A novel, rapid technique for clearing leaf tissues

Abstract

Premise Clearing leaves is a highly useful practice for many taxonomic, ecological, physiological, and eco‐physiological aspects of research. Using traditional methods, the procedure for clearing a leaf (referred to as diaphanization) can take several days or even weeks. In our laboratory we developed a technique, originally used for dissociating wood, that yields excellent epidermal and leaf venation preparations clearly showing the details of epidermal cells, hydathodes, trichomes, leaf margins, and leaf venation, in a maximum of three days. Methods and Results A solution originally used to macerate wood (Franklin’s solution) was used to accelerate the process of clearing leaves. Using this approach, it is possible to obtain clear images of the leaf adaxial and abaxial epidermal surfaces and leaf venation. Our technique works with both fragile and sturdy leaves, as well as with thin roots and stems. A few examples are presented. Conclusions Franklin’s solution is very simple to prepare and easy to manipulate. This new technique has the advantage of separating the leaf epidermal layers and venations, which can then be clearly observed.

Keywords

• diaphanization
• leaf clearing
• leaf epidermis
• leaf venation
• stomata
• trichomes