Protocol for localized macrophage stimulation with small-molecule TLR agonist via fluidic force microscopy

Autor: Elizabeth J. Mulder, Brittany Moser, Jennifer Delgado, Rachel Steinhardt, Aaron P. Esser-Kahn
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: STAR Protocols, Vol 5, Iss 1, Pp 102873- (2024)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2024.102873
Popis: Summary: Here, we present a protocol to deliver nanoliter volumes of Toll-like receptor (TLR) agonist onto a culture of nuclear factor κB (NF-κB) reporter macrophages using fluidic force microscopy and a micron-scale probe. We describe steps for quantifying the dose of agonist by modeling their diffusion with experimental inputs. We then detail procedures for quantifying and categorizing macrophage responses to individual and varied doses and combining agonist concentration and macrophage response to analyze the NF-κB response to localized TLR stimulation.For complete details on the use and execution of this protocol, please refer to Mulder et al. (2024).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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