Automatyczne metody analizy orientacji mikrotubul

Abstract

A microtubule is a fibrous structure of 25 nm diameter and 10—25 μm length, which is created as a result of the polymerization of the tubulin protein. The orientation of microtubules in relation to the cell axis changes due to both internal factors (associated with intracellular processes) and external ones (the gravitational field, light intensity, mechanical stresses, an electric field, etc.). In order to evaluate these changes, the inclination angle to the cell axis is usually measured. In this monograph three developed algorithms for measuring the inclination angle to the cell axis are presented. The algorithms allowed determining a histogram of microtubule inclination angles which takes into consideration the area of microtubules. Additionally, the work covers the employment of the developed algorithms to the analysis of inclination angle variability as a kind of a trend in a cell. The authors present a spatial, three-dimensional reconstruction of microtubules based on a sequence of images obtained with a fluorescence microscope, using the depth-of-focus change feature. The monograph also contains an analysis of the protofilaments present in cross-sectional images of microtubules. Various possible approaches to this issue are shown. The authors hope that their monograph will be particularly useful for biologists and confirm them in the belief that many laborious measurements can be performed with a profiled image analysis application, leaving to them the analysis of the obtained results only.