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

Felix Bruno Kleine Borgmann*, Celine Hoffmann (Main author), Anaïs Carpentier, Michel Mittelbronn (Main author), Clément Thomas

*Corresponding author for this work

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Cytotoxic lymphocytes, such as natural killer (NK) cells and cytotoxic T cells, can recognize and kill tumor cells by establishing a highly specialized cell-cell contact called the immunological synapse. The formation and lytic activity of the immunological synapse are accompanied by local changes in the organization, dynamics and molecular composition of the cell membrane, as well as the polarization of various cellular components, such as the cytoskeleton, vesicles and organelles. Characterization and understanding of the molecular and cellular processes underlying immunological synapse formation and activity requires the combination of complementary types of information provided by different imaging modalities, the correlation of which can be difficult. Correlative light and electron microscopy (CLEM) allows for the accurate correlation of functional information provided by fluorescent light microscopy with ultrastructural features provided by high-resolution electron microscopy. In this chapter, we present a detailed protocol describing each step to generate cell-cell conjugates between NK cells and cancer cells, and to analyze these conjugates by CLEM using separate confocal laser-scanning and transmission electron microscopes.

Original languageEnglish
Pages (from-to)96-106
JournalMethods in Cell Biology
Volume178
DOIs
Publication statusPublished - 28 Jul 2023

Keywords

  • Actin cytoskeleton
  • Cancer
  • Correlative light and electron microscopy (CLEM)
  • Cytotoxic lymphocytes
  • Immunological synapse
  • Natural killer cell

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