DisGUVery: A Versatile Open-Source Software for High-Throughput Image Analysis of Giant Unilamellar Vesicles

Abstract

Giant unilamellar vesicles (GUVs) are cell-sized aqueous compartments enclosed by a phospholipid bilayer. Due to their cell-mimicking properties, GUVs have become a widespread experimental tool in synthetic biology to study membrane properties and cellular processes. In stark contrast to the experimental progress, quantitative analysis of GUV microscopy images has received much less attention. Currently, most analysis is performed either manually or with custom-made scripts, which makes analysis time-consuming and results difficult to compare across studies. To make quantitative GUV analysis accessible and fast, we present DisGUVery, an open-source, versatile software that encapsulates multiple algorithms for automated detection and analysis of GUVs in microscopy images. With a performance analysis, we demonstrate that DisGUVery’s three vesicle detection modules successfully identify GUVs in images obtained with a wide range of imaging sources, in various typical GUV experiments. Multiple predefined analysis modules allow the user to extract properties such as membrane fluorescence, vesicle shape, and internal fluorescence from large populations. A new membrane segmentation algorithm facilitates spatial fluorescence analysis of nonspherical vesicles. Altogether, DisGUVery provides an accessible tool to enable high-throughput automated analysis of GUVs, and thereby to promote quantitative data analysis in synthetic cell research.

Synthetic control of actin polymerization and symmetry breaking in active protocells

“…The presented platform provides insight on the spatiotemporal localization of the molecular components and elucidates the design principles governing chemical signaling, actin assembly, and symmetry breaking. Our platform offers a previously unknown GUV manipulation technique ( 61 , 62 ) and a versatile imaging pipeline to track the locality-specific concentration of biomolecules in a dynamic GUV over time, complementing other realized modalities ( 63 , 64 ). Our image processing and statistical analysis scheme enables probing the biomolecular and geometric features of symmetry breaking, a pipeline that is applicable to other GUV- and cell-based studies.…”

Synthetic control of actin polymerization and symmetry breaking in active protocells

“…Our platform offers a novel GUV manipulation technique 52,53 and a versatile imaging pipeline to track the locality-specific concentration of biomolecules in a dynamic GUV over time, complementing other realized modalities. 54,55 Our developed image processing and statistical analysis scheme enables probing the biomolecular and geometric features of symmetry breaking, a pipeline that is is applicable to other GUV-and cell-based studies. By combining experiment and modeling, our results offer the ability to better understand macroscopic cellular processes, including morphogenesis, motility, and division, that build on fundamental biophysical and active matter principles to accomplish complex biological tasks.…”

Branched actin cortices reconstituted in vesicles sense membrane curvature

“…Quantitative confocal microscopy images were analyzed in an automated procedure. First, we located GUVs in the membrane images using the template-matching module in the open source DisGUVery toolbox ( 66 ) (see Fig. S4 C ).…”