Stephen D Jonhston
Australia
Research Article
Semi-Quantitative Assessment of Superoxide in the Human Neat Ejaculate Using a Nitro Blue Tetrazoilum Based Assay: A Comparison of Visual, Spectrophotometric and Image Analysis Approaches
Author(s): Moises de la Casa, José Luís Fernández, Vicente Badajoz, Carmen López-Fernández, Stephen D Jonhston and Jaime GosálvezMoises de la Casa, José Luís Fernández, Vicente Badajoz, Carmen López-Fernández, Stephen D Jonhston and Jaime Gosálvez
Purpose: To evaluate the efficacy of three colorimetric methods for detecting the presence of SuperOxide (SOx) in the neat human ejaculate based on the transformation of Nitro Blue Tetrazolium (NBT) to formazan.
Methods: The resultant color after reacting 230 neat semen samples for 30 min after ejaculation was assessed semi-quantitatively and the results compared using (1) a simple visual assessment, (2) spectrophotometry and (3) an image analysis methodology based on direct scanned digital images.
Results: After comparing visual classification with the spectrophotometric values and image analysis, 8 % and 6 %, of cases were misclassified, respectively. However, Pearson correlation calculated between values obtained by visual versus spectrophotometric or image analysis assessments were extremely high (P=0.000). The str.. View More»
