Correlative light and electron microscopy : strategies and applications

Leiden Repository

Correlative light and electron microscopy : strategies and applications

Type: Doctoral Thesis
Title: Correlative light and electron microscopy : strategies and applications
Author: Driel, Linda Francina van
Publisher: Department Molecular Cellbiology, Faculty of Medicine / Leiden University Medical center (LUMC), Leiden University
Issue Date: 2011-10-11
Keywords: Correlative microscopy
Cryo-electron microscopy
Cryo-electron tomography
Cryo-fluorescence microscopy
Regulated exocytosis
Weibel Palade Bodies
Abstract: Correlative light and electron microscopy (CLEM) refers to the observation of the same structures or ultrastructures with both light microscopy (LM) and electron microscopy (EM). LM provides an overview of the studied material, and enables the quick localization of structures that are fluorescently labeled, while EM allows high resolution imaging of structures. The thesis describes several technical developments that allow CLEM. In the first chapters, methods are described for room temperature CLEM. This includes tissue labeling with fluorescent dyes in ultrathin resin sections, and fluorescence imaging before further preparation for EM and electron tomography. The methods are applied to tissue biopsies, and to the study of regulated exocytosis of Weibel-Palade bodies. In the following chapters, CLEM developments are extended with CLEM of vitreous samples, i.e. cryo-CLEM. Vitrification allows easy maintenance of fluorescent probes, and it allows correlation with cryo-EM, a technique that preserves biological structures to atomic resolution. The thesis describes the design of a microscope stage for cryo-fluorescence microscopy. Following cryo-FM, samples are studied directly by cryo-EM and cryo-electron tomography. The technique is used to study mitochondrial ultrastructure, and the role of actin in the exocytosis of pancreatic zymogen granules.
Description: Promotor: A.J. Koster, Co-promotor: J.A. Valentijn
With summary in Dutch
Faculty: LUMC
Citation: Driel, L.F. van, 2011, Doctoral thesis, Leiden University
ISBN: 9789461910059
Sponsor: FEI Company, SEN (Stichting ter bevordering van de elektronenmicroscopie in Nederland).
Handle: http://hdl.handle.net/1887/17922
 

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application/pdf Title page_Table of contents_List of abbreviations 420.7Kb View/Open
application/pdf Chapter 1 Introduction 1.381Mb View/Open
application/pdf Chapter 2 1.683Mb View/Open Full text at publisher site
application/pdf Chapter 3 2.462Mb View/Open Full text at publisher site
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application/pdf Chapter 5 1.351Mb View/Open
application/pdf Chapter 6 Discussion and outlook 1.448Mb View/Open
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application/pdf Curriculum Vitae_List of publications 204.8Kb View/Open
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