Low-temperature Scanning Electron Microscopy of Frozen Hydrated Apple Tissues and Surface Organisms

Frozen hydrated buds and epicarp of `Golden Delicious' apple (Malus domestica Borkh.) were observed with a low-temperature, field emission scanning electron microscope (SEM). In addition to observing surface features of these specimens, holders were modified to observe fractured specimens. A modified hinged holder retained both halves of a fractured specimen for examination of the complementary faces of frozen hydrated tissues. Low-temperature SEM avoided artifacts, such as extraction, solubilization, and shrinkage, which are normally encountered with chemical fixation, dehydration, and drying, respectively. The technique allowed observations of well-preserved frozen hydrated structures, such as the platelets of epicuticular wax; loosely associated organisms on plant surfaces, such as spider-mite eggs; delicate structures, such as fungal hyphae; and partially hydrated tissues, such as fruit epicarp and winter bud scales.