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Published on: June 13, 2011
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Utilizing EdU to Track Leukocyte Recruitment to the Brain.
Zoie K Lipfert1, Erika Arias2, Ayush Batra1,2
1Ken and Ruth Davee Department of Neurology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.
Bio-Protocol
|December 12, 2025
Summary
Detecting cell proliferation using 5-ethynyl-2'-deoxyuridine (EdU) via click chemistry offers a simpler, versatile alternative to traditional methods. This protocol details EdU
Area of Science:
- Biotechnology
- Cell Biology
- Immunology
Background:
- Traditional methods for detecting cell proliferation, such as 5-bromo-2 '-deoxyuridine (BrdU) detection, involve harsh steps like DNA denaturation.
- Copper(I)-catalyzed azide-alkyne cycloaddition (click chemistry) offers a milder and more versatile approach for detecting cell proliferation.
Purpose of the Study:
- To detail a protocol for using EdU labeling to track leukocyte recruitment in an animal model of ischemic stroke.
- To demonstrate the adaptability of EdU detection for flow cytometry and fluorescence microscopy.
Main Methods:
- EdU injection into an animal model.
- Preparation of blood and tissue samples for analysis.
- Utilizing click chemistry for EdU detection without DNA denaturation.
Main Results:
- EdU labeling is simple, compatible with routine laboratory methods, and suitable for use with genetically encoded fluorophores like GFP and tdTomato.
- EdU incorporation varies among circulating leukocyte subtypes.
- The method effectively tracks leukocyte recruitment from the bloodstream.
Conclusions:
- EdU detection is a superior alternative to BrdU detection for cell proliferation studies due to its simplicity and versatility.
- This EdU labeling protocol can be adapted for various research models requiring extended cell tracking.
- The EdU method facilitates visualization and tracking of leukocyte recruitment in models like ischemic stroke.
