TY - JOUR
T1 - Correlative light and electron microscopy to explore the lytic immunological synapse between natural killer cells and cancer cells
AU - Kleine Borgmann, Felix Bruno
AU - Hoffmann, Celine
AU - Carpentier, Anaïs
AU - Mittelbronn, Michel
AU - Thomas, Clément
N1 - Funding Information:
CT's lab is supported by La Fondation Cancer (Luxembourg, ACTIMMUNE, FC/2019/02) and Think Pink Lux. The authors would like to thank the Roger de Spoelberch Foundation for funding the electron microscopy platform (LCSB/LNS).
Publisher Copyright:
© 2023 Elsevier Inc.
PY - 2023/7/28
Y1 - 2023/7/28
N2 - 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.
AB - 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.
KW - Actin cytoskeleton
KW - Cancer
KW - Correlative light and electron microscopy (CLEM)
KW - Cytotoxic lymphocytes
KW - Immunological synapse
KW - Natural killer cell
UR - http://www.scopus.com/inward/record.url?scp=85161006064&partnerID=8YFLogxK
UR - https://pubmed.ncbi.nlm.nih.gov/37516530
U2 - 10.1016/bs.mcb.2023.05.004
DO - 10.1016/bs.mcb.2023.05.004
M3 - Article
C2 - 37516530
AN - SCOPUS:85161006064
SN - 0091-679X
VL - 178
SP - 96
EP - 106
JO - Methods in Cell Biology
JF - Methods in Cell Biology
ER -