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| Title | Optimising Adjacent Membrane Segmentation and Parameterisation in Multicellular Aggregates by Piecewise Active Contours |
| Authors | Jorge Jara-Wilde, I. Castro, C.G. Lemus, Karina Palma, F. Valdes, Victor Castañeda, Nancy Hitschfeld, Miguel Concha, Steffen Haertel |
| Publication date | May 2020 |
| Abstract |
n fluorescence microscopy imaging, the segmentation of ad- jacent cell membranes within cell aggregates, multicellular samples, tissue, organs, or whole organisms remains a chal- lenging task. The lipid bilayer is a very thin membrane when compared to the wavelength of photons in the visual spec- tra. Fluorescent molecules or proteins used for labelling mem- branes provide a limited signal intensity, and light scattering in combination with sample dynamics during in vivo imaging lead to poor or ambivalent signal patterns that hinder pre- cise localisation of the membrane sheets. In the proximity of cells, membranes approach and distance each other. Here, the presence of membrane protrusions such as blebs; filopodia and lamellipodia; microvilli; or membrane vesicle trafficking, lead to a plurality of signal patterns, and the accurate localisation of two adjacent membranes becomes difficult. Several computational methods for membrane segmentation have been introduced. However, few of them specifically con- sider the accurate detection of adjacent membranes. In this ar- ticle we present ALPACA (ALgorithm for Piecewise Adjacent Contour Adjustment), a novel method based on 2D piecewise parametric active contours that allows: (i) a definition of prox- imity for adjacent contours, (ii) a precise detection of adjacent, nonadjacent, and overlapping contour sections, (iii) the def- inition of a polyline for an optimised shared contour within adjacent sections and (iv) a solution for connecting adjacent and nonadjacent sections under the constraint of preserving the inherent cell morphology. We show that ALPACA leads to a precise quantification of ad- jacent and nonadjacent membrane zones in regular hexagons and live image sequences of cells of the parapineal organ dur- ing zebrafish embryo development. The algorithm detects and corrects adjacent, nonadjacent, and overlapping contour sec- tions within a selected adjacency distance d, calculates shared contour sections for neighbouring cells with minimum al- terations of the contour characteristics, and presents piece- wise active contour solutions, preserving the contour shape and the overall cell morphology. ALPACA quantifies adjacent contours and can improve the meshing of 3D surfaces, the determination of forces, or tracking of contours in combina- tion with previously published algorithms. We discuss pitfalls, strengths, and limits of our approach, and present a guideline to take the best decision for varying experimental conditions for in vivo microscopy. |
| Pages | 59-75 |
| Volume | 278 |
| Journal name | Journal of Microscopy |
| Publisher | John Wiley & Sons (Hoboken, NJ, USA) |
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