Correlative fluorescence microscopy, transmission electron microscopy and secondary ion mass spectrometry (CLEM-SIMS) for cellular imaging

PLoS One. 2021 May 10;16(5):e0240768. doi: 10.1371/journal.pone.0240768. eCollection 2021.

Abstract

Electron microscopy (EM) has been employed for decades to analyze cell structure. To also analyze the positions and functions of specific proteins, one typically relies on immuno-EM or on a correlation with fluorescence microscopy, in the form of correlated light and electron microscopy (CLEM). Nevertheless, neither of these procedures is able to also address the isotopic composition of cells. To solve this, a correlation with secondary ion mass spectrometry (SIMS) would be necessary. SIMS has been correlated in the past to EM or to fluorescence microscopy in biological samples, but not to CLEM. We achieved this here, using a protocol based on transmission EM, conventional epifluorescence microscopy and nanoSIMS. The protocol is easily applied, and enables the use of all three technologies at high performance parameters. We suggest that CLEM-SIMS will provide substantial information that is currently beyond the scope of conventional correlative approaches.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells / cytology*
  • Cells / ultrastructure*
  • HeLa Cells
  • Humans
  • Microscopy, Electron, Transmission*
  • Microscopy, Fluorescence
  • Optical Imaging
  • Spectrometry, Mass, Secondary Ion*

Associated data

  • figshare/10.6084/m9.figshare.14416547

Grants and funding

The work was supported by the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG) through grants SFB1286/A05 to S.J., SFB1286/A03 to S.O.R. and SFB1286/B01 to N.T.N.P. The work was also supported by the Swedish Research Council through VR grant 2016-06800 to N.T.N.P.