Abstract
Leaf surfaces are highly complex functional systems with well defined chemistry and structure dictating the barrier and transport properties of the leaf cuticle. It is a significant imaging challenge to analyse the very thin and often complex wax-like leaf cuticle morphology in their natural state. Scanning electron microscopy (SEM) and to a lesser extent Atomic force microscopy are techniques that have been used to study the leaf surface but their remains information that is difficult to obtain via these approaches. SEM is able to produce highly detailed and high-resolution images needed to study leaf structures at the submicron level. It typically operates in a vacuum or low pressure environment and as a consequence is generally unable to deal with the in situ analysis of dynamic surface events at submicron scales. Atomic force microscopy also possess the high-resolution imaging required and can follow dynamic events in ambient and liquid environments, but can over exaggerate small features and cannot image most leaf surfaces due to their inherent roughness at the micron scale. Scanning ion conductance microscopy (SICM), which operates in a liquid environment, provides a potential complementary analytical approach able to address these issues and which is yet to be explored for studying leaf surfaces. Here we illustrate the potential of SICM on various leaf surfaces and compare the data to SEM and atomic force microscopy images on the same samples. In achieving successful imaging we also show that SICM can be used to study the wetting of hydrophobic surfaces in situ. This has potentially wider implications than the study of leaves alone as surface wetting phenomena are important in a range of fundamental and applied studies.
Highlights
The surfaces of all leaves are covered with a thin waxy layer known as the cuticle (Holloway, 1993)
These waxes can be within the cuticle, called the cuticular waxes (CW) or form a layer above the biopolymer framework, called the epicuticular waxes (EW) (Jeffree 2006)
It is worthy of note that a related area that has received significant attention recently is the phenomenon of super hydrophobic surfaces, many of which are based on studying the natural structures formed at leaf surfaces (Bargel et al, 2006, Marmur, 2004)
Summary
The surfaces of all leaves are covered with a thin waxy layer known as the cuticle (Holloway, 1993). The EW is known to form crystalline aggregates which can form different structures according to the chemical content of the waxes (Barthlott et al, 1998), and can be categorized in to six main groups (Jeffree, 2006); massive crusts, filaments, rods, tubules, plates and platelets. These crystalline structures range in size from 0.2 to 100 μm (Koch and Ensikat, 2008). It is worthy of note that a related area that has received significant attention recently is the phenomenon of super hydrophobic surfaces, many of which are based on studying the natural structures formed at leaf surfaces (Bargel et al, 2006, Marmur, 2004)
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