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Laboratory Soft X-Ray Microscopy with an Integrated Visible-Light Microscope-Correlative Workflow for Faster 3D Cell Imaging
Tech Univ Berlin, Inst Opt & Atomare Phys, Hardenbergstr 36, D-10623 Berlin, Germany.;Berlin Lab Innovat Xray Technol BLiX, Hardenbergstr 36, D-10623 Berlin, Germany..
Tech Univ Berlin, Inst Opt & Atomare Phys, Hardenbergstr 36, D-10623 Berlin, Germany.;Berlin Lab Innovat Xray Technol BLiX, Hardenbergstr 36, D-10623 Berlin, Germany..
Berlin Lab Innovat Xray Technol BLiX, Hardenbergstr 36, D-10623 Berlin, Germany.;Forschungsverbund Berlin eV, Max Born Inst MBI, Max Born Str 2A, D-12489 Berlin, Germany..
Charite Univ Med Berlin, Campus Mitte,Charitepl 1, D-10117 Berlin, Germany.;Humboldt Univ, Freie Univ Berlin, Campus Mitte,Charitepl 1, D-10117 Berlin, Germany.;Berlin Inst Hlth, Med Klin Kardiol & Angiol, Campus Mitte,Charitepl 1, D-10117 Berlin, Germany.;DZHK German Ctr Cardiovasc Res, Partner Site Berlin,Charitepl 1, D-10117 Berlin, Germany..
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2020 (English)In: Microscopy and Microanalysis, ISSN 1431-9276, E-ISSN 1435-8115, Vol. 26, no 6, p. 1124-1132Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
Cambridge University Press (CUP) , 2020. Vol. 26, no 6, p. 1124-1132
Keywords [en]
cell imaging, correlative microscopy, cryo microscopy, water-window X-ray microscopy, X-ray tomography
National Category
Medical Laboratory and Measurements Technologies
Identifiers
URN: urn:nbn:se:kth:diva-288464DOI: 10.1017/S1431927620024447ISI: 000596581400006PubMedID: 33023699Scopus ID: 2-s2.0-85095128159OAI: oai:DiVA.org:kth-288464DiVA, id: diva2:1514976
Note

QC 20210107

Available from: 2021-01-07 Created: 2021-01-07 Last updated: 2022-06-25Bibliographically approved

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Kördel, Mikael

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