Use of freeze-substitution and molecular distillation drying in the preparation of Dunaliella parva for ion localization studies by X-ray microanalysis

The determination of ion concentrations within cells and sub-cellular compartments remains a difficult procedure, as the volumes to be analyzed are rather small. X-ray microanalysis is sufficiently sensitive, and has adequate resolution, to measure these concentrations. The major difficulties are related to the preparation of material for analysis. We have compared the measurement of sodium, potassium, and chloride contents in a salt tolerant unicellular alga, Dunaliella parva, following either freeze-substitution (using two different resins) or molecular distillation drying. All three procedures gave similar results: after freeze substitution, ion contents were marginally (but not significantly) higher following embedding in Nanoplast MUV 116 resin than in Spurr resin. Since the Nanoplast can be polymerised at low temperatures, it has advantages over the Spurr resin.