Microfilament analyzer, an image analysis tool for quantifying fibrillar orientation, reveals changes in microtubule organization during gravitropism

Image acquisition is an important step to study cytoskeleton organization. As visual interpretations and manual measurements of the digital images are prone to errors and require a great amount of time, a freely available software package named MicroFilament Analyzer (MFA) was developed. The goal was to provide a tool that facilitates high-throughput analysis to determine the orientation of filamentous structures on digital images in a more standardized, objective and repeatable way. Here, the rationale and applicability of the program is demonstrated by analyzing the microtubule patterns in epidermal cells of control and gravistimulated Arabidopsis thaliana roots. Differential expansion of cells on either side of the root results in a downward bending of the root tip. As cell expansion depends on the properties of the cell wall, this could imply a differential orientation of cellulose microfibrils. As cellulose deposition is orchestrated by cortical microtubules, the microtubule patterns were analyzed. The MFA program detects the filamentous structures on the image and identifies the main orientation(s) within individual cells. This revealed four distinguishable microtubule patterns in root epidermal cells. The analysis indicated that gravitropic stimulation and developmental age are both significant factors that determine microtubule orientation. Moreover, the data show that an altered microtubule pattern does not precede differential expansion. Other possible applications are illustrated as well, including field emission scanning electron micrographs (FESEM) of cellulose microfibrils in plant cell walls and images of fluorescent actin.

Publication year:2013

Keywords:A1 Journal article

BOF-keylabel:yes

BOF-publication weight:10

CSS-citation score:1

Authors:International

Authors from:Higher Education

Accessibility:Closed