Electron Tomography and Correlative Approaches in Platelet Studies

Kasia B Engberts; Cor Seinen; Willie J C Geerts; Harry F G Heijnen

doi:10.1007/978-1-4939-8585-2_4

Electron Tomography and Correlative Approaches in Platelet Studies

Kasia B Engberts, Cor Seinen, Willie J C Geerts, Harry F G Heijnen

Laboratory of Clinical Chemistry and Hematology, University Medical Center Utrecht, Utrecht, The Netherlands.
Department of Cell Biology, Cell Microscopy Core, University Medical Center Utrecht, Utrecht, The Netherlands. [email protected].
Crystal and Structural Chemistry, Bijvoet Center for Biomolecular Research, Faculty of Sciences, Utrecht University, 3584 CH Utrecht, the Netherlands.

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

Abstract

Blood platelets play a central role in the arrest of bleeding and the development of thrombosis. Unraveling the complex processes of platelet biogenesis from megakaryocytes, platelet adhesion, aggregation, and secretory responses are important topics in the field of hemostasis and thrombosis. Analysis of the ultrastructural changes that occur during these processes is essential for understanding the rapid membrane dynamics and has contributed substantially to our present knowledge of platelet formation and functioning. Recent developments in real-time imaging, correlative light and electron microscopy imaging (CLEM), and 3D (cryo) electron microscopy and tomography offer exciting opportunities to improve studies of the platelet adhesive responses and secretion at the ultrastructural level in a close to native environment. In this chapter we discuss and illustrate cryo preparation techniques (high-pressure freezing, vitrification), correlative LM and EM workflows, and 3D cryo-electron tomography that we apply in our current research projects.

Original language	English
Title of host publication	Platelets and Megakaryocytes. Volume 4, Advanced Protocols and Perspectives
Publisher	Humana Press
Pages	55-79
Number of pages	25
ISBN (Electronic)	978-1-4939-8585-2
ISBN (Print)	978-1-4939-8584-5
DOIs	https://doi.org/10.1007/978-1-4939-8585-2_4
Publication status	Published - 2018

Publication series

Name	Methods in Molecular Biology
Volume	1812
ISSN (Print)	1064-3745

Keywords

TEM
Immuno-EM
Fixation and cryo-immobilization
High pressure freezing EMtomography
Cryo-EM
Correlative light and electron microscopy (CLEM)

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10.1007/978-1-4939-8585-2_4

Engberts2018_Protocol_ElectronTomographyAndCorrelatiFinal published version, 1.45 MBLicence: Taverne

Cite this

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title = "Electron Tomography and Correlative Approaches in Platelet Studies",

abstract = "Blood platelets play a central role in the arrest of bleeding and the development of thrombosis. Unraveling the complex processes of platelet biogenesis from megakaryocytes, platelet adhesion, aggregation, and secretory responses are important topics in the field of hemostasis and thrombosis. Analysis of the ultrastructural changes that occur during these processes is essential for understanding the rapid membrane dynamics and has contributed substantially to our present knowledge of platelet formation and functioning. Recent developments in real-time imaging, correlative light and electron microscopy imaging (CLEM), and 3D (cryo) electron microscopy and tomography offer exciting opportunities to improve studies of the platelet adhesive responses and secretion at the ultrastructural level in a close to native environment. In this chapter we discuss and illustrate cryo preparation techniques (high-pressure freezing, vitrification), correlative LM and EM workflows, and 3D cryo-electron tomography that we apply in our current research projects.",

keywords = "TEM, Immuno-EM, Fixation and cryo-immobilization, High pressure freezing EMtomography, Cryo-EM, Correlative light and electron microscopy (CLEM)",

author = "Engberts, {Kasia B} and Cor Seinen and Geerts, {Willie J C} and Heijnen, {Harry F G}",

year = "2018",

doi = "10.1007/978-1-4939-8585-2_4",

language = "English",

isbn = "978-1-4939-8584-5",

series = "Methods in Molecular Biology",

publisher = "Humana Press",

pages = "55--79",

booktitle = "Platelets and Megakaryocytes. Volume 4, Advanced Protocols and Perspectives",

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Electron Tomography and Correlative Approaches in Platelet Studies. / Engberts, Kasia B; Seinen, Cor; Geerts, Willie J C et al.
Platelets and Megakaryocytes. Volume 4, Advanced Protocols and Perspectives. Humana Press, 2018. p. 55-79 (Methods in Molecular Biology; Vol. 1812).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › Academic › peer-review

TY - CHAP

T1 - Electron Tomography and Correlative Approaches in Platelet Studies

AU - Engberts, Kasia B

AU - Seinen, Cor

AU - Geerts, Willie J C

AU - Heijnen, Harry F G

PY - 2018

Y1 - 2018

N2 - Blood platelets play a central role in the arrest of bleeding and the development of thrombosis. Unraveling the complex processes of platelet biogenesis from megakaryocytes, platelet adhesion, aggregation, and secretory responses are important topics in the field of hemostasis and thrombosis. Analysis of the ultrastructural changes that occur during these processes is essential for understanding the rapid membrane dynamics and has contributed substantially to our present knowledge of platelet formation and functioning. Recent developments in real-time imaging, correlative light and electron microscopy imaging (CLEM), and 3D (cryo) electron microscopy and tomography offer exciting opportunities to improve studies of the platelet adhesive responses and secretion at the ultrastructural level in a close to native environment. In this chapter we discuss and illustrate cryo preparation techniques (high-pressure freezing, vitrification), correlative LM and EM workflows, and 3D cryo-electron tomography that we apply in our current research projects.

AB - Blood platelets play a central role in the arrest of bleeding and the development of thrombosis. Unraveling the complex processes of platelet biogenesis from megakaryocytes, platelet adhesion, aggregation, and secretory responses are important topics in the field of hemostasis and thrombosis. Analysis of the ultrastructural changes that occur during these processes is essential for understanding the rapid membrane dynamics and has contributed substantially to our present knowledge of platelet formation and functioning. Recent developments in real-time imaging, correlative light and electron microscopy imaging (CLEM), and 3D (cryo) electron microscopy and tomography offer exciting opportunities to improve studies of the platelet adhesive responses and secretion at the ultrastructural level in a close to native environment. In this chapter we discuss and illustrate cryo preparation techniques (high-pressure freezing, vitrification), correlative LM and EM workflows, and 3D cryo-electron tomography that we apply in our current research projects.

KW - TEM

KW - Immuno-EM

KW - Fixation and cryo-immobilization

KW - High pressure freezing EMtomography

KW - Cryo-EM

KW - Correlative light and electron microscopy (CLEM)

U2 - 10.1007/978-1-4939-8585-2_4

DO - 10.1007/978-1-4939-8585-2_4

M3 - Chapter

C2 - 30171572

SN - 978-1-4939-8584-5

T3 - Methods in Molecular Biology

SP - 55

EP - 79

BT - Platelets and Megakaryocytes. Volume 4, Advanced Protocols and Perspectives

PB - Humana Press

ER -

Electron Tomography and Correlative Approaches in Platelet Studies

Abstract

Publication series

Keywords

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