An open-source computational tool to automatically quantify immunolabeled retinal ganglion cells.
A fully automated and robust method was developed to quantify β-III-tubulin-stained retinal ganglion cells, combining computational recognition of individual cells by CellProfiler and a machine-learning tool to teach phenotypic classification of the retinal ganglion cells by CellProfiler Analyst. In animal models of glaucoma, quantification of immunolabeled retinal ganglion cells is currently performed manually and remains time-consuming. Using this automated method, quantifications of retinal ganglion cell images were accelerated tenfold: 1800 images were counted in 3 h using our automated method, while manual counting of the same images took 72 h. This new method was validated in an established murine model of microbead-induced optic neuropathy. The use of the publicly available software and the method's user-friendly design allows this technique to be easily implemented in any laboratory.
Anesthesia Center for Critical Care Research, Department of Anesthesia, Critical Care, and Pain Medicine, Massachusetts General Hospital Research Institute, Harvard Medical School, Boston, MA, USA.
Ana C
AC
Dordea
Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Mark-Anthony
MA
Bray
Anesthesia Center for Critical Care Research, Department of Anesthesia, Critical Care, and Pain Medicine, Massachusetts General Hospital Research Institute, Harvard Medical School, Boston, MA, USA.
Kaitlin
K
Allen
Broad Institute of MIT and Harvard, Cambridge, MA, USA.
David J
DJ
Logan
Department of Ophthalmology, Schepens Eye Research Institute, Harvard Medical School, Boston, MA, USA.
Fei
F
Fei
Division of Cardiology, Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
Rajeev
R
Malhotra
Department of Ophthalmology, Schepens Eye Research Institute, Harvard Medical School, Boston, MA, USA.
Meredith S
MS
Gregory
Broad Institute of MIT and Harvard, Cambridge, MA, USA.
Anne E
AE
Carpenter
Anesthesia Center for Critical Care Research, Department of Anesthesia, Critical Care, and Pain Medicine, Massachusetts General Hospital Research Institute, Harvard Medical School, Boston, MA, USA. Electronic address: ebuys@mgh.harvard.edu.
Emmanuel S
ES
Buys
D000818:Animals; D002452:Cell Count / Q000379:methods; D019295:Computational Biology / Q000379:methods*; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D051379:Mice; D012044:Regression Analysis; D012165:Retinal Ganglion Cells / Q000166:cytology*; D012984:Software
Automated quantification; CellProfiler; CellProfiler analyst; Retinal ganglion cell