Taeyun Ku1,2,9, Justin Swaney3,9, Jeong-Yoon Park1,2,4,9, Alexandre Albanese1, Evan Murray1,5, Jae Hun Cho3, Young-Gyun Park1,2, Vamsi Mangena6, Jiapei Chen7 & Kwanghun Chung1–3,5,8
1Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA. 2Picower Institute for Learning and Memory, MIT, Cambridge, Massachusetts, USA. 3Department of Chemical Engineering, MIT, Cambridge, Massachusetts, USA. 4Department of Neurosurgery, Gangnam Severance Hospital, Spine and Spinal Cord Institute, Yonsei University College of Medicine, Seoul, Republic of Korea. 5Department of Brain and Cognitive Sciences, MIT, Cambridge, Massachusetts, USA. 6Harvard-MIT Division of Health Sciences and Technology, MIT, Cambridge, Massachusetts, USA. 7Department of Biological Engineering, MIT, Cambridge, Massachusetts, USA. 8Broad Institute of Harvard University and MIT, Cambridge, Massachusetts, USA.
9These authors contributed equally to this work.
Correspondence should be addressed to K.C..
Abstract
The biology of multicellular organisms is coordinated across multiple size scales, from the subnanoscale of molecules to the macroscale, tissue-wide interconnectivity of cell populations. Here we introduce a method for super-resolution imaging of the multiscale organization of intact tissues. The method, called magnified analysis of the proteome (MAP), linearly expands entire organs fourfold while preserving their overall architecture and three-dimensional proteome organization. MAP is based on the observation that preventing crosslinking within and between endogenous proteins during hydrogel-tissue hybridization allows for natural expansion upon protein denaturation and dissociation. The expanded tissue preserves its protein content, its fine subcellular details, and its organ-scale intercellular connectivity. We use off-the-shelf antibodies for multiple rounds of immunolabeling and imaging of a tissue's magnified proteome, and our experiments demonstrate a success rate of 82% (100/122 antibodies tested). We show that specimen size can be reversibly modulated to image both inter-regional connections and fine synaptic architectures in the mouse brain.