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Genetically encoded barcodes for correlative volume electron microscopy.

Felix Sigmund1,2, Oleksandr Berezin1,2, Sofia Beliakova1,2

  • 1Munich Institute of Biomedical Engineering, Department of Bioscience, TUM School of Natural Sciences and TUM School of Medicine, Technical University of Munich, Munich, Germany.

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|April 17, 2023
PubMed
Summary
This summary is machine-generated.

Researchers developed electron microscopy-compatible barcodes (EMcapsulins) using metal-interacting moieties within nanocompartments. These novel EMcapsulins enable multiplexed gene expression mapping in biological samples, advancing cellular imaging capabilities.

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Area of Science:

  • Biotechnology
  • Cell Biology
  • Microscopy

Background:

  • Genetically encoded reporters are crucial for fluorescence microscopy but limited for electron microscopy (EM).
  • Multiplexable gene reporters for EM are scarce, hindering correlative light and electron microscopy (CLEM).

Purpose of the Study:

  • To develop novel, multiplexable gene reporters for EM.
  • To create EMcapsulins, a suite of spherically symmetric and concentric barcodes readable by standard EM techniques.

Main Methods:

  • Engineered encapsulin nanocompartments with variable numbers of fixation-stable metal-interacting moieties.
  • Developed different sized EMcapsulins and arranged them using rigid spacers.
  • Expressed fluorescent EMcapsulins for monitoring subcellular structures and validated in Drosophila and mouse brains.

Main Results:

  • Created six classes of EMcapsulins that could be automatically segmented and differentiated by EM.
  • Increased coding capacity by arranging EMcapsulins in distinct patterns.
  • Enabled automatic identification of genetically defined cells in EM images of Drosophila and mouse brains.

Conclusions:

  • EMcapsulins are compatible with various EM techniques (TEM, SEM, FIB-SEM).
  • This expandable palette of EM-readable barcodes can augment anatomical EM images with multiplexed gene expression maps.
  • EMcapsulins offer a powerful tool for high-resolution, multiplexed gene expression analysis in biological systems.