A fully integrated, three-dimensional fluorescence to electron microscopy correlative workflow

Claudia S. López; Cedric Bouchet-Marquis; Christopher P. Arthur; Jessica L. Riesterer; Gregor Heiss; Guillaume Thibault; Lee Pullan; Sunjong Kwon; Joe W. Gray

doi:10.1016/bs.mcb.2017.03.008

A fully integrated, three-dimensional fluorescence to electron microscopy correlative workflow

Claudia S. López, Cedric Bouchet-Marquis, Christopher P. Arthur, Jessica L. Riesterer, Gregor Heiss, Guillaume Thibault, Lee Pullan, Sunjong Kwon, Joe W. Gray

Research output: Contribution to journal › Article › peer-review

13 Scopus citations

Abstract

While fluorescence microscopy provides tools for highly specific labeling and sensitive detection, its resolution limit and lack of general contrast has hindered studies of cellular structure and protein localization. Recent advances in correlative light and electron microscopy (CLEM), including the fully integrated CLEM workflow instrument, the FEI CorrSight with MAPS, have allowed for a more reliable, reproducible, and quicker approach to correlate three-dimensional time-lapse confocal fluorescence data, with three-dimensional focused ion beam–scanning electron microscopy data. Here we demonstrate the entire integrated CLEM workflow using fluorescently tagged MCF7 breast cancer cells.

Original language	English (US)
Pages (from-to)	149-164
Number of pages	16
Journal	Methods in cell biology
Volume	140
DOIs	https://doi.org/10.1016/bs.mcb.2017.03.008
State	Published - 2017

Keywords

3D FIB-SEM
CLEM
Correlative imaging technique
Electron microscopy
Fluorescence microscopy

ASJC Scopus subject areas

Cell Biology

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10.1016/bs.mcb.2017.03.008

Cite this

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title = "A fully integrated, three-dimensional fluorescence to electron microscopy correlative workflow",

abstract = "While fluorescence microscopy provides tools for highly specific labeling and sensitive detection, its resolution limit and lack of general contrast has hindered studies of cellular structure and protein localization. Recent advances in correlative light and electron microscopy (CLEM), including the fully integrated CLEM workflow instrument, the FEI CorrSight with MAPS, have allowed for a more reliable, reproducible, and quicker approach to correlate three-dimensional time-lapse confocal fluorescence data, with three-dimensional focused ion beam–scanning electron microscopy data. Here we demonstrate the entire integrated CLEM workflow using fluorescently tagged MCF7 breast cancer cells.",

keywords = "3D FIB-SEM, CLEM, Correlative imaging technique, Electron microscopy, Fluorescence microscopy",

author = "L{\'o}pez, {Claudia S.} and Cedric Bouchet-Marquis and Arthur, {Christopher P.} and Riesterer, {Jessica L.} and Gregor Heiss and Guillaume Thibault and Lee Pullan and Sunjong Kwon and Gray, {Joe W.}",

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year = "2017",

doi = "10.1016/bs.mcb.2017.03.008",

language = "English (US)",

volume = "140",

pages = "149--164",

journal = "Methods in cell biology",

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T1 - A fully integrated, three-dimensional fluorescence to electron microscopy correlative workflow

AU - López, Claudia S.

AU - Bouchet-Marquis, Cedric

AU - Arthur, Christopher P.

AU - Riesterer, Jessica L.

AU - Heiss, Gregor

AU - Thibault, Guillaume

AU - Pullan, Lee

AU - Kwon, Sunjong

AU - Gray, Joe W.

PY - 2017

Y1 - 2017

N2 - While fluorescence microscopy provides tools for highly specific labeling and sensitive detection, its resolution limit and lack of general contrast has hindered studies of cellular structure and protein localization. Recent advances in correlative light and electron microscopy (CLEM), including the fully integrated CLEM workflow instrument, the FEI CorrSight with MAPS, have allowed for a more reliable, reproducible, and quicker approach to correlate three-dimensional time-lapse confocal fluorescence data, with three-dimensional focused ion beam–scanning electron microscopy data. Here we demonstrate the entire integrated CLEM workflow using fluorescently tagged MCF7 breast cancer cells.

AB - While fluorescence microscopy provides tools for highly specific labeling and sensitive detection, its resolution limit and lack of general contrast has hindered studies of cellular structure and protein localization. Recent advances in correlative light and electron microscopy (CLEM), including the fully integrated CLEM workflow instrument, the FEI CorrSight with MAPS, have allowed for a more reliable, reproducible, and quicker approach to correlate three-dimensional time-lapse confocal fluorescence data, with three-dimensional focused ion beam–scanning electron microscopy data. Here we demonstrate the entire integrated CLEM workflow using fluorescently tagged MCF7 breast cancer cells.

KW - 3D FIB-SEM

KW - CLEM

KW - Correlative imaging technique

KW - Electron microscopy

KW - Fluorescence microscopy

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M3 - Article

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AN - SCOPUS:85017650996

SN - 0091-679X

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JO - Methods in cell biology

JF - Methods in cell biology

ER -

A fully integrated, three-dimensional fluorescence to electron microscopy correlative workflow

Abstract

Keywords

ASJC Scopus subject areas

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