RetFM-J, an ImageJ-based module for automated counting and quantifying features of nuclei in retinal whole-mounts. (May 2016)

• Record Type: Journal Article
• Authors: Hedberg-Buenz, Adam; Christopher, Mark A.; Lewis, Carly J.; Meyer, Kacie J.; Rudd, Danielle S.; Dutca, Laura M.; Wang, Kai; Garvin, Mona K.; Scheetz, Todd E.; Abràmoff, Michael D.; Harper, Matthew M.; Anderson, Michael G.
• Is Part Of: Experimental eye research. Volume 146(2016:May)
• Page Start: 386
• Page End: 392
• Publication Date: 2016-05
• Subjects: Retinal whole-mount -- Retinal ganglion cell layer -- Image analysis -- Quantitative analysis -- Cell counter -- ImageJ; Ophthalmology -- Periodicals; Eye -- Periodicals; Œil -- Périodiques; Ophthalmology; Periodicals; Electronic journals; 612.8405
• DOI: 10.1016/j.exer.2015.07.020
• Languages: English
• ISSNs: 0014-4835

Abstract

Abstract: The present article introduces RetFM-J, a semi-automated ImageJ-based module that detects, counts, and collects quantitative data on nuclei of the inner retina from H&E-stained whole-mounted retinas. To illustrate performance, computer-derived outputs were analyzed in inbred C57BL/6J mice. Automated characterization yielded computer-derived outputs that closely matched manual counts. As a method using open-source software that is freely available, inexpensive staining reagents that are robust, and imaging equipment that is routine to most laboratories, RetFM-J could be utilized in a wide variety of experiments benefiting from high-throughput, quantitative, uniform analyses of total cellularity in the inner retina. Highlights: RetFM-J counts and quantifies features for all nuclei in the inner retina. RetFM-J is packaged as a freely available plugin on the open ImageJ-based platform. Protocols for dissection and H&E staining of mouse retinal whole-mounts are detailed.