High data output and automated 3D correlative light-electron microscopy method

Giuseppe Vicidomini, Maria C. Gagliani, Michela Canfora, Katia Cortese, Fabio Frosi, Clara Santangelo, Pier Paolo Di Fiore, Patrizia Boccacci, Alberto Diaspro, Carlo Tacchetti

Research output: Contribution to journalArticlepeer-review

Abstract

Correlative light/electron microscopy (CLEM) allows the simultaneous observation of a given subcellular structure by fluorescence light microscopy (FLM) and electron microscopy. The use of this approach is becoming increasingly frequent in cell biology. In this study, we report on a new high data output CLEM method based on the use of cryosections. We successfully applied the method to analyze the structure of rough and smooth Russell bodies used as model systems. The major advantages of our method are (i) the possibility to correlate several hundreds of events at the same time, (ii) the possibility to perform three-dimensional (3D) correlation, (iii) the possibility to immunolabel both endogenous and recombinantly expressed proteins at the same time and (iv) the possibility to combine the high data analysis capability of FLM with the high precision-accuracy of transmission electron microscopy in a CLEM hybrid morphometry analysis. We have identified and optimized critical steps in sample preparation, defined routines for sample analysis and retracing of regions of interest, developed software for semi/fully automatic 3D reconstruction and defined preliminary conditions for an hybrid light/electron microscopy morphometry approach.

Original languageEnglish
Pages (from-to)1828-1838
Number of pages11
JournalTraffic
Volume9
Issue number11
DOIs
Publication statusPublished - 2008

Keywords

  • Confocal microscopy
  • Correlative microscopy
  • Cryosection
  • Electron microscopy
  • Immunofluorescence
  • Immunogold
  • Morphometry

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Structural Biology
  • Molecular Biology
  • Genetics

Fingerprint Dive into the research topics of 'High data output and automated 3D correlative light-electron microscopy method'. Together they form a unique fingerprint.

Cite this