Recent clearing methods have enabled labeling and imaging of intact tissues 6, mouse organs 7 and bodies 3, 8, 9, 10, 11, 12, 13, 14, 15, parts of human organs 16 and even human embryos 17.

However, such whole-body connectivity maps will be needed to understand the functional interdependence of organ systems and how a disease starting in one part of the body affects distal organs and tissues, for example, during neurodegeneration or systemic inflammation. Generating transgenic animals to map changes in the distribution of relevant proteins is usually prohibitively expensive and time consuming. In addition, most methods to image nerves or other cells in the context of whole bodies rely on transgenic animals 4, 5, which limits the flexibility of experimental design. For instance, although the nervous system pervades every region of the mammalian body, cellular-level maps depicting the intricate web of nerves linking various organs and connecting them to the central nervous system are still lacking 1, 2, 3. However, it remains challenging to map the distribution, connectivity and molecular makeup of cell types across the whole body for a given experimental condition. More than a century of dedicated work has provided a detailed understanding of the gross anatomy of the human body and the body of common model organisms and has produced detailed histological maps of many individual organs.

An atlas of high-resolution images showcasing mouse nervous, lymphatic and vascular systems is accessible at. We applied wildDISCO to map tertiary lymphoid structures in the context of breast cancer, considering both primary tumor and metastases throughout the mouse body. Additionally, we examined rare proliferating cells and the effects of biological perturbations, as demonstrated in germ-free mice. WildDISCO facilitates imaging of peripheral nervous systems, lymphatic vessels and immune cells in whole mice at cellular resolution by labeling diverse endogenous proteins. We identified heptakis(2,6-di- O-methyl)-β-cyclodextrin as a potent enhancer of cholesterol extraction and membrane permeabilization, enabling deep, homogeneous penetration of standard antibodies without aggregation.

We introduce wildDISCO, a new approach for whole-body immunolabeling, optical clearing and imaging in mice, circumventing the need for transgenic reporter animals or nanobody labeling and so overcoming existing technical limitations. Whole-body imaging techniques play a vital role in exploring the interplay of physiological systems in maintaining health and driving disease.

Brain neuron connections