Computational protocol: Glioprotection of Retinal Astrocytes After Intravitreal Administration of Memantine in the Mouse Optic Nerve Crush Model

Similar protocols

Protocol publication

[…] Immunostained whole-mount retinae were investigated by fluorescent microscopy using a Zeiss Axio Imager M2 (Carl Zeiss AG, Jena, Germany) (). Astrocyte counts for each image were performed manually using ImageJ 1.49m software (Wayne Rasband, National Institutes of Health, USA). Twelve randomly chosen images of 222×170 μm in size were taken from each retina for further evaluation (). Astrocytic somata were manually counted from each image. To achieve credible results, astrocyte somas were counted in the whole image, including left and lower borders. Astrocytic somata that crossed right and upper borders were excluded. Counts were taken manually and in a blinded manner by 2 examiners.Semi-thin sections of the optic nerves were analyzed by light microscopy (Zeiss Axio Imager M1, Carl Zeiss AG, Jena, Germany), under 100x magnification. We randomly acquired 10 images of each optic nerve and evaluated ONC damage by grading it as control, mild, moderate, or severe []. [...] IBM SPSS Statistics 22.0 Program Package (IBM Corporation, USA) was used for statistical analysis. The Kruskal-Wallis test was used to compare the 3 groups. A P value of p<0.05 was considered statistically significant. Inter-observer variability was examined using intra-class correlation coefficient (ICC) statistics. ICC cut-off values were defined as follows: <0.40 – poor agreement; 0.41–0.60 – moderate agreement; 0.61–0.79 – good agreement; and ≥0.80 – excellent agreement [,]. […]

Pipeline specifications

Software tools ImageJ, SPSS
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Mus musculus
Diseases Glaucoma, Ocular Hypertension, Soft Tissue Injuries, Neurodegenerative Diseases
Chemicals Memantine, Methylene Blue, N-Methylaspartate, Glutamic Acid