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Correlative light and electron microscopy to explore the lytic immunological synapse between natural killer cells and

Felix Bruno Kleine Borgmann1, Celine Hoffmann2, Anaïs Carpentier3

  • 1Department of Cancer Research (DoCR), Luxembourg Institute of Health (LIH), Luxembourg, Luxembourg; Luxembourg Center of Neuropathology (LCNP), Dudelange, Luxembourg.

Methods in Cell Biology
|July 29, 2023
PubMed

Insights

Natural killer (NK) cells and cytotoxic T cells form immunological synapses to kill tumor cells. Correlative light and electron microscopy (CLEM) enables detailed analysis of these crucial cell-cell interactions.

Area of Science:

  • Immunology
  • Cell Biology
  • Microscopy

Background:

  • Cytotoxic lymphocytes, including natural killer (NK) cells and cytotoxic T cells, engage tumor cells via specialized cell-cell junctions known as immunological synapses.
  • Immunological synapse formation involves dynamic changes in cell membrane organization, molecular composition, and the polarization of cellular components like the cytoskeleton and vesicles.
  • Understanding these intricate processes necessitates integrating functional and ultrastructural data, which is often challenging due to limitations of individual imaging techniques.

Purpose of the Study:

  • To present a detailed protocol for analyzing immunological synapses using correlative light and electron microscopy (CLEM).
  • To enable the accurate correlation of functional insights from light microscopy with ultrastructural details from electron microscopy.

Main Methods:

  • Generation of cell-cell conjugates between NK cells and cancer cells.
  • Analysis of these conjugates using correlative light and electron microscopy (CLEM).
  • Utilizing separate confocal laser-scanning and transmission electron microscopes for comprehensive imaging.

Main Results:

  • The protocol facilitates the detailed examination of immunological synapse formation and dynamics.
  • CLEM provides a powerful approach to correlate functional cellular events with ultrastructural features.
  • This method allows for a deeper understanding of the molecular and cellular mechanisms governing cytotoxic lymphocyte interactions with target cells.

Conclusions:

  • Correlative light and electron microscopy (CLEM) is an effective strategy for studying immunological synapses.
  • This technique bridges the gap between functional and ultrastructural analyses of cell-cell interactions.
  • The presented protocol offers a valuable resource for researchers investigating immune cell cytotoxicity and synapse biology.

Keywords:
Actin cytoskeletonCancerCorrelative light and electron microscopy (CLEM)Cytotoxic lymphocytesImmunological synapseNatural killer cell

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