4.5. Quantitative Image Analysis

For image analysis, data were collected from at least three sections of DRG tissue for each rat. At a minimum, three randomized microscopic fields (50 μm × 50 μm) were captured in one section, and averages were used for further analysis. Five of seven rats were randomly selected from each group at each point time for immunofluorescence staining. The number of positive cells in the images was blindly counted using Image J software (National Institutes of Health, Bethesda, MD, USA) [15] and averaged for further analysis. To determine the percentage of labeled neurons in the DRG, the number of positive neurons (three times the background signal) was divided by the total number of neurons [6].