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Visualization of Cellular Components in a Mammalian Cell with Liquid-Cell Transmission Electron Microscopy

Published online by Cambridge University Press:  31 January 2017

Stephanie Besztejan
Affiliation:
Chemistry Department, Institute for Biochemistry and Molecular Biology, University of Hamburg, Martin-Luther-King Platz 6, 20146 Hamburg, Germany The Hamburg Centre for Ultrafast Imaging, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany
Sercan Keskin
Affiliation:
Max Planck Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, Geb. 99 (CFEL), 22761 Hamburg, Germany
Stephanie Manz
Affiliation:
Max Planck Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, Geb. 99 (CFEL), 22761 Hamburg, Germany
Günther Kassier
Affiliation:
Max Planck Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, Geb. 99 (CFEL), 22761 Hamburg, Germany
Robert Bücker
Affiliation:
Max Planck Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, Geb. 99 (CFEL), 22761 Hamburg, Germany
Deybith Venegas-Rojas
Affiliation:
Institute of Microsystems Technology, Hamburg University of Technology (TUHH), Eißendorfer Straße 42, 21073 Hamburg, Germany
Hoc K. Trieu
Affiliation:
Institute of Microsystems Technology, Hamburg University of Technology (TUHH), Eißendorfer Straße 42, 21073 Hamburg, Germany
Andrea Rentmeister
Affiliation:
Institute of Biochemistry, Westfälische Wilhelms-Universität Münster, Wilhelm-Klemm-Strasse 2, 48149 Muenster, Germany
R. J. Dwayne Miller*
Affiliation:
The Hamburg Centre for Ultrafast Imaging, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany Max Planck Institute for the Structure and Dynamics of Matter, Luruper Chaussee 149, Geb. 99 (CFEL), 22761 Hamburg, Germany Departments of Chemistry and Physics, University of Toronto, 80 St George St, Toronto, ON, Canada M5S3H6
*
* Corresponding author. dwayne.miller@mpsd.mpg.de

Abstract

We present liquid-cell transmission electron microscopy (liquid-cell TEM) imaging of fixed and non-fixed prostate cancer cells (PC3 and LNCaP) with high resolution in a custom developed silicon nitride liquid cell. Fixed PC3 cells were imaged for 90–120 min without any discernable damage. High contrast on the cellular structures was obtained even at low electron doses (~2.5 e/nm2 per image). The images show distinct structures of cell compartments (nuclei and nucleoli) and cell boundaries without any further sample embedding, dehydration, or staining. Furthermore, we observed dynamics of vesicles trafficking from the cell membrane in consecutive still frames in a non-fixed cell. Our findings show that liquid-cell TEM, operated at low electron dose, is an excellent tool to investigate dynamic events in non-fixed cells with enough spatial resolution (few nm) and natural amplitude contrast to follow key intracellular processes.

Type
Biological Applications
Copyright
© Microscopy Society of America 2017 

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Footnotes

a

These authors contributed equally to this work.

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