Use of TIRF to Monitor T-Lymphocyte Membrane Dynamics with Submicrometer and Subsecond Resolution

Cellular and Molecular Bioengineering
Alexandre BrodovitchA Pierres

Abstract

A key step of adaptive immune responses is the T lymphocyte capacity to detect the presence of foreign antigens on specialized cells with high speed and specificity during contacts lasting a few minutes. Much evidence suggests that there is a deep link between the lifetime of molecular interactions between T cell receptors and ligands and T cell activation, but the precise mechanisms of bond formation and dissociation remain incompletely understood. Previous experiments done with interference reflection microscopy/reflection interference contrast microscopy disclosed transverse motions with several nanometer average amplitude of micrometer size membrane zones. More recently, total internal reflection fluorescence microscopy was used to show that the initial interaction between primary T lymphocytes and model surfaces involved the tip of microvilli (typically 0.2 µm(2) area) generating apparent contacts of a few seconds that allowed cells to detect ligands of their membrane receptors. Here we show that these microvilli displayed minimal lateral displacements but quantitative fluorescence measurement suggested the occurrence of spontaneous transverse fluctuations of order of 67 nm amplitude during 1-s observation periods. This ma...Continue Reading

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Citations

Jan 15, 2016·Proceedings of the National Academy of Sciences of the United States of America·Jing WangWanli Liu
Nov 28, 2015·Acta Biomaterialia·Peter G ChildsStuart Reid
Apr 12, 2015·Annual Review of Immunology·Bernard Malissen, Pierre Bongrand
Sep 21, 2016·Proceedings of the National Academy of Sciences of the United States of America·Yunmin JungGilad Haran
Aug 9, 2016·Annual Review of Cell and Developmental Biology·Michael L Dustin, Kaushik Choudhuri
Jul 19, 2019·Proceedings of the National Academy of Sciences of the United States of America·Laurent LimozinPhilippe Robert
Jul 29, 2019·The Journal of Membrane Biology·Laurent Limozin, Pierre-Henri Puech
Dec 22, 2020·Advanced Science·Rui LiWeiqiang Chen

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Methods Mentioned

BETA
fluorescence microscopy
light
density gradient centrifugation
electron microscopy

Software Mentioned

TIRFM
ImageJ

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