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  3. Soft X-ray tomography to map and quantify organelle interactions at the mesoscale

Soft X-ray tomography to map and quantify organelle interactions at the mesoscale

  • Structure. 2022 Apr 7;30(4):510-521.e3. doi: 10.1016/j.str.2022.01.006.
Valentina Loconte 1,  Jitin Singla 2,  Angdi Li 1,  Jian-Hua Chen 3,  Axel Ekman 3,  Gerry McDermott 3,  Andrej Sali 4,  Mark Le Gros 3,  Kate L White 5,  Carolyn A Larabell 6
Affiliations

Affiliations

  • 1 iHuman Institute, School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China.
  • 2 Department of Chemistry, Bridge Institute, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA.
  • 3 Department of Anatomy, University of California San Francisco, San Francisco, CA 94143, USA; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
  • 4 Department of Bioengineering and Therapeutic Science, Department of Pharmaceutical Chemistry, California Institute of Quantitative Bioscience, University of California San Francisco, San Francisco, CA 94158, USA.
  • 5 Department of Chemistry, Bridge Institute, USC Michelson Center for Convergent Bioscience, University of Southern California, Los Angeles, CA 90089, USA. Electronic address: [email protected].
  • 6 Department of Anatomy, University of California San Francisco, San Francisco, CA 94143, USA; Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA. Electronic address: [email protected].
PMID: 35148829 DOI: 10.1016/j.str.2022.01.006
Abstract

Inter-organelle interactions are a vital part of normal cellular function; however, these have proven difficult to quantify due to the range of scales encountered in Cell Biology and the throughput limitations of traditional imaging approaches. Here, we demonstrate that soft X-ray tomography (SXT) can be used to rapidly map ultrastructural reorganization and inter-organelle interactions in intact cells. SXT takes advantage of the naturally occurring, differential X-ray absorption of the carbon-rich compounds in each organelle. Specifically, we use SXT to map the spatiotemporal evolution of Insulin vesicles and their co-localization and interaction with mitochondria in pancreatic β cells during Insulin secretion and in response to different stimuli. We quantify changes in the morphology, biochemical composition, and relative position of mitochondria and Insulin vesicles. These findings highlight the importance of a comprehensive and unbiased mapping at the mesoscale to characterize cell reorganization that would be difficult to detect with other existing methodologies.

Keywords

3D cell mapping; mesoscale analysis; organelle interaction; pancreatic β cell; soft X-ray tomography.

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