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Laboratory Soft X-Ray Microscopy with an Integrated Visible-Light Microscope—Correlative Workflow for Faster 3D Cell Imaging

Published online by Cambridge University Press:  07 October 2020

Aurélie Dehlinger ,
Christian Seim Open the ORCID record for Christian Seim [Opens in a new window] ,
Holger Stiel ,
Shailey Twamley ,
Antje Ludwig ,
Mikael Kördel Open the ORCID record for Mikael Kördel [Opens in a new window] ,
Daniel Grötzsch ,
Stefan Rehbein  and
Birgit Kanngießer
Aurélie Dehlinger*
Affiliation:
Technische Universität Berlin, Institut für Optik und Atomare Physik, Hardenbergstraße 36, Berlin 10623, Germany Berlin Laboratory for Innovative X-ray technologies (BLiX), Hardenbergstraße 36, Berlin 10623, Germany
Christian Seim
Affiliation:
Technische Universität Berlin, Institut für Optik und Atomare Physik, Hardenbergstraße 36, Berlin 10623, Germany Berlin Laboratory for Innovative X-ray technologies (BLiX), Hardenbergstraße 36, Berlin 10623, Germany
Holger Stiel
Affiliation:
Berlin Laboratory for Innovative X-ray technologies (BLiX), Hardenbergstraße 36, Berlin 10623, Germany Max-Born-Institut (MBI) im Forschungsverbund Berlin e.V., Max-Born-Straße 2A, Berlin 12489, Germany
Shailey Twamley
Affiliation:
Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Medizinische Klinik für Kardiologie und Angiologie, Campus Mitte, Charitéplatz 1, 10117 Berlin, Germany DZHK (German Centre for Cardiovascular Research), Partner Site Berlin, Charitéplatz 1, 10117 Berlin, Germany
Antje Ludwig
Affiliation:
Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Medizinische Klinik für Kardiologie und Angiologie, Campus Mitte, Charitéplatz 1, 10117 Berlin, Germany DZHK (German Centre for Cardiovascular Research), Partner Site Berlin, Charitéplatz 1, 10117 Berlin, Germany Charité – Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Klinik für Radiologie, Charitéplatz 1, 10117 Berlin, Germany
Mikael Kördel
Affiliation:
Department of Applied Physics, KTH Royal Institute of Technology/Albanova, Stockholm 106 91, Sweden
Daniel Grötzsch
Affiliation:
Technische Universität Berlin, Institut für Optik und Atomare Physik, Hardenbergstraße 36, Berlin 10623, Germany Berlin Laboratory for Innovative X-ray technologies (BLiX), Hardenbergstraße 36, Berlin 10623, Germany
Stefan Rehbein
Affiliation:
Helmholtz Zentrum Berlin für Materialien und Energie GmbH, Wilhelm-Conrad-Röntgen Campus, Albert-Einstein-Str. 15, Berlin 12489, Germany
Birgit Kanngießer
Affiliation:
Technische Universität Berlin, Institut für Optik und Atomare Physik, Hardenbergstraße 36, Berlin 10623, Germany Berlin Laboratory for Innovative X-ray technologies (BLiX), Hardenbergstraße 36, Berlin 10623, Germany
*
*Author for correspondence: Aurélie Dehlinger, E-mail: [email protected]
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Abstract

Laboratory transmission soft X-ray microscopy (L-TXM) has emerged as a complementary tool to synchrotron-based TXM and high-resolution biomedical 3D imaging in general in recent years. However, two major operational challenges in L-TXM still need to be addressed: a small field of view and a potentially misaligned rotation stage. As it is not possible to alter the magnification during operation, the field of view in L-TXM is usually limited to a few tens of micrometers. This complicates locating areas and objects of interest in the sample. Additionally, if the rotation axis of the sample stage cannot be adjusted prior to the experiments, an efficient workflow for tomographic imaging cannot be established, as refocusing and sample repositioning will become necessary after each recorded projection. Both these limitations have been overcome with the integration of a visible-light microscope (VLM) into the L-TXM system. Here, we describe the calibration procedure of the goniometer sample stage and the integrated VLM and present the resulting 3D imaging of a test sample. In addition, utilizing this newly integrated VLM, the extracellular matrix of cryofixed THP-1 cells (human acute monocytic leukemia cells) was visualized by L-TXM for the first time in the context of an ongoing biomedical research project.

Keywords

cell imagingcorrelative microscopycryo microscopywater-window X-ray microscopyX-ray tomography
Type
Software and Instrumentation
Information
Microscopy and Microanalysis , Volume 26 , Issue 6 , December 2020 , pp. 1124 - 1132
Copyright
Copyright © Microscopy Society of America 2020

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