Assembly and Imaging Set up of PIE-Scope

Sergey Gorelick, David A. Dierickx, Genevieve Buckley, James C. Whisstock, Alex de Marco

Research output: Contribution to journalArticleOtherpeer-review

4 Citations (Scopus)

Abstract

Cryo-Electron Tomography (cryo-ET) is a method that enables resolving the structure of macromolecular complexes directly in the cellular environment. However, sample preparation for in situ Cryo-ET is labour-intensive and can require both cryo-lamella preparation through cryo-Focused Ion Beam (FIB) milling and correlative light microscopy to ensure that the event of interest is present in the lamella. Here, we present an integrated cryo-FIB and light microscope setup called the Photon Ion Electron microscope (PIE-scope) that enables direct and rapid isolation of cellular regions containing protein complexes of interest. The PIE-scope can be retrofitted on existing microscopes, although the drawings we provide are meant to work on ThermoFisher DualBeams with small mechanical modifications those can be adapted on other brands.

Original languageEnglish
Article numbere3768
Number of pages11
Journalbio-protocol
Volume10
Issue number19
DOIs
Publication statusPublished - 2020

Keywords

  • Correlative microscopy
  • Cryo-CLEM
  • Cryo-lamella
  • Cryo-light microscopy
  • PIE-scope

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