Combining multispectral and high-resolution 3D imaging for leaf vein segmentation and density measurement.

Accurate leaf vein segmentation and vein density (VLA) measurement are crucial for understanding plant physiology. Traditional 2D imaging techniques often require labor-intensive and destructive processes, such as leaf flattening or chemical clearing, thereby limiting their practicality for high-throughput applications. In this study, we present a novel framework that integrates multispectral and high-resolution 3D imaging to enhance leaf vein segmentation and VLA measurement. By leveraging digital fringe projection, our system captures grayscale, multispectral, and 3D topographical data within a unified coordinate system. The integration of 3D information improves vein detection, particularly in low-contrast regions, while also enabling direct and accurate measurements of leaf area, vein length, and VLA. However, this approach also introduces some false positives in vein segmentation due to mesophyll surface variability. Despite these challenges, our high-resolution 3D imaging method shows significant potential for non-invasive phenotyping and trait assessment in complex, unstructured environments.