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  Precise, Correlated Fluorescence Microscopy and Electron Tomography of Lowicryl Sections Using Fluorescent Fiducial Markers

Kukulski, W., Schorb, M., Welsch, S., Picco, A., Kaksonen, M., & Briggs, J. A. G. (2012). Precise, Correlated Fluorescence Microscopy and Electron Tomography of Lowicryl Sections Using Fluorescent Fiducial Markers. In T. MullerReichert, & P. Verkade (Eds.), Correlative Light and Electron Microscopy (pp. 235-257). San Diego: Elsevier Academic Press Inc.

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Genre: Book Chapter
Alternative Title : Methods Cell Biol.

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 Creators:
Kukulski, W., Author
Schorb, M., Author
Welsch, S., Author
Picco, A., Author
Kaksonen, M., Author
Briggs, John A. G.1, Author              
Affiliations:
1European Molecular Biology Laboratory, External Organizations, ou_3346677              

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Free keywords: light organization architecture cells tool
 Abstract: The application of fluorescence and electron microscopy to the same specimen allows the study of dynamic and rare cellular events at ultrastructural detail. Here, we present a correlative microscopy approach, which combines high accuracy of correlation, high sensitivity for detecting faint fluorescent signals, as well as robustness and reproducibility to permit large dataset collections. We provide a step-by-step protocol that allows direct mapping of fluorescent protein signals into electron tomograms. A localization precision of <100 nm is achieved by using fluorescent fiducial markers which are visible both in fluorescence images and in electron tomograms. We explain the critical details of the procedure, give background information on the individual steps, present results from test experiments carried out during establishment of the method, as well as information about possible modifications to the protocol, such as its application to 2D electron micrographs. This simple, robust, and flexible method can be applied to a large variety of cellular systems, such as yeast cell pellets and mammalian cell monolayers, to answer a broad spectrum of structure function related questions.

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Language(s): eng - English
 Dates: 2012
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: Other: WOS:000308189100014
DOI: 10.1016/b978-0-12-416026-2.00013-3
 Degree: -

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Title: Correlative Light and Electron Microscopy
Source Genre: Book
 Creator(s):
MullerReichert, T., Editor
Verkade, P., Editor
Affiliations:
-
Publ. Info: San Diego : Elsevier Academic Press Inc
Pages: - Volume / Issue: 111 Sequence Number: - Start / End Page: 235 - 257 Identifier: ISBN: 978-0-12-416026-2

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Title: Methods in Cell Biology
Source Genre: Series
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Publ. Info: -
Pages: - Volume / Issue: 111 Sequence Number: - Start / End Page: - Identifier: -