Modified protocol for culturing Drosophila S2 R+ cells and adult plasmatocytes to study actin cytoskeleton dynamics

Ceileigh M. Weaver; Stephanie Makdissi; Francesca Di Cara

doi:10.1016/j.xpro.2022.101588

Modified protocol for culturing Drosophila S2 R+ cells and adult plasmatocytes to study actin cytoskeleton dynamics

Ceileigh M. Weaver, Stephanie Makdissi, Francesca Di Cara

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5 Citas (Scopus)

Resumen

Here, we describe a protocol to culture Drosophila S2R+ cells and to extract plasmatocytes from adult flies. The modified seeding approach detailed here, in combination with coating of coverslips with concanvalin A, enables enhanced adhesion and spreading of cells. We describe the steps for confocal microscopy and a detailed quantification pipeline to evaluate changes in cortical actin cytoskeleton dynamics. The protocol can be applied to a variety of genetic or chemical perturbations. For complete details on the use and execution of this protocol, please refer to Nath et al. (2022).

Idioma original	English
Número de artículo	101588
Publicación	STAR Protocols
Volumen	3
N.º	3
DOI	https://doi.org/10.1016/j.xpro.2022.101588
Estado	Published - sep. 16 2022

Nota bibliográfica

Funding Information:
This work was funded by a Project Grant from the Canadian Institutes of Health Research to F.D., a Discovery Grant from the Natural Sciences and Engineering Research Council of Canada to F.D., a Canada Foundation for Innovation JELF equipment grant to F.D., and a Dalhousie Medical Research Foundation start-up fund to F.D.

Publisher Copyright:
© 2022 The Author(s)

ASJC Scopus Subject Areas

General Neuroscience
General Biochemistry,Genetics and Molecular Biology
General Immunology and Microbiology

PubMed: MeSH publication types

Journal Article
Research Support, Non-U.S. Gov't

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abstract = "Here, we describe a protocol to culture Drosophila S2R+ cells and to extract plasmatocytes from adult flies. The modified seeding approach detailed here, in combination with coating of coverslips with concanvalin A, enables enhanced adhesion and spreading of cells. We describe the steps for confocal microscopy and a detailed quantification pipeline to evaluate changes in cortical actin cytoskeleton dynamics. The protocol can be applied to a variety of genetic or chemical perturbations. For complete details on the use and execution of this protocol, please refer to Nath et al. (2022).",

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Modified protocol for culturing Drosophila S2 R+ cells and adult plasmatocytes to study actin cytoskeleton dynamics

Resumen

Nota bibliográfica

ASJC Scopus Subject Areas

PubMed: MeSH publication types

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