title: Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues.
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.
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA.|Picower Institute for Learning and Memory, MIT, Cambridge, Massachusetts, USA.
authors.forename: Taeyun
authors.identifier: 0000-0001-9447-7579
authors.initials: T
authors.lastname: Ku
authors.affiliation: Department of Chemical Engineering, MIT, Cambridge, Massachusetts, USA.
authors.forename: Justin
authors.initials: J
authors.lastname: Swaney
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA.|Picower Institute for Learning and Memory, MIT, Cambridge, Massachusetts, USA.|Department of Neurosurgery, Gangnam Severance Hospital, Spine and Spinal Cord Institute, Yonsei University College of Medicine, Seoul, Republic of Korea.
authors.forename: Jeong-Yoon
authors.initials: JY
authors.lastname: Park
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Albanese
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA.|Department of Brain and Cognitive Sciences, MIT, Cambridge, Massachusetts, USA.
authors.forename: Evan
authors.initials: E
authors.lastname: Murray
authors.affiliation: Department of Chemical Engineering, MIT, Cambridge, Massachusetts, USA.
authors.forename: Jae Hun
authors.initials: JH
authors.lastname: Cho
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA.|Picower Institute for Learning and Memory, MIT, Cambridge, Massachusetts, USA.
authors.forename: Young-Gyun
authors.initials: YG
authors.lastname: Park
authors.affiliation: Harvard-MIT Division of Health Sciences and Technology, MIT, Cambridge, Massachusetts, USA.
authors.forename: Vamsi
authors.initials: V
authors.lastname: Mangena
authors.affiliation: Department of Biological Engineering, MIT, Cambridge, Massachusetts, USA.
authors.forename: Jiapei
authors.initials: J
authors.lastname: Chen
authors.affiliation: Institute for Medical Engineering and Science, Massachusetts Institute of Technology (MIT), Cambridge, Massachusetts, USA.|Picower Institute for Learning and Memory, MIT, Cambridge, Massachusetts, USA.|Department of Chemical Engineering, MIT, Cambridge, Massachusetts, USA.|Department of Brain and Cognitive Sciences, MIT, Cambridge, Massachusetts, USA.|Broad Institute of Harvard University and MIT, Cambridge, Massachusetts, USA.
authors.forename: Kwanghun
authors.initials: K
authors.lastname: Chung
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism* / Q000648:ultrastructure; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D007089:Image Enhancement / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods; D021621:Imaging, Three-Dimensional / Q000379:methods*; D007118:Immunoassay / Q000379:methods; D008297:Male; D051379:Mice; D057054:Molecular Imaging / Q000379:methods*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D020543:Proteome / Q000378:metabolism* / Q000648:ultrastructure; D013569:Synapses / Q000378:metabolism* / Q000648:ultrastructure*; D014018:Tissue Distribution
chemical_list: D009419:Nerve Tissue Proteins; D020543:Proteome