Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas.
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...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4651327?pdf=render |
| _version_ | 1811225141113257984 |
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| author | Pablo de Gracia Beatriz I Gallego Blanca Rojas Ana I Ramírez Rosa de Hoz Juan J Salazar Alberto Triviño José M Ramírez |
| author_facet | Pablo de Gracia Beatriz I Gallego Blanca Rojas Ana I Ramírez Rosa de Hoz Juan J Salazar Alberto Triviño José M Ramírez |
| author_sort | Pablo de Gracia |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T09:01:54Z |
| format | Article |
| id | doaj.art-bac282e6aff44894941d2a64116343c3 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T09:01:54Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-bac282e6aff44894941d2a64116343c32022-12-22T03:39:13ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011011e014327810.1371/journal.pone.0143278Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas.Pablo de GraciaBeatriz I GallegoBlanca RojasAna I RamírezRosa de HozJuan J SalazarAlberto TriviñoJosé M RamírezProliferation 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.http://europepmc.org/articles/PMC4651327?pdf=render |
| spellingShingle | Pablo de Gracia Beatriz I Gallego Blanca Rojas Ana I Ramírez Rosa de Hoz Juan J Salazar Alberto Triviño José M Ramírez Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas. PLoS ONE |
| title | Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas. |
| title_full | Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas. |
| title_fullStr | Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas. |
| title_full_unstemmed | Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas. |
| title_short | Automatic Counting of Microglial Cells in Healthy and Glaucomatous Mouse Retinas. |
| title_sort | automatic counting of microglial cells in healthy and glaucomatous mouse retinas |
| url | http://europepmc.org/articles/PMC4651327?pdf=render |
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