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Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment
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TEM, SEM, and STEM-based immuno-CLEM workflows offer complementary advantages.

Viola Oorschot1,2, Benjamin W Lindsey3,4, Jan Kaslin5

  • 1Ramaciotti Centre for Cryo EM, Monash University, Melbourne, VIC, 3800, Australia.

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|January 14, 2021
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Summary
This summary is machine-generated.

Identifying tissue stem cells is crucial. Immuno-correlative light and electron microscopy (iCLEM) workflows effectively distinguish and characterize stem cells using ultrastructural morphology and cell markers.

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Area of Science:

  • Developmental Biology
  • Regenerative Medicine
  • Cell Biology

Background:

  • Identifying endogenous tissue stem cells is a significant challenge in developmental and regenerative biology.
  • Distinguishing stem cell populations in heterogeneous tissues requires combining cell markers with ultrastructural morphology.

Purpose of the Study:

  • To establish and compare immuno-correlative light and electron microscopy (iCLEM) workflows for stem cell identification.
  • To evaluate the practicality of iCLEM for characterizing heterogeneous cell populations in various biological contexts.

Main Methods:

  • Developed multi-resolution iCLEM workflows utilizing the Tokuyasu technique for antigenicity preservation.
  • Compared iCLEM combined with scanning electron microscopy (SEM), scanning transmission electron microscopy (STEM), and transmission electron microscopy (TEM).

Main Results:

  • Established robust iCLEM protocols suitable for plant, animal, and human tissues.
  • Demonstrated the identification, quantification, and characterization of heterogeneous cell populations, including neural stem and progenitor cells.
  • Highlighted the advantages of SEM, STEM, and TEM in conjunction with iCLEM for ultrastructural analysis.

Conclusions:

  • iCLEM workflows provide a powerful approach for molecularly characterizing stem cell populations.
  • These methods are broadly applicable for studying healthy and diseased tissues, particularly the vertebrate nervous system.
  • iCLEM facilitates detailed investigation of cell populations at multiple resolutions.