Rapid clearing for high resolution 3D imaging of ex vivo pancreatic cancer spheroids

Eliana Steinberg, Natalie Orehov, Katerina Tischenko, Ouri Schwob, Gideon Zamir, Ayala Hubert, Zakhariya Manevitch, Ofra Benny*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

The currently accepted imaging methods have been a central hurdle to imaging the finer details of tumor behavior in three-dimensional (3D) ex vivo multicellular culture models. In our search for an improved way of imaging tumor behavior in its physiological-like niche, we developed a simple, efficient, and straightforward procedure using standard reagents and imaging equipment that significantly enhanced 3D imaging up to a ~200-micron depth. We tested its efficacy on pancreatic spheroids, prototypes of high-density tissues that are difficult to image. We found we could both save time with this method and extract information about pancreatic tumor spheroids that previously was difficult to obtain. We were able to discern clear differences in the organization of pancreatic tumor spheroids generated from different origins, suggesting cell-specific, inherent, bottom-up organization with a correlation to the level of malignancy. We also examined the dynamic changes in the spheroids at predetermined time points, providing important information related to tissue morphogenesis and its metabolic state. Lastly, this process enabled us to assess a drug vehicle’s potential to penetrate dense tumor tissue by improving our view of the inert particles’ diffusion in the 3D spheroid. This clearing method, a simple procedure, can open the door to more accurate imaging and reveal more about cancer behavior.

Original languageAmerican English
Article number7703
Pages (from-to)1-18
Number of pages18
JournalInternational Journal of Molecular Sciences
Volume21
Issue number20
DOIs
StatePublished - 2 Oct 2020

Bibliographical note

Funding Information:
Funding: This study was kindly supported by the European Research Council (ERC-StG) under the European Union’s Horizon 2020 research and Innovation Programme (grant agreement number 0305260). We thank the Israel Ministry of Science and Technology (MOST) (grant agreement number 0394906), and the Israel Science Foundation (grants agreements number 0394883; 3011004240) for their funding.

Funding Information:
This study was kindly supported by the European Research Council (ERC-StG) under the European Union?s Horizon 2020 research and Innovation Programme (grant agreement number 0305260). We thank the Israel Ministry of Science and Technology (MOST) (grant agreement number 0394906), and the Israel Science Foundation (grants agreements number 0394883; 3011004240) for their funding.

Publisher Copyright:
© 2020 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

  • 3D imaging
  • Clearing
  • Pancreatic cancer
  • Spheroids

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