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Methods Cell Biol. 2017;140:149-164. doi: 10.1016/bs.mcb.2017.03.008. Epub 2017 Apr 19.

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

Author information

1
Oregon Health and Sciences University, Portland, OR, United States.
2
Thermo Fisher Scientific, Hillsboro, OR, United States.
3
Thermo Fisher Scientific, Hillsboro, OR, United States; Genentech, San Francisco, CA, United States.

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

PMID:
28528631
DOI:
10.1016/bs.mcb.2017.03.008
[Indexed for MEDLINE]

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