Three-Dimensional Single Molecule Localization Microscopy Reveals the Topography of the Immunological Synapse at Isotropic Precision below 15 nm.
Nano Lett
; 21(21): 9247-9255, 2021 11 10.
Article
em En
| MEDLINE
| ID: mdl-34709845
ABSTRACT
T-cells engage with antigen-presenting cells in search for antigenic peptides and form transient interfaces termed immunological synapses. Synapse topography affects receptor binding rates and the mutual segregation of proteins due to size exclusion effects. It is hence important to determine the 3D topography of the immunological synapse at high precision. Current methods provide only rather coarse images of the protein distribution within the synapse. Here, we applied supercritical angle fluorescence microscopy combined with defocused imaging, which allows three-dimensional single molecule localization microscopy (3D-SMLM) at an isotropic localization precision below 15 nm. Experiments were performed on hybrid synapses between primary T-cells and functionalized glass-supported lipid bilayers. We used 3D-SMLM to quantify the cleft size within the synapse by mapping the position of the T-cell receptor (TCR) with respect to the supported lipid bilayer, yielding average distances of 18 nm up to 31 nm for activating and nonactivating bilayers, respectively.
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1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Sinapses Imunológicas
/
Imagem Individual de Molécula
Idioma:
En
Revista:
Nano Lett
Ano de publicação:
2021
Tipo de documento:
Article
País de afiliação:
Áustria