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Barry D.J.,Dublin Institute of Technology | Barry D.J.,The Francis Crick Institute Lincolns Inn Fields Laboratory | Williams G.A.,Dublin Institute of Technology | Chan C.,Dublin Institute of Technology | Chan C.,University of Hong Kong
Biotechnology Progress | Year: 2015

The morphological quantification of filamentous microbes represents an important analytical technique in the optimization of bioprocesses involving such organisms, given the demonstrated links between morphology and metabolite yield. However, in many studies, much of this quantification has required some degree of manual intervention, if it has been conducted at all, burdening biotechnologists with a time-consuming process and potentially introducing bias into analyses. Here, software for the automated quantification of filamentous microbes is presented, implemented as a plug-in for the widely used, freely available image analysis package, ImageJ. The software, together with all related source code, documentation and test data, is freely available to the community via an online repository. © 2015 American Institute of Chemical Engineers. Source

Jenkins R.P.,The Francis Crick Institute Lincolns Inn Fields Laboratory | Hanisch A.,The Francis Crick Institute Lincolns Inn Fields Laboratory | Soza-Ried C.,The Francis Crick Institute Lincolns Inn Fields Laboratory | Sahai E.,The Francis Crick Institute Lincolns Inn Fields Laboratory | Lewis J.,The Francis Crick Institute Lincolns Inn Fields Laboratory
PLoS Computational Biology | Year: 2015

The somite segmentation clock is a robust oscillator used to generate regularly-sized segments during early vertebrate embryogenesis. It has been proposed that the clocks of neighbouring cells are synchronised via inter-cellular Notch signalling, in order to overcome the effects of noisy gene expression. When Notch-dependent communication between cells fails, the clocks of individual cells operate erratically and lose synchrony over a period of about 5 to 8 segmentation clock cycles (2–3 hours in the zebrafish). Here, we quantitatively investigate the effects of stochasticity on cell synchrony, using mathematical modelling, to investigate the likely source of such noise. We find that variations in the transcription, translation and degradation rate of key Notch signalling regulators do not explain the in vivo kinetics of desynchronisation. Rather, the analysis predicts that clock desynchronisation, in the absence of Notch signalling, is due to the stochastic dissociation of Her1/7 repressor proteins from the oscillating her1/7 autorepressed target genes. Using in situ hybridisation to visualise sites of active her1 transcription, we measure an average delay of approximately three minutes between the times of activation of the two her1 alleles in a cell. Our model shows that such a delay is sufficient to explain the in vivo rate of clock desynchronisation in Notch pathway mutant embryos and also that Notch-mediated synchronisation is sufficient to overcome this stochastic variation. This suggests that the stochastic nature of repressor/DNA dissociation is the major source of noise in the segmentation clock. © 2015 Jenkins et al. Source

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