Improved handling and embedding schemes for cultured murine neuroretinal explants

Abstract

ABSTRACT Traumatic, inherited, and age-related degenerative diseases of the retina, such as retinal detachment, retinitis pigmentosa, and age-related macular degeneration, are characterized by the irreversible loss of retinal neurons. While current treatments aim to prevent neuronal degeneration, there are no available treatments to restore neurons after loss. Cultured murine neuroretinal tissue explants model retinal injury and offer a high throughput approach to identify experimental interventions capable of regenerating neurons. Formalin-fixed paraffin-embedded (FFPE) preparations of murine neuroretinal explants can be used to identify cells throughout the retinal layers to provide information on proliferation and activity following exposure to therapeutics. However, retinal explants are friable, particularly after ex vivo culture, sample handling and FFPE processing steps can result in tissue loss and damage. Friability also prohibits bisecting samples post-culture to display more than one region of interest for analysis. We developed a sample handling and embedding technique for cultured murine neuroretinal explants using HistogelTM in combination with a post-processing trimming step that eliminates tissue loss, increases cross-sectional retinal representation, and captures proximal and central retina on one slide to facilitate analysis of explants subjected to neurotrophic compounds.