Microsurgical isolation of intact plant cell wall appositions for microanalyses

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Abstract
Little is known about the chemical components of the plant cell wall apposition as they relate to its structure and function. The small sizes (5–15 μm diameter) of the appositions, their low frequencies in the cells, and their intimate connections to the cell wall have almost precluded meaningful cytochemical and (or) biochemical analyses. With the development of analytical techniques using the electron microprobe it is now feasible to discover the elemental composition of minute cellular structures, such as the wall apposition, but problems in specimen preparation remain. As a necessary and critical first step in microprobe analysis we have found it best to microsurgically remove fresh wall appositions from their mother cells and then admit them directly into the scanning electron microscope (SEM) after air drying and gold coating. This paper describes the requisite technologies of specimen preparation, microtool fabrication, specimen selection and isolation which are involved. The technique described could find ready application in the microanalyses of many other subcellular structures.