Quantitative assessment of neutrophil phagocytosis using flow cytometry
Nordenfelt, Pontus LU
(2014)
In Methods in Molecular Biology
1124.
p.89-279
- Abstract
Neutrophils have an incredible ability to find and eradicate intruders such as bacteria and fungi. They do this largely through the process of phagocytosis, where the target is internalized into a phagosome, and eventually destroyed by the hostile phagosomal environment. It is important to study phagocytosis in order to understand how neutrophils interact with various pathogens and how they respond to different stimuli. Here, I describe a method to study neutrophil phagocytosis of bacteria using flow cytometry. The bacteria are fluorescently labeled before being introduced to neutrophils. After phagocytosis, both any remaining extracellular bacteria and neutrophils are labeled using one-step staining before three-color analysis. To... (More)
Neutrophils have an incredible ability to find and eradicate intruders such as bacteria and fungi. They do this largely through the process of phagocytosis, where the target is internalized into a phagosome, and eventually destroyed by the hostile phagosomal environment. It is important to study phagocytosis in order to understand how neutrophils interact with various pathogens and how they respond to different stimuli. Here, I describe a method to study neutrophil phagocytosis of bacteria using flow cytometry. The bacteria are fluorescently labeled before being introduced to neutrophils. After phagocytosis, both any remaining extracellular bacteria and neutrophils are labeled using one-step staining before three-color analysis. To assess phagocytosis, first the average time it takes for the neutrophils to internalize all bound bacteria is determined. Experiments are then performed using that time point while varying the bacteria-to-neutrophil ratio for full control of the analysis. Due to the ease with which multiple samples can be analyzed, and the quantitative nature of flow cytometry, this approach is both reproducible and sensitive.
(Less)
- author
- Nordenfelt, Pontus
LU
- publishing date
- 2014
- type
- Chapter in Book/Report/Conference proceeding
- publication status
- published
- keywords
- Flow Cytometry, Humans, Neutrophils, Phagocytosis, Journal Article, Research Support, Non-U.S. Gov't
- host publication
- Neutrophil Methods and Protocols
- series title
- Methods in Molecular Biology
- editor
- Quinn, Mark T. and DeLeo, Frank R.
- volume
- 1124
- pages
- 11 pages
- publisher
- Humana Press
- external identifiers
-
- pmid:24504959
- scopus:84899000167
- ISSN
- 1064-3745
- ISBN
- 978-1-62703-845-4
- 978-1-62703-844-7
- DOI
- 10.1007/978-1-62703-845-4_18
- language
- English
- LU publication?
- no
- id
- 5b8fee79-0626-451c-9d42-0273c618b6cc
- date added to LUP
- 2017-02-16 08:03:58
- date last changed
- 2024-01-13 14:36:51
@inbook{5b8fee79-0626-451c-9d42-0273c618b6cc,
abstract = {{<p>Neutrophils have an incredible ability to find and eradicate intruders such as bacteria and fungi. They do this largely through the process of phagocytosis, where the target is internalized into a phagosome, and eventually destroyed by the hostile phagosomal environment. It is important to study phagocytosis in order to understand how neutrophils interact with various pathogens and how they respond to different stimuli. Here, I describe a method to study neutrophil phagocytosis of bacteria using flow cytometry. The bacteria are fluorescently labeled before being introduced to neutrophils. After phagocytosis, both any remaining extracellular bacteria and neutrophils are labeled using one-step staining before three-color analysis. To assess phagocytosis, first the average time it takes for the neutrophils to internalize all bound bacteria is determined. Experiments are then performed using that time point while varying the bacteria-to-neutrophil ratio for full control of the analysis. Due to the ease with which multiple samples can be analyzed, and the quantitative nature of flow cytometry, this approach is both reproducible and sensitive.</p>}},
author = {{Nordenfelt, Pontus}},
booktitle = {{Neutrophil Methods and Protocols}},
editor = {{Quinn, Mark T. and DeLeo, Frank R.}},
isbn = {{978-1-62703-845-4}},
issn = {{1064-3745}},
keywords = {{Flow Cytometry; Humans; Neutrophils; Phagocytosis; Journal Article; Research Support, Non-U.S. Gov't}},
language = {{eng}},
pages = {{89--279}},
publisher = {{Humana Press}},
series = {{Methods in Molecular Biology}},
title = {{Quantitative assessment of neutrophil phagocytosis using flow cytometry}},
url = {{http://dx.doi.org/10.1007/978-1-62703-845-4_18}},
doi = {{10.1007/978-1-62703-845-4_18}},
volume = {{1124}},
year = {{2014}},
}