# Providing maceration protocols for xylem and phloem research

## TL;DR

This paper provides detailed maceration protocols for studying plant vascular tissues, using safer chemicals and techniques suitable for various microscopy methods.

## Contribution

The paper introduces flexible, detailed maceration protocols using Franklin’s solution for studying xylem and phloem in diverse plant species.

## Key findings

- P1 allows non-permanent slide preparation for observing xylem and phloem under multiple microscopy techniques.
- P2 and P3 produce permanent slides, with P2 effective for over 34 plant families and P3 requiring minimal substances and time.
- The protocols enhance the applicability of maceration for ecological and evolutionary research.

## Abstract

Maceration technique allows isolating and studying the structure of plant cells, providing essential insights into plant physiology and responses to environmental factors. Despite its importance, the methods sections of publications often lack sufficient detail to properly apply maceration technique, preventing broader applications beyond industrial and wood identification studies. Here we describe maceration protocols (P), as guidelines, to allow successful cell separation, in woody and herbaceous plants, for further studying the structure of vascular tissues (xylem and phloem), using Franklin’s solution instead of the traditional, more toxic Jeffrey’s. We included sample preparation, equipment and chemicals, recommended stains and means of slide preparation that ensure clear observation and imaging, and identified potential pitfalls and safety practices. P1 provides a simple xylem and phloem maceration technique, with non-permanent slides preparation, suitable for observation under light, fluorescence or confocal microscopy. P2 and P3 are suitable for producing permanent slides of macerated xylem cells. P2 has been successfully used for macerating xylem of angiosperms and gymnosperms from at least 34 families and 25 orders, with densities ranging widely. P3 uses minimal substances and time for dehydration and staining. These flexible protocols may contribute to unlocking the potential of maceration technique to advance research in ecology and evolution.