InFlow microscopy of human leukocytes: A tool for quantitative analysis of actin rearrangements in the immune synapse

Journal of Immunological Methods
Guido H WabnitzYvonne Samstag

Abstract

The actin cytoskeleton is a main component to preserve the cell shape. It represents a cellular machinery that enables morphological changes and orchestrates important dynamic cellular functions. Thereby, it supports T-cell migration, immune synapse formation, activation and execution of effector functions. The analysis of actin rearrangements in T-cells is therefore an important field of basic and clinical research. Actin reorganization is traditionally performed using flow cytometry or confocal microscopy. However, while flow cytometry lacks spatial and structural information, confocal microscopy is time consuming and not feasible for the characterization of rare events or of un-purified primary cell populations. Here we describe a methodology to analyze actin rearrangements using InFlow microscopy, which is a hybrid technique consisting of flow cytometric and microscopic features. We show that InFlow microscopy is a valuable tool for quantification of the amount and distribution of F-actin in human T-cells after stimulation with chemokines or antigen-presenting cells.

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Citations

Mar 8, 2016·Clinical Immunology : the Official Journal of the Clinical Immunology Society·Dixita I ViswanathJordan S Orange
Nov 30, 2015·Water Research·Anna M Karpinska, John Bridgeman
Nov 26, 2015·American Journal of Transplantation : Official Journal of the American Society of Transplantation and the American Society of Transplant Surgeons·S C JuvetA Bushell
Aug 24, 2016·The Analyst·Solaire A Finkenstaedt-QuinnChristy L Haynes
Sep 14, 2016·Methods : a Companion to Methods in Enzymology·Brian K McFarlin, Melody A Gary
Oct 4, 2016·Methods : a Companion to Methods in Enzymology·Emre BaltaGuido H Wabnitz
Sep 16, 2020·Journal of Leukocyte Biology·Claudia CantoniMassimo Vitale

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