Once samples have been frozen in the EM ICE, they can be freeze-substituted for regular TEM by replacing the water by a suitable solvent (acetone) at low temperature without ice recrystallization. Additional protocols include progressive lowering of the temperature (PLT). After freeze substitution, the samples are infiltrated with embedding resin, either epoxy (for structural studies) or acrylic (for immunolabeling). The FSP liquid handler allows automatic solvent and embedding media exchange in protocols that do not use osmium tetroxide.

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