Critical steps and troubleshooting in sample preparation for wood and phloem formation: from sampling to microscopic observation

Abstract

We present a technical note that supplements published procedures on optimal sample preparation for performing wood and phloem formation analyses. Before beginning sampling, it is important to learn about the characteristics of the tree or shrub species to be investigated. Some tips are given how to use the Trephor tool in the best way, how to remove the outer hard bark (periderm), how microcores should be handled after removal from the tree, and how they should be oriented for embedding in paraffin, and cutting thin sections for microscopy. Possible defects that may result from improper handling are illustrated and discussed. We also present optimal images to accurately identify different cell development stages in phloem and xylem, which is particularly challenging in hardwoods and Mediterranean tree and shrub species.

Keywords

• tissue sampling
• tissue preparation
• microscopic slides
• microscopy
• xylogenesis
• phloem formation