Quantification of Immune Mediator Levels in Human Mucosal Lining Fluid
Published: January 31, 2024
Abstract
Source: Wolsk, H. M. et al., Noninvasive Sampling of Mucosal Lining Fluid for the Quantification of In Vivo Upper Airway Immune-mediator Levels. J. Vis. Exp. (2017)
This video demonstrates the quantification of immune mediator levels in neonatal human nasal mucosal lining fluid extract. An electrochemiluminescence-based multiplex assay system measures these mediators by evaluating the intensity of emitted light when they are excited.
Protocol
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. Experimental Setup Use sheets of filter paper (fibrous hydroxylated-polyester sheets, see the Table of Materials). Cut out strips 3 x 15 mm in size for a 1-month-old child and in an L-shape for older children and adults (5 x 20 x 10 mm (short arm of the L)). Insert one piece…
Divulgations
The authors have nothing to disclose.
Materials
| Fibrous hydroxylatedpolyester sheets | Accuwik Ultra | SPR0730 | Filter paper |
| Milliplex Assay Buffer | Millipore | L-AB | Buffer |
| low-protein binding storage plates | Thermo Scientific | CLS8161 | Plates |
| Protease Inhibitor | Roche | 11873580001 | Complete EDTA-free Protease Inhibitor Cocktail |
| Reader of multi-spot plates | Mesoscale | NA | Sector imager 6000 |
| Assays | Mesoscale | Human 10-plex TH1/TH2 cytokine assay and 9-plex chemokine assay, and singleplex IL-17A, TGF-β1and TSLP. A description can be found online on www.mesoscale.com |
Tags
Citer Cet Article
Quantification of Immune Mediator Levels in Human Mucosal Lining Fluid. J. Vis. Exp. (Pending Publication), e21898, doi: (2024).