Description
Data related to: Implantation and long-term assessment of the stability and biocompatibility of a novel 98 channel suprachoroidal visual prosthesis in sheep
In Eggenberger et al., nine sheep of the Suffolk (N=2) and Dorper (N=7) breeds were implanted in the left eye with a suprachoroidal retinal stimulator (Bionic Eye) for durations up to 100 days. The surgical safety, implant stability and device biocompatibility were assessed.
All procedures were performed conducted in accordance with the Australian Code for the Care and Use of Animals for Experimental Purposes 8th edition (2013) and the ARVO standards for use of animals in ophthalmic research and approved by the Animal Care and Ethics Committee (ACEC) of UNSW Sydney (protocol number 14/155B) .
Animal identifier:
Animals were identified according to the experimental group to which they were assigned (duration of experiment 2 days, 1 month, 2 months and 3 months.
E.g.: 2D#1 was the first animal in the group implanted for a duration of 2 Days. 3M#3 was the third animal in the group implanted for 3 Months.
Heamatoxylin and eosin staining:
Sections were then stained with Harris Hematoxylin (7 min; prior to bluing in running tap water for 4 min) and eosin (4 min).
Fluorescence Immunohistochemistry:
10um paraffin sections. GFAP-Cy3 conjugate (mouse IgG; 1:800); cone photoreceptors: opsin-red/green (opsin R/G) (rabbit IgG; 1:500, 1:1000) (Merck Millipore) and macrophage/microglia: Iba1 (goat IgG; 1:200) (Abcam, USA).
Secondary antibodies to goat IgG, mouse IgG, or rabbit IgG, conjugated with Alexa-Fluor 488, 555, or 647 (Invitrogen, USA).
Nuclei: Hoechst 33258 (1:10,000)
Mounting in glycerol:PBS (80:20)
All images were captured using an LSM 700 Meta Confocal microscope system and ZEN Blue software (Carl Zeiss, Germany). Images were assembled in Photoshop CS software (Adobe Corporation, USA).
Iba1 immunoperoxidase and quantitative analysis:
4um paraffin sections. High pH buffer heat antigen retrieval (Target Retrieval Solution, S2367, DAKO®), blocking (Bloxall, SP6000, VectorLabs®), primary antibody (macrophage/microglia, clone Iba1, BioCare Medical®, rabbit polyclonal, 1:200), secondary (Envision K5007, DAKO®), chromogen ImmPACT Nova Red (SK4805, Vector Labs®), counterstained: haematoxylin (Whitlocks, VPDS Histo Lab Manual©, 2021). Negative controls processed identically with primary antibody omitted.
Sections scanned with Aperio Versa (Leica Biosystems®, UK) at 40x.
Quantitative analysis using ImageScope™ Positive Pixel Count algorithm with the following parameters: hue value 0, hue width 0.4, colour saturation threshold 0.25, weak positive pixels (high) 160, positive pixels (high) 140, strong positive pixels (high) 120, strong positive pixels (low) 0, negative pixels (high) -1. Retinas outlined manually in ImageScope™ .