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Specifically, we explain how to inject into animals exogenous markers of proliferation (i.e., Brd U) and prepare brain tissues to perform immunofluorescence reactions for neuronal markers in combination with Brd U staining.

As Brd U is incorporated in the DNA during the S-phase of the cell cycle of proliferating cells and is then inherited by daughter cells, by coupling Brd U-immunoreactivity together with the immunolabeling for neuronal markers, we provide the general procedures that can be applied to identify adult-born neurons and to characterize their specific phenotypes in different brain regions, under physiological condition or in pathological states.

In this two-part series on the thymidine analog Brd U, we tell the story of how Brd U revolutionized the field of neurogenesis as well as its recently identified limitations as a lineage tracer for neurogenesis studies.

Up until the mid-1960s, scientists believed that the generation of functional neurons from neural precursors (neurogenesis) only occurred during the embryonic and peri-natal stages in mammals.

Additionally, this technique is biased towards identifying microorganisms with A- and T-rich genomes.

Cell proliferation can be measured with the thymidine analog Brd U (5-bromo-2’-deoxyuridine) following its incorporation into newly synthesized DNA and its subsequent detection with an anti-Brd U antibody.

Subsequent sequencing of the labeled DNA can then be used to identify the microbial taxa that participated in the degradation of the added carbon source.The antibody used in this case will recognize and stain for cells that have incorporated Brd U into their genetic code.Any cell that stains positive for Brd U can be said to have undergone proliferation after the time of injection. Tritiated Thymidine The most important reason Brd U has become the predominant method to identify proliferating cells is its efficiency.When a scientist injects Brd U into the bloodstream of a test animal, the chemical becomes available to all cells – most importantly, those which are proliferating.As the cells build their new genetic strand, they pull either Thymidine nucleotides or Brd U analogues from their environment.Overall, Brd U allows for a significantly more efficient and cost effective method of identifying cells undergoing proliferation.