title: MicroFilament Analyzer, an image analysis tool for quantifying fibrillar orientation, reveals changes in microtubule organization during gravitropism.
abstract: Image acquisition is an important step in the study of cytoskeleton organization. As visual interpretations and manual measurements of 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 gravi-stimulated Arabidopsis thaliana roots. Differential expansion of cells on either side of the root results in downward bending of the root tip. As cell expansion depends on the properties of the cell wall, this may 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 also illustrated, including field emission scanning electron micrographs of cellulose microfibrils in plant cell walls and images of fluorescent actin.
authors.affiliation: Department of Biology, Plant Growth and Development, University of Antwerp, Groenenborgerlaan 171, 2020, Antwerpen, Belgium.
authors.forename: Eveline
authors.initials: E
authors.lastname: Jacques
authors.forename: Jan
authors.initials: J
authors.lastname: Buytaert
authors.forename: Darren M
authors.initials: DM
authors.lastname: Wells
authors.forename: Michal
authors.initials: M
authors.lastname: Lewandowski
authors.forename: Malcolm J
authors.initials: MJ
authors.lastname: Bennett
authors.forename: Joris
authors.initials: J
authors.lastname: Dirckx
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Verbelen
authors.forename: Kris
authors.initials: K
authors.lastname: Vissenberg
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D000199:Actins / Q000378:metabolism / Q000648:ultrastructure; D017360:Arabidopsis / Q000378:metabolism* / Q000502:physiology / Q000648:ultrastructure; D029681:Arabidopsis Proteins / Q000378:metabolism / Q000648:ultrastructure; D018522:Gravitropism; D018519:Meristem / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D018613:Microscopy, Confocal / Q000295:instrumentation* / Q000379:methods; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D019441:Plant Epidermis / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D018517:Plant Roots / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D018520:Plant Shoots / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins; D012984:Software
chemical_list: D000199:Actins; D029681:Arabidopsis Proteins; D011993:Recombinant Fusion Proteins