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

doi:10.1016/bs.mcb.2020.12.008

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 journal › Article (Academic Journal) › peer-review

5 Citations (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 language	English
Pages (from-to)	69-87
Number of pages	19
Journal	Methods in Cell Biology
Volume	162
DOIs	https://doi.org/10.1016/bs.mcb.2020.12.008
Publication status	Published - 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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title = "Refining a Correlative Light Electron Microscopy Workflow using Luminescent Metal Complexes",

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.",

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author = "Shewring, {Jonathan R} and Lorna Hodgson and Bryant, {Helen L} and Bullough, {Per A} and Weinstein, {Julia A} and Paul Verkade",

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: {\textcopyright} 2021 Elsevier Inc.",

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doi = "10.1016/bs.mcb.2020.12.008",

language = "English",

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AU - Shewring, Jonathan R

AU - Hodgson, Lorna

AU - Bryant, Helen L

AU - Bullough, Per A

AU - Weinstein, Julia A

AU - Verkade, Paul

N1 - 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.

PY - 2021/1/23

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N2 - 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.

AB - 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.

KW - CLEM

KW - luminscent metal complex

KW - finder dishes

KW - imaging dishes

KW - retracing

U2 - 10.1016/bs.mcb.2020.12.008

DO - 10.1016/bs.mcb.2020.12.008

M3 - Article (Academic Journal)

C2 - 33707023

SN - 0091-679X

VL - 162

SP - 69

EP - 87

JO - Methods in Cell Biology

JF - Methods in Cell Biology

ER -

Refining a Correlative Light Electron Microscopy Workflow using Luminescent Metal Complexes

Abstract

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Keywords

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