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Large-volume en-bloc staining for electron microscopy-based connectomics

Author

Listed:
  • Yunfeng Hua

    (Max Planck Institute of Brain Research)

  • Philip Laserstein

    (Max Planck Institute of Brain Research)

  • Moritz Helmstaedter

    (Max Planck Institute of Brain Research)

Abstract

Large-scale connectomics requires dense staining of neuronal tissue blocks for electron microscopy (EM). Here we report a large-volume dense en-bloc EM staining protocol that overcomes the staining gradients, which so far substantially limited the reconstructable volumes in three-dimensional (3D) EM. Our protocol provides densely reconstructable tissue blocks from mouse neocortex sized at least 1 mm in diameter. By relaxing the constraints on precise topographic sample targeting, it makes the correlated functional and structural analysis of neuronal circuits realistic.

Suggested Citation

  • Yunfeng Hua & Philip Laserstein & Moritz Helmstaedter, 2015. "Large-volume en-bloc staining for electron microscopy-based connectomics," Nature Communications, Nature, vol. 6(1), pages 1-7, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8923
    DOI: 10.1038/ncomms8923
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    Cited by:

    1. Carles Bosch & Tobias Ackels & Alexandra Pacureanu & Yuxin Zhang & Christopher J. Peddie & Manuel Berning & Norman Rzepka & Marie-Christine Zdora & Isabell Whiteley & Malte Storm & Anne Bonnin & Chris, 2022. "Functional and multiscale 3D structural investigation of brain tissue through correlative in vivo physiology, synchrotron microtomography and volume electron microscopy," Nature Communications, Nature, vol. 13(1), pages 1-16, December.
    2. P. J. Schubert & R. Saxena & J. Kornfeld, 2024. "DeepFocus: fast focus and astigmatism correction for electron microscopy," Nature Communications, Nature, vol. 15(1), pages 1-10, December.
    3. Gregg Wildenberg & Hanyu Li & Vandana Sampathkumar & Anastasia Sorokina & Narayanan Kasthuri, 2023. "Isochronic development of cortical synapses in primates and mice," Nature Communications, Nature, vol. 14(1), pages 1-13, December.

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