Botanical X-ray microanalysis in cryoscanning electron microscopy.

Modern microscopy in plant sciences has evolved in the direction of providing ultra-structural and analytical information simultaneously. Energy-dispersive X-ray microanalysis (EDX) is a powerful technique that allows the qualitative and quantitative measurement of many elements of physiological interest at the cellular and subcellular level. The most significant advance has been the development of freezing techniques to study cells in plant tissues by EDX in the cryoscanning electron microscopy. Cryofixation is fast enough to retain the original distributions of inorganic elements of tissue electrolytes sufficiently for microanalytical studies. This approach may have broad application for various types of localizations of relevance to plant physiology, environmental pollution, and plant-microbe interactions. In this chapter, the experimental procedure of analytical cryoscanning electron microscopy applied to botanical samples is outlined.