Ex-vivo-examination of ultrastructural changes in organotypic retina culture using near-infrared imaging and optical coherence tomography.
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.
Centre of Ophthalmology, University Eye Hospital Tübingen, Schleichstr. 12/1, D-72076, Tübingen, Germany. Electronic address: sven.schnichels@med.uni-tuebingen.de.
Sven
S
Schnichels
Centre of Ophthalmology, University Eye Hospital Tübingen, Schleichstr. 12/1, D-72076, Tübingen, Germany.
Tanja
T
Dorfi
Centre of Ophthalmology, University Eye Hospital Tübingen, Schleichstr. 12/1, D-72076, Tübingen, Germany.
Maximilian
M
Schultheiss
Centre of Ophthalmology, Institute for Ophthalmic Research Tübingen, Röntgenweg 11, D-72076, Tübingen, Germany.
Blanca
B
Arango-Gonzalez
Centre of Ophthalmology, University Eye Hospital Tübingen, Schleichstr. 12/1, D-72076, Tübingen, Germany.
Karl-Ulrich
KU
Bartz-Schmidt
Centre of Ophthalmology, University Eye Hospital Tübingen, Schleichstr. 12/1, D-72076, Tübingen, Germany; Eye Clinic Sulzbach-Saar, an der Klinik 10, 66111, Sulzbach-Saar, Germany.
Kai
K
Januschowski
Centre of Ophthalmology, University Eye Hospital Tübingen, Schleichstr. 12/1, D-72076, Tübingen, Germany; Department of Ophthalmology, University Medical Center Hamburg-Eppendorf (UKE), Martinistraße 52, 20251, Hamburg, Germany.
Martin S
MS
Spitzer
Centre of Ophthalmology, University Eye Hospital Tübingen, Schleichstr. 12/1, D-72076, Tübingen, Germany.
Focke
F
Ziemssen
D000818:Animals; D004195:Disease Models, Animal; D051381:Rats; D015203:Reproducibility of Results; D012160:Retina / Q000000981:diagnostic imaging* / Q000648:ultrastructure; D012164:Retinal Diseases / Q000000981:diagnostic imaging; D046509:Tissue Culture Techniques; D041623:Tomography, Optical Coherence / Q000379:methods*
Organ culture; Retinal thickness; SD-OCT; Structural integrity