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RetFM-J, an ImageJ-based module for automated counting and quantifying features of nuclei in retinal whole-mounts
Journal article   Open access   Peer reviewed

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

Adam Hedberg-Buenz, Mark A Christopher, Carly J Lewis, Kacie J Meyer, Danielle S Rudd, Laura M Dutca, Kai Wang, Mona K Garvin, Todd E Scheetz, Michael D Abràmoff, …
Experimental Eye Research, Vol.146, pp.386-392
05/2016
DOI: 10.1016/j.exer.2015.07.020
PMCID: PMC4753132
PMID: 26283021
url
http://doi.org/10.1016/j.exer.2015.07.020View
Open Access

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. •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.
ImageJ Image analysis Retinal whole-mount Retinal ganglion cell layer Cell counter Quantitative analysis

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