Freeze-fracture immunocytochemistry: fracture-label and label-fracture for the localization of membrane proteins

Freeze-fracture is a unique investigative tool for visualization of the en face topography of individual membrane leaflets of cell membranes at high resolution under the electron microscope. The development of a system of freeze-fracture cytochemical and immunocytochemical techniques has further advanced the utility of this methodological approach for high-resolution localization of specific membrane and intracellular macromolecules in tissues and cells. The unit focuses on description, in a step-by-step manner, of the experimental procedures for two specific freeze-fracture labeling techniques, namely fracture-label and label-fracture. Users are guided in a stepwise manner, starting from the preparation of tissue or cell samples to the final retrieval and mounting of fracture-label and label-fracture specimens for examination on the electron microscope.