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Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10
Flow cytometry is a method widely used to quantify suspended solids such as cells or bacteria in a size range from 0.5 to several tens of micrometers in diameter. In addition to a characterization of forward and sideward scatter properties, it enables the use of fluorescent labeled markers like anti...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MyJove Corporation
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5226252/ https://www.ncbi.nlm.nih.gov/pubmed/27911374 http://dx.doi.org/10.3791/54721 |
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author | Süring, Katrin Bach, Sabine Höflich, Conny Straff, Wolfgang |
author_facet | Süring, Katrin Bach, Sabine Höflich, Conny Straff, Wolfgang |
author_sort | Süring, Katrin |
collection | PubMed |
description | Flow cytometry is a method widely used to quantify suspended solids such as cells or bacteria in a size range from 0.5 to several tens of micrometers in diameter. In addition to a characterization of forward and sideward scatter properties, it enables the use of fluorescent labeled markers like antibodies to detect respective structures. Using indirect antibody staining, flow cytometry is employed here to quantify birch pollen allergen (precisely Bet v 1)-loaded particles of 0.5 to 10 µm in diameter in inhalable particulate matter (PM10, particle size ≤10 µm in diameter). PM10 particles may act as carriers of adsorbed allergens possibly transporting them to the lower respiratory tract, where they could trigger allergic reactions. So far the allergen content of PM10 has been studied by means of enzyme linked immunosorbent assays (ELISAs) and scanning electron microscopy. ELISA measures the dissolved and not the particle-bound allergen. Compared to scanning electron microscopy, which can visualize allergen-loaded particles, flow cytometry may additionally quantify them. As allergen content of ambient air can deviate from birch pollen count, allergic symptoms might perhaps correlate better with allergen exposure than with pollen count. In conjunction with clinical data, the presented method offers the opportunity to test in future experiments whether allergic reactions to birch pollen antigens are associated with the Bet v 1 allergen content of PM10 particles >0.5 µm. |
format | Online Article Text |
id | pubmed-5226252 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-52262522017-01-26 Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10 Süring, Katrin Bach, Sabine Höflich, Conny Straff, Wolfgang J Vis Exp Immunology Flow cytometry is a method widely used to quantify suspended solids such as cells or bacteria in a size range from 0.5 to several tens of micrometers in diameter. In addition to a characterization of forward and sideward scatter properties, it enables the use of fluorescent labeled markers like antibodies to detect respective structures. Using indirect antibody staining, flow cytometry is employed here to quantify birch pollen allergen (precisely Bet v 1)-loaded particles of 0.5 to 10 µm in diameter in inhalable particulate matter (PM10, particle size ≤10 µm in diameter). PM10 particles may act as carriers of adsorbed allergens possibly transporting them to the lower respiratory tract, where they could trigger allergic reactions. So far the allergen content of PM10 has been studied by means of enzyme linked immunosorbent assays (ELISAs) and scanning electron microscopy. ELISA measures the dissolved and not the particle-bound allergen. Compared to scanning electron microscopy, which can visualize allergen-loaded particles, flow cytometry may additionally quantify them. As allergen content of ambient air can deviate from birch pollen count, allergic symptoms might perhaps correlate better with allergen exposure than with pollen count. In conjunction with clinical data, the presented method offers the opportunity to test in future experiments whether allergic reactions to birch pollen antigens are associated with the Bet v 1 allergen content of PM10 particles >0.5 µm. MyJove Corporation 2016-11-19 /pmc/articles/PMC5226252/ /pubmed/27911374 http://dx.doi.org/10.3791/54721 Text en Copyright © 2016, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/ |
spellingShingle | Immunology Süring, Katrin Bach, Sabine Höflich, Conny Straff, Wolfgang Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10 |
title | Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10 |
title_full | Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10 |
title_fullStr | Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10 |
title_full_unstemmed | Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10 |
title_short | Flow Cytometric Analysis of Particle-bound Bet v 1 Allergen in PM10 |
title_sort | flow cytometric analysis of particle-bound bet v 1 allergen in pm10 |
topic | Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5226252/ https://www.ncbi.nlm.nih.gov/pubmed/27911374 http://dx.doi.org/10.3791/54721 |
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