Nanoscale imaging of the adhesion core including integrin β1 on intact living cells using scanning electron-assisted dielectric-impedance microscopy

PLoS One. 2018 Sep 20;13(9):e0204133. doi: 10.1371/journal.pone.0204133. eCollection 2018.

Abstract

The integrins are a superfamily of transmembrane proteins composed of α and β subunit dimers involved in cell-cell and cell-extracellular matrix interactions. The largest integrin subgroup is integrin β1, which contributes to several malignant phenotypes. Recently, we have developed a novel imaging technology named scanning electron-assisted dielectric-impedance microscopy (SE-ADM), which visualizes untreated living mammalian cells in aqueous conditions with high contrast. Using the SE-ADM system, we observed 60-nm gold colloids with antibodies directly binding to the focal adhesion core containing integrin β1 on mammalian cancer cells without staining and fixation. The adhesion core contains three or four high-density regions of integrin β1 and connects to the actin filament. An adhesion core with high-density integrin β1 is suggested to contain 10-20 integrin dimers. Our SE-ADM system can also visualize various other membrane proteins in living cells in medium without staining and fixation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism
  • Animals
  • Cell Adhesion
  • Cell Line, Tumor
  • Cell Membrane / metabolism
  • Cell Survival
  • Colloids / chemistry
  • Electric Impedance*
  • Female
  • Focal Adhesions / metabolism
  • Gold / chemistry
  • Image Processing, Computer-Assisted
  • Integrin beta1 / metabolism*
  • Mammary Neoplasms, Animal / pathology
  • Mammary Neoplasms, Animal / ultrastructure
  • Mice
  • Microscopy, Electron, Scanning*
  • Nanoparticles / chemistry*

Substances

  • Actins
  • Colloids
  • Integrin beta1
  • Gold

Grants and funding

This study was supported by JSPS KAKENHI Grant Numbers 15H04365, Toshihiko Ogura and a Grant-in-aid for scientific research on innovative areas from the MEXT of Japan (17H05829), Toshihiko Ogura. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.