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Differential expansion microscopy

Sebastian P. Pernal, Asiri Liyanaarachchi, Domenico L. Gatti, Brent Formosa, Rishika Pulvender, Eric R. Kuhn, Rafael Ramos, Akshata R. Naik, Kathleen George, Suzan Arslanturk, Douglas J. Taatjes, View ORCID ProfileBhanu P. Jena
doi: https://doi.org/10.1101/699579
Sebastian P. Pernal
1Department of Physiology, Wayne State University, Detroit, MI 48201, USA
2NanoBioScience Institute, Wayne State University, Detroit, MI 48201, USA
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Asiri Liyanaarachchi
1Department of Physiology, Wayne State University, Detroit, MI 48201, USA
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Domenico L. Gatti
4Biochemistry, Microbiology and Immunology, School of Medicine, Wayne State University, Detroit, MI 48201, USA
2NanoBioScience Institute, Wayne State University, Detroit, MI 48201, USA
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Brent Formosa
1Department of Physiology, Wayne State University, Detroit, MI 48201, USA
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Rishika Pulvender
1Department of Physiology, Wayne State University, Detroit, MI 48201, USA
2NanoBioScience Institute, Wayne State University, Detroit, MI 48201, USA
3Center for Molecular Medicine & Genetics, Wayne State University, Detroit, MI 48201, USA
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Eric R. Kuhn
1Department of Physiology, Wayne State University, Detroit, MI 48201, USA
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Rafael Ramos
1Department of Physiology, Wayne State University, Detroit, MI 48201, USA
2NanoBioScience Institute, Wayne State University, Detroit, MI 48201, USA
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Akshata R. Naik
1Department of Physiology, Wayne State University, Detroit, MI 48201, USA
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Kathleen George
1Department of Physiology, Wayne State University, Detroit, MI 48201, USA
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Suzan Arslanturk
5Department of Computer Science, College of Engineering, Wayne State University, Detroit, MI 48201, USA
2NanoBioScience Institute, Wayne State University, Detroit, MI 48201, USA
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Douglas J. Taatjes
6Department of Pathology and Laboratory Medicine, Microscopy Imaging Center, University of Vermont College of Medicine, Burlington, VT 05405, USA
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Bhanu P. Jena
1Department of Physiology, Wayne State University, Detroit, MI 48201, USA
2NanoBioScience Institute, Wayne State University, Detroit, MI 48201, USA
3Center for Molecular Medicine & Genetics, Wayne State University, Detroit, MI 48201, USA
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  • ORCID record for Bhanu P. Jena
  • For correspondence: bjena@med.wayne.edu
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Abstract

Expansion microscopy (ExM) involves the use of hydration-competent polymers to physically expand biological specimens approximately 4-fold linear increase to achieve 70 nanometer resolution using an ordinary diffraction limited optical microscope. Optimal conditions however for antigen retention during the expansion process and the relative expansion between organelles within cells has remained unclear. It is reported that different tissues expand to different extents, suggesting that although isotropic expansion is believed to occur, different subcellular compartments with different composition would undergo anisotropic or differential expansion (DiEx). Consequently, there would be distortion of the native shape and size of subcellular compartments upon expansion, parameters which are critical in assessing cellular states in health and disease. Here we report optimal fixation and expansion conditions that retain structural integrity of cells while exhibiting up to 8-fold linear and therefore 512-fold volumetric expansion. Anisotropic expansion is observed not just between tissues, but between different subcellular compartments and even within subcellular compartments themselves. Combining image analysis and machine learning, we provide an approach for the rapid and precise measurement of cellular and subcellular structures in expanded tissue. Using both manual and computation assessment of morphometric parameters, we demonstrate expansion to be anisotropic and therefore refer to this this method as differential expansion microscopy (DiExM).

Footnotes

  • http://veloce.med.wayne.edu/~gatti/image-processing.html

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted July 11, 2019.
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Differential expansion microscopy
Sebastian P. Pernal, Asiri Liyanaarachchi, Domenico L. Gatti, Brent Formosa, Rishika Pulvender, Eric R. Kuhn, Rafael Ramos, Akshata R. Naik, Kathleen George, Suzan Arslanturk, Douglas J. Taatjes, Bhanu P. Jena
bioRxiv 699579; doi: https://doi.org/10.1101/699579
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Differential expansion microscopy
Sebastian P. Pernal, Asiri Liyanaarachchi, Domenico L. Gatti, Brent Formosa, Rishika Pulvender, Eric R. Kuhn, Rafael Ramos, Akshata R. Naik, Kathleen George, Suzan Arslanturk, Douglas J. Taatjes, Bhanu P. Jena
bioRxiv 699579; doi: https://doi.org/10.1101/699579

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