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Lang, L. F.

Publications and source records attributed to Lang, L. F..

2 recordsLinked to original sources

Joint Motion Estimation and Source Identification using Convective Regularisation with an Application to the Analysis of Laser Nanoablations

We propose a variational method for joint motion estimation and source identification in one-dimensional image sequences. The problem is motivated by fluorescence microscopy data of laser nanoablations of cell membranes in live Drosophila embryos, which can be conveniently--and without loss of significant information--represented in space-time plots, so called kymographs. Based on mechanical models of tissue formation, we propose a variational formulation that is based on the nonhomogenous continuity equation and investigate the solution of this ill-posed inverse problem using convective regularisation. We show existence of a minimiser of the minimisation problem, derive the associated Euler-Lagrange equations, and numerically solve them using a finite element discretisation together with Newtons method. Based on synthetic data, we demonstrate that source estimation can be crucial whenever signal variations can not be explained by advection alone. Furthermore, we perform an extensive evaluation and comparison of various models, including standard optical flow, based on manually annotated kymographs that measure velocities of visible features. Finally, we present results for data generated by a mechanical model of tissue formation and demonstrate that our approach reliably estimates both a velocity and a source.

biophysics

Optical flow analysis reveals that Kinesin-mediated advection impacts on the orientation of microtubules

The orientation of microtubule networks is exploited by motors to deliver cargoes to specific intracellular destinations, and is thus essential for cell polarity and function. Reconstituted in vitro systems have largely contributed to understanding the molecular framework regulating the behavior of microtubule filaments. In cells however, microtubules are exposed to various biomechanical forces that might impact on their orientation, but little is known about it. Oocytes, which display forceful cytoplasmic streaming, are excellent model systems to study the impact of motion forces on cytoskeletons in vivo. Here we implement variational optical flow analysis as a new approach to analyze the polarity of microtubules in the Drosophila oocyte, a cell that displays distinct Kinesin-dependent streaming. After validating the method as robust for describing microtubule orientation from confocal movies, we find that increasing the speed of flows results in aberrant plus end growth direction. Furthermore, we find that in oocytes where Kinesin is unable to induce cytoplasmic streaming, the growth direction of microtubule plus ends is also altered. These findings lead us to propose that cytoplasmic streaming - and thus motion by advection - contributes to the correct orientation of MTs in vivo. Finally, we propose a possible mechanism for a specialised cytoplasmic actin network (the actin mesh) to act as a regulator of flow speeds; to counteract the recruitment of Kinesin to microtubules. HIGHLIGHT SUMMARYCytoskeletal networks do not exist in isolation, but experience crowded and dynamic intracellular environments. However, microtubule-environment interactions are not well understood, and such system-environment interactions are an unresolved question in biology that demands bridging across disciplines. Here we introduce an optical flow motion estimation approach to study microtubule orientation in the Drosophila oocyte, a cell displaying substantial cytoplasmic streaming. We show that microtubule polarity is affected by the regime of these flows, and furthermore, that the presence of flows is necessary for MTs to adopt their proper polarity. With these findings we are contributing to further understanding how microtubules organize in their impacting natural environment.

cell biology