Refining a Correlative Light Electron Microscopy Workflow using Luminescent Metal Complexes

Jonathan R Shewring, Lorna Hodgson, Helen L Bryant, Per A Bullough, Julia A Weinstein, Paul Verkade*

*Corresponding author for this work

Research output: Contribution to journalArticle (Academic Journal)peer-review

1 Citation (Scopus)

Abstract

The potential for increasing the application of Correlative Light Electron Microscopy (CLEM) technologies in life science research is hindered by the lack of suitable molecular probes that are emissive, photostable, and scatter electrons well. Most brightly fluorescent organic molecules are intrinsically poor electron-scatterers, while multi-metallic compounds scatter electrons well but are usually non-luminescent. Thus, the goal of CLEM to image the same object of interest on the continuous scale from hundreds of microns to nanometers remains a major challenge partially due to requirements for a single probe to be suitable for light (LM) and electron microscopy (EM). Some of the main CLEM probes, based on gold nanoparticles appended with fluorophores and quantum dots (QD) have presented significant drawbacks. Here we present an Iridium-based luminescent metal complex (Ir complex 1) as a probe and describe how we have developed a CLEM workflow based on such metal complexes.
Original languageEnglish
Pages (from-to)69-87
Number of pages19
JournalMethods in Cell Biology
Volume162
DOIs
Publication statusPublished - 23 Jan 2021

Bibliographical note

Funding Information:
We would like to thank the Wolfson Bioimaging Facility staff for their expert help in these studies with equipment used funded through BBSRC ALERT13 funding (BB/L014181/1), and Dr. L K McKenzie. We thank Chris Hill from the University of Sheffield's Faculty of Science Electron Microscopy Facility, and Prof. J.A.G. Williams from the University of Durham for collaboration. JRS was in receipt of a University of Sheffield Ph.D. studentship funded through the ?Imagine? initiative.

Publisher Copyright:
© 2021 Elsevier Inc.

Keywords

  • CLEM
  • luminscent metal complex
  • finder dishes
  • imaging dishes
  • retracing

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