Sorbonne Université, INSERM, CNRS, Institut de la Vision, 17 rue Moreau, F-75012, Paris, France.
CHNO des Quinze-Vingts, INSERM-DGOS CIC 1423, 28 rue de Charenton, F-75012, Paris, France.
Commun Biol. 2023 Sep 28;6(1):992. doi: 10.1038/s42003-023-05378-w.
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.
动态全场光学相干断层扫描（D-FFOCT）最近作为一种无标记成像工具出现，能够在 3D 活样本中分辨细胞类型和细胞器，同时以数十毫秒的分辨率监测它们的活性。这里提出了一种 D-FFOCT 模块设计，可以与带顶部孵育器的商用显微镜耦合，从而可以在同一样本上进行非侵入性无标记的纵向成像，时间从数分钟到数周不等。对人诱导多能干细胞衍生的视网膜类器官进行了长期的体积成像，突出了组织和细胞组织过程，如玫瑰花结形成和有丝分裂以及细胞形状和运动。对视网膜外植体的成像突出了单个 3D 锥体和杆状结构。演示了一种用于数据采集、后处理和保存的最佳工作流程，与之前的最先进技术相比，时间增益因子提高了 10 倍。最后，演示了一种提高 D-FFOCT 信噪比的方法，允许快速筛选类器官。
