Quantification of intracellular seed compounds of Moringa oleifera Lam. using digital image analysis

Histochemical staining of sectioned seed for the identification of various intracellular compounds has successfully been used in the past. The estimation of storage compound content through the quantification of a specific compound selecting stain through digital image analysis using Adobe (R) Photoshop (R) has however not been explored to date and therefore, became the main aim of this study. Moringa oleifera seeds of various developmental stages were prepared for microscopic analysis and stained with oil (Sudan III), protein (Light Green SF, Orange G) and starch (Periodic Acid Schiff) detecting stains. The percentage coverage of a specific stain in digital images of sectioned seed was determined using Adobe (R) Photoshop (R) and compared to analytical data. Protein stain: Orange G and oil detecting stain: Sudan III demonstrated the best correlation to the measured seed protein and oil content throughout seed development. For simple comparisons between storage compound contents at different developmental stages, image stain quantification proved reasonably effective.