conventional OCT by providing specific information related to flow dynamics, cell motility, and organelle metabolic activity. These biological phenomena can be detected with varying sensitivity depending on the OCT architecture parameters, including wavelength, numerical aperture, and implementation method (time domain or Fourier domain). Despite its potential, the field lacks standardization as various research groups have independently developed algorithms for specific applications. In this paper, we compare four widely used DOCT algorithms, each employing a distinct analytical approach: power spectral density moment analysis, frequency band visualization, logarithmic intensity variation evaluation, and motility-based analysis. These algorithms were originally optimized for different OCT technologies (full-field OCT, microscopic OCT, swept-source OCT, and spectral domain OCT), which vary in temporal and spatial resolution as well as susceptibility to motion artifacts. To conduct a fair evaluation, we perform comprehensive cross-wise comparisons using datasets acquired from each of these setups. Our findings reveal that each method exhibits unique advantages in specific imaging environments, thereby providing valuable guidance for algorithm selection based on particular application requirements.

Dynamic full-field optical coherence tomography (DFFOCT) has recently emerged as an invaluable label-free microscopy technique, owing to its sensitivity to cell activity, as well as speed and sectioning ability. However, the quality of DFFOCT images is often degraded due to phase noise and fringe artifacts. In this work, we present a new implementation, to the best of our knowledge, named rolling-phase (RP) DFFOCT, in which the reference arm is slowly scanned over magnitudes exceeding 2π. We demonstrate mathematically and experimentally that it shows superior image quality while enabling to extract both static and dynamic contrast simultaneously. We showcase RP-DFFOCT on a macaque retinal explant and demonstrate its ability to better resolve subcellular structures, including intranuclear activity.

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 $\sqrt{2}$-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 $\mathsf{\mu }$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 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.

Dynamic full-field optical coherence tomography (DFFOCT) has recently emerged as an invaluable label-free microscopy technique, owing to its sensitivity to cell activity, as well as speed and sectioning ability. However, the quality of DFFOCT images are often degraded due to phase noise and fringe artifacts. In this work, we present a new implementation named Rolling-Phase (RP) DFFOCT, in which the reference arm is slowly scanned over magnitudes exceeding 2$π$. We demonstrate mathematically and experimentally that it shows superior image quality while enabling to extract both static and dynamic contrast simultaneously. We showcase RP DFFOCT on monkey retinal explant, and demonstrate its ability to better resolve subcellular structures, including intranuclear activity.