In Vivo EdU Cell Proliferation Assay for Imaging

What is EdU Assay?

The EdU (5-ethynyl-2′-deoxyuridine) assay is a modern method for detecting cell proliferation by labeling newly synthesized DNA. During DNA replication, EdU is incorporated into DNA in a partial replacement of thymidine. Its specific chemical properties enable it to be detected via a gentle ‘click chemistry’ reaction, allowing proliferating cells to be visualized without the need for harsh treatments.

Compared to traditional BrdU assays, which require DNA denaturation for antibody access, EdU detection is faster, antibody-free, and preserves sample morphology. This makes it ideal for high-resolution imaging and multiplex applications in both cell culture and in vivo systems.

In vivo EdU Cell proliferation assays for imaging: What is it?​

Analyze cellular proliferation in whole organisms without harsh denaturation steps

 The ClickTech in vivo EdU Cell proliferation assays for imaging are an evolvement of EdU cell proliferation assays designed specifically for usage with whole organisms. The kit include increased EdU quantities and optimized reagents to support systemic labeling of dividing cells in vivo, enabling the analysis of biological processes such as tumor growth, neural and tissue development, and cellular regeneration.

On the contrary to BrdU, which requires DNA denaturation under harsh conditions (e.g. acid, heat, or enzymatic digestion), EdU assay enable chemistry preserves tissue morphology and antigenicity — offering a much gentler alternative for in vivo use.

EdU is incorporated into replicating DNA and can be administered through various methods, depending on the model organism:

• Injections: intraperitoneal, intramuscular or subcutaneous
• dissolved in drinking water: for continuous systemic delivery
• direct incubation e.g. zebrafish larvae

After incorporation, Edu labeled cells are visualized using a fluorescent dye through the highly specific copper-catalyzed click chemistry reaction CuAAC.

Difference between standard EdU cell proliferation kits for imaging and in vivo kits

Standard baseclick´s EdU cell proliferation assays are optimized for in vitro cell culture, where the concentrations of EdU and the protocols used are insufficient for labeling whole organisms. If researchers plan to follow cell proliferation in organism different protocols and higher amounts of EdU are needed. The ClickTech In vivo Kit addresses this with:

• Additional EdU for systemic administration
• Protocols adapted for in vivo labeling and tissue preparation
• Detection reagents tailored for animal or tissue sample analysis
• Format options for different experimental scales

baseclick is offering in vivo kits not only for EdU cell Proliferation kit for Imaging but also for other EdU cell proliferation variants as flow cytometry or high throughput screening for various wavelengths.

What does the user gain?​

ClickTech in vivo EdU Cell proliferation assays for imaging provide the customer with the advantage to have an all-in-one product in combination with protocols for both labeling of the organism and for analyzing the sample. EdU and its mild labeling conditions preserve tissue integrity.

Researchers also benefit from a reduced price when purchasing the in vivo variant instead of the ClickTech EdU Cell proliferation assay for imaging, when used in combination with additional EdU.

Features & Applications

• Suitable for imaging in mice, rats, zebrafish, and more
• Compatible with injection, water-based delivery, or incubation
• Antibody-free, fast detection (~30 minutes)
• Works with multiplex labeling and co-staining
• Analyzes S-phase activity in intact tissues

Optimization Tips

• Longer incubation → lower EdU concentration recommended
• If no EdU protocol exists, use BrdU-based references to optimize
• Lower EdU can yield comparable fluorescence to BrdU

LITERATURE

An efficient protocol for in vivo labeling of proliferating epithelial cells, C. Michel et al., 2018, J. Immunol. Methods, Vol. 457, p. 82-86.

https://doi.org/10.1016/j.jim.2018.03.015

Cisplatin-induced DNA double-strand breaks promote meiotic chromosome synapsis in PRDM9-controlled mouse hybrid sterility, L. Wang et al., 2018, eLife, Vol. 7, p. e42511.

https://doi.org/10.7554/eLife.42511

FXR Regulates Intestinal Cancer Stem Cell Proliferation, T. Fu et al., 2019, Cell, Vol. 176, p. 1098-1112.

https://doi.org/10.1016/j.cell.2019.01.036

Nuclear morphology is a deep learning biomarker of cellular senescence, I. Heckenbach et al., 2022, Nat Aging, Vol. 2, p. 742–755.

https://doi.org/10.1038/s43587-022-00263-3