@inbook{1554fcef8059465a9b2c65d6c69b74fc,
title = "Investigating IL-1β secretion using real-time single-cell imaging",
abstract = "The pro-inflammatory cytokine interleukin (IL)-1β is an important mediator of the inflammatory response. In order to perform its role in the inflammatory cascade, IL-1β must be secreted from the cell, yet it lacks a signal peptide that is required for conventional secretion, and the exact mechanism of release remains undefined. Conventional biochemical methods have limited the investigation into the processes involved in IL-1β secretion to population dynamics, yet heterogeneity between cells has been observed at a single-cell level. Here, greater sensitivity is achieved with the use of a newly developed vector that codes for a fluorescently labelled version of IL-1β. Combining this with real-time single-cell confocal microscopy using the methods described here, we have developed an effective protocol for investigating the mechanisms of IL-1β secretion and the testing of the hypothesis that IL-1β secretion requires membrane permeabilisation.",
keywords = "Confocal microscopy, IL-1β secretion, IL-1β Venus, Lentiviral transduction, Real-time single-cell imaging",
author = "Catherine Diamond and James Bagnall and Spiller, {David G.} and White, {Michael R.} and Alessandra Mortellaro and Pawel Paszek and David Brough",
note = "Funding Information: The authors are grateful to Prof Claire Bryant from the University of Cambridge for providing the immortalised BMDMs. P.P. holds a BBSRCBiotechnology and Biological Sciences Research Council David Phillips Research Fellowship (BB/I017976/1). This work was also supported by Medical Research Council Canada M.R.C. (MR/K015885/1) and Biotechnology and Biological Sciences Research Council (BB/K003097/1). The work leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2012-2017) under grant agreement n° 305564. Publisher Copyright: {\textcopyright} Springer Science+Business Media New York 2016.",
year = "2016",
doi = "10.1007/978-1-4939-3566-6_4",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "75--88",
booktitle = "Methods in Molecular Biology",
}