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Multiplex, quantitative cellular analysis in large tissue volumes with clearing-enhanced 3D microscopy (Ce3D).

Weizhe Li1, Ronald N Germain2, Michael Y Gerner2,3

  • 1Lymphocyte Biology Section, Laboratory of Systems Biology, National Institute of Allergy and Infectious Diseases/NIH, Bethesda, MD 20892.

Proceedings of the National Academy of Sciences of the United States of America
|August 16, 2017
PubMed
Summary

A new method called clearing-enhanced 3D (Ce3D) enables high-resolution 3D imaging of intact organs. This technique allows detailed mapping of cell types and their spatial organization within tissues for better biological understanding.

Keywords:
histo-cytometryimmune systemquantitative microscopytissue clearing

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Area of Science:

  • Cell Biology
  • Tissue Engineering
  • Microscopy

Background:

  • Cellular spatial organization is crucial for tissue function, homeostasis, and differentiation.
  • Understanding cell positioning, identity, and function within macroscale tissue architecture is essential for biological research.

Purpose of the Study:

  • To introduce clearing-enhanced 3D (Ce3D), a novel and user-friendly method for high-resolution 3D tissue imaging.
  • To demonstrate the compatibility of Ce3D with antibody-based immunolabeling and its ability to preserve cellular morphology and protein fluorescence.

Main Methods:

  • Developed clearing-enhanced 3D (Ce3D) for enhanced tissue transparency.
  • Utilized antibody-based immunolabeling for protein detection.
  • Employed 3D histo-cytometry for quantitative analysis of cell populations in intact tissues.

Main Results:

  • Achieved excellent tissue transparency across various organs using Ce3D.
  • Preserved cellular morphology and protein fluorescence, enabling high-quality imaging.
  • Successfully performed large-volume, high-resolution 3D microscopy and quantitative analysis of diverse cell types in lymphoid and nonlymphoid organs.

Conclusions:

  • Ce3D offers a robust strategy for volumetric quantitative imaging and analysis of complex tissues.
  • This method bridges the gap between traditional section imaging and dissociation-based techniques.
  • Ce3D combined with histo-cytometry provides a comprehensive approach to studying tissue architecture and cellular composition.