RT Journal Article
T1 Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas
A1 Gracia Pacheco, Pablo de
A1 Gallego Collado, Beatriz Isabel
A1 Rojas López, María Blanca
A1 Ramírez Sebastián, Ana Isabel
A1 Hoz Montañana, María Rosa De
A1 Salazar Corral, Juan José
A1 Triviño Casado, Alberto
A1 Ramírez Sebastián, José Manuel
AB Proliferation of microglial cells has been considered a sign of glial activation and a hallmark of ongoing neurodegenerative diseases. Microglia activation is analyzed in animal models of different eye diseases. Numerous retinal samples are required for each of these studies to obtain relevant data of statistical significance. Because manual quantification of microglial cells is time consuming, the aim of this study was develop an algorithm for automatic identification of retinal microglia. Two groups of adult male Swiss mice were used: age-matched controls (naïve, n = 6) and mice subjected to unilateral laser-induced ocular hypertension (lasered; n = 9). In the latter group, both hypertensive eyes and contralateral untreated retinas were analyzed. Retinal whole mounts were immunostained with anti Iba-1 for detecting microglial cell populations. A new algorithm was developed in MATLAB for microglial quantification; it enabled the quantification of microglial cells in the inner and outer plexiform layers and evaluates the area of the retina occupied by Iba-1+ microglia in the nerve fiber-ganglion cell layer. The automatic method was applied to a set of 6,000 images. To validate the algorithm, mouse retinas were evaluated both manually and computationally; the program correctly assessed the number of cells (Pearson correlation R = 0.94 and R = 0.98 for the inner and outer plexiform layers respectively). Statistically significant differences in glial cell number were found between naïve, lasered eyes and contralateral eyes (P<0.05, naïve versus contralateral eyes; P<0.001, naïve versus lasered eyes and contralateral versus lasered eyes). The algorithm developed is a reliable and fast tool that can evaluate the number of microglial cells in naïve mouse retinas and in retinas exhibiting proliferation. The implementation of this new automatic method can enable faster quantification of microglial cells in retinal pathologies.
PB Public Library of Science
SN 1932-6203
YR 2015
FD 2015-11-18
LK https://hdl.handle.net/20.500.14352/24694
UL https://hdl.handle.net/20.500.14352/24694
LA eng
NO Gracia Pacheco, P., Gallego Collado, B. I., Rojas López, M. B. et al. «Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas». PLOS ONE, vol. 10, n.o 11, noviembre de 2015, p. e0143278. PLoS Journals, https://doi.org/10.1371/journal.pone.0143278.
NO Ministerio de Economía, Comercio y Empresa (España)
NO Instituto de Salud Carlos III
NO Science Foundation Arizona
NO Fondo Europeo de Desarrollo Regional
DS Docta Complutense
RD 6 abr 2025