Protocol for total internal reflection fluorescence (TIRF) microscopy image preprocessing and radial analysis of LFA-1 in T cell immune synapses

This repository contains the raw TIRF microscopy images, Fiji/ImageJ macros, R scripts, and analysis outputs underlying the protocol “Protocol for total internal reflection fluorescence (TIRF) microscopy image preprocessing and radial analysis of LFA-1 in T cell immune synapses.” The workflow provides modular tools for background subtraction, segmentation, pSMAC center identification, center-cropping, and radial intensity analysis of LFA-1 distribution in T cell immune synapses. All data and code are provided to support transparency and reproducibility of the analysis pipeline, and to enable users to adapt the macros and scripts for related studies of immune synapse organization or other TIRF-based imaging assays.