Abstract
Imaging flow cytometry has been applied to address questions in infection biology, in particular, infections induced by intracellular pathogens. This methodology, which utilizes specialized analytic software makes it possible to analyze hundreds of quantified features for hundreds of thousands of individual cellular or subcellular events in a single experiment. Imaging flow cytometry analysis of host cell-pathogen interaction can thus quantitatively addresses a variety of biological questions related to intracellular infection, including cell counting, internalization score, and subcellular patterns of co-localization. Here, we provide an overview of recent achievements in the use of fluorescently labeled prokaryotic or eukaryotic pathogens in human cellular infections in analysis of host-pathogen interactions. Specifically, we give examples of Imagestream-based analysis of cell lines infected with Toxoplasma gondii or Mycobacterium tuberculosis. Furthermore, we illustrate the capabilities of imaging flow cytometry using a combination of standard IDEAS™ software and the more recently developed Feature Finder algorithm, which is capable of identifying statistically significant differences between researcher-defined image galleries. We argue that the combination of imaging flow cytometry with these software platforms provides a powerful new approach to understanding host control of intracellular pathogens.
| Original language | English |
|---|---|
| Pages (from-to) | 91-104 |
| Number of pages | 14 |
| Journal | Methods |
| Volume | 112 |
| DOIs | |
| Publication status | Published - Jan 1 2017 |
Fingerprint
Keywords
- Cellular heterogeneity
- Colocalization
- Feature Finder
- Fluorescent protein
- Imaging flow cytometry
- Intracellular pathogen
- Mycobacteria tuberculosis
- Phagosome maturation
- Rab5
- Rab7
- Toxoplasma gondii
ASJC Scopus subject areas
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
Imaging flow cytometry analysis of intracellular pathogens. / Haridas, Viraga; Ranjbar, Shahin; Vorobjev, Ivan A.; Goldfeld, Anne E.; Barteneva, Natasha S.
In: Methods, Vol. 112, 01.01.2017, p. 91-104.Research output: Contribution to journal › Review article
}
TY - JOUR
T1 - Imaging flow cytometry analysis of intracellular pathogens
AU - Haridas, Viraga
AU - Ranjbar, Shahin
AU - Vorobjev, Ivan A.
AU - Goldfeld, Anne E.
AU - Barteneva, Natasha S.
PY - 2017/1/1
Y1 - 2017/1/1
N2 - Imaging flow cytometry has been applied to address questions in infection biology, in particular, infections induced by intracellular pathogens. This methodology, which utilizes specialized analytic software makes it possible to analyze hundreds of quantified features for hundreds of thousands of individual cellular or subcellular events in a single experiment. Imaging flow cytometry analysis of host cell-pathogen interaction can thus quantitatively addresses a variety of biological questions related to intracellular infection, including cell counting, internalization score, and subcellular patterns of co-localization. Here, we provide an overview of recent achievements in the use of fluorescently labeled prokaryotic or eukaryotic pathogens in human cellular infections in analysis of host-pathogen interactions. Specifically, we give examples of Imagestream-based analysis of cell lines infected with Toxoplasma gondii or Mycobacterium tuberculosis. Furthermore, we illustrate the capabilities of imaging flow cytometry using a combination of standard IDEAS™ software and the more recently developed Feature Finder algorithm, which is capable of identifying statistically significant differences between researcher-defined image galleries. We argue that the combination of imaging flow cytometry with these software platforms provides a powerful new approach to understanding host control of intracellular pathogens.
AB - Imaging flow cytometry has been applied to address questions in infection biology, in particular, infections induced by intracellular pathogens. This methodology, which utilizes specialized analytic software makes it possible to analyze hundreds of quantified features for hundreds of thousands of individual cellular or subcellular events in a single experiment. Imaging flow cytometry analysis of host cell-pathogen interaction can thus quantitatively addresses a variety of biological questions related to intracellular infection, including cell counting, internalization score, and subcellular patterns of co-localization. Here, we provide an overview of recent achievements in the use of fluorescently labeled prokaryotic or eukaryotic pathogens in human cellular infections in analysis of host-pathogen interactions. Specifically, we give examples of Imagestream-based analysis of cell lines infected with Toxoplasma gondii or Mycobacterium tuberculosis. Furthermore, we illustrate the capabilities of imaging flow cytometry using a combination of standard IDEAS™ software and the more recently developed Feature Finder algorithm, which is capable of identifying statistically significant differences between researcher-defined image galleries. We argue that the combination of imaging flow cytometry with these software platforms provides a powerful new approach to understanding host control of intracellular pathogens.
KW - Cellular heterogeneity
KW - Colocalization
KW - Feature Finder
KW - Fluorescent protein
KW - Imaging flow cytometry
KW - Intracellular pathogen
KW - Mycobacteria tuberculosis
KW - Phagosome maturation
KW - Rab5
KW - Rab7
KW - Toxoplasma gondii
UR - http://www.scopus.com/inward/record.url?scp=84994494300&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84994494300&partnerID=8YFLogxK
U2 - 10.1016/j.ymeth.2016.09.007
DO - 10.1016/j.ymeth.2016.09.007
M3 - Review article
AN - SCOPUS:84994494300
VL - 112
SP - 91
EP - 104
JO - Methods
JF - Methods
SN - 1046-2023
ER -