Perforated-tape Epidermal Detachment (PED): A simple and rapid method for isolating epidermal peels from specific areas of Arabidopsis leaves

Abstract

Leaf epidermal stomata play pivotal roles in gas exchange and transpiration in higher plants. The regulatory mechanisms of the opening and closing of stomata (stomatal movement) and stomatal morphogenesis have been intensively studied. Such studies require the collection of detached epidermal cell layers ("peels") for the detailed observation of stomata under the microscope. However, there are drawbacks to the conventional techniques for preparing and handling epidermal peels, i.e., a certain level of skill or an apparatus (e.g., a Waring blender) is required. In this report, we present a simple and rapid method for preparing epidermal peels called Perforated-tape Epidermal Detachment (PED). For PED, Time Tape is adhered to the adaxial epidermis of an Arabidopsis leaf, and the abaxial epidermal layer is detached using Scotch Tape perforated with a small hole. The area inside the hole is suitable for microscopic observation because the Scotch Tape does not mask the detached epidermal layer there. PED can also be used to prepare epidermal peels from tobacco and legume leaves. The crucial advantage of PED over the conventional blender method is that the epidermal cells prepared by PED are rarely damaged, and we demonstrate that PED can be used to obtain material for physiological assays of stomatal responses to blue light and externally applied ABA. We thus believe that the PED method is suitable for preparing epidermal cell layers for physiological studies. © 2013 The Japanese Society for Plant Cell and Molecular Biology.