Dynamic full-field optical coherence tomography module adapted to commercial microscopes allows longitudinal in vitro cell culture study

Abstract

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.

Deep learning based characterization of human organoids using optical coherence tomography

“…In this study, we employed noninvasive, 3D OCT imaging for human organoids, highlighting its advantages over traditional histological methods (requiring tissue fixation, sectioning, and staining). OCT's non-destructive nature [ 24 , 41 ] is pivotal for observing dynamic biological processes and long-term organoid development [ 24 , 42 , 43 ]. Its capability for rapid, real-time, high-resolution, 3D imaging surpasses the time-consuming and labor-intensive nature of traditional 2D histology, providing a more complete understanding of whole organoid architecture.…”

Line-field dynamic optical coherence tomography platform for volumetric assessment of biological tissues

“…While OCT was initially used to study particle dynamics (Brownian motion) in microsphere suspensions [ 16 , 17 ], the idea of using temporal fluctuations in the OCT signal generated by light scattered from single cells to study cell apoptosis was first proposed by Van der Meer et al [ 4 ]. Over the past decade, the range of biomedical applications of dOCT has expanded from imaging cell cultures and spheroids for cancer research [ 9 , 18 – 20 ], to imaging retinal organoids for evolutionary development studies [ 14 , 21 ], to ex-vivo morphological imaging of healthy and pathological tissues with enhanced contrast to visualize the tissue’s cellular structure [ 22 – 24 ].…”