Understanding the building blocks of cellular machinery has dramatically advanced in recent years not only with cell cryo preparation but also with imaging and detection of proteins and particles in their near to native states. While cryo electron microscopy has allowed for the examination of single particles at unprecedented levels of resolution, the understanding of cellular machinery requires more than the knowledge of single particle structure but also a thorough analysis of the macromolecular elements as they exist in the cell and that represent the basis for their function. By cryo fixation of cells and/or tissues, it is possible to accurately, reproducibly and safely locate macromolecular networks as they exist in situ and prepare them in a way that is suitable for further electron microscopy examination. We show that high resolution light microscopy, under cryo conditions, is an effective method of locating particles of interest and via connected microscopy, relocating these exact structures with the cryo focused ion beam scanning electron microscope for cryo lamella preparation. Such frozen lamellae offer unique large windows into the cellular interior and therefore of macromolecular complexes in their native state or under specific physiological conditions, which are suitable for subsequent high-resolution electron microscope examination.