Multiplexed and scalable super-resolution imaging of three-dimensional protein localization in size-adjustable tissues

Nat Biotechnol. 2016 Sep;34(9):973-81. doi: 10.1038/nbt.3641. Epub 2016 Jul 25.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / metabolism*
  • Brain / ultrastructure
  • Female
  • Gene Expression Profiling / methods
  • Image Enhancement / methods
  • Image Interpretation, Computer-Assisted / methods
  • Imaging, Three-Dimensional / methods*
  • Immunoassay / methods
  • Male
  • Mice
  • Molecular Imaging / methods*
  • Nerve Tissue Proteins / metabolism
  • Proteome / metabolism*
  • Proteome / ultrastructure
  • Synapses / metabolism*
  • Synapses / ultrastructure*
  • Tissue Distribution

Substances

  • Nerve Tissue Proteins
  • Proteome