Freeze-substitution and conventional electron microscopy in the study of medically-important bacteria

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Abstract
Summary Electron microscopy (EM) has made important contributions to our knowledge of bacterial and host ultrastructure and of the changes to their structures during pathogenesis. Improvements in sample preparation and the development of new cryotechniques such as freeze-substitution, which is proving to be a more reliable method for preserving cell integrity and retaining antigenicity than conventional procedures, are rekindling an interest in the use of EM as a primary tool for investigating pathogenic interactions. Other techniques such as immunoelectron microscopy have emerged as powerful tools for identifying and localizing specific intracellular or cell surface molecules including virulence factors. When immunogold labelling is combined with freeze-substitution, even more precise molecular identification is possible. In this review, we discuss the significance of preparing clinical material for EM and describe the application and contribution of some new EM techniques to study medically-important bacteria.

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