Advantages of fast-freeze fixation followed by freeze-substitution for the preservation of cell integrity

Abstract

The use of fast-freeze fixation (FFF) followed by freeze-substitution (FS) has supplied new data in a variety of domains due to the rapidity of this fixation compared to the slow process of the conventional chemical fixation. FFF completely arrests all the cell's biological reactions, and physically immobilizes most molecules within a few milliseconds. This review deals with the main results we obtained concerning the preservation of organelles particularly sensitive to osmotic shock, the maintenance in situ of soluble or depolymerizable cell components, the visualization of brief transient events, and the opening up of new prospects in the field of immunocytochemical labeling.