A Modified Magnified Analysis of Proteome (MAP) Method for Super-Resolution Cell Imaging that Retains Fluorescence

Sci Rep. 2020 Mar 6;10(1):4186. doi: 10.1038/s41598-020-61156-2.

Abstract

Biological systems consist of a variety of distinct cell types that form functional networks. Super-resolution imaging of individual cells is required for better understanding of these complex systems. Direct visualization of 3D subcellular and nano-scale structures in cells is helpful for the interpretation of biological interactions and system-level responses. Here we introduce a modified magnified analysis of proteome (MAP) method for cell super-resolution imaging (Cell-MAP) which preserves cell fluorescence. Cell-MAP expands cells more than four-fold while preserving their overall architecture and three-dimensional proteome organization after hydrogel embedding. In addition, Optimized-Cell-MAP completely preserves fluorescence and successfully allows for the observation of tagged small molecular probes containing peptides and microRNAs. Optimized-Cell-MAP further successfully applies to the study of structural characteristics and the identification of small molecules and organelles in mammalian cells. These results may give rise to many other applications related to the structural and molecular analysis of smaller assembled biological systems.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cell-Penetrating Peptides / genetics
  • Cell-Penetrating Peptides / metabolism
  • Flow Cytometry
  • Fluorescence*
  • HEK293 Cells
  • Humans
  • Male
  • MicroRNAs / metabolism
  • Microscopy, Fluorescence
  • Proteome / metabolism*
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Cell-Penetrating Peptides
  • MicroRNAs
  • Proteome