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Related Experiment Video

Updated: Jul 30, 2025

Using Expansion Microscopy to Physically Enlarge Whole-Mount Drosophila Embryos for Super-Resolution Imaging
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Using Expansion Microscopy to Physically Enlarge Whole-Mount Drosophila Embryos for Super-Resolution Imaging.

Samia Parveen1, Nicolas W Jones1, Ian Millerschultz1

  • 1Department of Biological Sciences, University of Arkansas.

Journal of Visualized Experiments : Jove
|May 15, 2023
PubMed
Summary
This summary is machine-generated.

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Expansion microscopy (ExM) enables enhanced resolution of subcellular details in Drosophila embryos using standard confocal microscopes. This technique expands samples, revealing structures previously invisible with conventional imaging.

Area of Science:

  • Developmental Biology
  • Cell Biology
  • Microscopy

Background:

  • Confocal microscopy is crucial for 3D molecular localization but limited to ~200 nm resolution.
  • Subcellular biology demands resolutions of tens of nanometers, exceeding traditional confocal capabilities.
  • Hardware-based super-resolution microscopy offers higher resolution but requires specialized, inaccessible equipment.

Purpose of the Study:

  • To present a protocol for Expansion Microscopy (ExM) in whole-mount Drosophila embryos.
  • To enable visualization of subcellular details not resolvable by conventional confocal microscopy.
  • To demonstrate ExM's utility for studying molecular localization and cellular architecture.

Main Methods:

  • Expansion Microscopy (ExM) protocol for whole-mount Drosophila embryos.

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  • Utilized standard confocal microscopy for imaging expanded samples.
  • Employed antibodies against Par-3, myosin II, and fluorescently labeled streptavidin for labeling.

    • Main Results:

    • Achieved an approximate four-fold isotropic increase in sample size.
    • Resolved distinct pools of myosin-GFP between adjacent cell cortices.
    • Visualized fine details of the mitochondrial network architecture.

    Conclusions:

    • ExM provides an accessible method to enhance resolution with conventional confocal microscopes.
    • The protocol is compatible with common immunofluorescence techniques and reagents.
    • ExM significantly improves the ability to detect subcellular details in Drosophila embryos.