Skip to main content

Research Repository

Advanced Search

Fluorescence-activated cell sorting and quantitative real-time PCR to reveal VEGF-expressing macrophage populations in the Zebrafish larvae

Herman, Andrew; Greenhough, Alexander; Gurevich, David B.

Fluorescence-activated cell sorting and quantitative real-time PCR to reveal VEGF-expressing macrophage populations in the Zebrafish larvae Thumbnail


Authors

Andrew Herman

David B. Gurevich



Abstract

The transparent, genetically tractable zebrafish is increasingly recognized as a useful model to both live image and uncover mechanistic insight into cell interactions governing tissue homeostasis, pathology, and regeneration. Here, we describe a protocol for the isolation of macrophages from zebrafish wounds using fluorescence-activated cell sorting (FACS), and the identification of specific pro-angiogenic macrophage populations that express high levels of vascular endothelial growth factor (vegf) using quantitative real-time PCR (qPCR). The cell dissociation and FACS sorting techniques have been optimized for immune cells and successfully used to isolate other fluorescently marked populations within the wound such as neutrophils and endothelial cells. More broadly, this protocol can be easily adapted to other contexts where identification of pro-angiogenic immune cells is transformative for understanding, from development to pathologies such as infection, cancer, and diabetes.

Citation

Herman, A., Greenhough, A., & Gurevich, D. B. (2022). Fluorescence-activated cell sorting and quantitative real-time PCR to reveal VEGF-expressing macrophage populations in the Zebrafish larvae. In VEGF Signaling (325-337). Humana Press. https://doi.org/10.1007/978-1-0716-2217-9_24

Acceptance Date Apr 21, 2022
Online Publication Date Apr 21, 2022
Publication Date Apr 21, 2022
Deposit Date Jun 10, 2022
Publicly Available Date Apr 22, 2024
Publisher Humana Press
Pages 325-337
Series Title Methods in Molecular Biology book series (MIMB,volume 2475)
Book Title VEGF Signaling
Chapter Number 24
DOI https://doi.org/10.1007/978-1-0716-2217-9_24
Keywords Zebrafish; Wounding; VEGF; Immune cells; Macrophages; FACS; Quantitative; PCR
Public URL https://uwe-repository.worktribe.com/output/9621188
Publisher URL https://link.springer.com/protocol/10.1007/978-1-0716-2217-9_24

Files

Fluorescence-activated cell sorting and quantitative real-time PCR to reveal VEGF-expressing macrophage populations in the Zebrafish larvae (780 Kb)
PDF

Licence
http://www.rioxx.net/licenses/all-rights-reserved

Publisher Licence URL
http://www.rioxx.net/licenses/all-rights-reserved

Copyright Statement
This is a copy of the author’s accepted manuscript. The final published version is available here: https://link.springer.com/protocol/10.1007/978-1-0716-2217-9_24

© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature






You might also like



Downloadable Citations