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Related Concept Videos

Immunogold Electron Microscopy01:20

Immunogold Electron Microscopy

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Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
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Related Experiment Video

Updated: Mar 12, 2026

Expanding the Comprehension of the Tumor Microenvironment using Mass Spectrometry Imaging of Formalin-Fixed and Paraffin-Embedded Tissue Samples
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iDISCO: a simple, rapid method to immunolabel large tissue samples for volume imaging.

Nicolas Renier1, Zhuhao Wu1, David J Simon1

  • 1Laboratory of Brain Development and Repair, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.

Cell
|November 24, 2014
PubMed
Summary

We developed iDISCO, a fast and affordable method for 3D imaging of whole organs and embryos. This technique allows detailed visualization of molecular structures within large, cleared tissues, aiding neurobiology research.

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Last Updated: Mar 12, 2026

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ACT-PRESTO: Biological Tissue Clearing and Immunolabeling Methods for Volume Imaging
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ACT-PRESTO: Biological Tissue Clearing and Immunolabeling Methods for Volume Imaging

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Area of Science:

  • Neuroscience
  • Molecular Biology
  • Biotechnology

Background:

  • 3D visualization of molecularly labeled structures in large tissues is challenging.
  • Existing methods are often complex, time-consuming, or expensive.

Purpose of the Study:

  • To introduce iDISCO, a simple, rapid, and inexpensive method for whole-mount immunolabeling and 3D volume imaging of large cleared biological samples.
  • To demonstrate the compatibility of iDISCO with various antibodies and transgenic reporters.
  • To apply iDISCO for analyzing neuronal degeneration in complex tissues.

Main Methods:

  • iDISCO protocol for whole-mount immunolabeling and tissue clearing.
  • Volume imaging of cleared samples (perinatal mouse embryos, adult organs like brains and kidneys).
  • Compatibility testing with 28 antibodies and Green Fluorescent Protein (GFP) reporters.

Main Results:

  • iDISCO successfully enabled whole-mount immunolabeling and 3D volume imaging of diverse large tissues.
  • The method showed compatibility with a wide range of antibodies, including those targeting endogenous antigens and transgenic reporters.
  • Application to degenerating neurons revealed unexpected variability in apoptotic neuron numbers and allowed visualization of single degenerating axons and cleaved Caspase-3 in vivo.

Conclusions:

  • iDISCO is an effective and accessible technique for 3D imaging of immunolabeled structures in complex tissues.
  • The method facilitates the study of cellular and subcellular structures in intact organs and embryos.
  • iDISCO provides novel insights into neuronal degeneration processes.