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Related Experiment Video

Updated: Dec 22, 2025

4D Microscopy: Unraveling Caenorhabditis elegans Embryonic Development Using Nomarski Microscopy
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4D Microscopy: Unraveling Caenorhabditis elegans Embryonic Development Using Nomarski Microscopy

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Expansion microscopy of C. elegans.

Chih-Chieh Jay Yu1,2,3, Nicholas C Barry2,3, Asmamaw T Wassie1,3

  • 1Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, United States.

Elife
|May 2, 2020
PubMed
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Expansion microscopy (ExM) was adapted for C. elegans, overcoming cuticle challenges. The new Expansion of C. elegans (ExCel) method enables high-resolution imaging of proteins, RNA, and DNA in intact worms.

Area of Science:

  • Cell Biology
  • Microscopy Techniques
  • Developmental Biology

Background:

  • Expansion microscopy (ExM) provides nanoscale imaging by isotropically expanding hydrogel-embedded tissues.
  • The C. elegans cuticle presents a barrier to antibody penetration, challenging ExM application.
  • High-resolution imaging is crucial for understanding cellular structures and molecular localization in model organisms.

Purpose of the Study:

  • To develop a robust expansion microscopy method for intact C. elegans.
  • To enable simultaneous high-resolution imaging of multiple cellular components including proteins, RNA, and DNA.
  • To create adaptable ExCel protocols for various imaging needs, including epitope preservation and ultra-high expansion.

Main Methods:

  • Chemical fixation and hydrogel embedding of intact C. elegans.
Keywords:
C. elegansdevelopmental biologyexpansion microscopygene expressionimmunohistochemistryin situ hybridizationneurosciencesuper-resolution imagingsynapses

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  • Isotropic expansion of processed specimens to achieve nanoscale resolution.
  • Development of epitope-preserving and iterative ExCel protocols for advanced imaging.

    • Main Results:

    • ExCel achieves ~65-75 nm resolution with 3.3-3.8x linear expansion in intact C. elegans.
    • Simultaneous imaging of fluorescent proteins, RNA, and DNA locations was demonstrated.
    • Epitope-preserving ExCel enabled antibody staining of endogenous proteins, and iterative ExCel allowed imaging after 20x expansion.

    Conclusions:

    • ExCel is a versatile toolkit for high-resolution imaging in C. elegans.
    • The method facilitates detailed mapping of synaptic proteins and identification of novel proteins at cell junctions.
    • ExCel is valuable for gene expression analysis in individual neurons within the same animal.