New method to analyze cumulative BrdU labeling data

ROST FABIAN; CHARA OSVALDO; RODRIGO-ALBORS AIDA; DEUTSCH ANDREAS; BRUSCH LUTZ; TANAKA ELLY M

Dresden

Conferencia; Cytometry Across Disciplines; 2013

The cumulative Bromodeoxyuridine (BrdU) labeling protocol is a standard method to measure the cell cycle length and the S-Phase duration [Nowakowski et al., 1989]. Here, we overcome protocol limitations with a new data analysis method. This opens the possibility to apply the protocol to a wider range of experimental setups.The original method is only applicable in a limited number of cases where, firstly, all cells divide asymmetrically (implying a constant number of cells), secondly, all cells proliferate at the same rate. Moreover it depends on a ?good? initial guess of the cell cycle parameters and lacks rigorous error estimation.Here, we extend the standard method by providing a suitable fitting algorithm that overcomes the aforementioned limitations. We apply the new algorithm to a biological test case: ependymal cells of the spinal cord of the axolotl (A. mexicanum). Finally, we investigate how multiple sub-populations cycling at different cell cycle lengths would shape the BrdU data.Reference: Nowakowski , Lewin & Miller, 1989. J Neurocytol. 18: 311-318.