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

Elizabeth J. Mulder; Brittany Moser; Jennifer Delgado; Rachel Steinhardt; Aaron P. Esser-Kahn

doi:10.1016/j.xpro.2024.102873

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

Elizabeth J. Mulder, Brittany Moser, Jennifer Delgado, Rachel Steinhardt, Aaron P. Esser-Kahn

Research output: Contribution to journal › Article › peer-review

Abstract

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

Original language	English (US)
Article number	102873
Journal	STAR Protocols
Volume	5
Issue number	1
DOIs	https://doi.org/10.1016/j.xpro.2024.102873
State	Published - Mar 15 2024
Externally published	Yes

Keywords

AFM
Atomic Force Microscopy
Biophysics
Cell Biology
Cell culture
Cell-based Assays
Immunology
Microscopy
Single Cell

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2024.102873

Cite this

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abstract = "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",

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AU - Mulder, Elizabeth J.

AU - Moser, Brittany

AU - Delgado, Jennifer

AU - Steinhardt, Rachel

AU - Esser-Kahn, Aaron P.

PY - 2024/3/15

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AB - 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

KW - AFM

KW - Atomic Force Microscopy

KW - Biophysics

KW - Cell Biology

KW - Cell culture

KW - Cell-based Assays

KW - Immunology

KW - Microscopy

KW - Single Cell

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

Abstract

Keywords

ASJC Scopus subject areas

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