RetFM-J, an ImageJ-based module for automated counting and quantifying features of nuclei in retinal whole-mounts.
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.
VA Center for the Prevention and Treatment of Visual Loss, Iowa City VA Health Care System, Iowa City, IA, USA; Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA 52242, USA.
Adam
A
Hedberg-Buenz
Department of Biomedical Engineering, University of Iowa, Iowa City, IA 52242, USA.
Mark A
MA
Christopher
Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA 52242, USA.
Carly J
CJ
Lewis
Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA 52242, USA.
Kacie J
KJ
Meyer
VA Center for the Prevention and Treatment of Visual Loss, Iowa City VA Health Care System, Iowa City, IA, USA.
Danielle S
DS
Rudd
VA Center for the Prevention and Treatment of Visual Loss, Iowa City VA Health Care System, Iowa City, IA, USA; Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52242, USA.
Laura M
LM
Dutca
Department of Biostatistics, University of Iowa, Iowa City, IA 52242, USA.
Kai
K
Wang
VA Center for the Prevention and Treatment of Visual Loss, Iowa City VA Health Care System, Iowa City, IA, USA; Department of Electrical and Computer Engineering, University of Iowa, Iowa City, IA 52242, USA.
Mona K
MK
Garvin
Department of Biomedical Engineering, University of Iowa, Iowa City, IA 52242, USA; Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52242, USA.
Todd E
TE
Scheetz
VA Center for the Prevention and Treatment of Visual Loss, Iowa City VA Health Care System, Iowa City, IA, USA; Department of Biomedical Engineering, University of Iowa, Iowa City, IA 52242, USA; Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52242, USA; Department of Electrical and Computer Engineering, University of Iowa, Iowa City, IA 52242, USA.
Michael D
MD
Abràmoff
VA Center for the Prevention and Treatment of Visual Loss, Iowa City VA Health Care System, Iowa City, IA, USA; Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52242, USA.
Matthew M
MM
Harper
VA Center for the Prevention and Treatment of Visual Loss, Iowa City VA Health Care System, Iowa City, IA, USA; Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA 52242, USA; Department of Ophthalmology and Visual Sciences, University of Iowa, Iowa City, IA 52242, USA. Electronic address: michael-g-anderson@uiowa.edu.
Michael G
MG
Anderson
D000818:Animals; D002452:Cell Count / Q000379:methods*; D002467:Cell Nucleus; D003936:Diagnosis, Computer-Assisted; D003941:Diagnostic Techniques, Ophthalmological; D007091:Image Processing, Computer-Assisted; D051379:Mice; D008810:Mice, Inbred C57BL; D008853:Microscopy / Q000379:methods; D023421:Models, Animal; D012160:Retina / Q000000981:diagnostic imaging*; D012165:Retinal Ganglion Cells / Q000166:cytology*
Cell counter; Image analysis; ImageJ; Quantitative analysis; Retinal ganglion cell layer; Retinal whole-mount