Automated Image-Based Quantification of Neutrophil Extracellular Traps Using NETQUANT
(2019) In Journal of visualized experiments : JoVE- Abstract
Neutrophil extracellular traps (NETs) are web-like antimicrobial structures consisting of DNA and granule derived antimicrobial proteins. Immunofluorescence microscopy and image-based quantification methods remain important tools to quantitate neutrophil extracellular trap formation. However, there are key limitations to the immunofluorescence-based methods that are currently available for quantifying NETs. Manual methods of image-based NET quantification are often subjective, prone to error and tedious for users, especially non-experienced users. Also, presently available software options for quantification are either semi-automatic or require training prior to operation. Here, we demonstrate the implementation of an automated... (More)
Neutrophil extracellular traps (NETs) are web-like antimicrobial structures consisting of DNA and granule derived antimicrobial proteins. Immunofluorescence microscopy and image-based quantification methods remain important tools to quantitate neutrophil extracellular trap formation. However, there are key limitations to the immunofluorescence-based methods that are currently available for quantifying NETs. Manual methods of image-based NET quantification are often subjective, prone to error and tedious for users, especially non-experienced users. Also, presently available software options for quantification are either semi-automatic or require training prior to operation. Here, we demonstrate the implementation of an automated immunofluorescence-based image quantification method to evaluate NET formation called NETQUANT. The software is easy to use and has a user-friendly graphical user interface (GUI). It considers biologically relevant parameters such as an increase in the surface area and DNA:NET marker protein ratio, and nuclear deformation to define NET formation. Furthermore, this tool is built as a freely available app, and allows for single-cell resolution quantification and analysis.
(Less)
- author
- Mohanty, Tirthankar LU and Nordenfelt, Pontus LU
- organization
- publishing date
- 2019-11-27
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of visualized experiments : JoVE
- issue
- 153
- article number
- e58528
- publisher
- JoVE
- external identifiers
-
- pmid:31840654
- scopus:85076554865
- ISSN
- 1940-087X
- DOI
- 10.3791/58528
- language
- English
- LU publication?
- yes
- id
- 03fd1948-042a-4f4e-93a3-ad8bbadd3aa3
- date added to LUP
- 2019-12-22 07:08:35
- date last changed
- 2024-03-20 03:06:59
@article{03fd1948-042a-4f4e-93a3-ad8bbadd3aa3, abstract = {{<p>Neutrophil extracellular traps (NETs) are web-like antimicrobial structures consisting of DNA and granule derived antimicrobial proteins. Immunofluorescence microscopy and image-based quantification methods remain important tools to quantitate neutrophil extracellular trap formation. However, there are key limitations to the immunofluorescence-based methods that are currently available for quantifying NETs. Manual methods of image-based NET quantification are often subjective, prone to error and tedious for users, especially non-experienced users. Also, presently available software options for quantification are either semi-automatic or require training prior to operation. Here, we demonstrate the implementation of an automated immunofluorescence-based image quantification method to evaluate NET formation called NETQUANT. The software is easy to use and has a user-friendly graphical user interface (GUI). It considers biologically relevant parameters such as an increase in the surface area and DNA:NET marker protein ratio, and nuclear deformation to define NET formation. Furthermore, this tool is built as a freely available app, and allows for single-cell resolution quantification and analysis.</p>}}, author = {{Mohanty, Tirthankar and Nordenfelt, Pontus}}, issn = {{1940-087X}}, language = {{eng}}, month = {{11}}, number = {{153}}, publisher = {{JoVE}}, series = {{Journal of visualized experiments : JoVE}}, title = {{Automated Image-Based Quantification of Neutrophil Extracellular Traps Using NETQUANT}}, url = {{http://dx.doi.org/10.3791/58528}}, doi = {{10.3791/58528}}, year = {{2019}}, }