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

Research output: Contribution to journalArticlepeer-review


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 languageEnglish
Article number7703
Pages (from-to)1-18
Number of pages18
Issue number20
StatePublished - 2 Oct 2020


  • 3D imaging
  • Clearing
  • Pancreatic cancer
  • Spheroids

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry


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