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bioRxiv · 10.1101/2025.07.14.664660

Optimizing scheduling in dual-pulse nucleoside labeling experiments for cell cycle analysis

Phelan, A.·Pospori, C.·Lo Celso, C.·Lee, C. F.
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Abstract

All eukaryotic cells go though a universal sequence of phases during their division cycle, where the phase timings vary according to cell type and state. Dual-pulse nucleoside labeling (DPNL) is a standard, widely applicable experimental technique to probe cell cycle kinetics at the population level, including in living organisms. In such an experimental protocol, a key scheduling parameter is the choice of waiting time between the two labeling pulses. Here, we use simulation to show that the waiting time can be optimized with regards to the signal-to-noise ratio of the cycle parameters as inferred from experimental outputs, which is of particular importance in DPNL experiments with small numbers of cells and repeats. We further discuss the procedure to perform such a task in an experimentally relevant setting.

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Phelan, A., Pospori, C., Lo Celso, C., Lee, C. F.. 2025-07-18. Optimizing scheduling in dual-pulse nucleoside labeling experiments for cell cycle analysis. https://doi.org/10.1101/2025.07.14.664660

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