Ex-vivo-examination of ultrastructural changes in organotypic retina culture using near-infrared imaging and optical coherence tomography

•OCT-measurements were adapted to organotypic cultures.•in-vitro in-depth analysis of the retina and its layers are possible.•an easy to use protocol with commercially available OCT devices is given.•Animal experiments can be reduced with this technique.

Optical coherence tomography (OCT) dramatically changed the way of diagnostic assessment in retinal diseases during the last years. Using this technique in-vivo in-depth analysis of the retina and its layers is possible. Since animal research is changing by intrinsic and extrinsic pressure to animal-(in-vivo)-free methods, we adapted OCT-measurements to organotypic cultures. An easy to use protocol was generated to assess standardized OCT assessments in organotypic culture. First, two custom-made devices need to be made to change any commercially available OCT for examinations in humans into a device allowing ex-vivo analyses of organotypic culture. The modification is feasible within seconds. After OCT measurement of the ex-vivo tissues, quantitative evaluation of the retinas were performed via ImageJ software. OCT pictures of ex-vivo retinas were obtained for time periods of seven days and the thickness of retinal tissue was evaluated. The reproducibility of the pictures and measurements was very high (SD < 15%). In conclusion, an easy to use protocol for the investigation of different effects on retinal cultures with commercially available OCT devices was successfully established.