Quantitative live-cell imaging of Candida albicans escape from immune phagocytes

Françios A.B. Olivier; Ana Traven

doi:10.1016/j.xpro.2023.102737

Quantitative live-cell imaging of Candida albicans escape from immune phagocytes

Research output: Contribution to journal › Article › Other › peer-review

Abstract

Population-level dynamics of host-pathogen interactions can be characterized using quantitative live-cell imaging. Here, we present a protocol for infecting macrophages with the fungal pathogen Candida albicans in vitro and quantitative live-cell imaging of immune and pathogen responses. We describe steps for detailed image analysis and provide resources for quantification of phagocytosis and pathogen escape, as well as macrophage membrane permeabilization and viability. This protocol is modifiable for applications with a range of pathogens, immune cell types, and host-pathogen mechanisms. For complete details on the use and execution of this protocol, please refer to Olivier et al.¹

Original language	English
Article number	102737
Number of pages	30
Journal	STAR Protocols
Volume	4
Issue number	4
DOIs	https://doi.org/10.1016/j.xpro.2023.102737
Publication status	Published - 15 Dec 2023

Keywords

Immunology
Microbiology
Microscopy
Model Organisms

Access to Document

10.1016/j.xpro.2023.102737Licence: CC BY-NC-ND

Cite this

@article{81c6804a255a4e09a142d27eee91803a,

title = "Quantitative live-cell imaging of Candida albicans escape from immune phagocytes",

abstract = "Population-level dynamics of host-pathogen interactions can be characterized using quantitative live-cell imaging. Here, we present a protocol for infecting macrophages with the fungal pathogen Candida albicans in vitro and quantitative live-cell imaging of immune and pathogen responses. We describe steps for detailed image analysis and provide resources for quantification of phagocytosis and pathogen escape, as well as macrophage membrane permeabilization and viability. This protocol is modifiable for applications with a range of pathogens, immune cell types, and host-pathogen mechanisms. For complete details on the use and execution of this protocol, please refer to Olivier et al.1",

keywords = "Immunology, Microbiology, Microscopy, Model Organisms",

author = "Olivier, \{Fran{\c c}ios A.B.\} and Ana Traven",

note = "Funding Information: This work was supported by funding from the Australian National Health and Medical Research Council (NHMRC) project grant APP1158678 (to A.T.) and Australian Research Council (ARC) Future Fellowship FT190100733 (to A.T.). F.A.B.O. was funded by an Australian Government RTP PhD scholarship. We thank Mihalis Lionakis for sharing the dTomato C. albicans strain with us. We thank Volker Hilsenstein for consultation over development of the CellProfiler image analysis pipeline and the Monash Microimaging Facility for technical support, training, and maintenance of equipment. Conceptualization, F.A.B.O. and A.T.; methodology, F.A.B.O.; formal analysis, F.A.B.O.; visualization, F.A.B.O.; resources, A.T.; writing – original draft, F.A.B.O.; writing – review and editing, F.A.B.O. and A.T.; supervision, A.T.; project administration, A.T.; funding acquisition, A.T. The authors declare no competing interests. Funding Information: This work was supported by funding from the Australian National Health and Medical Research Council (NHMRC) project grant APP1158678 (to A.T.) and Australian Research Council (ARC) Future Fellowship FT190100733 (to A.T.). F.A.B.O. was funded by an Australian Government RTP PhD scholarship. We thank Mihalis Lionakis for sharing the dTomato C. albicans strain with us. We thank Volker Hilsenstein for consultation over development of the CellProfiler image analysis pipeline and the Monash Microimaging Facility for technical support, training, and maintenance of equipment. Publisher Copyright: {\textcopyright} 2023 The Author(s)",

year = "2023",

month = dec,

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doi = "10.1016/j.xpro.2023.102737",

language = "English",

volume = "4",

journal = "STAR Protocols",

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AU - Olivier, Françios A.B.

AU - Traven, Ana

N1 - Funding Information: This work was supported by funding from the Australian National Health and Medical Research Council (NHMRC) project grant APP1158678 (to A.T.) and Australian Research Council (ARC) Future Fellowship FT190100733 (to A.T.). F.A.B.O. was funded by an Australian Government RTP PhD scholarship. We thank Mihalis Lionakis for sharing the dTomato C. albicans strain with us. We thank Volker Hilsenstein for consultation over development of the CellProfiler image analysis pipeline and the Monash Microimaging Facility for technical support, training, and maintenance of equipment. Conceptualization, F.A.B.O. and A.T.; methodology, F.A.B.O.; formal analysis, F.A.B.O.; visualization, F.A.B.O.; resources, A.T.; writing – original draft, F.A.B.O.; writing – review and editing, F.A.B.O. and A.T.; supervision, A.T.; project administration, A.T.; funding acquisition, A.T. The authors declare no competing interests. Funding Information: This work was supported by funding from the Australian National Health and Medical Research Council (NHMRC) project grant APP1158678 (to A.T.) and Australian Research Council (ARC) Future Fellowship FT190100733 (to A.T.). F.A.B.O. was funded by an Australian Government RTP PhD scholarship. We thank Mihalis Lionakis for sharing the dTomato C. albicans strain with us. We thank Volker Hilsenstein for consultation over development of the CellProfiler image analysis pipeline and the Monash Microimaging Facility for technical support, training, and maintenance of equipment. Publisher Copyright: © 2023 The Author(s)

PY - 2023/12/15

Y1 - 2023/12/15

N2 - Population-level dynamics of host-pathogen interactions can be characterized using quantitative live-cell imaging. Here, we present a protocol for infecting macrophages with the fungal pathogen Candida albicans in vitro and quantitative live-cell imaging of immune and pathogen responses. We describe steps for detailed image analysis and provide resources for quantification of phagocytosis and pathogen escape, as well as macrophage membrane permeabilization and viability. This protocol is modifiable for applications with a range of pathogens, immune cell types, and host-pathogen mechanisms. For complete details on the use and execution of this protocol, please refer to Olivier et al.1

AB - Population-level dynamics of host-pathogen interactions can be characterized using quantitative live-cell imaging. Here, we present a protocol for infecting macrophages with the fungal pathogen Candida albicans in vitro and quantitative live-cell imaging of immune and pathogen responses. We describe steps for detailed image analysis and provide resources for quantification of phagocytosis and pathogen escape, as well as macrophage membrane permeabilization and viability. This protocol is modifiable for applications with a range of pathogens, immune cell types, and host-pathogen mechanisms. For complete details on the use and execution of this protocol, please refer to Olivier et al.1

KW - Immunology

KW - Microbiology

KW - Microscopy

KW - Model Organisms

UR - https://www.scopus.com/pages/publications/85177491746

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DO - 10.1016/j.xpro.2023.102737

M3 - Article

C2 - 37980567

AN - SCOPUS:85177491746

SN - 2666-1667

VL - 4

JO - STAR Protocols

JF - STAR Protocols

IS - 4

M1 - 102737

ER -

Quantitative live-cell imaging of Candida albicans escape from immune phagocytes

Abstract

Keywords

Access to Document

Other files and links

The escape of Candida albicans from macrophages is enabled by the fungal toxin candidalysin and two host cell death pathways

An investigation into metabolic competition between host and pathogen in infection.

Monash Micro Imaging (MMI)

Cite this

Quantitative live-cell imaging of Candida albicans escape from immune phagocytes

Abstract

Keywords

Access to Document

Other files and links

Research output

The escape of Candida albicans from macrophages is enabled by the fungal toxin candidalysin and two host cell death pathways

Projects

An investigation into metabolic competition between host and pathogen in infection.

Equipment

Monash Micro Imaging (MMI)

Cite this