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Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids

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, efficie...

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Autores principales: Steinberg, Eliana, Orehov, Natalie, Tischenko, Katerina, Schwob, Ouri, Zamir, Gideon, Hubert, Ayala, Manevitch, Zakhariya, Benny, Ofra
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7589457/
https://www.ncbi.nlm.nih.gov/pubmed/33081011
http://dx.doi.org/10.3390/ijms21207703
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author Steinberg, Eliana
Orehov, Natalie
Tischenko, Katerina
Schwob, Ouri
Zamir, Gideon
Hubert, Ayala
Manevitch, Zakhariya
Benny, Ofra
author_facet Steinberg, Eliana
Orehov, Natalie
Tischenko, Katerina
Schwob, Ouri
Zamir, Gideon
Hubert, Ayala
Manevitch, Zakhariya
Benny, Ofra
author_sort Steinberg, Eliana
collection PubMed
description 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.
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spelling pubmed-75894572020-10-29 Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids Steinberg, Eliana Orehov, Natalie Tischenko, Katerina Schwob, Ouri Zamir, Gideon Hubert, Ayala Manevitch, Zakhariya Benny, Ofra Int J Mol Sci Article 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. MDPI 2020-10-18 /pmc/articles/PMC7589457/ /pubmed/33081011 http://dx.doi.org/10.3390/ijms21207703 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Steinberg, Eliana
Orehov, Natalie
Tischenko, Katerina
Schwob, Ouri
Zamir, Gideon
Hubert, Ayala
Manevitch, Zakhariya
Benny, Ofra
Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids
title Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids
title_full Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids
title_fullStr Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids
title_full_unstemmed Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids
title_short Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids
title_sort rapid clearing for high resolution 3d imaging of ex vivo pancreatic cancer spheroids
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7589457/
https://www.ncbi.nlm.nih.gov/pubmed/33081011
http://dx.doi.org/10.3390/ijms21207703
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