A protocol for full-rotation soft X-ray tomography of single cells

Jian-Hua Chen; Bieke Vanslembrouck; Valentina Loconte; Axel Ekman; Mirko Cortese; Ralf Bartenschlager; Gerry McDermott; Carolyn A Larabell; Mark A Le Gros; Venera Weinhardt

doi:10.1016/j.xpro.2022.101176

A protocol for full-rotation soft X-ray tomography of single cells

STAR Protoc. 2022 Feb 10;3(1):101176. doi: 10.1016/j.xpro.2022.101176. eCollection 2022 Mar 18.

Authors

Jian-Hua Chen^{1

2}, Bieke Vanslembrouck^{1

2}, Valentina Loconte^{1

2}, Axel Ekman², Mirko Cortese³, Ralf Bartenschlager^{3

4

5}, Gerry McDermott^{1

2}, Carolyn A Larabell^{1

2}, Mark A Le Gros^{1

2}, Venera Weinhardt^{1

6}

Affiliations

¹ Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
² Department of Anatomy, University of California, San Francisco, San Francisco, CA 94143, USA.
³ Department of Infectious Diseases, Molecular Virology Heidelberg University, Heidelberg 69120, Germany.
⁴ German Center for Infection Research, Heidelberg Partner Site, Heidelberg 69120, Germany.
⁵ Division Virus-Associated Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg 69120, Germany.
⁶ Centre for Organismal Studies, Heidelberg University, Heidelberg 69120, Germany.

Abstract

The protocol describes step-by-step sample preparation, data acquisition, and segmentation of cellular organelles with soft X-ray tomography. It is designed for microscopes built to perform full-rotation data acquisition on specimens in cylindrical sample holders, such as the XM-2 microscope at the Advanced Light Source, LBNL; however, it might be generalized for similar sample holder designs for both synchrotron and table-top microscopes. For complete details on the use and execution of this profile, please refer to Loconte et al. (2021).

Keywords: Cell Biology; Microscopy; Single Cell.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Imaging, Three-Dimensional* / methods
Microscopy / methods
Rotation
Synchrotrons
Tomography, X-Ray* / methods

Abstract

Publication types

MeSH terms

Grants and funding